Annual Report 2009-2010 - Department of Biotechnology

Annual Report2009-2010Department of BiotechnologyMinistry of Science & TechnologyGovernment of India

CONTENTSChapter – 1 : Overview 1- 21Chapter – 2 : Human Resource Development 23-30Teaching 23Training 26Fellowships 27DBT Biology Scholarships & Awards 28Biotechnology Overseas Associateship 29Rapid Grant for Young Investigator (RGYI) Scheme 29Biotechnology Entrepreneurship Student Teams 30Biotech Popularization 30Support for Publications 30Chapter – 3 : Biotech Facilities, Technology Platforms & Research Resources 31-37Centres of Excellence 31Programme Support 33Long Term R&D Projects 35Biotechnology Facilities and Research Resources 35BUILDER Schemes 37Chapter – 4 : Research and Development 39-112Agricultural Biotechnology 39Biofertiliser Biotechnology 52Biopesticides & Crop Management 54Bioresource Development and Utilization 57Plant Biotechnology 63Medicinal and Aromatics Plants 67Animal Biotechnology - Production 69Animal Health 73Annual Report 2009-2010

ContentsAquaculture and Marine Biotechnology 78Seribiotechnology 82Basic Research in Modern Biology 85Medical Biotechnology 86Stem Cell Research 93Bioengineering 94Human Genetics and Genome Analysis 97Environmental Biotechnology 98Nano Science and Nanobiotechnology 105RNAi Technology Platforms 108Energy Bioscience Programme 110Chapter – 5 : Biotechnology Programmes for Societal Development 113-125Programmes for SC and ST Population 113Programmes for Rural Development 117Programmes for Women 122Chapter – 6 : Biotech Product & Process Development 127-137Small Business Innovation Research Initiative 127Biotechnology Industry Partnership Programme 128Biotechnology Industry Research Assistance Programme 129Biotechnology Industry Research Assistance Council 129Product and Process Development - R & D 130Biotechnological Approaches for Food and Nutritional Security 131Biosafety Issues 133Patent Facilitation Cell 137Chapter – 7 : Biotechnology Information System Network 139-145Chapter – 8 : Biotechnology Parks, Incubators & Industry Platforms 147-150Chapter – 9 : International Collaboration 151-160Chapter – 10 : Autonomous Institutions 161-177National Institute of Immunology, New Delhi 161National Centre for Cell Sciences, Pune 162Annual Report 2009-2010

ContentsNational Brain Research Centre, Gurgaon 163Centre for DNA Fingerprinting and Diagnostics,Hyderabad 164National Institute of Plant Genome Research, New Delhi 166Institute of Life Sciences, Bhubaneswar 168Institute of Bioresources and Sustainable Development, Imphal 170Rajiv Gandhi Centre for Biotechnology, Thiruvanthapuram 171Up-Coming Institutions/Centers 173Bioclusters 176Chapter – 11 : Public Sector Undertakings 179Chapter – 12 : International Centre for Genetic Engineering & Biotechnology 181-184Chapter – 13 : Promotion of Biotechnology in North Eastern States of India 185-188Chapter – 14 : New Legislations and Regulations 189-191Chapter – 15 : Administration and Finance 193-196Annexures I – VI 197-210Abbreviations 211-218Annual Report 2009-2010

CHAPTER 1OVERVIEWPreambleThe development of biotechnology in the countryhas opened up unprecedented opportunities for bioentrepreneurshipdevelopment and commercialization.The Department of Biotechnology, over the yearshas provided unstinted support in fostering andnurturing the various sectors of biotechnology in anintegrated manner. Though it is customary to reviewevery year the developments that have taken place,linking annual review to past and future provides aperspective and helps build a strategy, provide a scopefor redesign, while preserving core values and emphasison all effective programs.With the drawing up of the National BiotechnologyDevelopment Strategy, the DBT established aroadmap for the coming years. The initial thrust ofDBT was on strengthening Master’s level trainingand Ph.D programmes and to promote the breadthand depth of biological science in the country. Inthe bio-tech development strategy, while maintainingthe push for producing quality people and makingIndia more competitive in life sciences, additionalfocus is now being given to innovation andentrepreneurship. Our efforts to promote innovationis based on the encouraging early translation researchin the public sector institutions, universities andproduct development through the market innovationsystem. In the later, two important approaches arethe public sector led public private partnership tofollow up leads from discovery and early translationprograms in the public sector institutions andsecondly, through industry led academic partnerships;facilitated by novel funding schemes like SBIRI andBIPP. Through this approach, the industry is coinvestingwith DBT in challenging innovationprograms. It is only with such a balanced R&Dpush in the academic and industrial sector thatsustained economic success in the biotechnologysector can take place. Another approach adopted byDBT is around technology development thoughsocial contracts, involving the Government, smallcompanies and the many not-for-profit fundingpartners focused on public-good technologies aroundaffordable health care, agriculture and bio-energy.The social contract approach links productdevelopment and diffusion of developed products.This linkage is essential for social impact. DBTtoday is a major partner of such innovationinstitutions like Wellcome Trust, Gates Foundationand bi-lateral international programs.The base of academic bio-technology is still suboptimal in size, range and depth. DBT is thereforecreating additional institutional capacity by buildingsix new institutions with total innovation valuechain ability, and by establishing long termpartnership centres with existing universities andinstitutions around focused priorities. This approachenables capacity expansion with limited and targetedinvestments to address opportunities in affordablehealth care, food security, bio-energy andenvironmental conservation.Annual Report 2009-2010 1

OverviewAccess to technology platforms has been identifiedas a predominant weakness. Some of the investmentsin the past, for infrastructure, are too much centeredaround individual scientists. Our emphasis is nowto create centralized independently managed,accessible, technology platforms in the country. Theaccess is required not only in basic sciencesinstitutions but also in medical and agriculturalcolleges and for small and medium industryresearchers. DBT is committed to strengtheningresearch in the places of education i.e. in theuniversity system and substantial progress is beingmade in strengthening the 20 best universities forinter-disciplinary life science research.Our current research institutes do good basicresearch, and they are being helped to developtransitional capacity. A major thrust is plannedfor genomics at the highest level in plant, animaland bio-medical areas; including technologyhelpful for plant genomics and phenomics, andcancer genome analyses. Creation of quality Ph.Dsin large numbers is critical for the expansion.The encouraging news is that the top 80universities and institutes are providing 18 percentmore Ph.Ds annually during the last few years.This augurs well for the future. Finally, reformin the governance of science is critical to dealwith basic science, science education and training,early translation research and prompt developmentand commercialization. The DBT is engaged inbuilding a unique innovation agency called BIRACfor this purpose.The overview presented here touches upon some ofthe highlights of the various sectoral programmes.It is intended to provide the reader a flavour of theresearch and developments undertaken, and a senseof the likely trends in the various projects, and thenew initiatives planned.Human Resource DevelopmentThe importance of highly skilled manpower forthe success of research in biotechnology and bio -enterprise cannot be over emphasized. Recognizingthis paramount need, the DBT continued to accordprime importance to HRD. Postgraduate teachingprogrammes in biotechnology continued to be aplatform for selecting the best quality studentsand imparting high quality training with emphasison ‘hands on’ training. Industrial training ofpostgraduate students emerged as a successfulmodel with a large number of industriesparticipating in the programme. More than 3000applications were received and 440 trainees wereselected and placed in 110 companies. Over 30%of the trained students were absorbed by theindustry. The department also nurtured aninnovative ‘Star College’ scheme at 35 collegesselected from across the country, with emphasison curriculum reform, experimental design, handson laboratory work and industrial exposure. Thevarious fellowships and awards instituted by theDBT have become increasingly popular withstudents, and scientists. Selection is highlycompetitive as out of over 5000 applicants, only250 are offered the fellowship. Short term trainingprogrammes supported by DBT provide a uniqueopportunity to mid career scientists and faculty tointeract with leaders in the field and upgrade theirskills in niche areas. The Young BiotechnologyInnovation Award including a research grant hasbeen a successful young leadership buildingprogramme, a kind of post doctoral and mid2Annual Report 2009-2010

Overviewcareer scheme. The Ramalingaswami Fellowshipis a major and effective scheme to bring Indianscientists back home. Post doctoral meetings havebeen held in USA, Europe and India to connectScientists to the changing opportunities in sciencein India. While there are many recognitions forbasic science excellence, we have introduced theTata Innovation Fellowship programme to rewardthose who create translational knowledge of highquality and useful products.Centres of Excellence and BiotechfacilitiesStrengthening of institutional research capabilitiesfor promotion of excellence in interdisciplinaryscience and innovation in specific areas ofbiotechnology is a major priority for DBT. Theapproach provided flexible long-term support forhighly innovative research i.e both basic andtranslational in nature, which results not only inhigh-quality publications and intellectual propertybut also in translational outputs through mid andhigh end innovation. Institutional programmesupport was given to strengthen a multi prongedapproach to address a research topic. The platformsfor translational research on transgeneic crops andmolecular cryatallography clearly filled a long feltneed. The BUILDER programme was novel in thesense that it was crafted to boost research in lifesciences in selected universities. The Centre ofExcellence Scheme marked a significant departurein our approach to problem oriented research.Among the portfolio of research undertaken at thecentres, there were interesting leads. Using theRNAi approach 8 transgenic silkworm lines resistantto baculovirus (BmNPV) were developed. A smallstructured hepta – peptide derived from a hostfactorwas found to inhibit translational replicationof HCV RNA. Bioreactors were developed for tissueengineering of liver and cartilage tissue engineering.Several transgenic lines of rice that were developedusing various stress related genes such as Rab7,Glyoxalase with NHX, PDH45, SOD with APXshowed enhanced tolerance to salinity. Thesetransgenics are under evaluation. It must also bementioned that good leads were seen in thedevelopment of fish vaccines, nano composites forantimicrobial activity and proteomics related toneurological disorders.Research and DevelopmentAn integrated approach comprising a large numberof institutes, universities, NGOs and internationalagencies has woven together a national effort thathas truly benefited the cause of biotechnology andbioentrepreneurship while strengthening thefoundation for human resource development. Thisis amply borne out by the fact that there are morethan 1600 on going research projects.The department provided a major thrust to R&Dacross a wide range of sectors in basic biology andbiotechnology. A robust monitoring mechanism hasbeen put in place and the constant endeavour is toidentify leads and take them forward on the roadto process/product development.Agricultural BiotechnologyResearch activities spanned a broad canvas but laidemphasis on crops like rice, wheat and maize.Genomics of agricultural crops formed a base forapproaches that targeted improvement of importantAnnual Report 2009-2010 3

Overviewtraits, and modifying the response to biotic andabiotic stresses. Research on transgenesis remained akey tool in furthering these objectives.The study ofnovel genes and their functions have been extendedto a number of species.In a project on identification of molecular markerslinked to quality parameters in rice, their validationand utilization in marker-assisted selection (MAS)at DRR, Hyderabad a total of 239 markers werefound polymorphic out of the 575 markers tested.Another study revealed the association of SSS1-SSR (linked to soluble starch synthase 1 enzyme)and SBE1-SSR1 (starch branching enzyme 1) withintermediate amylose content; SBE1-STS1(starchbranching enzyme 1) with low ASV. At JamiaHamdard University, 110 rice genotypes wereanalyzed at various levels of nitrogen treatmentsfor selecting nitrogen efficient and nitrogeninefficient genotypes. Based on their growthperformance, nitrogen distribution and nitrogenefficiencies three categories of rice genotypes wereidentified.A study was undertaken at UDSC, New Delhi foridentification and characterization of differentiallyexpressed genes during heat stress in wheat (Triticumaestivum cv. CPAN 1676) A total of 4500 ESTswere generated and about 3000 of them submittedto Genbank. A large number of high temperatureresponsive genes were identified and characterized.Vivek QPM9, an early maturing quality proteinmaize hybrid was developed through marker assistedselection (MAS) at Vivekanand Parvatiya KrishiAnusandhan Sansthan (VPKAS), Almora. VivekHybrid 9 that was earlier released for commercialuse is considered to be one of the best maizehybrids developed by VPKAS.The DBT’s Programme Support provided thenecessary impetus for bringing about excellence inagricultural biotechnology. Four SAUs weresupported to take up work on specific researchthemes/projects pertaining to the management ofbiotic and abiotic stresses. The department isimplementing projects to bring together, throughthe transgenic approach, important traits, and alsospeed up the selection process through MAS.Transgenes viz. OsglyI, OsglyII and ZAT12 wereintroduced into the commercial rice variety PR 116for developing abiotic stress tolerance. A number oftransgenic plants were grown to maturity. Fordevelopment of drought tolerant rice, 22 QTLswere identified and efforts were made to developIR20 and IR64 NILs by introgressing three QTLsassociated with root traits and yield under stress. Aset of 400 BC 4F 3IR20 NILs developed throughmarker assisted breeding were in the field forassessing drought tolerance. Through participatorybreeding, a superior drought tolerant genotype,Paramakudi 4 was developed and released.BiofertilisersOur constant need to enhance food production hasdrawn us into the increased use of fertilizers andchemicals. The down side to this is the resultantpollution and contamination of soil that has emergedas a major concern. Use of biofertilizers has emergedas an environmentally friendly option that is nowadopted in many countries that can reduce the useof chemical fertilizers by 25 to 30 percent.Moreover, the use of vermicomposts can increasethe health of soil. Recognising the immense potentialof this twin option the DBT supported thedevelopment of technology packages for scaling upthe production of biofertilisers and also focused on4Annual Report 2009-2010

Overviewconversion of agricultural wastes into vermicompost.Development of sulphur oxidizing bacteria asbiofertilizers and integrated nutrient managementpackages for high value plantation crops areimportant outcomes.BiopesticidesEcofriendly approaches to pest management leanrather heavily on the use of microbial bio controlagents. The challenges lie in standardization ofprotocols and upscaling the production of variousmicrobial pesticides and natural enemies and ensuringtheir wide application.Research done at TNAU, Coimbatore, showed thatbacterial antagonists for potato cyst nematodes weremore in soils having low cyst densities. Among 73isolates examined, eight were found to be effectivein reducing potato cyst nematodes and enhancingplant growth. At IIHR, Bangalore, noteworthymortality of hoppers was noticed by the applicationof M. anisopliae and Verticillium lecanii. Biologicalcontrol of mango hopper Idioscopus nitidulus by M.anisoplaie revealed a 76.4 % reduction in mangohopper population. Work done at IIPR, Kanpur,demonstrated the bio-efficacy of Steinernema andHeterorhabditis against H. armigera. Mention mustbe made of the two novel heat tolerant EPN viz.Steinernema masoodi and S. seemae that weresuccessfully field tested. S. seemae was found to killH. armigera larvae within two days. In order tofocus on the current status biopesticides, the 5thInternational Conference on Biopesticides:Stakeholders’ perspectives” was organized under theauspices of “Society for Promotion and Innovationof Biopesticides” in collaboration with TERI fromApril 26-30, 2009 at New Delhi. The conferencewas marked by the release of a “Strategy Documenton Biopesticides” .BioresourcesThe National Bioresource Development Board hastaken up a number of interagency programmes forbioresource development and utilization. Majorprogrammes are on bioprospecting, inventorizationand characterization, value addition and sustainableutilization of bioresources together with a strongcomponent for training, capacity building andawareness generation. In the microbial prospectingprogramme, nearly 1,75,000 isolates were collectedfrom different ecological niches and 480000 extractsscreened for bioactivity. This huge repository is atreasure trove that needs careful characterization forpotential drug molecules. A microbial culturecollection centre was established at NCCS, Pune.Primary data on distribution and population statusof plant resource of Western Ghats and EasternGhats were completed. The inventorization ofresources and the increasing use of bar coding wasaccomplished. Nearly 550 DNA clubs were launchedin 26 states and UTs targeting about 50,000 schoolchildren. The immense reach of these initiativeswill certainly make a difference in the way theyounger generation will appreciate the value of ourbioresources.Plant BiotechnologyThis major theme includes key initiatives in basicresearch, tissue culture and forestry, horticulture andplantation crops and the Solanaceae genomeinitiative. Basic research focused on signaltransduction, root development, flowering and hostpathogen interaction. A study conducted at JNU,Annual Report 2009-2010 5

OverviewNew Delhi resulted in chilli leaf curl virus isolatesbeing cloned and classified into 3 species ofbegomoviruses. At MKU, Madurai, following theidentification of C4 and âC1 of Bhendi yellow veinmosaic virus (BYVMV) as strong suppressors ofgene silencing, and C2 as a mild suppressor, theireffects on a number of plants, like bhendi, chilli,tobacco and tomato were studied. An internationalconsortium of 10 countries ie., USA, Korea, China,UK, India, Netherlands, France, Japan, Spain andItaly took up the sequencing of the tomato genome.India was assigned the task of sequencing 12 Mb ofthe euchromatic region of chromosome 5. Transgenictomato lines resistant to leaf curl virus weredeveloped.Medicinal and Aromatic PlantsThese plants are coming under increasing focus dueto their potential for development of health careproducts. The emphasis was on isolation andcharacterization of bioactive compounds.Morphological, phytochemical and molecularcharacterization studies were conducted on differentaccessions of Adhatoda zeylanica, Coleus aromaticus,Hemidesmus indicus, Saraca asoca, Tinospora cardifoliaand Terminalia chebula from different agro-climaticregions of South India. Identifcation of high yieldingsources of metabolites such as galanthamine andcoumarins from herbarium material as well as fromWestern Ghats was pursued. Hippeastrumsolandrifarum and Hymenocalis caribeae were foundto have the highest galanthamine content.Construction of a DNA molecular genetic linkagemap of Catharanthus roseus was taken up at NIPGR,New Delhi. QTLs for vindoline and catharanthinehyper-accumulation were identified and mapped.Eight known genes of the morphine biosyntheticpathway in opium poppy were cloned andsequenced. A small-scale microarray chip containingall the significant alkaloid biosynthesis-related genesin P. somniferum was developed.A concoction of three extracts developed fromAzadirachta indica, Withania somnifera and W.somnifera was found to be most efficacious againstStaphylococcus aureus of Bovine mastitis during invivo testing. The highest galanthamine content(> 1% Dw) was found in Hippeastrumsolandrifarum, Hymenocalis caribeae, where as Menthapiperita, Lavandula pinnata var. bipinnate, Melilotusparviflora and Cinnamomum camphora were foundpromising for their coumarin content.Animal BiotechnologyThe recognition of the important contribution ofour animal resources was underscored by a rangeof projects undertaken by the department forenhancing production as well as modification ofimportant genetic traits. Transgenic mice modelswere generated using the technique of in vivoelectroporation of the gene. The stability of thetransgene integration and its propagation wasconfirmed upto the F3 generation. To enhance thereproductive efficiency of buffalo, the mainstay ofIndian dairy industry, estrus-specific pheromoneswere analyzed to confirm pregnancy at an earlystage. Studies were undertaken to understand thegenetic structuring of various breeds of cattle. Atotal of 483 mitochondrial DNA (mtDNA) D-loop sequences were generated to analyze themtDNA diversity. High haplotype and nucleotidediversity indicated substantial mtDNA diversity inIndian cattle. Enhancement of the nutritional value6Annual Report 2009-2010

Overviewof feeds was an important aspect of research. Thisneeds to be viewed against the under utilization ofthe huge agro residues in the country. Effortswere made to use non conventional oil cakes as analternate source of protein in fodder. Anaerobicfungii with superior fibre degrading qualities wereisolated and characterized. Biomaterial developmentfrom animal sources holds promise for a numberof applications. A collagen based scaffold of bovineorigin that was prepared showed the necessarymechanical strength to sustain beatingcardiomyocytes. A decellularisation technique wasdeveloped for bovine pericardium and jugular vein,for cardiovascular application.The programmes on vaccines and diagnostics werereprioritized and oriented towards translationalresearch. Apart from supporting open-endedprojects, network programmes on classical swinefever virus and brucellosis were supported. Thekey elements of research on classical swine feverincludes surveillance, epidemiology, developmentof a repository and development of new generationvaccines. Special focus has been accorded to theNorth East region. The other projects were onidentification of novel proteins expressed bySalmonella spp. in biofilm and their utility indiagnosis; seroprevalence and diagnostics of caninemonocytic ehrilichiosis in various regions of India;and development of diagnostic tools for avianmycoplasma infections. Development of specificassays with reference to Mycobacterium bovisinfection in animals and man, transmission oflivestock derived pathogenic Mycobacteria and itsprevalence among humans also received attention.The programmes on vaccines, included a biomarkerapproach for differentiating natural infection andvaccination of chicken, development of RNAreplicon-based vaccine vector for enhanced immuneresponse, development of recombinant vaccine forleptospira, production and evaluation of microparticle coated DNA vaccine for FMD,development of biofilm based vaccine againstmastitis and a DNA based vaccine againstClostridium perfringens.Aquaculture & Marine BiotechnologyThe aquaculture and marine biotechnologyprogrammes showed a shift towards the nonfood sector. The portfolio of research was targetedtowards bioactive molecules, development ofimproved feed, molecular characterization of species,development of cell lines etc. A number of peptidetoxins were isolated from various species of conesnails and an inventory comprising 50 toxinpeptides and their sequences was prepared. Morethan 20 marine organisms, harbouring potentialantimalarial bio active compounds, were identified.The purified compounds were found to arrest thegrowth of parasite at the trophozoite stage andone of the promising molecules was found toinhibit invasion of merozoites into fresh red cells.A multi centric programme was launched for themuch needed development of cell lines fromvarious fresh water and marine fishes. A diagnostictest was developed for the identification of whitetail disease of Macrobrachium rosenbergii usingreverse transcriptase loop mediated isothermalamplification (RT-LAMP). In an effort to developtechniques for induced maturation and earlyspawning techniques in shrimp (P. monodon), tworecombinant proteins viz. the crustaceanhyperglycaemic hormone (CHH) and moultinhibiting hormone (MIH) were expressed in PichiaPastoris.Annual Report 2009-2010 7

OverviewSeribiotechnologyThe major areas of focus were on improvement ofproductivity, enhancement of the quality of silkand improvement of the host plants. A DNAmarker associated with resistance to powdery mildew(Phyllactania corylea) was identified in mulberry.The first generation HVA-I mulberry transgenicplants with constitutive and inducible promoterswere developed for drought and salinity toleranceand transferred to the Sericulture Institute forevaluation. QTLs were identified in mulberry forwater use efficiency and related root traits. A pilotlevelprocess for the recovery and production ofsericin powder from industrial waste degummingliquour was taken up for up scaling with an industry.Basic Research in Modern BiologyBasic research continued to receive support in orderto enhance our understanding of principles andtheories that would help in developing appropriatestrategies for research. A synthetic polymer hydrogelbased on N-vinyl pyrrolidone and hydroxy ethylmethacrylate was synthesized at BAIT,Sathyamangalam and its characteristics weredetermined and correlated with its swellability anddrug release properties. This polymer seemed to bea promising carrier for delivering drugs withproperties similar to paracetamol and wasbiodegradable. Work carried out at IICB, Kolkata,succeeded in generating a mouse model forinflammation with many features of RheumatoidArthritis (RA). Cells isolated from lymph glands ofmice showed the presence of macrophage like cellsinvolved in inflammation. A series of glycolipidspresenting sugar residues in the form ofoligosaccharides of M. tuberculosis were synthesizedand their structural properties established at IIScBangalore. The study demonstrated that syntheticglycolipids inhibited the growth of Mycobacteriumand also affected the bio-film necessary for itssurvival.Medical Biotechnology– Vaccine Research and DevelopmentConsidering the huge disease burden, vaccine researchhas been accorded top priority through a largenumber of programmes involving multiple partners.The initiatives of the various expert groups and theoutcomes of the Jai Vigyan Mission are culminatingin new vaccines. Several candidate vaccines haveemerged from these concerted efforts. Some of themare into pre-clinical or clinical trial stages such asvaccines for Rotavirus, Cholera, Typhoid, Rabies(DNA based), Anthrax, Malaria, Dengue,Tuberculosis, Japanese encephalitis, etc. The projectson rotaviral diarrhoea were implemented at AIIMS,New Delhi in collaboration with CDC, Atlanta;and IISc, Bangalore in collaboration with StanfordUniversity, USA. The first rotaviral diarrhoea vaccine116E developed in India has completed phase-IIhuman clinical trials. The vaccine induced immuneresponse was reported in about 90% of therecipients.The notable work on Dengue vaccine carried out atICGEB was innovative and seems to be the onlysubunit vaccine approach in the world today and ofrelevance to the pediatric population. Efforts areunderway for the development of a tetravalentDengue vaccine and initiation of clinical trials. TheVaccine Grand Challenge Programme has alsofocused on Dengue, Cholera, Tuberculosis,8Annual Report 2009-2010

OverviewRotavirus, Typhiod, Malaria and Rabies. The progressof the Indo-US Vaccine Action Programme (VAP)was reviewed by Joint Working Group (JWG) inNovember, 2009 at NIH, Washington DC, USA.A Vaccine & Infectious Disease Research Centre(VIDRC has been established as a niche centre ofthe newly established THSTI, with the overallobjective to conduct multi disciplinary basic andclinical research on vaccines and biologics.– Infectious DiseasesResearch on Tuberculosis has remained a frontlinearea for the last two decades The department hasbeen funding basic, applied and translational researchprojects. Under a project at the UDSC, New Delhi,an ELISA based test system for confirming MTBcomplex for treatment decision using MPT64 alongwith another antigen (patented) was developed. Thedepartment continued its support for developmentof a live oral Cholera vaccine. The candidate vaccinestrain VA1.3 developed by IMTECH, Chandigarhhad undergone Phase I/II studies at SAS, Kolkata;SGPGIMS, Lucknow and CMC, Vellore and itwas found to be safe and significantly immunogenic.A new strain VAI.4 developed for Cholera vaccinewould undergo toxicology studies and phase I/IItrials.A study on the virological and immunologicalaspects associated with opportunistic viralinfections in HIV-1 infected individuals in SouthIndia, showed that individuals with < 100 CD4+ T cell counts had comparatively moreopportunistic viral types that were detectable. Fordevelopment of Chikungunya vaccine candidates,CHIKV isolates were tested for growth in verocells. One isolate which showed a titer of 108pfu/ml in vero cells was purified. Towardscloning, expression and characterization of filarialAcetylcholinesterase (AchE), two isozymes offilarial AchE were found to be biochemically andimmunochemically similar but different from thehost enzyme. It was seen that one of themonoclonal antibodies produced against theparasite AchE inhibited the enzyme activity ofboth isoforms of parasite AchE indicating the useof same antifilarial agent to target both forms ofthe parasite AchE. Work on designing stable foldedfragments of HIV env for use as immunogens,showed that a gp120 based fragment CD4Bs wasfound to elicit a better neutralization than thatseen with gp120.Stem Cell ResearchThe exciting developments in stem cell researchand the emerging therapeutic applications are many.The challenges that need to be addressed pertainto expertise, infrastructure, interdisciplinarynetworking of researchers and clinicians for welldefined basic research leading to translationalresearch. Efforts were made to generate a criticalmass in the country. During the year, about 30proposals were received and 20 were sanctionedon different aspects of basic and translationalresearch. Programmes were taken up for thegeneration of human autologous embryonic stemcell lines by somatic cell nuclear transfer, assays fortoxicity testing and drug discovery usingcardiomyocytes differentiated from hESCs, andsomatic cell nuclear transfer using sheep eggs.Generation of induced pluripotent stem cells(iPSCs) from human somatic cells, in-vitrodifferentiation of hESC lines to hepatocytes andthe role of serotonin in its expansion anddifferentiation were also studied. Cell lines fromAnnual Report 2009-2010 9

Overviewthe stem cell bank in UK were made available toresearchers. The initiation of clinical trials on acutemyocardial infarction using bone marrow cellsneeds mention.BioengineeringIntegration of various disciplines to developprocesses, systems or models for diagnosis andtreatment of diseases is indeed promising. Workdone at IIT, Kanpur has shown that hybrid polymerceramicbiocomposites can be developed usingtailored processing conditions on a compressionmolding machine. Biocompatibility tests on mousefibroblast cells showed good cell adhesion, cell-cellinteraction and cellular bridge formation among theadhered cells. Another piece of research work doneat IIT, Kanpur led to the development of a dualdrug delivery system for the treatment of lungcancer which can be used as a therapeutic optionfor other solid tumors also. Work done at CMC,Vellore established an animal model of physealinjuries in goat and the treatment was standardizedwith autologous chondrocytes. A measurementsystem suitable for body pressure distribution byusing MEMS based pressure sensors was developedat IISc, Bangalore.The Stanford-India Biodesign (SIB) programme wasinitiated by the Department in July 2007 incollaboration with the Stanford University, USAand the Indo-US S&T Forum. The programmeoffers a multi disciplinary team based fellowshipfor developing cost effective solutions for clinicalneeds in India. The SIB fellows developed anautomated device for cardiac resuscitation, a novelmethod to deliver fluids to critically ill patients anda critical care product to manage patients in intensivecare. The interns developed multiple prototypes andcompleted their clinical validation. They alsoconducted live demonstrations of the novel devices.Human Genetics and Genome AnalysisVery early on itself, the DBT accorded priority tothe programme on human genetics keeping in viewthe national priorities and international efforts toidentify, map and characterize genes associated withgenetic disorders and more so to use the knowledgecreated, to develop molecular medicine tools forhealth management strategies. A study ofpolymorphisms on the physiological anti -hypertensive peptide catestatin conducted on 600subjects at IIT, Chennai, showed naturally occurringgenetic variations in 159 of them. Work done onpharmacogenetics at SGPGIMS, Lucknow showedthat 155 renal transplant patients could be categorizedbased on SNPs in drug metabolising and drugtransporter gene. The results suggested a significantassociation of CYP3A5*3 with CsA absorption at 1month post transplantation. The India specificproject on oral cancer, under the International CancerGenome Consortium was taken up at IndianStatistical Institute, Kolkata and ACTREC,Mumbai. A pilot study was initiated with samplesfrom 100 oral cancer patients at the NationalInstitute of Bio-medical Genomics (NIBMG),Kolkata.Environmental BiotechnologyThe growing awareness on the unsustainable practicesand the need to push towards ecofriendly approacheshave defined many of the programmes in this sector.Biotechnological tools were used to mitigateenvironmental problems and conserve natural10Annual Report 2009-2010

Overviewresources. Interactions were organized for assessingnew technologies for environmental monitoring, ecorestoration and use of renewable resources.Technological options that were developed throughvarious projects, were taken up for adoption. Thedemonstration of technologies for environmentalmonitoring, restoration of environmental quality,oil spill remediation and hydrocarbon recovery fromoily sludges were issues addressed. Desulphurizationof coal, substitution of non-renewable resources withrenewable ones, and conversion of lignocellulosicsinto value added products strengthened theecofriendly approaches adopted for mitigation ofvarious problems. Studies on carbon sequestrationhave shown promising leads towards mitigating theeffects of climate change.Nanoscience and NanobiotechnologyThe potential of nanotechnology to dramaticallychange the contours of health care, consumerdurables, textiles, materials, communicationtechnology, and energy is clear. Severalnanotechnology based products are in the market.Keeping in view the immense potential, thedepartment supported the development of productsemploying nanotechnology. More than 90 projectswere sanctioned, in the last two years.Nanobiotechnology approach was used in designingfunctionalized biodegradable nanoparticulate drugdelivery system (NPDDS) of Doxorubicinhydrochloride (DOX) for application in hepaticcancer therapy. An NPDDS with a high entrapmentefficiency and drug loading capacity for the drugDOX in a hydrophobic polymer PES, was designed.A study conducted on the role of nano silverthrombosis biology demonstrated that nano silverinhibited platelet functions. A number of otherprojects showed interesting leads. The thrust givento the key sectors of agriculture and medicine forthe use of nano technology enabled options is verytimely.RNAi Technology PlatformsThe epoch making discoveries of RNA interferenceand RNA-mediated post transcriptional gene silencinghave provided novel approaches for the regulationand knocking down of gene expression in organismsand cells. These are leading towards novel approachesfor human therapeutics. Keeping in view theemerging potential of RNAi technology, thedepartment started the programme in 2007.Altogether 55 projects are being implemented acrossa range of topics. RNAi technology in medicineand agriculture has received a high priority.Expression of miRNA in cancer, mi RNA alterationsin diabetes and use of modified siRNAs for targetingmalaria are the important investigations beingundertaken. RNAi intervention in agriculturetowards improvement of traits and resistance topathogens are important initiatives.Energy Bioscience ProgrammeProgrammes on biofuels and bioenergy have largelyfocused on using alternate feed stocks as a rawmaterial for ethanol and biodiesel production. ACentre for Energy Biosciences (CEB) was set up byDBT and Institute of Chemical Technology for anintegrated approach for basic and translationalscience capability. The key areas identified for researchwere bioethanol, biodiesel, biohydrogen andbiomethane. The centre has developed an industryreadytechnology for producing ethanol fromAnnual Report 2009-2010 11

Overviewlignocellulose. Marker genes for selection of highoil content genotypes of Jatropha were identified atJP University, Solan. A number of ESTs weregenerated for identifying genes for oil biosynthesispathway at NBRI, Lucknow, MSSRF, Chennai,and SRM University, TamilNadu. Out of the 6000ESTs generated, nearly 3000 were sequenced. Workdone at the Osmania University, Hyderabad resultedin the isolation of thermotolerant yeasts from varioushigh temperature regions of India. The Saccharomycescerevisiae strain, OC21 showed the maximumfermentation efficiency.Biotechnology Programmes forSocietal DevelopmentProgrammes for SC/ST and Rural PopulationIn order to empower the members of the ScheduledCaste, Scheduled Tribes, the under privileged andthe rural population, various training anddemonstration programmes in biotechnology wereorganised. Proven technologies in sericulture,vermicomposting, integrated pest management,mushroom cultivation, aquaculture, apiculture, valueaddition, bio-control, agri waste management, andbio-resource management were transferred for thebenefit of SC/ ST populations. Programmes foradoption of piggery, goat rearing, poultry, biofertilizers, medicinal and aromatic plants, organicfarming, integrated crop management wereimplemented. A suite of activities were undertakento improve the livelihood conditions of ruralpopulation. The range of options includedapiculture, mushroom cultivation, bio-fertilizers,vermicomposting, sericulture, organic farming, goatrearing, banana cultivation, use of medicinal andaromatic plants, aquaculture, agri waste management,bio pesticides, provision of mobile veterinary clinics,and setting up of rural bio resource complexes,These activities have significantly contributed toenhancing the livelihood conditions of the SC/STand rural populations.Programmes for WomenThe programme is designed for the benefit ofwomen by way of improving their income,entrepreneurship skills and health status throughthe use of various soft technology options inbiotechnology. The programme includes projectsin areas such as organic farming, mushroomcultivation, floriculture, aquaculture, livestockfarming, vermicomposting, and production ofvalue added marketable products. In addition tothese, projects on key aspects like health, hygiene& nutrition received attention. Specialprogrammes on bioresource complexes includinga network programme on ‘Prasad Kit’ werepursued. Through all these initiatives, emphasiswas laid on training, demonstration, extensionactivities, enterprise development, and marketlinkages for the benefit of women from differentbackgrounds. Around 12,000 women werebenefited through the 65 projects supportedduring the year.Biotech Product and ProcessDevelopmentINDUSTRY SCHEMESSmall Business Innovation Research Initiative (SBIRI)The Small Business Innovation Research Initiative(SBIRI) has funded nearly 60 projects to the industryin the various sector of biotechnology. The scheme12Annual Report 2009-2010

Overviewhas been well received by the industry and a numberof interesting developments were seen. Thecommercialization of a recombinant folliclestimulating hormone (RFSH), the development ofa silk protein film for burn wound management, anitrifying bioreactor for shrimp hatcheries and anauto dispenser for diagnostic applications are amongthe noteworthy achievements. The SBIRI, within aspan of three years has emerged as a flagshipprogramme of the department in promoting aculture of innovation in the biotech industry inIndia.Biotechnology Industry Partnership Programme(BIPP)The BIPP scheme was approved in 2008 forpartnership with industries on a cost sharing basiswith the key emphasis on (i) path-breaking researchin frontier, futuristic technology areas with majoreconomic potential and (ii) development ofappropriate technologies in the context of recognizednational priorities in the area of agriculture, health,bioenergy, green manufacturing etc. In response tothe 5 rounds of call for proposals, more than 250proposals were received. Altogether, 20 proposalswere cleared and 11 projects have been supportedso far.Biotechnology Industry Research AssistanceProgramme (BIRAP)The pilot BIRAP was established in 2008 topromote and nurture innovation in the biotechindustry especially SMEs. The first programmethat was operationalized from the pilot BIRAPwas the BIPP scheme. The BIRAP recruited thenecessary staff and they executed programmes forcapacity building in partnership with ABLE andBCIL.Biotechnology Industry Research AssistanceCouncil (BIRAC)It is proposed to set up a Biotechnology IndustryResearch Assistance Council (BIRAC) as anindependent autonomous organization to deal withindustry related activities of DBT. The BIRACwould handle the schemes for ignition grant, pre-SBIRI, SBIRI, BIPP, Incubation centre, Biotech Park,Research Resource Facilities, Industry Platform etc.R & D SCHEMESThe thrust of the scheme was on scaling-up ofresearch leads and development of products andprocesses for utilization by industries. Researchhighlights include the purification of recombinantchitinase to near homogeneity using His-Tag affinitychromatography. The recombinant chitinaseproduced medicinally important N-acetyl D-glucosamine and chitobiose from swollen chitin.The gene encoding phytase gene ( phyA ), isolatedfrom A. ficuum and A. niger were sequenced. Thegrowth parameters for pigment producingmicrofungi were optimized. Shades were developedfrom Pestalotia sp. and Sclerotinia sp. A xylanaseproducer thermophilic Scttalidium thermophilumstrain was isolated and used for solid statefermentation of deoiled Jatropha seed cake.Fermentation of hemicellulose hydrolysate of sweetsorghum bagasse was achieved with Pichia stipitisand Debaryomyces hansenii sp. A number of projectswere implemented for enhancing food andnutritional security. Other projects pertained toneutraceuticals from unconventional sources, omega-Annual Report 2009-2010 13

Overview3-fatty acid, monitoring of iron deficiency anemiaand process optimization for production of zinctablets for management of diarrhea in children.BIOSAFETY ISSUESThe programmes on biosafety ensured theimplementation of biosafety procedures, rules andguidelines under the Environment (Protection) Act1986 and Rules 1989 to ensure safety from the useof Genetically Modified Organisms (GMOs) andtheir products in research, to the users as well as tothe environment. A three tier mechanism has beenestablished for granting approval for research anddevelopment activities on recombinant DNAproducts, environmental release of transgenic cropsand monitoring and evaluation of research activitiesinvolving recombinant DNA technology. A total of412 IBSCs were set up in the various researchinstitutions and industrial R&D units in the country.The RCGM took initiatives for the promotion ofrDNA technology. These included the revision ofthe IBSC Handbook and Guidelines, and Guidelinesfor Net House Trials. Though cotton is the onlytransgenic crop approved for commercial release inthe country, several others like brinjal, cabbage,castor, cauliflower, corn, groundnut, mustard, okra,potato, rice, tomato etc. are in the pipeline.Biotechnology Information SystemNetworkBioinformatics has clearly emerged as a pivotaldiscipline for research and various industrialapplications. The need to handle large biologicaldata sets without errors calls for large memoryspace and huge computation time. TheBiotechnology Information System Network(BTISnet) program of DBT has promoted thedevelopment of specialized manpower and facilitatedthe taking up of challenging research activities. Thesystem promoted the creation of a valuable database,software tools, and infrastructure facilities. TheBTISnet has emerged as a major distributedscientific network in the world. To day it comprises155 Bioinformatics centers spread across the country.More than 25 of these centers are located in theNorth Eastern states of India. A North EastBioinformatics network (NEBInet) has beenspecially designed for these states to speed up growthin the region. Significant progress has been registeredby the centers in terms of research publications andhuman resource development. The CoEs inbioinformatics have been set up at six leadinguniversities. A major initiative taken by the DBTwas the launching of the e-Library Consortium(DeLCON) in January 2009, which provided theautonomous institutions of DBT full access to 600journals. This facility was upgraded to include morethan 900 journals for online access.Biotechnology Parks, Incubators andIndustry PlatformsThe purpose of setting up biotechnology parks isto facilitate innovation through the developmentof biotechnology industrial clusters and create askill pool of biotechnologists and entrepreneurswho have a strong foundation in research andinnovation. The incubators help the start upcompanies, provide support to young entrepreneurswho are not able to invest in facilities but havethe capability to develop and design new biotechproducts and processes by making use of thefacilities in incubators.14Annual Report 2009-2010

OverviewIndustry PlatformWith research becoming increasingly expensive owingto the growing complexity, there is a need forintegration of research efforts at the national level.This necessitates joint efforts through various publicprivatepartnership models. Such partnerships wouldhave a catalytic effect on the innovative capabilityof the industry and the academia. The incubationof start up companies and SMEs in a multidisciplinaryresearch setting is possible through theestablishment of a framework and infrastructuredevelopment through Industry Platforms. Theseplatforms would promote research, anddevelopmental programmes through a strategicresearch agenda. The department has set up a numberof parks and incubators which are fosteringbioentrepreneurship in the country. The biotechparks and incubators in Lucknow, Chennai andHyderabad are operational. The others beingestablished in Kochi, Bangalore, Guwahatiand Bhubneswar are in various stages ofoperationalization. In order to meet the growingneeds of biotech companies in the country we wouldsee the emergence of more incubators in the publicprivate partnership mode.International CollaborationCollaborations with other countries provide a greatopportunity to enlarge the scope of work, engagein partnerships that are mutually beneficial and alsoprovide for inflow of technical skills and knowhow. The DBT has endeavored to open up newavenues for international collaboration for takingresearch leads forward. The numerous collaborationsinitiated by the DBT resulted in interesting leads indifferent areas. Currently there about 60 projectswith various countries across a broad spectrum ofbiotechnologies. The department has establishedsuccessful collaborations with strategic partners likeAustralia, Canada, Denmark, Finland, Norway,Sweden, Switzerland, UK, EU and USA. It is alsoworking very closely with non-governmentalfunding organizations such as Wellcome Trust, UK.The bilateral collaborations were actively pursuedand significant achievements were seen in theprogrammes. The DBT collaborated with the firstEra-net project (European Research Area) namedNEW INDIGO, aimed at fostering andcoordinating scientific cooperation between EuropeanResearch Area and India. The project was launchedwith the announcement of a joint call for proposalsin the area of biomarkers and diagnostics;bioinformatics for health; and structural biology forhealth. The Wellcome Trust-DBT India Alliancewas established to nurture talent of the highestinternational standards, in cutting-edge biomedicalresearch in India, leading to important scientificbreakthroughs and applications to society. The mainfunction would be to award fellowships toscientists. The University of Wisconsin-Madison(UW), the DBT, and the Indo-US Science andTechnology Forum (IUSSTF) have partnered tosupport the prestigious Khorana Program forScholars named in honour of Dr. Har GobindKhorana.Autonomous InstitutionsThe established institutions of DBT and the ‘newgeneration’ institutions being established are creatinga new synergy in taking forward the research leadsto their end points in terms of processes andproducts. It marks a rather bold experiment inAnnual Report 2009-2010 15

Overviewmoving towards translational research and designinga portfolio of research to attain excellence in thevarious sectors.National Institute of Immunology, New DelhiAmong the various projects taken up during theyear, mention must be made of the focus onapoptosis-based therapies for various tumors usingestrogen-induced modulation of death pathways inhuman macrophages; modulation of acetyltransferaseactivity in M. tuberculosis; presence of novel HIV-1 LTR and VPr sequences in North Indianpopulations and siRNA-mediated knockdown ofSPAG9 expression in reducing cancer cell growth.A new invention was the Sustained Insulin ReleaseFormulation (SIRF) for treatment of diabetes. Itwas found promising in studies in diabetic modelsof mice, rats and rabbits. The method has thepotential to be a long lasting treatment for diabetesand halting the advance of secondary diabeticcomplications. The PhD programme continued tobe a very attractive one and contributed significantlyto generation of trained manpower. Fifty six peerreviewed papers were published.National Centre for Cell Science, PuneIn addition to serving as the National CellRepository, NCCS focused on human resourcedevelopment and R & D in the areas of cell biology,signal transduction, cancer biology, diabetes, infection& immunity, gene regulation, stem cells andregenerative biology. The NCCS provided 1980cell lines of various types to scientific institutions inthe country. The centre developed novel methodsto maintain viable stem cells and also succeeded indifferentiating these into specific lineages, forpotential applications in cell/tissue replacementtherapy. The researchers at NCCS identified aspecific non-coding RNA that is important fortumorigenesis. In recent years, the focus has beenon cancer as a stem cell disease. Studies have shownthe presence of a subset of cancer stem cells thatcontribute to tumour dormancy and resistance tochemotherapy. The Culture Collection at NCCSgreatly aided the conservation and utilization ofbiodiversity of India. The microbial distribution invarious habitats was studied using the metagenomicapproach. NCCS published 60 papers in peerreviewed national and international journals.National Brain Research Center, GurgaonThe NBRC continued to demonstrate its excellencein brain research. The major areas identified forresearch include computational neuroscience,cognitive neuroscience, stem cell research, anddevelopmental neurobiology. An increasing numberof students with a background in biological sciencespursued research in computational neuroscience areas,and a good number from the physical or engineeringsciences background took up research inneurobiology. Work at the centre has shown thatthe expression of mutant Huntington’s diseaseresulted in massive transcriptional induction of severalchemokines as MCP-1 and KC. In another study,the methodology and mechanism of restoration ofblood-brain barrier integrity by minocycline-basedtreatment of Japanese encephalitis was elucidated.Investigations carried out on retinal prosthesisshowed that photoreceptor degeneration led to risein synaptic proteins in amacrine cells, suggesting anenhanced inhibitory neurotransmission. Harnessingof adult neural stem cell/progenitor cell bymodulating the opioid system by naloxone was16Annual Report 2009-2010

Overviewdemonstrated through studies in animal systemsand zebra finches. NBRC and the Salk Institute,San Diego organised an International Workshop onCognitive Neuroscience at NBRC which wassponsored by the International Brain ResearchOrganization (IBRO). The DELCON centre atNBRC functioned as the digital resource centre forall institutions of DBT.Centre for DNA Fingerprinting andDiagnostics, HyderabadThe centre continued to provide its valuable servicesto assist law enforcement agencies in addition toimplementing its research portfolio. A total of 91cases were received for DNA fingerprintingexamination. Out of these, 49 cases were related topaternity/maternity disputes while the others werelargely related to the identification of the deceased.A total of 1664 patients attended the clinic forgenetic evaluation and counselling. At the instanceof APEDA, the APEDA-CDFD centre wasestablished for quantification of adulteration ofBasmati rice. Based on the novel technology ofCDFD, methods were developed for thequantification.National Institute of Plant Genome Research,New DelhiThrough its research agenda the institute contributedvery significantly to nurturing research in plantgenomics and various aspects of plant molecularbiology. Isolation of genomic SSRs led to thedevelopment of about 300 novel STMS markers inchickpea. Two full-length cDNA libraries of chickpeawere generated and the cDNA libraries that wereconstructed yielded more than 1500 unigenes. Inwheat, three phenologically plastic lines were intercrossedto pyramid the genes favourable for tandemcropping. F 2generations were developed from thesecrosses for testing. Two ripening modifying enzymesN-glycoprotein, α–mannosidase and α-D-Nacetylhexosaminidasewere used for enhancing fruitshelf life by reducing the rate of softening intransgenic tomato RNA interference lines. The shelflife of the transgenic tomato was seen to be enhancedupto 30 days.Institute of Life Sciences, BhubaneswarThe institute’s focus included infectious diseases,aspects of gene regulation with emphasis ontranslational aspects. Research on the biology ofinfectious disease found an association betweenmannose binding lectin mutations and susceptibilityto filariasis. Studies on V. cholerae indicated aphylogenetic relationship among non toxigenic V.cholerae O1 biotype E1 Tor strains isolated fromIndia, Guam and Brazil. A screening programmewas undertaken for bio-molecules frommicroorganisms collected from different ecologicalniches. A thermophilic bacterium was isolated froma hot spring and another one isolated from a sulfurspring. Gene mining for salt stress toleranceindicated the association of glycinebetaine and prolinein salt tolerance in S. maritima.Institute of Bioresources and SustainableDevelopment, ImphalThe IBSD continued its endeavor towards utilizationof the rich bioresources of the North East regiontowards development of products and processes.The institute joined the DBT & Electronic JournalConsortia (DELCON) which gave it access to moreAnnual Report 2009-2010 17

Overview800 international research journals. A germplasmrepository of the family Zingiberaceae was enhancedwith new collections. The institute is involved in anetwork programme on screening of bacteria froma wide range of environments for biomolecules oftherapeutic value. Induced breeding technique forBengana devdevi (Khabak) was standardized andtransferred to fish farmers.Rajiv Gandhi Centre for Biotechnology,ThiruvananthapuramThe emergence of RGCB as a national instituteunder DBT’s umbrella has led to a redefining ofits R&D programmes into two major streams i.e.disease biology and plant biotechnology. The missionof the disease biology programme is to understandthe underlying causes of human diseases. Cancer,atherosclerosis & heart disease, tuberculosis, viralinfections, cholera, and neurological disorders arebeing studied. Application of modern technologiessuch as high throughput sequencing, DNA barcoding,synthetic biology, and nano-biotechnologyare focussed towards delivery systems, andunderstanding the fundamentals of cellular functionsduring disease. RGCB has set up an IARC-WHOsponsored vaccine efficacy centre for clinical trial ofthe vaccine for Human Papilloma Virus, supportedby the Bill and Melinda Gates Foundation.UPCOMING INSTITUTION /CENTRESInstitute for Stem Cell Biology, BangaloreThe institute (InStem) is being established inBangalore in partnership with CMC, Vellore. Theprogrammatic linkages are being developed withhospitals and academic institutions. The first brainstorming session organized by the institute was oncardiac hypertrophy and stem cells. A collaborationhas been developed among InStem, NCBS andStanford University. The institute has established anindustry interaction unit, and the Centre for Cellularand Molecular Platforms with NCBS.Translational Health Science & TechnologyInstitute, FaridabadThe activities of the Translational Health Science &Technology Institute, an autonomous institution ofDBT, were launched after the necessary approvalswere obtained. Discussions were held with nationaland international experts to conceptualize the designof the cluster at Faridabad and steps were taken toinitiate the activities of the Health Science Technology(HST) component with MIT. Niche centres are beingestablished under the THSTI. These are Vaccine &Infectious Disease Research Centre (VIDRC),Paediatrics Biology Centre (PBC) and NationalDevelopment (Clinical) Service Agency (NDSA).National Institute of Biomedical Genomics,Kalyani, West BengalThe institute is established as an autonomousinstitution under the DBT, at Kalyani, West Bengal.The major objectives of the institute are to promotebasic research through biomedical genomicapproaches and establish state-of-the-art infrastructurefor genomic and proteomic analyses. The GoverningBody, Scientific Advisory Committee, FinanceCommittee and Building Committee have beenconstituted. The Government of West Bengal hasprovided 30 acres of land for building the institutein addition to 120,000 sq. ft. of quality space atKalyani, as an interim facility. The activities on theInternational Cancer Genome Consortium-IndiaProject are being launched.18Annual Report 2009-2010

OverviewRegional Centre for Biotechnology, Faridabad,HaryanaThe center would act as the focal point forcooperation amongst the member countries of theAsian Region in biotechnology. A Board ofGovernors with representatives from India,UNESCO and the member countries would provideoversight for the programmes. The research anddevelopment activities have been initiated towardsstrengthening knowledge based drug discoveryapproaches and the study of complex diseases foridentification of intervention points. Recruitmentof the faculty, technical staff and administrativestaff is in progress. The interim facility of thecentre is being set up in Gurgaon, Haryana.National Agri-Food Biotechnology Institute,MohaliThe NABI would focus on a designer crop breedingprogramme and address areas of stress tolerance andproblems related to drought, post harvest losses andnutrition enhancement. With a focus on novel foodprocesses and products it will use the locationaladvantages. The Bio Processing Unit and the AgrifoodBiotech Park will generate a synergy througha cluster approach. The NABI will have a partnershipwith the BPU and the park to promote bioentepreneurship and start up companies. TheExecutive Director of NABI has been selected andother positions are being filled in. An interim facilityhas been set up for NABI and the BPU in Mohali.BioclustersThe Bangalore bio-cluster is being set up in theGKVK campus of the UAS, Bangalore. The clusterwill comprise the National Centre for BiologicalSciences (NCBS), the Institute for Stem Cell Scienceand Regenerative Medicine (InStem) and the Centrefor Cellular and Molecular Platforms (C-CAMP).InStem which is adjacent to the NCBS, willencourage inter-disciplinary work with specialemphasis on translational and clinical stem cellresearch. It will build on the existing links with theCMC-DBT Centre for Stem Cell Research at CMC,Vellore. The, C-CAMP will act as the provider anddeveloper of technology. The Health Science clusteris being set up at Faridabad. The Faridabad clusterwill have multiple institutions like THSTI and itsniche centres ie., the Vaccine & Infectious DiseaseResearch Centre (VIDRC), Paediatrics Biology Centre(PBC) and National Development (Clinical) ServiceAgency (NDSA). The Faridabad cluster will be anew model of educational institutions comingtogether with advanced research centres for productdevelopment.Public Sector UndertakingsThe Bharat Immunologicals and BiologicalsCorporation Limited, (BIBCOL) has set up itsnew production facilities for manufacturing zincdispersible tablets, employing the technologytransferred from M/s Nutiset, France. After obtainingthe manufacturing license and GMP certification,BIBCOL is engaged in the manufacturing andsupplying of zinc dispersible tablets to the StateGovernments under a project “Large scaledemonstrative studies of zinc in the management ofdiarrhea as an adjunct to oral rehydration solutionin high burden Indian States”. The restructuringprocess of IVCOL was undertaken to effectivelyutilize the assets created. Accordingly, a facility ofthe National Brain Research Centre (NBRC) wasestablished in a portion of the IVCOL premises.Annual Report 2009-2010 19

OverviewInternational Centre for GeneticEngineering and BiotechnologyICGEB through its various programmes continuedto promote biotechnology and genetic engineeringresearch in the member countries. The centretransferred a technology on ‘Dengue specificantigens and monoclonal antibodies’ to an industrialpartner in India and filed new patent applicationsincluding PCT applications. A genome-wide siRNAscreening was done to identify host factors thatregulated pathogen load in human macrophagesinfected with a virulent strain of Mycobacteriumtuberculosis. About 275 such molecules wereidentified to be functionally associated with eachother through a dense network of interactions.Under a program on development of a vaccine forPlasmodium vivax malaria, methods to producerecombinant PvRII were developed. Clinical gradeJAIVAC-1 was manufactured under cGMP byBharat Biotech, Hyderabad. Proteomics basedapproach was used to investigate the mechanismof action of antimalarial drug, artemisnin. Anantibiotic marker free system based on theglyoxalase system for the selection of transgenicsin rice was developed. These efforts resulted in‘engineered’ rice over-expressing glyoxalase I andII, vacuolar antiporter and PDH45 genes. A fewtransgenic events were transferred to seedcompanies for validation.Promotion of Biotechnology in NorthEastern States of IndiaDBT has earmarked 10% of its total budgetamounting to Rs.100/- crores for the year 2009-10to promote these activities in the North EasternRegion. The region is endowed with immenselyrich bioresources which hold a lot of promise forprocess and product development that can contributeto the development of the region. The establishmentof Regional Research Centre of IBSD in Sikkim,the biotechnology incubator in Assam, a programmebased support to the Bioresources DevelopmentCentre in Shillong, the establishment of 25 centersof North Eastern Bioinformatics Network(NEBINet) and the NER- DelCON E-Journalsaccess to NER institutions are important initiativesthat were taken up for the development of theregion.New Legislations and RegulationsRecognizing the need to create an appropriatelegislation to regulate various issues relating toDNA profiling and establishment of DNA databanks, quality control, obligations and accreditationof DNA laboratories, the DBT set up a DNAProfiling Advisory Committee (D-PAC) inDecember 2003 to make recommendations forenacting a suitable legislation. The draft Bill wascirculated to the relevant Ministries andDepartments of the Central Government to obtaintheir comments. It was revised keeping in viewthose comments. The draft Bill will be sent to theDepartment of legal Affairs, Ministry of Law &Justice for their vetting. The Protection andUtilization of Public Funded Intellectual PropertyBill, 2008 approved by the Cabinet on October30, 2008, was introduced in the Rajya Sabhaon 15 th December, 2008. The Bill is nowunder consideration of the Department RelatedParliamentary Standing Committee on Science &Technology, Environment & Forests.20Annual Report 2009-2010

OverviewAdministration, Establishment andFinanceThe administration, establishment and financedivisions of the department rendered support inthe implementation of various programmes. Thedivisions also provided support for various scientificprogrammes and meetings. Modern multimediaequipment were procured and installed for effectivepresentation. To keep pace with the growth of thedepartment, new positions of Scientists, JAs andJTAs were created and action was taken to fillthem. Staff were also deputed for various trainingprogrammes to enhance their skills in relevant areas.The department also participated in extra curricularactivities and performed well in various events.Hindi fortnight was organized in the Departmentfrom 14 th -30 th September, 2009. Altogether 60officers/officials were imparted training in fourHindi workshops organized by the Hindi Section.The department was allocated an amount ofRs.1000.00 crores under the Plan head andRs. 24.00 crores under the Non-Plan head forthe year 2009-10. This was revised to Rs.902.00crores and Rs.23.90 crores respectively at theR.E. stage.Annual Report 2009-2010 21

CHAPTER 2HUMAN RESOURCE DEVELOPMENTAvailability of quality human resource holds thekey to success of R&D, production ormanufacturing in biotechnology. Realizing theimportance of well trained manpower, an integratedprogram of human resource development inbiotechnology comprising of postgraduate, doctoraland post-doctoral teaching, specialized training ofmid-career scientists, faculty involved inundergraduate and postgraduate teaching in leadingnational and overseas laboratories has been underimplementation for more than two decades.Considering the specialized needs of the biotechindustry, emphasis has been laid on involving thebiotech industry in imparting training topostgraduate students. To improve ‘hands-on’training to students, provision of ‘thesis grants’ toacademic institutions and bench fee to trainerindustry has been incorporated. The department hastaken special steps to improve ‘hands-on’ trainingduring undergraduate education by providing supportto selected colleges under ‘Star College Scheme’.Significant achievements under different programmesare given below:TeachingM.Sc. and M. Tech Teaching ProgrammePostgraduate teaching programmes were initiated insix universities in 1985-86 to generate a criticalmass of trained manpower and these have sincebeen expanded to 72 universities offering generalbiotechnology courses and specialized courses in areasof medical, agricultural, marine, veterinary, industrialand pharmaceutical biotechnology. Universities areselected for support by DBT on the basis of corestrength of their faculty, existing infrastructuralfacilities, R&D grants received on a competitivefunding basis, proximity to other institutionsengaged in biotechnology R&D etc. DBT providesliberal grants for establishment of specializedlaboratory infrastructure, equipment, recurring grantsfor consumables, studentship, publications, travel,contingency, thesis grant to ensure intensive ‘handson’training etc.Students for these courses are selected through aCommon Entrance Test (CET) conducted by JNU,New Delhi at 53 centers spread across the country.The selection is stringent with only one out of 70applicants getting selected. Students passing out ofDBT funded teaching programs are seen to do wellin competitive exams conducted by CSIR, UGC,DBT and ICMR for doctoral fellowships. Theprogram has resulted in retaining many students inthe country. More than 50% of M.Sc. students optfor Ph.D. within the country and 25% studentsopt for jobs in teaching, research and industry. Onthe other hand, more than 50% of the M.Tech.students were seen to join industries.During the year, the department supported anM.V.Sc. programme in animal biotechnology atAssam Agricultural University, Guwahati; GuruAngad Dev Veterinary and Animal SciencesAnnual Report 2009-2010 23

Human Resource DevelopmentUniversity, Ludhiana; Maharashtra Animal & FisheryScience University, Nagpur; and M.Tech.Biotechnology at IIT, Guwahati. A one timesupport was provided for the M.Sc. programme inMarine Biotechnology at Algappa University,Chennai. Altogether 16 new proposals received forconsideration of financial support for initiating PGteaching programmes in biotechnology were peerreviewed.Star CollegesAltogether 35 colleges offering undergraduateeducation in life sciences to improve the teachingof biotechnology were supported. These collegeswere provided a one time non recurring grant forequipment for all participating science departmentsand a recurring grant for a period of 2 years initially.The department has so far recommended 35 collegesunder ‘Star College Scheme’ (Table 1). The emphasisof the scheme is on improving knowledge andskills of teachers, provide access to specializedinfrastructure and make available consumables tostudents to improve hands on training, and alsofacilitate industrial exposure to students.Table 1 : Star College/InstituteS.No.Star College/Institute1. St Anthony College, Shillong, Meghalaya2. St. Edmund’s College, Shillong, Meghalaya3. Lady Keane College, Shillong, Meghalaya4. Goalpara College, Goalpara, Assam5 Kamla Nehru College For Women, Kapurthala, Punjab6 Govt. Mohindra College, Patiala, Punjab7 Govt. College of Science Education & Research, Jagraon, Punjab8 DAV College, Amritsar, Punjab9 Khalsa College, Amritsar, Punjab10 DAV College, Jalandhar, Punjab11 Govt. College, Bilaspur, H.P.12 Govt. College, Dharmashala, H.P.13 D. A. V. College, Chandigarh24Annual Report 2009-2010

Human Resource DevelopmentS.No.Star College/Institute14 Goswami Ganesh Dutta Sanatam Dharma College, (GGDSD), Chandigarh15 Gargi College, New Delhi16 Maitreyi College, New Delhi17 University Arts & Science College, Warangal, A. P.18 Nizam College, Hyderabad, A. P.19 Madras Christian College (Autonomous), Chennai, T.N.20 Stella Maris College (Autonomous), Chennai, T.N.21 PSG College of Arts & Science, Coimbatore, T.N.22. Vidyasagar College, Kolkata, W.B.23 Presidency College, Kolkata, W.B.24 Khallikote College, Berhampur, Orissa25 St. Xavier’s College, Ahmedabad, Gujarat26 Govt. Vidarbha Institute of Science and Humanities, Amravati, Maharastra27 Shree Manibhai Virani & Smt. Navalben Virani Science College, Rajkot, Gujarat28 College of Veterinary Sciences & Animal Husbandry, Srinagar, J&K29 Loyola College, Chennai, T.N.30 Lady Doak College, Madurai T.N.31 Thiagarajar College, Madurai, T.N.32 M.G. Science Institute, Ahmedabad, Gujarat33. Vidya Pratishthan’s, Arts, Science and Commerce College, Pune, Maharashtra34. S.S.V.P.S’s L.K. Dr. P. R. Ghogrey Science College, Dhule, Maharashtra35. Vallabh Government College, Mandi, H.P.Annual Report 2009-2010 25

Human Resource DevelopmentTraining ProgrammesShort term training courses for mid careerscientists and facultyThe proposals received for conducting short termtraining courses are evaluated by an expert committee.This programme has played a crucial role inupgrading the skills of mid career scientists fromresearch laboratories as well as the faculty involvedin undergraduate and postgraduate teaching.During the year, 35 proposals were received. Ofthese 24 proposals were considered and 19 proposalsrecommended. So far, 17 courses have beensanctioned.the past experience, choice and availability oftrainees, these trainees were placed in 110companies. A stipend of Rs. 6500/- p.m. is aavailable for the trainees and Rs. 50000/- to thecompany towards bench fee. The programme isbeing implemented through the BiotechConsortium India Ltd. An industry meet‘Bioindustry Conclave’ was organized on 9 th April,2009 which was attended by representatives fromindustry, academia, BCIL and DBT.Biotech Industrial Training ProgrammeThe Biotech Industrial Training Programme (BITP)has been supported for providing practical industrialtraining to fresh B.E./B.Tech./M.Sc./M.Tech.students in biotechnology for a duration of sixmonths. The programme is mutually beneficial tothe students and the industry as it providesindustrial exposure to the students and anopportunity to industry to select manpower fortheir requirements. Of the 501 trainees placed indifferent companies during 2008-09, 131 foundpermanent placements in the industries. Sectorwisedistribution of companies involved in BITPtraining is depicted in the pie chart. During 2009-10, 3062 applications were received from studentsfrom different universities all over India. Altogether,1537 candidates were shortlisted for interview and748 candidates selected. A total of 440 traineeswere placed for training in 110 companies.Requisition for placement of trainees has beenreceived from 172 biotech companies. Based onSector wise distribution of industries offering training in biotechnologyEntrepreneurship Development Programme inBiotechnologyTo train prospective entrepreneurs on differentaspects of business management and to promotesetting up of new commercially viable enterprisesrelated to biotech products, 8 entrepreneurshipdevelopment programmes are being organized atManipur, Shillong, Arunachal Pradesh, Sikkim,Assam, Mizoram, Nagaland and Tripura. These willbe organized in association with the respective StateS&T councils. It is proposed to impart training onbusiness management covering human resources,finance, marketing, management, sourcing ofcommercially viable technologies etc.26Annual Report 2009-2010

Human Resource DevelopmentTechnologists Training ProgrammesThe department has taken this new initiative toaugment the training of technicians and technologists,to meet the emerging needs of the industry andresearch institutes. Institutions and courses supportedunder the programme include the Advanced Diplomain Plant Genetic Engineering at MKU, Madurai;Post M.Sc. Advanced Diploma in MolecularDiagnostics at Alagappa University, Karaikudi;Advanced Diploma (post M.Sc.) in Plant TissueCulture and Micropropagation at Aligarh MuslimUniversity, Aligarh; Post M.Sc. Advanced DiplomaCourse in Molecular Diagnostics at BharathidasanUniversity, Tiruchirapalli and B.Sc. Diploma in PlantTissue Culture and Micropropogation at NorthLakhimpur College, Assam. The participants carriedout independent research projects in differentlaboratories and made notable research contributions.One of the successes of this scheme is the developmentof PCR based technique to detect the deadly humanpathogen Streptococcus pyogenes within a few hoursfrom a throat swab instead of 3-4 days. In order tomake the course industry-oriented, the participantswere encouraged to visit other research centres andhospitals for clinical-lab internship and project work.FellowshipsDBT Junior Research Fellowship (DBT- JRF)ProgrammeThe DBT JRF programme was implementedthrough the University of Pune. BiotechnologyEligibility Test (BET) was conducted at 10 centres.Altogether 8493 applications were received and 5100students appeared for the exam. A total of 100students were selected in the first category i.e. withthe option to join any institute and 150 studentsselected in the second category i.e. with the optionto join DBT supported teaching institutes or DBTautonomous institutes.DBT Postdoctoral Fellowship (DBT- PDF)ProgrammeThe DBT-PDF programme initiated in 2001 hasbecome hugely popular with students as evidentfrom the increase in applicants over the last fewyears. The programme is being implemented bythe IISc, Bangalore. Selection of students is basedon All India advertisement followed by presentationby the prospective candidates to the expertcommittee. The profile of selected candidatesrevealed that students from all parts of country getselected and are placed in premier institutions inthe country.Ramalingaswami FellowshipThe “Ramalingaswami Fellowship” scheme wasinstituted during 2007-08 as a re-entry grant forscientists of Indian origin working abroad in variousfields of biotechnology and life sciences, andinterested in taking up research positions in India.Under this scheme, a fellowship of Rs. 75,000 permonth is provided for a period of five years, inaddition to, a contingency grant of Rs. 5.00 lakhsper annum. The awardee can also apply for R&Dgrants from funding agencies through the hostinstitution. During 2009-10, about 130 applicationswere received and 27 scientists were selected. So far20 scientists joined various R&D laboratories/academic institutions.Tata Innovation FellowshipTo recognize the contribution of scientists in theAnnual Report 2009-2010 27

Human Resource Developmentarea of life sciences, agriculture, biomedical sciencesand other fields of biotechnology, who are engagedin discovery, innovation and invention, the DBTinstituted the scheme “Tata Innovation Fellowship”.Interdisciplinary work of high quality withemphasis on translation and innovation forms thebasis of the award. The award is given for aperiod of three years, and is extendable by twoyears. Each awardee receives a fellowship ofRs. 20,000/- per month, in addition to the regularsalary with a contingency grant of Rs. 5.00 lakhsper annum. During 2009-10, five candidates wereselected for the award. So far, 14 scientists haveavailed this fellowship.Distinguished Biotechnology ResearchProfessorshipThe DBT initiated the scheme “DistinguishedBiotechnology Research Professorship” in 2008-09to recognize eminent scientists who hadsuperannuated and have made outstandingcontribution in their respective fields. The schemeis intended to honour those scientists who are activein the country’s forum of senior eminent scientists.The professorship comprises of an award of Rs.60,000 per month, a research grant upto Rs. 20.00lakhs for a period of five years or till the awardeeattains the age of 70 years whichever is earlier.During 2009-10, four distinguished scientists wereselected for the award.DBT Biology Scholarships and AwardsDBT Biology ScholarshipThe DBT Biology Scholarships are awarded everyyear to students from the combined merit list ofBiology/Biotechnology at Higher Secondary/Intermediate/10+2 level to encourage them to pursuestudies in biological sciences. From the year 2007-08, the amount of scholarship was enhanced to alumpsum @ Rs.20,000/- per scholarship and thetotal number of students increased to 100 i.e. 44students from Central Board of Secondary Education(CBSE), 4 from the Council for Indian SchoolCertificate Examinations (CISCE) and 52 from StateBoards, (2 each from 26 State Boards). The awardalso carries a medal and a Certificate of merit.During the year 2008-09, 80 Students were selectedfor the scholarship.Innovative Young Biotechnologists Awards(IYBA)The scheme is intended, to identify and nurtureoutstanding young scientists with innovative ideasand desire pursuing research in frontier areas ofbiotechnology. The BTIC, DBT coordinates theprogramme. After an initial screening of submittedapplications by IYBA team, the applications arereviewed online by the expert committee followedby presentations by the short listed candidates.Awardees with a regular employment receive a cashaward of Rs. 1.00 lakh every year during the courseof the project in addition to a grant of Rs. 40.00lakhs for a project. A maximum of 25 awards aregiven away every year. Fourteen candidates havebeen selected this year for is award.Biotech Product, Process Development andCommercialization AwardsUpto five Biotech Product, Process Developmentand Commercialization awards are given each yearin recognition of outstanding contributions of28Annual Report 2009-2010

Human Resource Developmentscientists / innovators / entrepreneurs/ Indianinstitutions & companies, both in the public aswell as private sector towards a new process, productdevelopment and commercialization of a technology.Each award carries a cash amount of Rs.1.00 lakhalong with a citation. The awardee should be acitizen of India, and Indian institution or an Indiancompany should be involved in the developmentof biotechnology processes / products andcommercialization of technologies based onindigenous research. Out of 23 nominationsreceived, one awardee was selected.National Women Bioscientist AwardThe National Women Bioscientists Awards are givenevery year for two categories. These are (i) NationalWomen Bioscientist Award - senior category (one) tobe awarded to a senior women biologist for life timecontributions and (ii) National Women BioscientistsAward - young category (two) to be given foroutstanding contributions of women scientists below45 years of age in basic and applied research inbiosciences and biotechnology. Each award carries acash prize of Rs. 1.00 lakh along with citation anda medal. Out of 19 nominations in the senior categoryone was awarded and out of 20 nominations underthe young category, two were selected.National Bioscience Award for CareerDevelopmentThe National Bioscience Awards for CareerDevelopment are given in recognition ofoutstanding contributions of young scientists below45 years of age in basic and applied research in theareas of biosciences and biotechnology withpotential for application/product and technologydevelopment. Each award carries a cash prize ofRs.1.00 lakh and a research grant of Rs.9.00 lakhsfor three years. Out of 50 nominations received,10 were selected for the year 2008.Biotechnology Overseas AssociateshipBiotechnology Overseas Associateship andAssociateship for Specialized Training of YoungScientists in Niche Areas of BiotechnologyThere is a need to meet the ever-increasing demandfor skilled manpower and capacity building inbiotechnology. Towards capacity building andgeneration of a pool of technically trained humanresources, the Biotechnology Overseas AssociateshipProgramme was initiated. For nearly 25 years, theprogramme has helped to create a resource base thathas emerged as one of India’s major strengths andhas contributed significantly to the development ofthe Indian biotech sector.Visiting Scientists from Abroad ProgrammeUnder the Visiting Scientists from AbroadProgramme (VSAP), eminent scientists working infrontier areas of biotechnology in research laboratoriesabroad are invited for a period upto three monthsto research institutions in India to take upcollaborative research, conduct training courses andparticipate in teaching. Four scientists visited Indiafrom Japan, Canada and USA and worked withNBRC, CDFD, UAS and IISc.Rapid Grant for Young Investigator(RGYI) SchemeThe Rapid Grant for Young Investigator (RGYI)Annual Report 2009-2010 29

Human Resource Developmentscheme was under implementation since 2005-2006in order to give young scientists (below 40 years) achance to get their first grant as principal investigatorand take forward innovative ideas in the areas ofbiotechnology. The scheme was initiated to releasefunds in a fast track mode i.e. within eight weeksof the last date of submission of the proposal.More than 750 proposals were received in threebatches under this scheme, and about 240 projectswere implemented.Biotechnology EntrepreneurshipStudent TeamsDBT initiated the Biotechnology EntrepreneurshipStudent Teams (BEST-India) Programme aimed atencouraging young postgraduate and doctoralstudents in developing biotechnologyentrepreneurship by exposing them to issues incommercialization of bio-science. Altogether, 130concept notes in the area of life sciences that havea potential for commercialization were received. Fromthese, 20 teams were selected to attend a workshopdesigned to develop business skills and getexposed to finance and funding, IP protection,communication and HR management. Theworkshop was conducted by Association ofBiotechnology Led Enterprise (ABLE) India. Theproposals from 20 participating teams wereevaluated for their techno-economic feasibility/commercialisation potential; innovativeness; marketpotential; and level of motivation. Three teamswere adjudged as winners to receive the prize moneyof Rs. 5 lakhs, Rs. 3 lakhs and Rs. 2 lakhs for thefirst, second and the third team respectively. Thefirst prize went to the team from Stanford IndiaBio Design, the second prize to the team from IITKharagpur and the third prize to the team fromIIT Chennai (Fig).The wining team from Stanford India Bio Design, comprising ofMr. Shitij Malhotra, Dr. Mansi Aggarwal and Pooja Singh receivingthe award in the presence of Dr. Kiran Majumdar Shaw.Biotech PopularizationThe objective of biotechnology popularizationprogramme is to promote biotechnology and itsapplication amongst students, scientific communityand the general public. During the year 2009-10,the department participated in two exhibitions inIndia and one international event held in USA.Sixty four national and international seminars/symposia/conferences were supported and onehundred and nineteen scientists were recommendedfor international travel.Support for PublicationsThe Department of Biotechnology supported theWaterfall Institute of Technology Transfer, NewDelhi for publication of bimonthly issues of IPRbiotechnology and All India Biotech Associationfor publication of a bimonthly newsletter.30Annual Report 2009-2010

CHAPTER 3BIOTECH FACILITIES, TECHNOLOGYPLATFORMS AND RESEARCH RESOURCESCentres of ExcellenceThe scheme was designed to augment andstrengthen existing institutional capacity forpromotion of excellence in interdisciplinarysciences and innovation in specific areas. Theprogramme provided a flexible model of longtermsupport for highly innovative research, bothbasic and translational to create high-qualitypublications and intellectual property. The salientachievements of some of the centers supported sofar are as follows:Genetics and genomics of silkmoths at CDFD,HyderabadEight transgenic silkworm lines resistant tobaculovirus (BmNPV) were developed.Transgenic silkworms had already crossed sevengenerations in laboratory trials carried out incollaboration with APSSR&DI, Hindupur. Theselines were used as ‘donor parents’ to introgressthe transgenes to productive silkworm varietieswhich are currently popular among sericulturefarmers (Fig).Coordinated research on tuberculosis:Development of alternate strategies jointly atICGEB, New Delhi and UDSC, New Delhi.Host-specific siRNA screening was completed againstall kinases and phosphatases in murine macrophagesinfected with H37Rv. The hits were validated andSchematic diagram showing transfer of baculovirus resistance gene fromtransgenic Nistari lines to commercial CSR2 linesAnnual Report 2009-2010 31

Biotech Facilities, Technology Platforms and Research Resourcesscreened in cell infected with two separate multipledrugresistant strains of Mycobacterium tuberculosis.It has led to the identification of those host targets,the depletion of which led to elimination of bothdrug-sensitive and drug-resistant strains of M.tuberculosis and promises a new approach to TBchemotherapy.Novel paradigms of inhibitor design againstkey metabolic pathways to decimate infectiousagents at NII, New DelhiWork on the biochemical and biophysicalcharacterization of the M. tuberculosisdephosphocoenzyme A kinase (CoAE), an enzymeessential for the pathogens survival was completed.It was shown that the CoAE may be used as apotential drug target. Biotin protein ligase and biotincarboxyl carrier protein genes were cloned, expressedand characterized (Fig).pearl millet, sorghum and groundnut for developingand enhancing genetic maps of these species. Morethan one million SSR data points were generatedon several crops for diversity analysis / fingerprints,linkage map construction, QTL identification andmarker-assisted breeding. A total of 137 Indianscientists were trained on application of molecularmarkers and genomics for crop improvement.Tuberculosis research at IISc, Bangalore; BITS,Pilani and the Foundation for MedicalResearch, MumbaiThe role of magnesium ions in Mycobacteriumsmegmatis topoisomerase I activity was elucidated.A new microtitre plate assay was developed formycobacterial topoisomerase I for rapid screeningand identification of inhibitory monoclonalantibodies. The structures of three new crystal formsof RuvA from M. tuberculosis were determined.Research on Hepatitis C Virus at IISc, Bangalore,ICGEB, New Delhi, UDSC, New Delhi andACTREC, Navi MumbaiMycobacterial CoAE interacting with DCoA and the docked ligandHigh-throughput allele determination formolecular breeding at ICRISAT, Patancheru,Andhra PradeshThe first set of arrays were developed and used forA small structured hepta – peptide derived from ahost-factor was found to inhibit translationalreplication of HCV RNA. A patent application hasbeen filed in US. Some herbal extracts and purenatural compounds inhibiting HCV RNAreplication in cell culture system were identified.An efficient delivery of anti-viral agents into mouseliver using sendai-virosome to block HCVreplication was achieved (Fig).Stem Cell Research: Basic and Translational atAIIMS, New DelhiProtocols for isolation, expansion and32Annual Report 2009-2010

Biotech Facilities, Technology Platforms and Research Resourcessusceptibility markers/genes and their putativeinteraction.The picture represents a 7mer structured peptide entering liver derivedcell. It is thematically indicated that the peptide derived from humanLa protein when tagged with hexa-arginine can enter Huh7 cellsand block IRES mediated translation and subsequently inhibitreplication of hepatitis C virus RNA. (The illustration is adaptedfrom the picture published in cover page of Journal of VirologyDecember 2008 and described in the article by Mondal et al in thesame issue.)cryopreservation of adult stem cells were standardizedand validated. Procedures for mesenchymal stemcell isolation from human adult bone marrow andumbilical cord were optimized. Variousbiodegradable polymers for culture of stem cellswere prepared.Genome Science and Predictive Medicine atUDSC, New Delhi; NII, New Delhi; Universityof Delhi, Delhi; IASRI, New Delhi; AIIMS,New Delhi; Dayanand Medical College &Hospital, Ludhiana and S.V College, New DelhiEfforts were initiated to unravel the genetic basisalong with translation of knowledge to developnovel diagnostic and therapeutic tools forrheumatoid arthritis (RA) and ulcerative colitis (UC).The recruitment of over 2500 well-characterizedpatients was completed. Genotyping of ~670,000SNP/CNV markers in 1500 samples (500 each ofRA, UC and controls) using the Illumina Infiniumtechnology was done for identification ofMultidisciplinary approaches aimed atinterventions against M. tuberculosis at UOH,Hyderabad; ILS, Hyderabad; CDFD,Hyderabad; NIN, Hyderabad; LEPRA-BPRC,Hyderabad and Bhagwan Mahavir Hospital &Research Centre, HyderabadBased on the molecular docking studies, around100 structures were identified as hits which can besynthesized as promising inhibitors of chorismatemutase and these were subjected to further biologicalassays. Mtb IciA protein was shown to inhibit theDnaA-mediated in vitro helix opening by specificallybinding to A+T rich region of ori-C.Programme SupportInstitutional programme support is given to aninstitution/department with multiple investigatorsin various disciplines to strengthen researchcapabilities for a focus on a common researchproblem. The salient achievements of the ongoingactivities are:• For the first time, a soluble acyl-coenzyme-Adependent lysophosphatidic acid acyltransferasewas identified at the IISc Bangalore. Thisenzyme was shown to possess triacylglycerollipase as well as phosphatidylcholinehydrolyzing enzymatic activities.• Bioreactors were developed for tissueengineering of liver and cartilage tissueengineering at the Sree Chitra Tirunal Institutefor Medical Sciences & Technology,Thiruvananthapuram. An electrospinningAnnual Report 2009-2010 33

Biotech Facilities, Technology Platforms and Research Resourcesfacility was set up for the fabrication ofpolymer nanofibres. An oesteochondralscaffold was designed which is cost effective,easy to use and ensured immobilization atthe surgical site.• An antigen detection method wasdeveloped using monoclonal antibiodiesagainst filarial candidate antigen SXPexpressed both in baculovirus and E. coliat Anna University, Chennai. A simple andrapid virulence based colorimetric assay fordetecting Shigella and entero-pathogenic E.coli in stool was developed and tested withclinical samples.• Clinical trial (Phase I/II) of G-CSF therapyin patients with acute ischemic stroke wascarried out in 10 patients (5 interventions, 5controls) at AIIMS, New Delhi. A statisticallysignificant rise in the mean leucocyte countand alkaline phosphatase level was seen in thepatients when compared to the baseline andcontrol.• Polymorphism in homocystein (Hcy) pathwaygenes (MTHFR and CSF) were shown to begenetic risk factors for nonsyndromic cleft lipand palate through a study carried out atBanaras Hindu University, Varanasi. Screeningof the Hcy pathway genes (MTHFR, MTRR,CPS and RCF), levels of Hcy and the cofactorsVitamin B12 and folate showed highhamocyteinemia in more than 40% of thepopulation studied.• Investigations on the proteomics ofneurological disorders were initiated jointlyat Institute of Bioinformatics, Bangaloreand NIMHANS, Bangalore. Out of over800 proteins identified, 213 proteins werefound to be differentially expressed in thebrain of patients with tuberculosismeningitis, when compared to non-infectedbrain. Three potential biomarker proteinswere identified.• A programme on computational biology wasinitiated at the IIT, New Delhi. A multi-terafacility had become fully functional and twosoftwares were developed for drug design andgenome study.• Development of DNA vaccines for bacterialinfection in marine fish and white spotsyndrome virus (WSSV) in shrimp was takenup at the College of Fisheries, Mangalore.Four coat proteins of WSSV were cloned andtheir immunological response studied.• Under a programme on translational researchon transgenic rice at University of Calcutta,Kolkata, Bt-rice was developed with fused cry1Ab/Ac in the cultivars – IR64, Swarna andBR29, and insect bioassays were underway.Homozygous transgenic lines of IR64 andSwarna with -chi 11 gene were developed forresistance to sheath blight (Rhizoctonia solani).• Several transgenic lines of rice developed atICGEB, New Delhi using various stressrelated genes such as Rab7, Glyoxalasewith NHX, PDH45, SOD with APX haveshown enhanced salinity tolerance. Thesetransgenics were being evaluated for theirrelative drought tolerance. Efforts wereunderway on generating markers and reporterfreetransgenic rice plants.34Annual Report 2009-2010

Biotech Facilities, Technology Platforms and Research Resources• A programme covering the areas ofrecombinant antibodies, nanomaterial – cellinteraction, bio-electronic devices and heavymetal tolerance in plants has been initiatedat IIT, Guwahati. Gold-nanoparticlepolythiophenenanocomposite as well asiodinated chitosan silver nanoparticles that weresynthesized have demonstrated broad-spectrumanti-microbial activity. An enzyme-basedbiosensor fabricated with carbon nano-tubehas been developed for total cholesterolestimation in human serum.Long-Term R&D ProjectsIt aims to provide support to outstandinginvestigators of high scientific caliber withpublications and / or patents record. Three longtermR&D projects were supported during the year.The salient achievements of ongoing projects are:• More than 950 stool samples from childrensuffering from acute diarrhea were examinedfor non-polio enteroviruses at IISc., Bangalore.Between 12.5 to 15% of the children withdiarrhea were found to be positive forenteroviruses.• The Sendai virosome system was developedas a delivery vehicle for gene therapy ininherited jaundice in rat models which have adefect in the UGT1A1 gene jointly at AIIMS,New Delhi and UDSC, New Delhi.• The IdeR target genes associated with wellknown process of iron uptake transport andstorage in Mycobacterium were identifiedjointly at CCMB, Hyderabad and LEPRASociety, Hyderabad.Biotechnology Facilities and ResearchResourcesThe National Biotechnology Development Strategybeing implemented by the department recognizesthat public investment in major equipment/infrastructure required for research and developmentin biotechnology should be so directed that itsaccess and utility are maximized. It is also recognizedthat for translational research of both health carebiotech products and agri-biotech products,sophisticated infrastructure is required in the formtechnology platforms and research resources forregulatory tests, product scale-up, formulation,validation and development.To develop drugs and vaccines, non-human primatemodels are often required because it is safe toextrapolate their results to humans. The departmentis setting up a colony of genetically defined nonhumanprimates for conducting experiments onwhole animal systems using species that are closestto humans, at the NII campus II in Faridabad. Theplan will include open enclosures for macaques thatare gregarious animals and live in social groups.One type will be for the breeding units of macaques.Another will be the rehabilitation units to acclimatizebefore release into the wild those animals which areno longer suitable for research. The third categorywill be for housing the captivated bred primateswhich cannot be released into the wild. The primateanimal platform will provide services to other DBTinstitutions like NIPGR, New Delhi, NBRC,Manesar, and ICGEB, New Delhi.The Research Resources set up comprises humanresources and institutional components. Trained andspecialized scientists, well-equipped labs, experimentalAnnual Report 2009-2010 35

Biotech Facilities, Technology Platforms and Research Resourcesfarms, and other support services are woven togetherin institutional and organizational structures. Thereinclude the flowcytometer sorter facility at ILS, theP3 facility at JNU; Automatic DNA Sequencerand controlled environment growth chamber facilityat NIPGR; National Facility for Microarray Geneticsand Cell Imaging at IIT, Kanpur; facilities forstructural biology of proteins and nucleic acidfragments at University of Madras; and highthroughputgenomic analysis system at JNU. Anaerosol containment facility for slow growing andfast growing pathogens and microscopy cum flowcytometery unit were set up at NII, New Delhi.The DBT has jointly established with CSIR theInternational Depository Authority at IMTECH,Chandigarh. Besides IDA deposits, cultures forregular patent purposes and safe deposits aremaintained in the IDA. MTCC has more than12000 cultures and 38000 records on microbialdiversity of India. IDA has more than 200 patentdeposits. During the year 2009-10, about 30 patentdeposits were added.A GM product toxicology facility was set up atShriram Institute for Industrial Research, NewDelhi. The facility is expected to produce modulesfor toxicological evaluation for safety assessment onrets, mice, rabbit, and guinea pigs and large animals.The facility provides a 50% rebate to DBT funded/recommended projects for toxicological evaluation.So far about 16 users from various DBT projectshave availed the facilities.A repository for filarial parasites and reagents wasset up at Mahatma Gandhi Institute of MedicalSciences (MGIMS), Sevagram, Maharashtra. Theactivities of the facility include maintenance of B.malayi filarial infection in rodents, storage ofdifferent stages of B. malayi parasite, isolation of W.bancrofti microfilariae from field samples fromdifferent regions, immunoscreening of filarial cDNAlibraries to identify, clone and express genes ofdiagnostic and prophylactic importance. Analysis ofgene polymorphism and development of in vitroand in vivo anti-filarial drug screening are alsoundertaken. The facility also holds a filarial parasitebank with about 40 million B. malayi mf, 500male and female adult worms (from infectedanimals) and 1000 infective larva (collected frominfected mosquitoes) and more than 2000 W.bancrofti mf. The filarial serum bank has a collectionof 504 bancroftian filarial sera of different patientgroups. The sera are supplied for diagnostic work.Platform for Translational Research onTransgenic Crops (PTTC)To provide an effective interface between the laband land, the DBT set up this innovative“Technology Platform at ICRISAT, Hyderabad.The platform leverages ICRISAT’s existingexcellence in the areas of transgenic research oncrop plants, molecular plant sciences and plantbreeding, to improve its ability to enhance thedelivery of transgenic crops in agriculture. Themain mission of this platform is to translatetransgenic technology and harness its potential tomeet the growing needs of agriculture.The major objective of the PTTC is to developsynergistic links with different research institutes,universities and industry offering various servicesincluding contract research on tissue culture andtransformation, product development along the valuechain, consultancy services, intellectual propertyadvice and management, and guidance on biosafety36Annual Report 2009-2010

Biotech Facilities, Technology Platforms and Research Resourcesissues. The design and construction of the buildingfor the platform will be completed by April 2010.Subject matter specialists have been recruited forthe different programmes. Various types of contractresearch agreements have been developed inconsultation with stakeholders. Projects contractson insect resistant chickpea, drought tolerant peanuthave been finalized. The platform is expected to befully operational in 2010.Platform Facility for MacromolecularCrystallographyThe DBT negotiated cost/time based sharingmechanism at BM14 beamline of EuropeanSynchrotron Research Facility (ESRF), Grenoble,France in partnership with Medical Research CouncilU.K and European Molecular Biology Laboratories(EMBL). The proposal aims to distribute beamtime to researchers who need access to synchrotronx-ray beam line for macromolecular crystal datacollection/structure determination.Boost to University Interdisciplinary LifeScience Departments for Education andResearch (BUILDER) SchemesThe department launched a new scheme to supportadvanced research and education in universitiesknown as DBT-University Interdisciplinary Schoolof Life Sciences for advanced research and education(DBT-ISLS) programme. The specific aim is toupgrade post-graduate teaching, and the laboratoriesin terms of infrastructure and equipment. There isalso a provision for new faculty, fellowships, newtraining programmes, and for promotion academiaindustryinteraction. The following initiatives havebeen supported.UoH-DBT Centre for research and education inbiology and biotechnology was funded at Universityof Hyderabad. The salient features include theestablishment of the state-of-art laboratories, andthe strengthening and upgradation of labs. Theestablishment of facilities include biologicalcontainment facility, animal house, culture facilitiesand central facility for research. Six state-of-artlaboratories for M.Sc and M.Tech programmes wereestablished. There was steep increase in the numberof students admitted to the courses of M.Sc.,M.Tech and Ph.D., besides post doctoral fellows.Sixteen workshops were conducted. Six technicianswere trained on the use of the sophisticated researchfacilities. State-of-the-art facilities have been createdin the areas of 2-D gel electrophoresis, imagingsystems, proteomics, functional genomics, microarrayand radioactivity measuring facility. There was anincrease in the number of research projects (from

CHAPTER 4RESEARCH AND DEVELOPMENTA sector wise approach to R&D ensures the fosteringof research initiatives across a wide range of topicsthat have immense potential in generating usefulleads. A suite of options are employed in carryingout these researches. This chapter captures theessentials of the largest thematic efforts of thedepartment i.e. Research and Development. Theseare tailored towards production and productivityenhancement in commodity species, developmentof technologies with a strong underpinning ofmanpower development and capacity building.Agricultural BiotechnologyCROPSRiceA project on identification of molecular markerslinked to quality parameters in rice, validation andutilization in marker-assisted selection (MAS) wastaken up at DRR, Hyderabad. The interestingdevelopments were in the polymorphic surveyamong parents. A total of 239 markers were foundpolymorphic out of 575 markers tested.Phenotyping for kernel length, kernel length aftercooking and alkali spreading value was completed.Genetics of kernel length and kernel length aftercooking revealed a polygenic mode of inheritancewhile alkali spreading value showed monogenicrecessive. Association study revealed association ofSSS1-SSR (linked to soluble starch synthase 1enzyme) and SBE1-SSR1 (starch branching enzyme1) with intermediate amylose content; SBE1-STS1(starch branching enzyme 1) with low ASV. Alarger set of population will be used to validate theabove promising markers. Ten crosses were madebetween parents with contrasting grain and cookingquality traits and the F 2were grown to developmapping populations.Under a project on cytological screening andmolecular characterization of thermo sensitive genicmale sterile lines for developing inter-subspecifichybrids in rice at TNAU, four TGMS lines werefound promising for TGMS expression along withcompact plant type, desirable grain quality and wellexerted purple stigma with medium durationtype. In order to assess the critical fertilitytemperature (CFT)/ critical sterility temperature(CST) of promising TGMS lines, sowing was doneat monthly intervals. In another project onidentification of DNA markers for enhanced proteincontent and development of QTL introgressedgenotypes for high protein content in rice, a highprotein local genotype from germplasm with 14percent protein was crossed to BPT 5204 andprogenies developed for developing RILs. Fieldevaluation of RILs was carried out in dry and wetseasons-2009 and several high yielding fine graingenotypes were identified with varying proteincontent. Total protein content of selected populationsvaried from 5 to 22 percent.Annual Report 2009-2010 39

Research and DevelopmentAt AAU, Jorhat a set of 203 indigenous glutinousrice cultivars were collected from various agriculturalstations, farmers’ fields, including four japonica varietiesfrom IRRI, Philippines. Two different crosses weremade in order to develop a mapping population forunderstanding the genetic base of waxiness inindigenous glutinous rice of Assam. At JamiaHamdard University, 110 rice genotypes were analyzedat various levels of nitrogen treatments for selectingnitrogen efficient and nitrogen-inefficient genotypes.Three levels (low, medium and high) and two sources(nitrate and ammonia) of nitrogen were supplied tohydroponically grown rice genotypes, and growthperformance, nitrogen distribution in various plantparts and nitrogen efficiencies were measured. Onthe basis of these parameters, three categories of ricegenotypes were identified.At TNAU, Coimbatore, molecular tagging ofthermo-sensitive genic male sterile gene fordevelopment of new TGMS lines in rice wasundertaken and identified markers viz., RM 3476and RM 341 were used for surveying the individualplants of F2/F3 population. RM 3476 markers cosegregatedwith the phenotypic observations recordedunder the field condition. The tight linkage ofmarker with TGMS gene was identified by RM3476 a marker located adjoining tms 6 gene inchromosome 5 of rice. The stability of TGMSsegregant lines in F4 generation was ascertained bycrossing with tester parents (Fig).Bring is in centre Field View of two line hybridsP1P2SterileFertileIndividual plant (F3) screening with RM 3476TGMS segregant (F4) withgood floral traits favouringoutcrossingA single new superior TGMSsegregant (F4)40Annual Report 2009-2010

Research and DevelopmentWheatA project was implemented at GNDU, Amritsaron transgenic analysis for the role of wheat gene(TaVAP) encoding for a vesicle-associated membraneprotein-associated protein in abiotic stress tolerance.The TaVAP full length gene sequence was found tobe around 997 bp and ORF of the TaVAP genearound 717bp long. For successful isolation of theprotein with MBD and His-Tag labels, cloning ofthe PCR-amplified TaVAP sequence was carried outin pMAL-p4X and pET23a vector. TaVAP proteinwith His-tag label was successfully induced in theheterologous E. coli system by IPTG. Floral-Dipwas carried out for transformation. The seed fromtransgenic plants germinated but the seedlings diedon the selection medium containing kanamycin.The vector transformed control seedlings havingonly the native pBI121 vector survived at 1-2 %frequency.In another project on identification andcharacterization of differentially expressed genesduring heat stress in wheat at UDSC, New Delhi,wheat plants (Triticum aestivum cv. CPAN 1676)were given heat shock at 37 o C and 42 o C for twohrs, and responsive genes identified through PCRselectsubtraction technology. Three subtractivecDNA libraries were constructed from two differentdevelopmental stages. A total of 4500 ESTs weregenerated and about 3000 ESTs submitted toGenbank. A large number of high temperatureresponsive genes have been identified andcharacterized. Reverse subtraction analysis indeveloping grains showed extensive transcriptionalchanges upon heat stress as revealed by comparativeanalysis with forward subtraction. Heat tolerancerelated gene transcripts were identified based ontheir putative functions helping to unravel thecomplexities of heat stress response in wheat. Inan ongoing project at DWR, Karnal onimprovement of biscuit making quality of Indianwheat, a back cross programme was initiated totransfer Glu-D1 double null of Nap Hal intoPBW 502, PBW373 and HD2687 backgroundsusing a molecular approach. PCR analysis ofdifferent backcross populations were used inselecting desirable segregants for soft grain texture.Desirable recombinants were identified foradvancement to next generation. Micro level testsof gluten strength showed very weak gluten inplants with Glu-D1 double null. Marker assistedselection showed very promising result with respectto improving traits for biscuit making.MaizeAt PAU, Ludhiana genetic diversity amongQPM germplasm and non-QPM lines in maizewas studied using SSR markers. Also a surveyof polymorphism was done between QPM andnon-QPM lines for background selection, andback-cross population generated for transferringQPM trait into elite maize lines using MAS. AtUAS, Mandya, two sterility mosaic disease(SMD) resistant maize inbred lines (MAI 105and SKV 50) were selected and crossed to highlysusceptible inbred line CM 500 to sterility mosaicdisease to develop backcross mapping populations(CM 500 X (CM 500 X MAI 105 and CM500 X (CM 500 X SKV 50). The resistantparents, MAI 105 and SKV 50, were also crossedto another susceptible inbred line CM 202 andforage maize, African tall to incorporate resistanceto SDM employing foreground and backgroundselection.Annual Report 2009-2010 41

Research and DevelopmentAt CSK HP Agricultural University, Palampur aproject was implemented on molecularcharacterization of hill maize inbreds andprediction of heterosis using SSR and RAPDmarkers where RAPD, InDel and SSR markerswere able to assign the 82 maize inbreds todifferent groups. However, these markers showedrelatively limited usefulness in predicting singlecross hybrid performance and/or heterosis owing,primarily, to low genetic diversity among theparental inbreds.Vivek QPM9, an early maturing quality proteinmaize hybrid was developed through MAS atVivekanand Parvatiya Krishi Anusandhan Sansthan(VPKAS), Almora. Vivek Hybrid 9 is consideredto be one of the best maize hybrids developed byVPKAS and was earlier released for commercialcultivation in Uttarakhand, Himachal Pradesh,J&K, Andhra Pradesh, Karnataka, Tamilnadu andMaharashtra (Fig.). In order to popularize thenew QPM hybrid, seed production of VivekQPM 9 was undertaken on a large scale. Thetechnology has been licensed to private companiesfor large scale seed production (Fig).Vivek Maize Hybrid 9 and Vivek QPM 9Pigeon peaAt IARI, New Delhi, mapping population ofpigeon pea was developed and parentalpolymorphism study undertaken and a total of275 EST-SSR markers screened for polymorphism.Altogether, 27 SSR markers were foundpolymorphic between Pusa Dwarf and H 2001-4. Concerted efforts were made by Agrobacteriumtumefaciens-mediated genetic transformation ofchickpea and pigeonpea for resistance to thelegume pod borer, Helicoverpa armigera using sixdifferent gene constructs of cry1Ac with differentpromoters (ubiquitin or CaMV 35S). Over 200independent transgenic events of chickpea wereproduced.Oilseeds & other cropsIn a project on molecular marker assistedintrogression of Aspergillus flavus resistance andaflatoxin production into farmer preferred varietiesof groundnut, screening of 100 lines interspecificderivatives of groundnut for A. flavus infection andaflatoxin production was carried out by growingthe material in A. flavus sick plot. In anotherapproach, synthetic groundnuts (new sources of A.hypogaea), were developed at ICRISAT. These wereused in the crossing program to transfer A. flavus –aflatoxin resistance.In a project which aims at introgression of resistanceto late leaf spot from wild species to cultivatedgroundnut, eleven synthetic amphidiploids usingtwenty-one available inter-specific hybrids weredeveloped. Among them, ISAT 278-18 (A.duranesisICG 8138 X A.batizocoi ICG 13160), ISAT 1212(A.duranesis ICG 8123 X A.ipaensis ICG 8206)and ISATGR 265-5A (A.magna ICG 8966 X42Annual Report 2009-2010

Research and DevelopmentA.batizocoi ICG 8209) were found resistant to lateleaf spot (Fig.).Utilization of amphidiploid ISATR 265-5Towards achieving the objective of raising transgenicsfor abiotic stress tolerance, OsRLCK253 andOsSAP11 genes were functionally validated by overexpressionin Arabidopsis. These transgenics showedchanges in gene expression and tolerance to waterdeficit,salt and freezing stress, with decrease inyield penalty as well. In rice, 23 genes encodingsHSPs were identified by reiterative database search.These sHSPs represent proteins localized to differentcellular compartments and at least 19 of them areinduced by high temperature stress. To understandthe network of genes regulating somaticembryogenesis, the leucine-rich repeat receptor-likekinase (LRR-RLK) genes were identified from rice.To alter the agronomic traits like plant height andflowering time, the genes encoding blue light sensingphotoreceptors, CRY1 and CRY2, were overexpressedin Brassica juncea. These transgenics arebeing subjected to microarray analyses to identifygene networks involved in regulation of plantdevelopment by light and plant hormones(Fig.).XUtilization of amphidiploidISATR 265-5A. hypogaeaPods of F1 PlantsPlant Molecular Biology ProgrammesSurvival of wild-type (WT) and transgenic plants under high saltand water-deficit stress : (A), effect of salt stress on OsRLCK253plants and WT. (B), effect of salt stress on OsSAP11 plants andWT. (C), effect of water-deficit stress on OsRLCK253 plants andWT. (D), effect of water-deficit stress on OsSAP11 plants and WT.Three-week-old seedlings were treated with 600 mM NaCl solutionfor two hours or water was withdrawn from two-week-old potgrownplants. Plants were photographed after two weeks of recovery.WTUS, unstressed WT plants; WTS, stressed WT plants;OsRLCK253-24, stressed transgenic plants of line 24 of OsRLCK253over-expressing plants; OsSAP11-3, stressed transgenic plants of line24 of OsSAP11 over-expressing plantsProgramme Support for Research andDevelopment in Agricultural BiotechnologyProgramme Support was given to the followingState Agricultural Universities to bring about overallAnnual Report 2009-2010 43

Research and Developmentexcellence in agricultural biotechnology. Thehighlights of the progress are as follows:PAU, LudhianaThe programme support at PAU involves projectson heat tolerance in wheat and enhancement ofwater use efficiency in rice, resistance to stem borer,tolerance to abiotic stresses in maize, and developingresistance to Phytophatora & tolerance to salinity inrough lemon.Towards developing heat tolerance in wheat, oneentry (PBW657=C273/PBW 534) was taken tonational trials for late sown conditions. Doubledhaploid approach was used to generatehomozygous lines from selected crosses. A yieldtrial of 28 doubled haploids and an observationnursery of about 200 lines were planted in thecurrent season. BC 2F 1progenies derived from thecrosses of the elite durum cultivars and heattolerant Ae. speltoides accessions were phenotypedfor various morphological parameters. Polymorphicand SSR markers identified are beingamplified on the BC-RIL population. In rice, fordeveloping abiotic stress tolerance, transgenes viz.OsglyI, OsglyII and ZAT12 were introduced intothe commercial rice variety PR 116 throughparticle bombardment. A number of independent(PCR positive) transgenic plants have been grownto maturity. Further, introgression lines of Pusa44(21), PR 114(6) background and one IMBLline Feng-AI-ZAN were field evaluated. It wasfound that the performance of the introgressionlines was better under irrigated conditions ascompared to aerobic conditions (Fig). In citrus,for developing resistance against Phytophthora, achitinase gene has been cloned from Trichoderma.A gene construct with chitinase ORF under thecontrol of CaMV 35S promoter was developedfor transformation. Further, root stockRice transformationA. Scutceller-derived embryogenic callus with seedB. Embryogenic callusC. Traget plate used for bombardmentD. Selection of bombarded calli on kanamycinE. GUS expression 48 hrs after bombardmentRice transformationA. Hardening of putative transgenic plantsB. Transfer of putative transgenic plants in transgenic glass houseC-D. Establishment of putative plants in transgenic glass houseE.-F. Putative plants showing normal seed set.G. PCR amplification of Osgly II in transgenic rice plantsH. PCR amplification of Osgly I in transgenic rice plants44Annual Report 2009-2010

Research and Developmenttransformation was achieved at cellular/shoot budlevel and entire plants were generated.TNAU, Coimbatore:This programme includes nine sub-projects underdifferent thematic areas. For engineering resistanceagainst banana bunchy top virus (BBTV), 140putative transgenic plants were generated using RNAivectors prepared with replicase gene of BBTV genomeand 18 of them transferred to greenhouse and theremaining ones were in the regeneration and rootingmedium. For development of drought tolerant rice,22 QTLs linked to various drought tolerance relatedtraits were identified and efforts made to developIR20 and IR64 NILs by introgressing three QTLsassociated with root traits and yield under stress. Aset of 400 BC 4F 3IR20 NILs developed throughmarker assisted breeding were in the field for testingdrought tolerance. Through participatory breeding aSomatic embryoShoot and root developmentGermination of somatic embryosRooting of banana plantletRegeneration of somatic embryo bananasuperior drought tolerant genotype, Paramakudi 4was developed and released.For the purpose of developing micronutrient richmaize cultivars meant for poultry feed, a total of2725 immature embryos of maize inbred UMI29were bombarded with the construct pZphycontaining phytase coding sequence driven by zeinpromoter. A total of 27 putative transgenic plantswere developed.GBPUA & T, Pantnagar:The programme support covers 8 sub-projects underthree theme areas (i) manipulation of signaltransduction cascades to achieve disease resistanceagainst Alternaria blight in rapeseed mustards(ii) characterization of aroma and use of molecularmarkers to develop aromatic rice as per consumers’need (iii) development of Madua (ragi) asneutraceutical crop through partial structural andfunctional genomics. In order to determine the roleof MAP kinase-3 in early and late defense in Brassica,three varieties of Brassica (Sinapsis alba, Divya andVaruna) were infected with Alternaria brassicae, andit was found that MAPK-3 acts as an early defensivegene.Studies on rice accessions showed that Kalanamakrice accession 3212 was found to be the most saltsusceptible accession. It was found that the trendof selected Kalanamak accession was similar toPokkali for most of the parameters underinvestigation. Genetic diversity analysis of fingermillet genotypes from Uttarakhand indicated thatcalcium accumulation is a genetically governedphenomenon. Based on the differential spatialdistribution of calcium in five varieties of fingermillets due to differential calcium ion delivery toAnnual Report 2009-2010 45

Research and Developmentthe seed coat, aleurone layer and endosperm bycalmodulin mediated transporters, an attempt wasmade to clone and sequence the CaM gene offinger millet along with other cereals and millets.In order to understand the molecular regulation ofnitrogen responsiveness of finger millet genotypes,Dof 1 transcription factor was identified based onin silico and PCR based approaches. The expressionstudy under different nitrogen inputs with Dof1indicates that the expression was higher with Ragibrown (PRM-1) and golden (PRM-701) genotypesof finger millet as compared to white (PRM-801)genotype.UAS, Dharwad:The programme has projects pertaining todevelopment of transformation protocol andproduction of transgenics for insect/fungal diseasetolerance in important crop plants; cloning andexpression of agriculturally important genes fromnative soil microbes; development of genetic linkagemaps, identification of markers and marker assistedselection in sorghum and pigeonpea; and genediscovery and functional genomics in sorghum usingiac/ds mutagenesis.In pigeon pea, events previously generated wereforwarded to the next generation and confirmed forthe presence of cry1Ac gene. Selected progenies weresubjected to insect bioassay and a mean Helicoverpaarmigera mortality of 68-70% was recorded. Levelsof Cry1Ac protein in young leaf tissues of selectedprogenies varied from 30 to 665 ppb. A total of565 new co-cultivated plants were regenerated(ICPL8863) and screened for the presence of genein T 1generation progenies. A set of 5 new putativeevents (23 plants) were identified; cry1Ac proteinlevel was found to vary between 32 to 71 ppb withan insect mortality of 55 to 68%.A total of 68 polymorphic SSR markers wereidentified in sorghum. The entire sorghum ESTresource (2.75 lakhs) was mined for SNPs andSNP-CAPS markers. A total of 2326 high qualitySNP and 1597 SNP-CAPS markers were identifiedand annotated. Three charcoal rot resistance QTLswere further backcrossed and the progenies in BC 3F 1harbouring all the three QTLs were marked throughforeground selection and selfed. BC 3F 2progenies inthe background of M35-1 and SPV86 are in field.Two new populations segregating for root traits,Basavanapada/E36-1 and SPV570/E36-1 wereforwarded to F 6and F 7generations respectively.Multi-institutional projectsFunctional genomics programme on riceBased on whole-genome microarray expressionanalyses in rice, major regulatory genes from theclasses of MADS box, bZIP, homeobox, CDPK,Mediator and methyltransferase genes were identifiedfor functional validation studies in rice andArabidopsis. From the MADS box class, OsMADS1,that was previously shown to control thedevelopment of all floret organs and also floraldeterminacy, was examined for its downstreamtargets by microarray analysis of panicles in theOsMADS1 knock-down transgenics. The dataindicates involvement of various classes oftranscription regulators, specifically those that regulatedifferent steps in auxin and cytokinin signalingpathways. Another seed-specific MADS box gene isbeing characterized for its involvement in theregulation of embryo/endosperm development in46Annual Report 2009-2010

Research and Developmentknock-down and over-expressor transgenics.Preliminary analysis to analyze the function of seedspecificCDPK encoding genes, OsCPK31, inoverexpression and RNAi-based silencing linesindicated its involvement in flowering time andseed maturation. The over-expression lines ofOsbZIP genes were developed and phenotyping isbeing carried out. The PA-1823 was found to bethe strongest promoter construct followed by PA”1and other deletion constructs, suggesting that therecould be a series of positive elements all along the1823 bp promoter region (Fig).OsiPP3 promoter driven GUS expression in mature pollen grains.(a) Anthers from wild-type and (b) a transgenic plant.Identification and functional analysis of genesrelated to yield and biotic stresses in riceThis network project is aimed at identification andfunctional analysis of genes related to yield andfour biotic stresses like blast, bacterial blight (BB)diseases and insects pests gall midge (GM) andbrown planthopper (BPH) in rice. At NRCPB,New Delhi, one of the major yield QTL qGN4-1for high grain number as identified. Differentiallyexpressed genes in the qGN4-1 QTL region wereidentified by using the combined mapping andexpression profiling approach. At the Directorate ofRice Research (DRR), Hyderabad, the major effectQTL yld2.1 from O. rufipogon was narrowed toone sub-QTL spanning just 62 Kb region for genecontent analysis. Many of these sub-QTLs wereintorgressed in KMR3 background and these lineshave performed well.A novel dominant bacterial blight resistance gene,Xa33(t) at DRR was identified from the O. nivaraaccession with the flanking SSR markers. This genehas been introgressed into Samba Mahsuribackground through MAS. Another new recessivegene, xa34t, (introgressed from O. rufipogon) wasmapped on Chr. 1 at ~ 26 Mb in the vicinity ofthe SSR markers. Besides, functional markers weredeveloped and validated for Xa5 and Xa13. AtPAU, Ludhiana, a novel gene Xa30(t) has beenintrogressed from O. nivara (acc. 81825) andmapped on the long arm of chromosome 4 betweenthe markers RM17499 and RM17502 and finemapped within 38.4 kb region. Another recessivegene Xa-g has been intorgressed from O. glaberrimaand mapped on 5S chromosome between themarkers RM548 and RM593. These genes conferreda high level and broad spectrum of resistance againstBLB. At CCMB, Hyderabad, innate immuneresponse of rice was studied to understandmechanistic basis for induction and suppression ofinnate immunity in the interaction between riceand Xanthomonas oryzae pv. oryzae (Xoo). Transientoverexpression of OsAP2/ERF, one of theupregulated transcription factor genes, induced innateimmune response in rice that enhanced resistantagainst Xoo infection.At NRCPB, New Delhi, molecular cloning andfunctional characterization of the blast resistancegene, Pik h , was carried out. Transgenic linescontaining Pik h gene expressed resistance againstthe pathogen. At DRR, Hyderabad markers forPi1, Pi2 and Pikh genes were validated and theAnnual Report 2009-2010 47

Research and Developmentthree genes introgressed singly into the backgroundof BPT5204 and Swarna using MAS andintercrosses to generate two or three gene pyramidsfor durable blast resistance. Identification andfunctional analysis of gall midge resistance geneshas been the focus of study at DRR, Hyderabadand IGKVV, Raipur. Two flanking linked markerswere identified for the gall midge resistance genes.Studies on genes related to brown planthopper(BPH) were conducted at TNAU, Coimbatore;DRR, Hyderabad and CRRI, Cuttack usingmapping populations derived from the crosses ofBasmati 370/ASD16, TN1/ARC10550 and TN1/Salkathi, respectively.Generation, characterization and use of EMSinduced mutants of upland variety Nagina-22for Functional Genomics in RiceSub-sets of some lines in this network projectwere phenotyped for different traits and this hasled to the identification of putative mutants. Atotal of 347 mutants with visible phenotypicalterations isolated from the first batch of 4309lines were grown in the mutant garden in theIARI farm and further characterized. Mutantphenotypes such as dwarf plant height, hightillering habit, small grain size, root length/mass,BLB resistance, salt tolerance and blast resistancereported in the previous year were confirmed byprogeny testing. Thirty-five different mutantshaving salt tolerance, blast resistance, phosphoroususe efficiency, drought tolerance, dwarf height,short grains, shorter roots etc were crossed withNagina 22 to generate segregating populationsfor studying inheritance pattern of the mutantphenotype.CROP BIOFORTIFICATIONBiofortification of wheat for micronutrientsthrough conventional and molecular breedingapproachesDuring the year, non-progenitor genomes (U andS) were found to confer higher micronutrientconcentration in experiments conducted at IIT,Roorkee. At IARI, Tritcum spelta accessions, syntheticderivatives and a few landraces have been confirmedas additional sources of high micronutrient content.A set of 600 durum varieties in two replicationssent by IARI, Indore is being screened for zinc andiron content at ARI, Pune. For estimating variabilityfor beta carotene in durum wheat, IARI, regionalstation, Indore analyzed a total of 120 durum wheataccessions having more than 5.4 ppm b-carotene.Towards GX E interaction studies, in collaborationwith Harvest Plus project, agronomic biofortificationtrials were raised in which effect of Znsupplementation on grain Zn content was studiedin twenty genotypes.On mapping for high grain Fe and Zn content, agenome linkage map was generated in a diploid A-genome recombinant inbred line (RIL) populationby crossing T. boeoticum acc. pau5088 with a springtype T. monococcum acc. pau14087. Micronutrientanalysis of the parental accessions showed that T.boeoticum had higher grain Fe and Zn than T.monococcum. The population showed a near normaldistribution indicating that grain Fe and Zn weregoverned by QTLs.Development of micronutrient-enriched maizethrough molecular breedingBased on molecular profiling of selected lines with48Annual Report 2009-2010

Research and Developmentphenotypic contrast with respect to kernel Fe- andZn-concentrations, parental lines for generatingmapping populations were chosen. Three F 2:3mapping populations were developed.Polymorphism survey of the parental lines using aset of 800 SSR markers was completed. The analysishas led to the identification of several individualswith transgressive segregation for kernel-Fe and theseprogenies would be grown during Kharif-2010 forobtaining stable lines with high kernel iron content.With regard to MAS for low phytate content, twocrosses were generated and a total of 961 plantswere screened through foreground selection, whilebackground selection was done employing 30polymorphic SSR markers. A set of 94 inbredsdeveloped at the different maize breeding centreswere evaluated at Maize Winter Nursery, Hyderabadduring Rabi 2008-09. Analysis of this set of diverseinbreds, besides 62 Indian maize landraces, forcarotenoid content at IARI led to the identificationof some promising inbred lines with >25 ug/g totalcarotenoid.Rice bio-fortification with enhanced iron andzinc in high yielding non basmati cultivarsthrough marker assisted breeding andtransgenic approachesIron content in dehusked and powdered grains of183 samples & 86 samples were assessed in bothwetland and aerobic conditions and it ranged from2-21 ppm and 7-25 ppm respectively. In general,the genotypes grown under wetland conditionrecorded lower iron content than in aerobiccondition. Phenotypic evaluation of mappingpopulations and tagging factors contributing toacquisition, content and distribution of iron andzinc in the rice grain and their bioavailability inwhite rice as influenced by content of phytic acid iscurrently under F4 generation. Mapping populationsare being developed using several ‘high’ and ‘low’iron content genotypes. The resulting progeny is atF 4(as of Kharif 2008) generation.Introgression of transgenes with Ferritin into localvarieties of indica rice was undertaken. From thepopulation of average 200 BC4F1 plants in eachvariety (ADT43, White Ponni, IR 20 and IR 64),the positive plants (50 %), which possess thetransgene, were identified. These were screened forrecovery of the respective parental characterphenotypically and a single plant with highestrecovery of parental characters was identified foreach variety. Twelve rice cultivars were selected forgene expression studies. The gene expression oftwo high grain zinc rice lines was found to showhigher expression of most of the metal related genescompared to that of rice lines with low levels ofmicronutrients.Generation of virus-resistant rice for India:Diversifying transgenic resistance to popularvarietiesThe project deals with back-crossing of a transgenicrice line (RTBV-O-Ds2), showing resistance to Ricetungro bacilliform virus (RTBV) to a number ofpopular rice varieties in Tamil Nadu and WestBengal. The project collaborators are BidhanChandra Krishi Viswavidyalaya, Kalyani (BCKV),Tamil Nadu Agricultural University, Coimbatore,(TNAU) and University of Delhi South Campus,New Delhi (UDSC). BCKV has taken the backcrossingline to BC 3stage using two recurrentparents, Khitish and Shatabdi, highly popular butAnnual Report 2009-2010 49

Research and Developmenttungro-susceptible varieties. During each stage, theprogeny has been screened for the presence of theresistant gene (RBV-O-Ds2) by PCR. Eighteen SSRmarkers linked to various agronomic traits wereused to study the introgression of the recurrentparent genome into the BC 2progeny. Agronomicperformance and gain quality was also determined.Results indicate greater resemblance of the progenyto the recurrent parents. Similarly, at TNAU, theback-crossing was performed using three recurrentparents, ASD-16, CR-1009 and BPT-5204 till BC 2generation. In all the progenies, the introgression ofthe resistance gene was confirmed by PCR. AtUDSC, the estimation of RTBV DNA in inoculatedplants was standardized using real-time PCR as ameans of estimating resistance. Currently, some ofthe BC 2lines are being tested at UDSC underglasshouse conditions for resistance against RTBV.Use of Molecular Marker Technology Approachin Wheat Quality BreedingIn the network project the PAU, Ludhiana andCCSU, Meerut centres introgressed HGPC geneGpc-B1 to elite Indian bread wheat using MAS.The CCSU, Meerut centre developed frameworkmaps for three mapping populations, viz. WL711 xPH 132, HD 2329 x SPR 8198 and Rye Selection111 x CS. Another framework map using Ryeselection 111 x CS population was developed atNCL, Pune. NCL, Pune used a mapping populationfor bread quality, HI 977 x HD 2329 developedby DWR, Karnal. The mapping population HI977 x HD 2329 was tested in 3 locations.Development and application ofbiotechnological tools for millet improvementPopulation mapping has been developed for droughtand diseases in millets. Marker assisted breeding forshootfly resistance in rabi sorghum hybrids resultedin many promising progenies. Efforts on transgenicapproach for Indian sweet sorghum genotypes withBT constructs resulted in 100 transgenic plantswhere twenty shoots are regenerated in the genotypeRSSV9 with Bt cry1Aa after passing through 3cycles of stringent selection on medium containingbar gene as selection marker.Development of virus resistant transgenic cropsFor developing citrus transgenics resistant to CTV,an efficient plant regeneration and Agrobacteriummediated transformation for Citrus sinensis weredeveloped. Transgenic approaches in transformationof water melon determined the diversity intospoviruses which severely affect cucurbits.Transgenic plants of Vigna unguiculata carryingMYMV rep-hairpin RNA were developed.Transformation of V. unguiculata With rep-hpRNAbinary plasmidA- Untransformed explants regenerated on non-selection mediumB- Untransformed explants regenerated on selection mediumC- transformed explants regenerated on selection mediumD- transformed shoot rooted on selection MediumE- Transformed plant established in potF- transgenic plants growing in green house50Annual Report 2009-2010

Research and DevelopmentIndo-US Collaborative Project in AgriculturalBiotechnologyThere are two ongoing network projects under Indo-US Programme and their salient achievements areas follows:Development and evaluation of transgenicgroundnut for resistance to the groundnut stemnecrosis disease (GSND) caused by the tobaccostreak virus (TSV)At ICRISAT Hyderabad, Agrobacterium tumefaciensmediatedgene transfer into mature cotyledonexplants of peanut was carried out using binaryvector, pCAMBIA 2300 carrying the coat proteingene of TSV (TSVcp) driven by the CaMV 35Spromoter. Over 112 transgenic events were developedand 80% of the selected plants in T0 generationwere confirmed for integration and expression ofthe transgenes. ELISA/western analysis showedsignificant expression of the TSVcp in at least 32transgenic events. The transgenic events in T1generation were screened in a contained greenhouse.Altogether 12 events were found to be promisingand showed no symptoms where the virus wasrestricted only to the inoculated leaves without anysystemic infection. These selected transgenic eventsare being advanced to subsequent generations (T3)in containment for seed multiplication and to studythe inheritance pattern and further evaluations.Additionally, over 50 marker-free transgenic eventscarrying the TSVcp gene have been developed andcharacterized.Development and evaluation of salt and droughttolerant transgenic riceThe ICGEB, New Delhi, DRR, Hyderabad andCentral Soil Salinity Research Institute (CSSRI), Karnalpursued the project. Agrobacterium strains were usedfor co-transformation of pLHW-TPSP gene constructalong with pJS17 received from Cornell Universityand Green gene to mobilize TPSP genes into IR64rice. From ICGEB and DRR, 11 independenttransgenic events that showed enhanced salt tolerancewere transferred to the CSSRI for evaluation.At CSSRI, a total of 14 genotypes including 11transgenic events, control (IR64), tolerant check(CSR36) and sensitive check (VSR156) werereplicated twice under control as well as salinity andsodic stress conditions. The observations indicatedthat event DR8-19 followed by ICG A18 seemmore promising than the control IR64 in both salinityand normal environments, and DR8-19 performedbetter than over tolerant check CSR36. Under sodicitystress, ICG A 18 and ICG Ta7-R1-3 seemed betterthan both control (IR64) and check (CSR23).National Plant Gene RepositoryThe Department set up the National Plant GeneRepository (NPGR) at National Institute of PlantGenome Research, for (i) storage and distribution ofgenes, promoters and ESTs in form of DNA clone,library, etc. and (ii) developing a database for stocks.Next Generation Challenge Programme onChickpea GenomicsThis programme has been sanctioned to NIPGR,New Delhi to develop tools and genomic resourcesfor national/international chickpea community. Theoutcome of the project is expected to serve as a keycomponent of future research on chickpea breeding,gene mapping, functional genomics and geneticenhancement for important agronomic traits.Annual Report 2009-2010 51

Research and DevelopmentBiofertiliser BiotechnologyPollution and contamination of soil have emergedas a major concern. The use of chemical fertilizersand pesticides has harmed the environment. Ananswer to this is the use of biofertilizer. Variousprojects have been reported to develop biofertilizerformulations and technology for mass production.Besides, efforts were made to develop integratednutrient management packages for plantation crops.Some of the salient findings are:Sulphur oxidising bacterial (SOB) formulation wasdeveloped for enhancing the productivity andquality of groundnut and black gram. SOB isolateBGS2 was found to be compatible with Rhizobium,Pods/plant of GroundnutSOB(st),(sa)+RhizobiumSOB(st)+RhizobiumcontrolField trial to test the efficacy of SOB in groundnut : Lignite basedSOB Formulation(St-Seed treatment; sa-Soil application)Azospirillum and phosphobacteria. A field trial thatwas conducted showed that SOB seed treatment@2.5 Kg/ha and soil application on the 45th day@5 Kg/ha registered the highest plant growthincluding, nodule number, nodule dry weight andpod yield (Fig).A study was supported at TERI, New Delhi todevelop a broad spectrum next generationbiofertilizer for multiple applications. Studies onclonal selection of host, determination of moreefficient hosts and multiple species of AMF,Agrobacterium rhizogenes mediated transformationand putative transformants of brinjal was undertaken.These putative root lines were maintained forstabilization. Development of ectomycorrhiza dualcultures was also taken up.At Anand Agricultural University, Anand work donehas been successful in revival of native isolatesusing appropriate production broth in laboratory,including one recent phosphate solubilizing isolateBacillus nr. polymyxa. All the proven native PGPR(five) were mass multiplied to achieve best growthand based on it a consortium formulation isunderway.In continuation of the ongoing work ondocumentation and characterization ofectomycorrhizal fungi associated with Pinuswallichiana and Cedrus deodara in the KashmirHimalaya, field surveys that were conducted resultedin the identification of hitherto uncollected ECM,namely Paxillus, Suillus, Laccara and Amanitaspecies. This was in addition to 96 sporocarp typesidentified earlier. In vitro mycorrhization andperformance of conifer seedlings under nurseryconditions is underway.52Annual Report 2009-2010

Research and DevelopmentA network project involving nine specializedinstitutions has been launched on tea, coffee, cashewnut, rubber, mandarin orange, rosewood, blackpepper and cardamom. The crop-based specialtycentres were engaged in field testing the efficacyand adoptability of a combination of differentbiofertilizers as a partial substitute for standard dosesof nitrogen and phosphorus fertilizers. A centralfacility was developed to provide quality assuranceof the candidate microorganisms.The use of bio consortium like Azospirillum, PSBand Pseudomonas in black pepper increased thegrowth and total biomass in trials conducted atKerala, Karnataka and Sikkim. Among the twophosphorus sources, bio inoculant PSB was foundto be more promising (Fig).INM black pepper nursery I poly bags10- INM influence on black pepper growth(4 months after imposition)INM influence on black pepper root development(4 months after impositionIBlack Pepper Nursery trial (Karnataka & Kerala)Annual Report 2009-2010 53

Research and DevelopmentStudies were carried out on mandarin orange onphosphate solubilisation and nitrogen fixation,wherein the emphasis was on exploring thecompatibility of Trichoderma harzianum(biopesticides), Azotobacter (nitrogen fixer) andAspergillus awamorii (phosphate solubiliser) whenused in combined manner for sustainable nutrientmanagement in barren land. Initial studies revealedthat Trichoderma sp. could act as a promisingbiocontrol agent against many phytopathogenicorganisms.Biopesticides and Crop ManagementBiocontrol agents/biopesticides have been a verypromising option to conventional methods forcontrol of pests and diseases. They confer severaladvantages including reduced environmental damageand reduced risks to human health. A number ofresearch projects have been supported for thecontrol of pests and diseases of economicallyimportant crops, thereby enhancing agriculturalproductivity. The highlights of some of the projectsare as follows:Microbial Pesticides and Natural EnemiesIn a study at TNAU, Coimbatore, the bacterialantagonists for the potato cyst nematodes werefound to be more in soils having low cyst (adultdead female nematodes) densities. Among 73isolates, eight were found to be effective in reducingpotato cyst nematodes and increasing the plantgrowth parameters. The strains ULTV112(Trichoderma longibrachiatum) and EPPF238 (Bacilluspumilus) were found performing well under fieldconditions. In another study, field experimentsconducted on blackpepper revealed a significantreduction in the population of root knot andburrowing nematode in the vines treated with theconsortial formulation of Pseudomonas spp. – (Pfbv)and Bacillus spp (Bbv) @ 10g/vine. The survivalpopulation was 42.5 X10 9 on the initial day and9.25 X10 7 , 180 days after inoculation.At IIHR, Bangalore, noteworthy mortality ofhoppers was noticed by application of M. anisopliaeand Verticillium lecanii. Biological control of mangohopper Idioscopus nitidulus by M. anisoplaie revealeda 76.4 % reduction in mango hopper population.Biochemical studies in infected mango hoppersindicated changes in protein and carbohydrateconcentration. Molecular characterization of M.anisopliae strain was also completed.At IHBT, Palampur, cold tolerant Trichoderma speciesexhibiting antifungal activity were identified againstFusarium spp. inciting rhizome rot of ginger. In invitroassessment of compatibility among antagonisticplant growth promoting rhizobacteria (PGPR), amixture of Bacillus spp. i.e., S2BC1+ GIBC- Jamogand a mixture of Bacillus spp. with a fluorescentpseudomonad i.e., S2BC1+TEPF- showedmaximum antagonism against Fusarium and Pythiumspp. inciting rhizome rot of ginger.Botanical PesticidesAt PDBC, Bangalore LC 50value of multipleinsecticides tolerant (resistant) strain (MIRS) ofTrichogramma chilonis was determined as 13 and4.5 folds higher than the susceptible strain forendosulfan. Out of the 45 random primers used inRAPD analysis of MIRS and susceptible strain of T.chilonis, 15 showed consistent and polymorphicbands. The resistant factor was 13.3 folds in thetolerant strain over susceptible strain. Partial sequence54Annual Report 2009-2010

Research and Developmentof hsp70 in the tolerant strain of T. chilonis wasamplified by RT- PCR.At VMSRF, Bangalore, selected botanicals werescreened in-silico for activity against neural cytoskeletalproteins, actin and tubulin. Azadirachtin, nimbin(neem), karanjin (Pongemia) and melianotriol(Dharek) were found to target insect actin whilethey were found safe to bovine (mammalian) actin.Molecular studies of interaction of azadirachin onthe disruption of insect actin and tubulin, in-vitro,showed pronounced effect of azadirachtin on actinpolymerization, while it was minimal in tubulincytoskeleton. Azadirachtin (30ppm) as a watersoluble powder formulation caused 87 per centmortality but pongemia did not cause larvalmortality in three days.PheromonesThrough a project implemented at IICT,Hyderabad, the existence of female based sexpheromone communication system was confirmedin groundnut bruchid, Caryedon serratus throughrelevant electrophysiological and behavioral bioassaystudies. Existence of male based aggregationpheromone system in C. serratus was confirmedthrough bioassays. At VMSRF, Bangalore,identification and profiling of plant volatiles offive tomato cultivars (2 varieties-PKM, Pusa Ruby;3 hybrids – Shaktiman, Lakshmi and all rounder)were done and diurnal and nocturnal responses intomato plants to the herbivory by Spodoptera lituraand Helicoverpa armigera studied. The in-vitro tissueregeneration and transformation protocol fortomato (cultivar–PKM) was standardized. Thetomato was transformed with linalool synthase(LS) gene to enhance linalool emission in tomatoplants.Molecular Studies on Biocontrol AgentsThrough studies conducted at ANGRAU, Tirupati,twenty isolates of Sclerotium rolfsii causing stem rotof groundnut were isolated, characterized and theirdiversity was studied. The results revealed that theculture filtrates of four potential bacterial antagonists(GSE 36, GSE 37 GGS 8 and GRE 2) proved100% effective in inhibiting the sclerotialgermination of S.rolfsii than the fungicide,carbendazim. The potential seed endophyte (GSE37) and root endophyte (GRE 2) were given asseed treatment to farmers, to evaluate their efficacyagainst S.rolfsii during Rabi crop.In a project funded at NCL, Pune, specificity ofthe monomeric and dimeric alpha-amylase inhibitorsfrom wheat towards the alpha-amylases from S.incertulas were studied and the genes encoding themonomeric and dimeric alpha-amylase inhibitor(GeneBank Accession Number GQ374443) werecloned. Homology modeling and comparativemodeling was done. At TNAU, Coimbatore, rearingof thrips in cucumber fruit/field bean wasstandardized. The PCR-RFLP analysis of thripspopulations of Thrips tabaci and Scirtothrips dorsaliswere done. There were no significant variationsamong the populations of both thrips collectedfrom different locations.Entomopathogenic Nematodes (Epn)-Insecticidal NematodesAt MPAUT, Udaipur, isolation and massmultiplication of desiccation tolerantentomopathogenic nematodes (Steinernema sp. andHeterorhabditis sp.) was done. Survival and infectivitywere tested and bioassay studies conducted.Bioefficacy of different strains of Steinernema spAnnual Report 2009-2010 55

Research and Developmentwas tested. Powder based formulations (viz; chalkpowder and inert clay material) of the promisingstrains of Steinernema sp. with different proportionsand varying moisture levels (5, 10, 15, and 20percent moisture ) were developed. At IIPR, KanpurThe bio-efficacy of promising populations ofSteinernema and Heterorhabditis against H. armigerawas established. Two novel heat tolerant EPN viz.Steinernema masoodi and S. seemae were identifiedand successful field trials conducted to establishtheir efficacy. S. seemae was found more potentialin killing H. armigera larvae within two days.Storage Pests and their Eco-FriendlyManagementAt PDBC, Bangalore, FAT3 and FAT6 fat oilswere screened based on their good stability of invertemulsionformulation i.e. 95% upto 9 months andtested for their inhibitory effects on growth andsporulation of T. harzianum. Phytotoxicity tests withchickpea, tomato and groundnut proved that theinvert emulsion formulation were non toxic to thesecrop plants. The seed treatment with standardizedinvert emulsion formulations at the rate of 10ml/kg of seed resulted in good yield as well as reductionof groundnut root rot incidence.Referral LabsAt AAI, Allahabad, efforts were made to popularizethe referral laboratories among various departmentsof State Government. Samples were receivedregularly for testing from the Directorate ofAgriculture, Directorate of Horticulture and others.A total of 502 samples were received which consistsof the common bio-pesticides being used in biocontrolprogram in UP viz. Trichoderma viride,Trichoderma harzianum, Beauveria bassiana,Metarhizium anisopliae, Verticillium lecani, Bacillusthuringiensis, Pseudomonas florescence, NPV etc.Popularization of Biopesticide Technology:Extension and Training ProgrammesWide publicity of the biocontrol technology underIPM was given through farmers’ field school, krishimelas/ exhibition cum information-cell andconsultancy/ biocontrol stalls, agro-clinics, agri fairs,regional workshops and through mass media whichincluded newspaper, magazines/popular articles/radiotalks and video films etc. Besides, press meets,Farmers’ Goshthis were also organized and thesignificance of biocontrol under IPM washighlighted. Various centres were identified forimparting training regularly on the production andmanagement strategies of biocontrol agents toprogressive farmers, small entrepreneurs, scientistsfrom private and public sectors and unemployedyouth.About 90,000 farmers’ benefited and over 1800handouts in various regional languages were releasedand distributed among the farmers and other users.This awareness programme has evoked the interestof the policy makers, extension workers, smallentrepreneurs and farmers throughout the country.Other InitiativesAn interactive meeting was organized on 9th-10thOctober, 2009 at University of Mysore, Mysore(Karnataka). Two potential biopesticides(Trichoderma spp. and Pseudomonas fluorescens) forthe management of pests and diseases in the northeastregion were identified. A Comprehensive anddedicated website on “Biocontrol Strategies & Eco-56Annual Report 2009-2010

Research and Developmentfriendly Pest Management in India” was developedand officially launched on 23rd June, 08 by thedepartment. The URL of the website is http://www.dbtbiopesticides.nic.in.The 5th International Conference onBiopesticides : Stakeholders’ perspectives” (ICOB-V) has been organized under the auspices of“Society for Promotion and Innovation ofBiopesticides” in collaboration with TERI fromApril 26-30, 2009 at New Delhi. A “StrategyDocument on Biopesticides” was released by theSecretary, DBT, Government of India duringthe inauguration.Mission Programme on Microbial Prospecting:First Public-Private Project for Screening ofMicrobesA public-private-partnership mission programme wasunderway for prospecting of drugs from microbes.The academia partners aimed at isolating about7000 bacteria per month with the support of PiramalLife Sciences Limited for screening of these isolatesfor potential anti-cancer, anti-diabetic, anti-infectiveand anti- inflammatory activities. More than1,75,000 isolates were collected and almost 4,80,000extracts generated to screen for bioactivity. Thenumber of hits in various categories are shown infigure.Bioresource Development and UtilizationNational Bioresource Development Board(NBDB) focuses on: (i) Bioresourceinventorisation and characterization both spatialand non-spatial (ii) Prospecting of bioresourcesfor novel genes, gene products, biomolecules andcompounds (iii) Improvement of economicallyimportant bioresources and (iv) Capacity buildingincluding strengthening of infrastructure.Microbial Culture Collection CentreA Microbial Culture Collection Facility wasestablished at NCCS, Pune to preserve, characterizeand authenticate microbial resources from acrossthe country. There are approximately 50,000cultures and more than 30,000 were processed forlong term preservation while 25,000 cultures wereretained as frozen stocks. Efforts are on to take itto the status of International Depository Authority(IDA).A promising plant extract based biopesticideformulation ‘TERI-DBT-Bollcure’, against cottonbollworm (Helicoverpa armigera), affecting cottonwas prepared. The technology has been receivedwell by Indian and international companies. It wastransferred to two industries viz Sri Biotech,Hyderabad and Pasura Biotech, Jammu.Annual Report 2009-2010 57

Research and DevelopmentIn a project aimed at mining for novel terpenoidmolecules using marine purple bacteria and theirbioprospecting, a library of 39 carotenoids of purplebacteria was prepared, 15 purple bacteria were minedfor carotenoids and 3 novel carotenoid glycosideswere characterized as mannitol glycoside, rhodopinaldi and mono glycoside, and rhodopinal bisdiglycoside.In another project, 475 cyanobacterialisolates from ecological habitats of NE India,belonging to 28 genera were characterized andmaintained as unialgae in the fresh watercyanobacterial repository at IBSD, Imphal, Manipuras reference materials. One hundred fiftycyanobacterial isolates were evaluated and screenedfor total soluble proteins and some of them wereon par to the commercially available isolates.In a project aimed at bioprospecting and preclinicaldevelopment of peptide antibiotics from frog skinsecretions, skin secretion of frog Rana curtipes/R.temporalis were subjected to HPLC purification andthose that showed activity against gram negativeand positive bacteria were characterized.Transcriptomic library produced six peptides havingpredicted antimicrobial activity and flow cytometricantimicrobial assay confirmed their activity.CHARACTERIZATION AND DIGITIZEDINVENTORIZATIONThe Indian Bioresource Information Network(IBIN) was a key initiative of the NBDB, tobring all the databases and information systems onthe bioresources and biodiversity under oneumbrella web site. The IBIN currently has twonodes – Spatial Node, Jeeva Manchitra at NRSC,Hyderabad and Species node, Jeeva sampada atUAS, Bangalore,The Species DatabaseThe filed data on the plant resources of EasternGhats and Western Ghats was uploaded at a scaleof 6.3X6.3 km with Google and Yahoo maps asbackground, The entire plant resource (2700 species)and insect pests (700 species) pollinators (1500 files),predators (300 species) and parasitoids (730 species)data was updated.The Department of Biotechnology and Departmentof Space initiated a joint initiative on biodiversitycharacterization at landscape level using RS and GIS.The phase I & II work have been completed in NEregion, Western Ghats, Western Himalayas,Andaman & Nicobar Islands, Eastern Ghats & EastCoast and Central India. The phase III of the projecthas been launched for the rest of India which includesparts of Deccan Peninsula, North West India andHimalayan Cold Desert (15.06 lakh km 2 totalgeographical area with 1.06 lakh km 2 of forestcover).The programmes on mapping andquantitative assessments of geographic distributionand the population status of plant resources ofWestern Ghats, Eastern Ghats and EasternHimalayan Regions showed significant achievements.The biodiversity at the ground level wascharacterized.DNA BarcodingA total of 130 plant species belonging to 11 generawere analyzed for barcoding purposes using differentbarcode loci and their combinations were explored.Ten different loci and their combinations wereexplored. The internal transcribed spacer, (ITS) wasfound to the best loci, identifying 49 species intested 65 species. The matK was identified as thesecond best barcode loci identifying 66 speciesamong 118 tested species followed by trnH-psbA,58Annual Report 2009-2010

Research and Developmentidentifying 41 species in 108. The percent speciesresolution by the best three loci is shown in Figure.and a transgenic Cauvery (Arabica) plant with ricechitinase gene were confirmed to have single copiesof the integrated T-DNA. Six transgenic arabica(Cauvery) plants with rice chitinase gene werehardened and grown in soil under containmentconditions. The plants showed semi-dwarfmorphology of donor Cauvery cultivar.TeaUnder the network programme on TeaBiotechnology, 225 novel microsatellite primers weredesigned from the tea genomic libraries enrichedwith 9 different repeat motifs. Of these, 129 primerswere successfully validated in selected accessions oftea. These primers also reported a wide range ofcross species amplification (59 % to 73 %) inrelated Camellia species. Sequencing of a few allelesfrom related Camellia species revealed the presenceof repeats.RESOURCE SPECIFIC NETWORKSCoffeeThe main objectives were the development of 100new SSR markers, making a framework linkagemap, creation of EST databases from normal (2000ESTs) and stressed tissues, profiling of differentialgene expression between arabica and robusta seedsand transformation of arabica coffee for fungalresistance and producing coffee for lowered caffeinecontent. Altogether, 90 new validated microsatellitemarkers of coffee were developed. One transgeniccoffee plant (Robusta) with tobacco osmotin geneIn order to identify, develop and implement MASin tea, whole transcriptome polymorphism analysisapproach was initiated to study drought stress-relatedgene activity. Whole genome transcript profilingwas done using cDNA-AFLP technology and EST-SSR analysis. Two genotypes i.e., drought tolerant(TV-23) and drought susceptible (S3/A.3) wereRepresentative picture of amplification validation and cross speciesamplification of newly developed TGMS (tea genomic microsatellite).M= 20 bp ladder standard; (a) TGMS 16; (b) TGMS11.Annual Report 2009-2010 59

Research and Developmentselected. SSR analysis at different stages of droughtstress showed that the number of alleles changedwith different stages of drought suggesting anexpansion or contraction of SSR repeats intranscripts related to drought stress.ZingibersEight wild, endemic and elite taxa and 150 cultivarsof ginger were collected from different places inOrissa. Morphological and molecularcharacterization of 150 cultivars were completed.Morphological characterization of six rare, endemicgingers collected from the Western Ghats werecompleted and protocols optimized for isolation ofessential oils and oleoresins. A ginger house hasbeen established at TBGRI, Trivandrum.Multilocation trials for turmeric were completed inSikkim, Assam, Meghalaya and Manipur. Lakadongturmeric was successfully promoted as a cash cropamong self help groups. A technology for microand mini rhizome production was optimized fromginger, turmeric and Kaempheria sp. The technologyfor Kaempheria was transferred to AVS, Kottakkalfor mass production for medicine manufacture (Fig).Eight EST SSR markers and 165 genomicmicrosatellite containing sequences were isolatedfrom Curcuma. Biologically active peptide turmerinwas isolated from 13 Indian Curcuma and 15turmeric varieties and cultivars.Honey BeeThrough a pre-screening of 80 microsatellite markersfor optimal genetic properties, a set of 15 primerpairs each for Apis dorsata, A. cerana and A. floreawere identified and the information was shared withall participating centers for their further use inrespective populations of target area. Geneticcharacterization of Apis dorsata samples of Karnatakawas completed. Primer pairs targeting a hypervariable300 bp region of COX 2 gene weredesigned and PCR amplification was optimized.SugarcaneUnder a network programme the emphasis was onSSR markers using EST database of sugarcane, foridentification of linked markers for red rot resistancein sugarcane. A total of 67 EST-SSR markers weredesigned and developed from general and subtractivecDNA library specific to red rot disease. Of the103 primers from genomic cDNA library, 82 showedpolymorphism. A new class of functional markers,M 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Minirhizomes produced from microhizomes in Kaempferia rotundaDNA profiling pattern of different sugarcane genotypes as generated byEST-SSR 13 — M:50 bp DNA ladder; 1-CoS 96268, 2-CoLk94184, 3-CoLk 97147, 4-CoLk 05202, 5-CoJ 64 somaclone, 6-ISH 288, 7-ISH 07, 8-ISH 133, 9-ISH 164, 10-ISH 38, 11-CoJ64, 12-CoC 671, 13-CoS 8432, 14-CoLk 7701, 15-Co 997, 16-Co7717, 17-CoS 2399, 18-Khakai60Annual Report 2009-2010

Research and DevelopmentConserved Intron Scanning Primer (19 CISP)markers were used for genomic analysis of 38genotypes of sugarcane consisting of varieties,interspecific hybrids, species and related genera (Fig).Network Programme on bamboo and someother resources.A field demonstration and evaluation of tissueculture raised bamboo species continued over 800ha with eight species. Over 150 accessions belongingto 25 species collected across India were genotypedusing AFLP markers. Genetic diversity of majorspecies of bamboos namely, B. bambos, B. balcooaand B. tulda was found to be very low. Speciesspecificbanding profiles for 8 species namely, B.bambos, B. balcooa, B. tulda, B. vulgaris, B. nutans,P. stocksii, G. angustifolia and D. strictus weredeveloped for identification of species from leafsamples. (Fig)nutritive traits revealed wide diversity. AlthoughDichanthium, Sehima, Heteropogon, Panicum sp areconsidered to be polyploids and apomictic, existenceof diversity indicates presence of facultative sexuality.The genetic diversity was independent of theirgeographical origin. Screening of genotypes forbioethanol production potential indicated widediversity for lignocellulosic content, which can beexploited further.SeabuckthornFor the isolation of bioactive phytochemicals fromseabuckthorn, the leaves of seabuckthorn (SBL) wereextracted sequentially with judiciously selectedsolvents of different polarities. One of the mainField trial of tissue culture raised Bambusa bambos plants growing atForest Department land, Yamunanagar (2.5-years-old).In a programme on evaluation of genotypes ofperennial forage, grasses collected from different partsof the country for morphological, agronomical andSeabuckthron: A. Female Flowers, B. Male Flowers, C. Chromosomespread for a male plant D. Ripe berries, E. Male Plant, F.Hermaphrodite Plant.Annual Report 2009-2010 61

Research and Developmentconstituent of SBL, ursolic acid, was isolated fromthe non-polar extract. It is a potential high-valueproduct for anti-tumour, anti-bacterial, and antiinflammatoryactivities. Protocols for isolation ofpolar constituents such as flavonoid glycosides,tannins and phenolics were developed. Flavonoidglycosides were separated and isolated. The resultsindicate that some of the phenolic compounds/complexes show a higher bioactivity than thosereported for flavones.Capacity BuildingThe Department launched DNA clubs (DBT’sNatural Resources Awareness clubs) in schoolsacross the country in 2007 -08. These clubsoperate as a nucleus for a suite of activities andhands-on learning opportunities focusing onbioresources that seamlessly blend with formalcourse curricula. Till date, 195 clubs have beenlaunched across the country. Another 361 clubswere launched exclusively for the NE region inthe current year. The earlier Vacation TrainingProgramme for the children of class X wasintegrated with the DNA club programme.Rural Bioresource ComplexesDBT has established five Rural BioresourceComplexes at University of Agricultural Sciences(UAS), Bangalore; Marathwada AgriculturalUniversity (MAU), Parbhani; Haryana AgriculturalUniversity (HAU), Hisar; G.B. Pant University ofAgriculture and Technology (GBPUA&T), PantNagar and Orissa University Agriculture andTechnology (OUAT), Bhubaneswar.DBT’s natural resource awareness (DNA) club programme – students in various activities62Annual Report 2009-2010

Research and DevelopmentThe summary of achievements of Rural Bioresource complexes is as follows:-UAS, HAU, OUAT, MAU, GBPUA&,Bangalore Hisar Bhubaneswar Parbhani Pant NagarNumber of interventions 20 8 17 06 4Number of beneficiaries 8340 2621 1058 2394 6744familiesTraining programmes 399 336 48 64 59conductedNumber of direct 13737 776 3462 1716 3402beneficiariesNumber of indirect 20263 1845 15350 675 3342beneficiariesPlant BiotechnologyKey initiatives continued in the area of basic research,tissue culture and forestry, horticulture and plantationcrops and the Solanaceae genome initiativeBasic ResearchThe studies in basic research, focused on signaltransduction, root development, flowering, hostpathogen interaction etc. To understand the roleof stress induced proteins in plant development,studies were supported at GNDU, Amritsar. Saltinducible calmodulin binding proteins (CaMBPs)(HSp85) were purified from Sorghum bicolorbelonging to HSp90 family. It further providedevidence that HSP85 in plants exhibited cambindingproperty and in vivo expression of HSP85 and HSp 87 in Sorghum was modulated byCa2 + /CaM. At TERI, studies were conducted toisolate and characterize flowering locus and leafyorthologs from Brassica sps. Two molecular variantseach from B. juncea cv. Varuna; B. nigra cv. IC-247 and B. napus cv. GSC-5, were identified andone ortholog was cloned and its sequence wascharacterized. The Brassica FT was over-expressedin B. juncea cv. Varuna and the transgenics arebeing regenerated. Expression of a rose petalabscission related gene, RbFBL, encoding an F-boxlike protein led to early flowering in transgenicArabidopsis plants. Flowering was advanced byabout 15 days.Host - Pathogen InteractionIn a study conducted at JNU, New Delhi, Chillileaf curl virus (Chi LCuV) isolates were cloned andclassified into 3 distinct species of begomovirusesviz. chilli leaf curl Multan virus, Tomato leaf curlJoydebpur virus and Pepper leaf curl Bangladeshvirus. DNA-β from different geographical regionsof chilli growing area were cloned. At present, partialAnnual Report 2009-2010 63

Research and Developmenttandem repeats of selected species are being carriedout to study the interaction.At MKU, Madurai, following the identification ofC4 and βC1 of Bhendi yellow vein mosaic virus(BYVMV) as strong suppressors of gene silencing,and C2 as a mild suppressor, their effects studiedon a number of plants, like bhendi, chilli, tobaccoand tomato. C2 was found to produce the strongestof hyper-sensitive responses in all the plants tested.Diagnostic tools were developed for peach leafdecline disease at NBRI, Lucknow and YS ParmarUniversity, Solan. Phytoplasma was detected atdifferent growth stages of infected peach treesthrough peach phytoplasma specific PCR .BAC by BAC sequencing method and Sangerstechnology. So far 96 BAC clones, have been mappedon tomato chromosome 5. The next generationsequencing of the whole genome of tomato hasbeen achieved to 5X level, in collaboration with theNetherlands groups using both 454 sequencing &SOLID technology. The consortium has succeededin sequencing 85% of the genome (Fig).RNAi approach was used to characterize plantparasitic nematode (PPNs) genes at IIT, Kanpur. Atotal of 8 genes were identified and the host deliverymethod of RNAi in PPNs was functionallyvalidated. The study revealed that the genes encodingsecretory gland products are essential for nematodedevelopment and survival, and hence may be anexcellent target for developing strategies fornematode control.SOLANACEAE GENOME INITIATIVEStructural genomicsAn international consortia of 10 countries includingUSA, Korea, China, UK, India, Netherlands, France,Japan, Spain and Italy was set up for sequencing ofthe tomato genomes. India has been assigned theresponsibility of sequencing 12 Mb of euchromaticregion of chromosome 5. This task has been takenup by the UDSC, New Delhi. (5 Mb region),IARI, New Delhi (5Mb) and NIPGR, New Delhi(2 Mb). So far 8 Mb has been sequenced usingGenome sequence of Tomato chromosome 5* Sequenced by China64Annual Report 2009-2010

Research and DevelopmentUnder target induced local lesions in genomes(TILLING) program at University of Hyderabad, alarge number of mutant lines were developed whichwill act as resource centre for identifying desiredtraits using forward genetics approach.Functional GenomicsFunctional genomics studies were supported intomato to address important issues of (i) diseaseresistance (ii) shelf life and (iii) nutritional quality.singlet conformation or in combination whileothers do not show any Ty marking. Thusthe ToLCV resistance could be a quantitativetrait.• Transgenic lines in T-2 & T-3 stage againstToLCV showed ~ 80% resistance to leaf curlvirus. Toxicity and allergenicity assays wereperformed on mice. Protocols for tomatotransformation and gene silencing with RNAiconstruct were developed using conserved repSome of salient achievements are:-Disease ResistanceThe main focus was on tomato leaf curl virusdisease - a major disease of tomato caused by whitefly borne virus of family geminiviridae. The focuswas on screening the available tomato germplasmto develop mapping populations, identify resistantlines for developing markers to segregate susceptibleand resistant varieties, identifying genes impartingresistance to the leaf curl virus. Some of salientachievements are:• About five mapping populations (at F2 stageand beyond) were developed. Mapping/taggingwork is in progress with those populations,which are being phenotyped using the localToLCV pathovars. A few resistant lines werebackcrossed with the elite varieties to developresistant lines with market-demand-fulfillingagronomic characters.• More than one hundred markers (SSR, ISSRetc) were identified to distinguish betweensensitive and resistant parents A few resistantlines showed Ty1/2/3/4 markers either in(A)(B)(C)Two transgenic lines carrying replicase gene sequences fromTomato leaf curl virus , maintained under phytotronHistological analysis of gut tissues: hematoxylin and eosinstaining of gastric mucosa: (a) PBS, (b) wild type tomato,(c) GE tomato fed mice: sections showing jejunum villiDual resistant transgenic plant, (a) & (b) Schematicpresentation of chimeric construct) (c) Primary transformantsAnnual Report 2009-2010 65

Research and Developmentgene sequences of ToLCV at IARI, NewDelhi. Molecular analysis of primarytransformants for dual and multiple resistanceusing chimeric construct for CMV, GBNVand ToLCV was done. The transgenicconstructs are being validated to check theirefficacy. (Fig)Shelf life and nutritional qualityEfforts are on to improve shelf life of tomato andincrease its nutritional quality.Using mutant approach, two putative mutants viz.,atr-1 that appears to be defective in ethyleneperception and kin-27 defective in ethylenebiosynthesis were isolated. Efforts are underway toclone the candidate genes using map-based cloning.Improvement of agronomically important traits suchas increase in total soluble solids, ascorbic acids,flavonoids, lycopenes, beta carotene and folate intomato fruits was observed. Carotenoid biosynthesispathway genes were cloned and are being validatedat IARI, New Delhi (Fig).ABTop panel: Transgenic tomato plant over-expressing the polyamine biosynthesis gene, ornithine decarboxylase under the control of fruit-specificpromoter (2A11) showing deep red colour fruits because of increase in lycopene content (right) as compared to the fruits of an untransformedtomato plant (left).Bottom panel: Increase in the levels of lycopene pigment in different lines of transgenic tomato expressing the polyamine biosynthesis genes(ODC, ADC & SAMDC ) during the fruit development as compared to control fruits.C66Annual Report 2009-2010

Research and DevelopmentInitiatives on MetabolomicsBased on the leads obtained under the ‘SOL’programme, a national network on metabolomicswas initiated on various important crop plants liketomato, rice, potato, maize, Arnebia, Curcuma etc.involving 13 institutes across the country with afocus to develop the complete metabolomics profilein potato, understanding metabolomics of tomatowith special reference to fruit quality; aroma,nutritional quality; metabolomics changes inducedby nutrient uptake (maize), secondary metabolitepathway related to Shikonin synthase (Arnebia);metabolomics related to stress (rice) and metabolomicsof therapeutic properties of Curcuma.FORESTRY, HORTICULTURE ANDPLANTATION CROPSStudies continued in tissue culture forestry,horticulture, plantation crops etc. A protocol wasdeveloped for micropropagation of Crocus sativus L.through a study at Kashmir University. Vegetativegrowth was observed in 90% of the transferredcormlets and flowering was observed in 70% of thetransferred cormlets. Cost effective method forgermination of seeds and rapid regeneration ofcomplete plantlet regeneration of Dendrobiumlituiflorum was achieved by using banana extract.Complete plantlet regeneration of two native species– Cymbidum aloifolium and Cymbidium bicolor wasachieved by using banana extract.Large scale production of virus free citrus reticulatecv Nagpur Mandarin planting material throughmicrografting was developed and transferred togreen house with 80% success at NRC Citrus,Nagpur. A total of 72,864 certified disease freeNagpur mandarin budgrafts were released togrowers / nursery owners of Maharashtra, MP,Bihar, Orissa & southern states and the technologywas transferred to farmers. A PCR based kit fordetection of RNA and DNA pathogens in citruswas developed and validated at PAU, Ludhianaand NRC Citrus. Attempts were made to developonion resistant to purple leaf blotch.Agrobacterium mediated transformation of onionembryos was attempted using the Trichodermaharzianum chitinase (Th-chi) gene constructs.Transgenic tobacco plants with Trichodermaharzianum chitinase (Th-chi) gene construct underthe expression of 35 S promoter were generatedfor validation of the transgene against Alternariaporri.Studies continued to address the cold inducedsweetening in potato at CPRI, Shimla. Seventransgenic lines of Kufri Chipsona – 1 and KufriJyoti expressing Nt-Inhh was developed. Twotransgenic lines were found promising based onglass house evaluation. In another project, 7transgenic lines developed excellent cold- chippingperformance after 90 days of cold storage, and onetransgenic line developed excellent chip colour evenafter 180 days of cold storage. Tubers of theseseven transgenic lines are now being evaluated underconfined field conditions at CPRS, Jalandhar afterobtaining permission from GEAC/ RCGM.Medicinal and Aromatic PlantsBiotechnological interventions on medicinal andaromatic plants (MAPs) continued for conservation,characterization, micropropagation, development ofherbal products, isolation of novel bioactive agents,genomics and biosynthetic pathway studies. Thesalient achievements are summarized below:Annual Report 2009-2010 67

Research and DevelopmentEx-situ Conservation, Characterization andDocumentationMorphological, phytochemical and molecularcharacterization studies were carried out in differentaccessions of Adhatoda zeylanica, Coleus aromaticus,Hemidesmus indicus, Saraca asoca, Tinospora cardifoliaand Terminalia chebula from different agro-climaticregions of South India at Arya Vaidyashala,Kottakkal. Superior lines of C. aromaticus withincreased amount of carvacrol (74.25%) wereidentified. Higher percentage of rutin (up to74.66%) was observed in three accessions of H.indicus. A digitized database on medicinal plantsused as food by ethnic groups of North-East regionwas developed jointly at Rajiv Gandhi University,Itanagar and Chaiduar College, Gohpur, Assam.Ninety seven species belonging to 77 genera and 57families were recorded.Herbal FormulationUnder a network project, a concoction of threeextracts (bark of Azadirachta indica; stem ofWithania somnifera and leaf of W. somnifera in theratio of 2:1:1, respectively) was developed. It wasfound to be most efficacious against Staphylococcusaureus of Bovine mastitis during in vivo testing.The concoction was also found to minimize theexpression of inflammatory cytokines. Work ondeveloping a herbal formulation against diabetesfrom leaf extract of Annona squamosa continuedjointly at Bombay College of Pharmacy, Mumbaiand TN Medical College, Mumbai. Polyphenolicextract of A. squamosa at a dose of 50 mg/kgshowed a maximum fall in fasting blood sugarlevels in rat model of streptozotocin inducedhyperglycemia. Efforts continued to identify highyielding plant sources for metabolites such asgalanthamine and coumarins from herbariummaterial as well as Western Ghats jointly at UAS,Bangalore and BSI, Kolkata. Highest galanthaminecontent (> 1% Dw) was found in Hippeastrumsolandrifarum and Hymenocalis caribeae where asfour species vi3. Mentha piperita, Lavandula pinnatavar. bipinnate, Melilotus parviflora and Cinnamomumcamphora were found promising for their coumarincontent. Asiatic acid, medacassic acid, medacassosideand asiaticoside isolated from Centella asiaticashowed anti-leishmanial activities in BALB/c miceat IICB, Kolkata. The n-hexane fractions ofNyctanthes arbortristis and Artemisia annua showedcytotoxicity on Leishmania donovani promastigotesat Jamia Hamdard, New Delhi.Genomics and Biosynthetic PathwaysConstruction of a DNA molecular genetic linkagemap of Catharanthus roseus was taken up atNIPGR, New Delhi. QTLs for vindoline andcatharanthine hyper-accumulation in leaves and /or roots were identified and mapped. A leaftrichome cDNA library of Artemisia annua alongwith 427 ESTs was generated at CIMAP, Lucknow.Four full-length genes from the artemisininbiosynthetic pathways were cloned. A small scalemicroarray chip (with 750 target genes) of A.annua has also been developed. About 300 ESTswere generated from the capsule and leaf cDNAlibraries of opium poppy (Papaver somniferum).Eight known genes of the morphine biosyntheticpathway in opium poppy (cv. Sampada) werecloned and sequenced. A small-scale microarraychip containing all the significant alkaloidbiosynthesis-related genes in P. somniferum wasdeveloped.68Annual Report 2009-2010

Research and DevelopmentAnimal Biotechnology - ProductionThe emphasis of the various programs was aimedat enhancing production and productivity oflivestock through biotechnological interventions.The salient achivements of the on going keyprogrammes were:TransgenicsA novel technique for in vivo testicularelectroporation of a gene, to generate transgenicanimals, was standardized at NII, New Delhi. Fourlines of transgenic mice carrying different constructswere developed and the stability of the transgenicintegration and its propagation were confirmed inF2 and F3 generations. The transgenisis in micewas further confirmed by RT-PCR & the spermproduced retained their variability. Further attemptshave been made to develop a construct expressingcell surface protein in sperm to enable selection oftransgenic sperm.Animal Reproduction and related areasTo understand the development and function ofovarian follicle and corpus luteum in buffalo(Bubalus bubalis) a project was funded at IISc,Bangalore. Model systems for identification of largeovarian follicles from the first wave of thefollicular growth and induction of luteolysis ofthe functional corpus luteum were standardized Anumber of genes, belonging to various functionalgroups, were identified and these were found tobe expressed differentially. A few selected geneswere further validated by real time RT-PCRanalysis. Molecular cloning studies of buffaloprostaglandin F2 alpha receptor cDNA from corpusluteum and expressions of genes considered asapoptotic in granulosa cells from folliclesundergoing atresia were done. The results suggestedthat induced ovulation protocol could be usedsuccessfully for artificial insemination of buffalocows.In an another study on in vitro development ofbuffalo preantral follicles (PFs) at IVRI, Izatnagar, asignificant increase in total DNA concentrationcorresponded to an increased cell number anddiameter of the follicle. Presence of insulin-likegrowth factor binding protein-3 and 4 (IGFBP-3& 4) genes in ovarian tissue, PFs, antral follicles(AFs), granulosa cells, immature and mature oocytesindicated constant role in follicle growth anddevelopment. Immunocytochemical localization ofthe Insulin-like growth factor-I (IGF-I), IGF-II theirreceptors and IGFBP-2, 3 and 4 proteins in thePFs and AFs confirmed their role during mitogenesisand steroidogenesis.At NIRRH, Mumbai, studies were conducted tounderstand the involvement of imprinting in themale germ line in early embryo loss. The microarrayanalysis revealed aberrant expression of imprintedgenes viz. Dlk1, Lot1/Zac1, Peg3, Peg5, Osbpl5,Ascl2 etc. in the resorbing embryos which could bea combined effect of methylation defects inpaternally expressed genes viz. Dlk1 and Lot1/Zac1coupled with coordinated regulation of imprintedgenes through the imprinting gene network (IGN).Differential gene expression profile revealed an effecton genes involved in placental development. Thestudy suggests that IGN could be contributing tothe overall process of embryo loss.Estrus-specific pheromones are being analyzed fortheir possible role in enhancing reproductiveAnnual Report 2009-2010 69

Research and Developmentefficiency in buffaloes at Bharathidasan University.Twenty six types of volatiles have been identified inbuffalo fecal samples during the estrus cycle, therebyindicating a relationship between the fecal compoundsin estrus and the occurrence of ovulation.A novel technique for the detection ofgonadotropin-releasing hormone (GnRH) receptorin live buffaloes for its presence in lymphocyte wasdeveloped at IVRI, Izatnagar. The GnRH type Ireceptor in normal buffaloes and those sufferingfrom reproductive disorders were observed. Thenormal buffalo receptor content showed pulsatilevariation whereas the infertile buffalo receptorcontent remained constant. Some infertile buffaloesshowed persistently elevated levels of FollicleStimulating Hormone (FSH) and luteinisinghormone (LH) and buffaloes showing poor receptorexhibited very low levels of FSH and LH in theirperipheral circulation. A direct correlation wasobserved between the level of FSH and LH innormal buffalo & those with reproductive disorders.The receptor binding capacity was very low inbuffalo with reproductive disorders & high innormal buffaloes thereby indicating that the geneencoding Gn-RH type-II was inactive.Genetic Characterization and GenomicsMolecular marker based genetic structuring ofvarious breeds of native cattle (Bos indicus) wascarried out at NBAGR, Karnal. A total of 483mitochondrial DNA (mtDNA) D-loop sequenceswere generated across 20 breeds to analyze mtDNAdiversity. Sequence analysis revealed a total of 134Indian cattle haplotypes (ICH) with a highproportion of singleton haplotypes (92, 68.8%).High haplotype and nucleotide diversity indicatedsubstantial mtDNA diversity in Indian cattle.Phylogenetic analysis showed a major clad of ICHwith several subclads. Median-network revealed acomplex network of subnets connecting differentICH and also distinct dichotomy and independentdomestication events for the taurine and indicuscattle types. Single-nucleotide polymorphism (SNP)genotyping of toll-like receptor-1 gene in a panel of11 Indian cattle breeds revealed a total of 17 SNPs,of which 71% were novel.Molecular analysis of the mRNA transcripts fromgermline of buffalo (Bubalus bubalis) tagged withminisatellites and assesment of their functional statusin somatic tissues was done at NII, New Delhi.The 33.15-tagged mRNA transcripts showedmaximum expression in testis and spermatozoaimplicating their involvement in various testicularfunctions and possibly post fertilization events. Allthe transcripts uncovered from spermatozoa weredistinct from those of somatic tissues. This studywould help in understanding the role of repeattagged mRNA transcripts in paternal genome duringpost fertilization events as well as in different stagesof embryonic development.The study of gene expression pattern in buffaloudder during lactogenic pathways was studied atAAU, Anand. A total of 142 expressed sequencetags (ESTs) were sequenced out of which 32 werenovel ESTs. Relative quantification analysis by RT-PCR assay confirmed the differential nature of theseESTs.Animal NutritionShea nut cake (SNC) based concentrate mixturesupplemented with probiotics was tested as a sourceof protein rich feed and as a suppressor of methane70Annual Report 2009-2010

Research and Developmentemission in buffalo calves at SVVU, Hyderabadand NIANP, Bangalore. SNC expeller and solventextraction samples were analyzed for their cell wallconstituents, amino acid content, mineral contentand anti nutritional factors. The degradation kineticsof the SNC cake indicated that the minimum gasproduction (Y min) was 0.81 ml and maximumwas 18.93 ml. The study indicates its potential asa source of energy and also in reducing methaneemission.Detoxification and utilization of key agro-forestrybased non-conventional oil cakes viz. Jatropha,karanj and neem oil as a possible source of proteinrich fodder was investigated at IVRI, Izatnagar. Thechemical composition of seeds / oil cakes of thesenon-conventional oil cakes varied from region toregion. Laboratory scale detoxification techniqueswere developed and efforts are on to scale up thedetoxification process at commercial level. Alaboratory animal study with Wistar rats indicatedthe possible use of detoxified Jatropha oil cake asan alternate to soybean meal upto 25% withoutany adverse effects.Production of xylooligosaccharides from agriculturalby-products and their evaluation as prebiotic wasattempted at NIANP, Bangalore. Chemical analysisof crop residues revealed that corn cob and cornhusk contained higher quantities of hemicellulosesfollowed by ragi straw, paddy straw etc. The xylanspresent in the byproducts or crop residues wereextracted and steam application was found to beeffective for higher recovery of xylan.Studies on the effect of tannin containing feeds onrumen microbial ecosystem and its manipulationfor improving their utilization as quality feed weresupported at IVRI, Izatnagar. A total 62 tannindegrading / tolerant bacteria were isolated from therumen liquor of goats fed on tannin rich diet.These isolates were tested for their ability to degradethe tannin rich pakar leaves using in vitro gasproduction test. The degradability of pakar leaveswas increased with 12 isolates ranging from 2-11%.The trials showed that the population size of rumenprotozoa, total bacteria and fungi were unaffectedbut tannin degrading/tolerating bacterial numberswere increased due to feeding of specific isolate.In an effort to enhance nutrient utilization fromfibrous / non starch polysaccharide rich feedingredients / crop residue, a fibrolytic enzymemixture was developed at NDRI, Karnal. In vitrotrials were conducted to identify the minimumconcentration of non starch polysaccharidase enzymesviz. cellulase, xylanase and pectinase that caused themaximum sugar release from sorghum stover, ragistraw and groundnut haulm individually and invarious combinations. The customized non starchpolysaccharidase enzyme mixture was developed andits feeding trials were carried out successfully inbroiler chicken which showed promising results.Superior fibre degrading anaerobic fungus wasisolated, characterized and its internal transcribedspacer region sequenced. The weekly dosing of200ml culture of the potent isolate in liquid andencapsulated form significantly improved growth(20-30%), feed intake and digestibility of growingbuffaloes.Mitigation of methane emission through rumenmanipulation for increased productivity in buffaloeswas carried out at NDRI, Karnal. A total of 108clones of buffalo rumen protozoa were analysedwhich revealed 17 different 16S rRNA geneAnnual Report 2009-2010 71

Research and Developmentsequences or phylotypes. Methanomicrobium was themost dominant component of methanogenpopulation in Murrah buffloes (Bubalus bubalis).The largest group of clones (102 clones) was morethan 98% similar to Methanomicrobium mobile.Basic local alignment search tool analysis (BLAST)of the rumen contents from individual animalsrevealed 17 different phylotypes with a range of 3-10 phylotypes per animal. The results indicatedthat Methanomicrobium phylotype might be adominant methanogen phylotype present in Murrahbuffaloes.Animal ByproductsA collagen based scaffold of bovine origin wasprepared using a novel microbial method atSRMCRI, Chennai and CLRI, Chennai. Thescaffold was found to have the mechanical strengthto sustain the beating of cardiomyocytes. Thecardiomyocytes were seeded in the scaffold whichstarted beating upto a period of 17 days. Efforts areon to quantify the beating of cardiomyocytes in thescaffold and also to record their electrical impulses.An Indian and an US patent has been jointly filedfor this novel method. In another study atSCTIMST, Thiruvananthapuram, a non-detergent180 days explant of decellularised bovine jugular vein implanted inpig aorta. Healing response under (A) load condition in 0.2PEGprocess, (B) compared to 0.2Dcl process.based decellularisation technique of bovinepericardium and jugular vein was developed forcardiovascular application. In vivo studies of postdecellularized tissue on animal models under staticand load bearing conditions were undertaken (Fig).At IVRI, Izatnagar, protocols for conversion ofporcine urinary bladder into bladder acellular matrixgraft was optimized. Crosslinking with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)resulted in decreased degradation and increasedbiocompatibility of the graft and these were usedfor cell seeding. Experimental studies in a rabbitmodel revealed that EDC cross-linked cell seedingbladder acellular matrix grafts (BAMG) resulted inbetter healing compared to unseeded grafts.PoultryMolecular evaluation of forced moulting in whiteleghorn hens and development of its alternative isbeing studied at CARI, Izatnagar. The forcedmoulting by feed withdrawal (FW) and high ZincFeeding (ZnF) reduced the weight of reproductiveorgans. The regression of reproductive organsappeared to be regulated by both extrinsic andintrinsic pathways of apoptosis in the FW. Cytokineswere also involved in the regression of reproductivesystem during forced moulting by both FW andZNF methods. A few critical genes were identifiedbased on the differential expression pattern of various(apoptotic and cytokine) genes in the regression ofreproductive system. The expression pattern of thecritical genes is being further investigated to developan effective alternative method of forced moulting.In another study, efforts are on to enhance posthatchimmunocompetence and growth of broilerchickens through in ovo approaches. In ovo72Annual Report 2009-2010

Research and Developmentadministration of glucose enhanced the chickengrowth hormone (cGH) expression duringembryonic stage whereas, chicks with riboseadministration showed higher expression during earlypost-hatch period. Fructose enhanced both Insulinlikegrowth factor-I (IGF-I) and IGF-II geneexpression during pre and post-hatch period, whereas,ribose increased IGF-II expression only in theembryonic stage. In ovo glucose enhanced theexpression of candidate genes for humoral immunity,whereas, ribose enhanced expression of cell mediatedimmunity gene.Animal HealthAnimal Vaccines & DiagnosticsThe focus of the programme is the promotion oftranslational research on veterinary vaccines anddiagnostics. A special Task Force was reconstitutedand two Translational Research Network Projects,on Classical Swine Fever and Brucellosis have beentaken up, apart from supporting open-endedprojects. Some of the salient achievements are given:Network project on Classical Swine Fever (CSF)Establishment of Classical Swine Fever ReferralLaboratory and Development of Nucleic Acidbased Diagnostics for CSFCSF outbreaks were confirmed in Rajasthan,Himachal Pradesh, and Uttar Pradesh. Aslaughterhouse based study aimed to detect CSFVinfection in meat carcasses, indicated 29% positivityby the real time -RT-PCR assay. Pathological studieswere conducted on pigs from a farm where CSFoutbreak occurred despite vaccination. Real-time RT-PCR for quick diagnosis of CSF was developedusing self designed primers and probes targeting5’UTR of Indian isolates of CSFV. Variouscombinations of primers and probes were then testedin a multiplex format to detect CSFV isolates fromNorth India available in the lab. This assay alsohelped detection of a new strain of CSFV in anoutbreak that occurred despite CSF vaccination.Surveillance and monitoring in North EasternStates and Establishment of DBT ClassicalSwine Fever Virus RepositoryThe seroprevalence of CSFV antibody was studiedin 8 districts of Assam. Also, sera samples werereceived from Meghalaya, Arunachal Pradesh, Sikkim,Maharashtra and Kerala. A total of 427 sera sampleswere collected and 282 were found to be positivefor CSFV antibody. These samples have beenarchived in the repository. A total of 174 tissuesamples collected from Assam, Meghalaya,Arunachal Pradesh, Manipur, Mizoram andMaharashtra were archived in the repository.Currently, the repository has 455 tissue samples intotal, of which 302 are from domestic pigs, 146from Pygmy hog and 7 wild pigs. A total of 107tissue samples were found positive for CSFV. Atpresent, 14 CSFV isolates are being maintained andare available for future studies on CSF.Epidemiological studies on Classical Swine Feverin Mizoram, Manipur, Tripura and NagalandThe Regional Referral Laboratory for CSF diagnosiswas established by installing equipment in thedesignated laboratories. The recruited staff was trainedon various diagnostic techniques and some suspectedfield outbreaks of CSF in Mizoram wereAnnual Report 2009-2010 73

Research and Developmentinvestigated. An outbreak of CSF in a vaccinatedherd was confirmed. Investigations are being carriedout to study the cause of vaccine failure.Establishment and maintenance of DBTRepository of Classical Swine Fever Virus(CSFV), serum and cell linesA RT-PCR based test for detection of four genes ofCSFV (5’UTR, E2, Erns and NS5B) wasstandardized. A total of 30 clones having differentCSFV genes were sent for sequencing. Diagnosticservices were provided to IVRI, Izatnagar and BiharVeterinary College, Patna. During the period, 131serum samples and 70 tissue samples were archivedin the repository. A total of 14 tissue samples, outof 30, were found positive for CSFV genome byRT-PCR.Epidemiological studies on Classical Swine Feverin Central India and GoaThe DBT CSF Referral Laboratory was setup andlinkages established with farms/farmers/diseaseinvestigation laboratories in Maharashtra and MadhyaPradesh. Serum (229) and tissue samples (310) werecollected from pig farms in central India and Goa.A total, of 220 serum samples were found positivefor CSFV antibodies. Seropositivity was very highin unvaccinated animals (69.66%) from Nagpur,Mumbai, Dahiwad, Gondia, Ballarshah, Varora,Kankawali and Goa.Development of user-friendly, discriminatorydiagnostics and new generation Vaccines forClassical Swine FeverSamples were obtained from 7 different outbreaks,i.e. 4 from Andhra Pradesh and 3 from TamilNadu. Three passages were given in PK15 cellsbefore confirming the virus by indirect FAT andRT-PCR. Two CSFV isolates were recovered fromoutbreaks in AP (CSFV/AP/IND/09-1 and CSFV/AP/IND/09-2). RT-PCR amplification of 5’ UTR(138 bp) and E2 (1044 bp) genes of these twoisolates was done for sequencing these.Two conventional diagnostic tests (AGID andindirect FAT) for routine use in the laboratory weredeveloped by raising CSFV specific hyperimmuneserum in rabbits inoculated with CSFV vaccinevirus propagated in PK15 cells.Computational bioinformatics for design ofoptimum vaccine candidates and developmentof Quantum Dot based diagnostics for ClassicalSwine Fever VirusTo predict optimum vaccine candidates for CSFusing computational biology aided reversevaccinology approaches, complete genome sequencesof CSFV were used for the analysis. Multiplesequence alignment (MSA) of the polyprotein codingframe revealed a high degree of similarity withidentities ranging from 95% to 98% between strains.The Indian strain (CSFV lapinized) showedmaximum similarity (98%) with the C/HVRI strainfrom China.The structural protein regions within the polyproteinshowed little variability, with most residuescompletely conserved. Following the preliminarysequence analysis, the list of peptides within theCSFV polyprotein was prioritized using theNetMHCpan-2.0 server, for predicting SLA bindingpeptides. The top binders to the various SLA alleleswere filtered and a final list of fifty peptidespredicted to have excellent SLA binding efficacy74Annual Report 2009-2010

Research and Developmentwere short-listed. These will be validated throughin vitro and in vivo experiments.DIAGNOSTICSIdentification of Novel proteins expressed bySalmonella spp. in biofilm and their utility indiagnosis of SalmonellosisIn order to explore possibility of using biofilmproteins as diagnostic markers, biofilm formationability of different Salmonella serovars wasdetermined by adhesion assay (microtiter platemethod) with some modification. A total of 156strains of Salmonella belonging to 55 serotypeswere examined for biofilm production. The majority(56.41%) of the strains were found to be moderatebiofilm producers, while 20.51% strains were strongand the rest were weak biofilm producers. All 156strains tested for BapB gene yielded positive results.Similarly, 67 strains tested for BapA gene gavepositive amplification indicating the conserved natureof these genes in Salmonella spp. In silico analysisof the sequence of BapA gene (Salmonella enteritidis,E-2478) revealed 99.5% homology with previouslyreported BapA gene of Salmonella typhimurium LT2and the highest percentage (99.8%) of homologywas obtained with S. enteritidis str. P125109. TheSDS-PAGE analysis of whole-cell protein of nineSalmonella serotypes revealed expression of uniquebands in the region 25 – 30 kda in the biofilmcells.Seroprevalance and molecular biologicalapproach for diagnostics of Canine MonocyticEhrilichiosis in IndiaA total of 375 blood/serum samples werescreened for the presence of E. canis antibodiesand 327 animals were found positive. A nestedPCR, targeting 16s rRNA gene and amplifyinga product of 389 bps was standardized fordetection of E. canis from blood and tissues ofaffected animals. The PCR products generatedby the nested PCR (amplicon size 389 bps)was cloned and sequenced is available online atNCBI with Accession No.FJ793810. The clonedPCR product of the Morula Membrane Proteinhas been forwarded for sequencing. Clinicopathologicalstudies in E. canis positive animals arein progress. In vitro-cultivation of the organism isto be initiated.PCR-based molecular discrimination ofparagonimiasis and other crustacean-bornetrematode zoonoses in North East IndiaWith a view to investigate the zoonotic potential,of the helminth faunal diversity and food-bornezoonoses in the region, edible crustacean specieswere surveyed to ascertain the occurrence ofParagonimus and other trematode infections. Of thesix species of crabs surveyed, only Barytelphusalugubris masoniana showed metacercarial infection.The metacercaria were identified as those ofParagonimus species. The species was identified as P.westermani. For comparing with the other plausibletrematode infections, PCR amplification of ITSregions of Fasciolopsis buski, Fasciola sp., Gastrodiscoidshominis and Artyfechinostomum sufrartyfax was alsodone. ITS sequences of Fasciola isolate revealed itsmatch with Fasciola gigantica. ITS sequences ofGastrodiscoides hominis revealed a similarity with thefamily Paramphistomidae. ITS and those ofArtyfechinostomum sufrartyfax showed a similaritywith the family Echinostomatidae.Annual Report 2009-2010 75

Research and DevelopmentDevelopment of specific assays with specialreference to histopathology of Mycobacteriumbovis infection in animals and manTissue samples were collected / obtained fromvarious slaughterhouses/ Veterinary colleges/ Institutesat Puducherry, Chennai, Namakkal, Bangalore,Thiruvanthapuram, Jabalpur and Kanpur. Out of7200 cattle and buffalo carcasses examined, 88 cases(1.22%) were suspected for tuberculosis.Histopathological examination, of 88 cases, 24 casesshowed granulomatous reactions. In two cases,calcified nodular lesions were seen throughout thepleura and were adherent to the rib cage.Study of transmission of livestock derivedpathogenic Mycobacteria and its prevalenceamong humans (Zoonotic Tuberculosis)Altogether 1094 samples were processed duringthe period of study. Out of 207 isolates obtained,163 were of pulmonary and 44 were of extrapulmonaryorigin. The predominant species wasM. tuberculosis, followed by M. chelonae, M.fortuitum, M intracellulare, M. gastri and M. bovis.Molecular characterization was done by variablenumber of tandem repeat, spoloigotying, IS6110copy number, mutation at codon 463 of katGgene and at nucleotide 285 of the OxyR gene.The largest variation by VNTR was seen in ETR-A, D, E & C loci in pulmonary isolates. Nodifferences in IS6110 copy number betweenpulmonary and extra-pulmonary isolates were seen.Differences in response of immuno-competent cellsof patients and healthy contacts to mycobacterialantigens were seen. CD4, CD8 T cells, NK andNKT cells showed the highest proliferation incontacts compared to patients when co-culturedwith bacilli. Highest proliferative index was seenwith the HupB antigen among patients with theCD4 T cell subset. However, the proliferativeresponse of NK and NKT cells among the patientsand contacts differed. NK and NKT cells derivedfrom healthy contacts showed the highestproliferation when co-cultured with HupB antigencompared to patients.VACCINESBiomarker approach for differentiating naturalinfection and vaccination of chickensThe expressed and purified enhanced greenfluorescent protein (EGFP) from a prokaryoticvector, pRSETa and a commercial eukaryotic plasmid(pEGFPC1) containing the EGFP gene were usedas a potential candidate biomarkers in chicken alongwith the Newcastle disease virus (NDV) vaccine.By screening random sera samples (n=225), it wasshown that antibodies to EGFP were not presentin the target population of chicken. The results ofthe immunogenicity trials showed that the EGFPas protein elicited an early but short-lasting responsewhile as plasmid DNA it elicited a delayed butlonger response. The biomarker inoculated chickenwere apparently normal and their feed consumptionwas not altered. Thus it has been seen that EGFPcould be used as a biomarker along with existingvaccines to differentiate vaccinated chickens frominfected ones.Production and the evaluation of micro particle(Cationic PLG) coated DNA vaccines for Footand Mouth Disease (FMD)To study the duration of immunity provided by76Annual Report 2009-2010

Research and DevelopmentDNA vaccine construct delivered by PLGmicro particles, guinea pigs were immunizedwith the construct with the appropriate controls.Tissue distribution and expression studieswith ID-pVAC-A DNA vaccine construct inguinea pigs showed the expression of thegene insert in the tissues in the PLG coatedDNA vaccine group. CMI response as measuredby MIT and lympho proliferation assay hasshown good CMI index up to 150 days in thePLG coated DNA vaccine group. PLG-DNAvaccine indicated 100% protection againstthe challenge with homologous virus (100GPID50) at 42 days dpv. 75% at 180 dpv and50% at 270 dpv. Enhanced CD4/CD 8 ratio(measured by Flow Cytometry) was seen at 14,21 and 28 days in guinea pigs vaccinated withPLG coated DNA vaccines compared to thenaked DNA vaccine and controls.Development of a multivalent oral vaccineagainst chicken coccidiosis using local isolatesField isolates were collected from 270 broiler and90 layer farms of South India. Eimeria tenella wasthe most predominant species both in broiler (37.4per cent) and Layer (34.4 percent) farms.Propagation of 150 field isolates of six Eimeriaspecies and 10 E. tenella isolates were done in 650chicks to obtain sufficient quantity of oocysts forfurther attenuation trials. Molecular Speciation ofEimeria by SCAR PCR showed a strong positivecorrelation with the morphometry results. Aneconomic field diagnostic was standardized.Attenuation trials were carried out by passaging sixEimeria species in 576 birds. Precocious linesdeveloped after 13 th attenuation showed progressiveresults with reduced oocyst out put. As per theEuropean Commission guidelines of Coccidiosisresearch for development of vaccine, 7 moreattenuation trials and immunization trials areunderway.Development of DNA vaccine against rabiesusing targeted vectors for veterinary useIn the project, four DNA vaccine candidates wereconstructed, wherein the glycoprotein gene ofrabies virus was genetically fused to signalsequences for targeting of fusion proteins torespective intracellular locations. Expression andlocalization of these DNA vaccine candidates waschecked by transient transfection in BHK-21 cells,followed by sub-cellular fractionation andimmunoblotting. All the targeted DNA vaccineselicited superior immune response to the native(untargeted) construct. A lysosomal targeted DNAvaccine showed promising results. Its immuneparameters were optimized to attain maximalimmunogenicity. This optimized DNA vaccineformulation was found to be highly potent forprophylactic as well as therapeutic applicationsand conferred complete protection upon lethalrabies challenge. Bioprocess optimization of theDNA vaccine construct was carried out.Development of recombinant E-toxin andDNA based vaccine against ClostridiumperfringensFor the development of recombinant proteinbased vaccine, gene encoding epsilon toxin ofClostridium perfringens type D was cloned inpQE60 expression vector. High level expressionand single step purification of recombinantepsilon toxin was achieved. The constructs wereAnnual Report 2009-2010 77

Research and Developmentevaluated for their immunogenic potential inmice. It was seen, that the constructs were ableto raise good immune response and the antiserathus raised had the neutralization ability of thetoxic effect of Etx in the MDCK cells. TherEtx was also evaluated for its immunogenicand protective potential in rabbits and sheep atIIL, Hyderabad.Production and the evaluation self replicatingDNA vaccines for foot and mouth disease(FMD)The project is designed to develop self replicatingDNA vaccine as an alternative to conventionalFMD vaccine by achieving higher expression,secretion and anchoring the protein secreted out.Two Sindbis polymerase gene based vectors forhumoral immune response were constructed. Thelinked polyvalent protein genes of FMDVserotype A, O and Asia 1 were cloned into thevector. The functionality of the constructed DNAvaccine (pVac Self Rep 990) was confirmed byWestern blot analysis. Guinea pigs vaccinated withthe polyvalent vaccine construct and challengedwith 100 GPID 50of each serotype separatelyshowed protective response. This approach ofconstructing self replicating DNA vaccine forhumoral response is the first report. The vaccineconstructs were evaluated in bovine calves. FourSiRNA (D1,D2,D3 and D4) producing cassetteswere constructed and confirmed by sequenceanalysis. In vitro studies on these cassettes werecarried out in BHK cells. Protective responsestudies were carried out in guinea pigs. One ofthe constructs (D2) showed 100% protectionagainst challenge virus.Development of biofilm based vaccine againstmastitis causing S. aureus and E. coli organismsin bovinesStaphylococcus aureus and Escherichia coli werefound to be the predominant isolates from thecases of bovine mastitis. The prevalence of intercellular adhesion A (ica A) and Biofilm associatedprotein (Bap) genes among S. aureus isolates byPCR studies, suggested its role in biofilmproduction. In vitro growth conditions werestandardized for optimum production of bentoniteclay based biofilms by S. aureus and E. coli.Immunoblotting of BF and Free Cell (FC) OMPsconfirmed the cross reactivity among strains/serotypes. Vaccination trials in rabbits indicatedthe low values of mastitis indicators in bothhomologous and heterologous challenged groupsamong S. aureus and E. coli BF vaccinates. Cattlein early lactation were subjected to vaccinationtrials using benetonite clay based S. aureus and E.coli BF and FC vaccines. In S. aureus and E. coliBF vaccinated cattle, significant enhancement inCD4 and CD8 cells by Flow cytometry andenhanced serum lgG levels by indirect ELISA weredetected when compared to FC and control groups.The outcome of the study indicted the supremacyof BF vaccine over FC vaccine.Aquaculture & Marine BiotechnologyThe aquaculture and marine biotechnologyprogrammes were targeted towards, identificationof novel bioactive molecules, development ofimproved feed, molecular characterization ofimportant species, development of cell lines etc.Some of the important achievements are asfollows:78Annual Report 2009-2010

Research and DevelopmentBiomoleculesA number of peptide toxins were isolated fromvarious species of cone snails and an inventory ofnearly fifty toxin peptides with confirmed sequenceswas developed. The three-dimensional structure oftoxin Ar1447 and Lo 959 was also determined.Contryphan Lo959, a toxin isolated from Conusloroisii, was found to increase the high-voltageactivated Ca current on rat dorsal root ganglionneuron while another toxin, Am975 isolated fromConus amadis, caused inhibition. The modulationof the high threshold Ca current was confirmedusing a two-pulse protocol. The analgesic effect ofthese peptides confirmed their potential therapeuticvalue. Active biomolecules isolated from variousspecies of marine sponges were screened for anticataractpotential and their ability to inhibit cataractmediated through Indoleamine 2,3 dioxygenase(IDO), a first rate limiting enzyme of tryptophancatabolism. One of the compounds screened viz.aldrin was found to be a potent in normalizingbiochemical changes/alterations induced by UVexposure in lenticular tissue of eyes. Mutationscreening of IDO gene in cataract cases were carriedout.Marine actinomycetes were screened for bioactivecompounds like antibiotics, enzyme inhibitors, anticholesterolimic,insecticidal and bioherbicidalcompounds. The physiological and biochemicalcharacterization of two potent isolates from coralreefs, showed significant activity against multi-drugresistant Staphylococcus aureus and Pseudomonasaeruginosa. A few of the isolates showed significantamylase inhibitory and insecticidal activities.More than 20 marine organisms, harbouringpotent antimalarial bioactive compounds, wereidentified. The purified compounds arrested thegrowth of parasite at trophozoite stage and oneof the molecules was found to inhibit invasionof merozoites into fresh red cells (Fig). Thechemical structure of these antimalarials wasdetermined to facilitate chemical synthesis. Thesecondary metabolites of endophytic fungalisolates, isolated from marine algae and sea-grasses,were tested for antibacterial, anti-fungal, antialgal,anti-oxidant and anti-insecticidal activities.Seven endophytic fungi showed antialgal activityand one isolate exhibited antifungal activity whilefour showed insecticidal activities. About 90% ofthe endophytic fungal isolates showed antibacterialactivities (Fig).Malaria parasite infected red blood cells have been stained with SYBRGreen dye. Parasitemia was varied by differential dilutions of theinfected culture with uninfected red blood cells. The linear relationshipbetween % parasitemia (abscissa) and Relative fluorescence (RF)(Ordinate) allows HTP evaluation of the IC 50of test drugs.Annual Report 2009-2010 79

Research and DevelopmentCell Line DevelopmentA multi centric programme on the development ofcell lines from various fresh water and marine fisheswas initiated. Cell lines from eye muscle, braintissue and heart tissue of Catla, eye muscle ofLabeo rohita, eye muscle of grouper (Epinephelus)and fin tissue of Indian catfish were established andcharacterized. These cell lines were sub-culturedcontinuously and showed characteristics of eitherepithelial cells or fibroblast cells. The grouper eyecell line was found to be susceptible to nodavirusand the infection was confirmed by cytopathic effectand reverse RT - PCR. Clown fish cell lines weredeveloped from various tissues and used for thequantification of viral agents from marineornamental fishes.Diagnostics and VaccinesA diagnostic test was developed for the identificationof white tail disease (WTD) of Macrobrachiumrosenbergii using reverse transcriptase loop mediatedisothermal amplification (RT-LAMP). The diagnosticmethod was more sensitive than the RT-PCRmethod, currently used to detect thehepatopancreatic parvovirus (HPV), in shrimp. Theloop mediated isothermal amplification assay(LAMP) was developed for the detection ofparvovirus. The detection limit of LAMP was 0.1pg.Efforts continued to develop a DNA based vaccineagainst Aeromonas spp. The lamB gene of A.hydrophila was amplified and cloned into the DNAvaccine vectors targeting the expression of the antigeninto cytosolic, endosomal and secreted endosomalexpression. Immunization of mice with differentdoses of plasmid DNA expressing cytosolic lamBporin was done to check immune response.Immunization studies in Labeo rohita with pEGFP/lamB DNA construct was initiated. The challengedose of A. hydrophila for Labeo rohita wasoptimized.Genetic CharacterizationMolecular markers were developed for theidentification of various species of Tor andNeolissochilus hexagonolepis at NBFGR, Lucknow.The two rDNA families (45S and 5S) were preciselymapped on the chromosomes of Tor spp. The 5SrDNA was found to be localized on a single pair ofchromosomes in T. tor, T. putitora, T. progeneius, T.chelynoides and N. hexagonolepis and one additional5S rDNA signal was found in T. mosal mahanadicus,which can be used as a molecular marker for thissubspecies. EST libraries of rohu were constructedand characterized. About 470 clones were sequencedfrom the brain library which resulted in 329 uniquesequences. A cDNA library was constructed fromthe spleen of Clarias batrachus and approximately5000 clones were screened. So far, 1014 cDNAclones were sequenced. Analysis of 208 cDNAsequences revealed similarity of clones with annotatedproteins including 4 immune relevant genes.Molecular characterization of Artemia, was carriedout. ITS-1 sequence of the 27 specimens belongingto eight geographical strains was PCR amplifiedand sequenced along with the reference strain ofArtemia franciscana. The study confirmed the closerelation of Indian strains to A. franciscana. Selectivebreeding of various strains up to five generationswere carried out and the naupliar size was reducedfrom 494 ìm in the base generation to 470 ìm inthe fifth generation. Nutritional profiles showedthem to be suitable as live food.80Annual Report 2009-2010

Research and DevelopmentMolecular characterization of white spot syndromevirus (WSSV) was carried out using variable numbertandem repeat (VNTR) based genome analysis.Pathogenicity studies of 12 WSSV isolates of shrimpcarried out by oral challenge in juvenile Penaeusmonodon showed variation in induced mortality.Fourteen WSSV isolates were analysed for thefunctional ORFs for the detection of possiblevirulence associated genes. The anti-apoptosis geneis likely to be one of the genes causing mortality inshrimps (Fig).ORF 94 based VNTR specific banding pattern of various WSSV isolatesobserved in 1.2% agarose gel.Lane 1- M128,lane 2- M135,lane 3-M136,lane 4- M137,lane 5- M140, lane 6-100bp marker,lane 7-M32,lane 8- M57,lane 9- M77P1,lane10-NCORF125 based VNTR specific banding pattern of various WSSV isolatesobserved in 1.2% agarose gel. Lane 1- M128,lane 2- M135,lane 3-M136,lane 4- M137,lane 5- M140, lane 6-100bp marker,lane 7-M32,lane 8- M57,lane 9- M77P1,lane10-NCORF 14/15 based VNTR specific banding pattern of various WSSVisolates observed in 1.2% agarose gel. Lane 1- M128,lane 2-M135,lane 3- M136,lane 4- M137,lane 5- M140, lane 6-100bpmarker, lane 7- M32,lane 8- M57,lane 9- M77P1,lane10-NCMultiplex PCR targeting ORF94, ORF125, and ORF14/15 region ofvarious WSSV isolates observed in 1.2% agarose gel. Lane 1-M128,lane 2- M135,lane 3- M136,lane 4- M137,lane 5- M140,lane 6-100bp marker, lane 7- M32,lane 8- M57,lane 9-M77P1,lane10-NCAnnual Report 2009-2010 81

Research and DevelopmentReproductionIn an effort to develop techniques for inducedmaturation and early spawning techniques in shrimp(P. monodon), two recombinant proteins viz.crustacean hyperglycaemic hormone (CHH) andmoult inhibiting hormone (MIH) were expressedin Pichia pastoris using the genomic DNA derivedCHH and MIH genes of P. monodon.Administration of the crude protein of CHH genein P. monodon and in haemolymph was found toincrease their glucose levels up to 74%. Thephenotypic expression of MIH gene silencing wasfound to reduce the duration of the moulting cycle(6.83±1.47 days) in comparison to the control(11.5±1.04 days). In another study, a novel approachto inhibit the ERK signaling pathway, using specificinhibitor (PD98059) for the blocking ofvitellogenesis-inhibiting hormone expression wasdeveloped. These techniques will help in earlymaturation and spawning of shrimp.Regeneration and NeurogenesisThe zebra fish was used as a model animal toelucidate cellular and molecular mechanisms ofregeneration. Studies on cellular proliferationindicated a low level of neurogenesis in uninjuredadult spinal cord of zebra fish and high proliferativeresponse after crush injury. Neurogenesis enabledzebra fish to recover anatomically and functionallyfrom spinal cord injury. A proliferating precursorviz. radial glia and other restricted precursors likeastrocyte, oligodendrocyte etc. were found in theadult spinal cord. The study confirmed axonal regrowthafter spinal cord injury as shown by itsfunctional recovery and reappearance of axonalmarkers.SeribiotechnologyWork on application of biotechnology forimproving the productivity, enhancing the qualityof silk, improvement of host-plants, improving thepost-harvest processing of silk along with utilizationof by-products in sericulture continued during theyear. A “Consortium on mulberry genomics andcrop improvement” in a network mode is beinginitiated with the aim to identify genes and QTLsfor crop improvement through well focusedmolecular breeding and transgenic approaches.Significant achievements during the year aresummarized:Development of improved Races of Silkwormfor Increased ProductivityA network project implemented at CDFD,Hyderabad; Seribiotech Research Laboratory (SBRL),Bangalore; Central Sericulture Research and TrainingInstitute (CSR&TI), Mysore; Andhra Pradesh StateSericulture Research and Development Institute(APSSR&DI), Hindupur and Karnataka StateSericulture Research and Development Institute(KSSR&DI), Bangalore led to the identification ofa few putative DNA markers associated withBmNPV resistance in silkworm (Bombyx mori).Studies have been initiated on mapping andvalidation of these markers with the aim to introgressthem into the desired breeds of B. mori jointly atSBRL, Bangalore and APSSR&DI, Hindupur.Efforts continued to identify cDNA markers linkedto genes controlling cocoon and post-cocoon traitsin B. mori at CSR&TI, Mysore. The EST cDNAclones (five each) pertaining to all the 28 linkagegroups have been amplified through transformation82Annual Report 2009-2010

Research and Developmentof E. coli strain. Plasmid DNA along with ESTswas extracted, purified and quantified.Studies continued to identify the physical,environmental and genetic factors responsible forpupal diapause and mechanism in tasar silkworm(Antheraea mylitta) at CTR&TI, Ranchi. Duringearly and mid-aged pupae in diapausing generations,accumulation of four diapause specific proteins wasobserved. The role of these proteins in the fecundityand reproduction in A. mylitta is being studied.Work has been recently initiated on identificationof genes involved in the diapause process in diapauseinduced eggs of mulberry silkworm (B. mori) atSBRL, Bangalore.Development of Control Measures for MajorDiseases and Pests of SilkwormStudies on epidemiology, spatial and temporaldynamics of important muga silkworm (Antheraeaassama) diseases continued jointly at AssamAgricultural University, Jorhat and CMER&TI,Jorhat. Analysis of bacterial diseases (toxicosis andsepticemia) with the weather data showed that thedisease incidence was increased by the maximumtemperature and rainfall. The incidence of fungaldiseases caused by Baeuveria bassiana, Aspergillusniger and Fusarium moniliforme were found to besignificantly affected by the maximum temperatureand relative humidity. Work on identification ofproteins involved in host-parasite interactionfollowing uzi fly infestation in the mulberrysilkworm (B. mori) and eri silkworm (Samia cynthiaricini) was initiated at SBRL, Bangalore. Studies ondifferent microsporidians infecting mulberry andnon-mulberry silkworms with respect to theiridentification, life cycle features, cross infectivity,pathogenecity, tissue specificity and transovarialtransmission have been recently initiated at SBRL,Bangalore.Improvement of Host-PlantsA network project implemented at Centre forCellular and Molecular Biology (CCMB),Hyderabad, CSR&TI, Mysore, KSSR&DI,Bangalore; CSR&TI, Berhampore and CentralSericultural Germplasm Resource Centre (CSGRC),Hosur led to identification of a DNA markerassociated with resistance to powdery mildew(Phyllactinia corylea). Work on validation andutilization of SCAR marker(s) for powdery mildewresistance in mulberry has been recently initiatedjointly at CCMB, Hyderabad and CSR&TI,Berhampore.The first generation HVA-I mulberry transgenicplants with constitutive and inducible promotershave been developed for drought and salinitytolerance at UDSC, New Delhi. Eleven HVA-Itransgenic lines have been transferred to CSR&TI,Mysore for evaluation of the agronomic traits.Additional mulberry transgenics with bch (betacarotene hydroxylase) gene have also been generatedand are being evaluated for tolerance against hightemperature,high irradiation and UV-stress.Efforts were continued to develop mulberrytransgenics over-expressing stress-specific transcriptionfactor (DREB2A) to increase drought tolerance andreduced post harvest loss jointly at University ofAgricultural Sciences (UAS), Bangalore andCSR&TI, Mysore. Full-length SHN-3 like genewas successfully cloned from mulberry cDNA andused for constructing suitable binary vector for over-Annual Report 2009-2010 83

Research and Developmentexpression. Transformation protocols in elite varietiesof mulberry (V-1 and S-36) are being standardized(Fig).the gene pool of tropical tasar silkworm host plantswith respect to yield and quality of tasar silk hasbeen carried out at CTR&TI, Ranchi. Indexdescriptor of 50 accessions each of Terminaliaarjuna and T. tomentosa and estimation of geneticdiversity based on 36 phenotypic parameters hasbeen completed. All the 100 accessions studiedwere grouped in nine clusters based on leaf qualityand cocoon characteristics.Improving Post-Harvest Processing of SilkStandardization of multiple shoot induction protocol from mulberry(variety V1) 1 & 2: Establishment of mulberry cultures from nodalexplants, 3: Multiple shoots emerging from the basal region ofexplant, 4: Root initiation from plantlets, 5: Hardening of culturedplantQTLs for water use efficiency (WUE) and relatedroot traits in mulberry were identified jointly atUAS, Bangalore and CSR&TI, Mysore. A fewpromising lines of mulberry with high growth ratewere identified.Work on characterization of transcriptome ofmulberry was taken up at UDSC, New Delhi.EST sequencing has been done from the matureleaf and root cDNA libraries and also from thesubtracted cDNA libraries from the mature leaffor drought tolerance. Functional characterizationof genes of interest is in progress. Evaluation ofWork on production of microbial dyes and theirapplication to silk and other fibres has beencontinued jointly at Bhavan’s College, Mumbai andInstitute of Chemical Technology (ICT), Mumbai.Six pigment producing strains namely N-1 (bluishviolet),A-10 (red), A-11 (rose pink), L-1 (yellow),E-1 (orange) and MF-1 (green) were shortlistedbased on the consistency of their production andcharacterization. Dyeing with extracts of thesepigments namely V-1, A-10 and A-11 was carriedout on silk, wool and cotton fibres. The dye wasfound to have an affinity for all the three fibrestested. The test for fastness properties, washing andlight showed poor rating but the rubbing fastnessproperties were good.Utilization of By-ProductsStudy on up-scaling of the process at pilot-level forthe recovery and production of sericin powder fromthe industrial waste degumming liquor was takenup at IIT, New Delhi, jointly with an industrialpartner – M/s Nath Brothers Exim Ltd., Noida.The design of the pilot plant of 1000 liter/daycapacity along with centrifuge unit of capacity 100litre has been finalized.84Annual Report 2009-2010

Research and DevelopmentBasic Research in Modern BiologySupport to basic research was continued to increasethe understanding of fundamental principles andtheories that would help in evolving appropriatestrategies for research. The salient achievements ofsome of the on-going projects are summarized:Drug DeliveryA synthetic polymer hydrogel based on N-vinylpyrrolidone and hydroxy ethyl methacrylate wassynthesized at BAIT, Sathyamangalam and itsstructure and composition were determined andcorrelated with its swellability and drug releaseproperties. This polymer seems to be a promisingcarrier for delivering drugs with structure andproperties similar to paracetamol and wasbiodegradable.Molecular CardiologyStudies conducted on in vitro model of cellularhypoxia confirmed that hypoxia retards G1-Stransition in cardiac fibroblasts. Significant reductionsin cyclins D1 and A were observed in hypoxiccardiac fibroblasts. The cyclin-dependent kinaseinhibitor, p27 in these cells under hypoxic conditionsshowed that p38 MAPK-mediated p27 inductiontrigger hypoxic regulation of G1-S transition incardiac fibroblasts.Molecular BiologyIn a project on nuclear factor of activated T-antigen(NFAT), a state of art facility in mammalian cellculture was created at Panjab University,Chandigarh. The raw material required tounderstand the pathophysiology associated withAutosomal Polycystic Kidney Disease (ADPKD)was generated. The findings revealed that activationof the NFAT was a consequence of polycystin-1and Gá12 crosstalk resulted in developmentaldefects in kidney and cardiac tissues in ADPKDpatients.A comprehensive examination of the regulatorynetwork required for iron homeostasis in thegenome of human pathogenic fungus Candidaalbicans was carried out at ICGEB, New Delhi. Ina mouse model of systemic candidiasis, cap2-/-mutant was impaired for virulence, to establish thecritical role of CAP2 in vivo.Study conducted at IICB, Kolkata, has generated amouse model for inflammation which has manyfeatures of Rheumatoid Arthritis (RA). The culturedcells isolated from lymph glands of the mice showedthe presence of macrophage like cells that are knownto promote inflammation (Fig). Analysis ofmessenger RNA (mRNA) from these cells showedincreased expression of the growth modulatorWnt5A when compared to control cells harvestedfrom the lymph glands of normal mice.ExperimentalControlRepresentation of macrophages like cells cultured from the lymph nodeof arthritic (experimental) mice as pointed out by arrows. Such cellsare not present in similar abundance in lymph node cell culture ofcontrol mice.Annual Report 2009-2010 85

Research and DevelopmentBiophysical StudiesA series of glycolipids presenting sugar residues inthe form of oligosaccharides of M. tuberculosis weresynthesized and their structural properties establishedat IISc., Bangalore The studies have shown thatsynthetic glycolipids not only inhibit the growth ofMycobacterium, but also inhibit the motilities ofgrowing bacteria and affected the bio-film, whichprovides protection for mycobacterial survival. Workdone at IISER, Chandigarh resulted in computingthe full chemical shift anisotropy tensor using DFTlevel theory for protein backbone nuclei participatingin different secondary structures. 2D and 3D CSAparameter plots were constructed for differentbackbone nuclei and the role of such correlations inidentifying secondary structure elements in proteinswas investigated.straight in normal state, form clusters and becomewavy from the seventh decade of life. From thesixth decade onwards, the aging retinas prominentlyexpressed the biomarkers of oxidative stress such as4-hydroxy nonenal (Fig.) and nuclear respiratoryfactors-1. The endogenous antioxidants glutathione–S- transferase ð-1 and superoxide dismutase-2, onthe other hand, showed a decreased expression withaging.Reproductive BiologyThe male reproductive tract specific antimicrobialgene expression in rodents was studied at Universityof Hyderabad, Hyderabad. Experimental rats treatedwith bacterial toxic compound lipopolysaccharide,showed increased antimicrobial gene expression inthe male reproductive tract. Induction of proinflammatorycytokine expression precededantimicrobial induction, suggesting the involvementof inflammatory processes during reproductive tractinfections. The study will help in developing peptideantibiotics to treat these infections.NeurobiologyIn an effort to investigate the impact of oxidativestress on photoreceptor changes in the aging humanretina at AIIMS, New Delhi, it was found that thephotoreceptor microtubules, which are disperse andNuclear respiratory factor-1immunoreactivity is localised in outer andinner nuclear layer (onl, inl)Medical BiotechnologyVACCINE RESEARCH & DEVELOPMENTThe department has made concerted efforts todevelop human vaccines through Task Force onMedical Biotechnology and also through NationalJai Vigyan Mission on S&T for generation of newvaccines during the past two decades. Vaccines remainthe most cost effective means to combat infectiousdiseases. Several candidate vaccines emerged fromthese efforts. Some of them are into pre-clinical orclinical trial stages e.g. vaccines for Rotavirus,86Annual Report 2009-2010

Research and DevelopmentCholera, Typhoid, Rabies (DNA based), Anthrax,Malaria, Dengue, Tuberculosis, Japanese Encephalitis,etc.The major achievements during the year on thedevelopment of vaccines are summarized.RotavirusThe projects on rotaviral diarrhoea wereimplemented at AIIMS, New Delhi in collaborationwith CDC, Atlanta; and IISc, Bangalore incollaboration with Stanford University, USA. Thefirst rotaviral diarrhoea vaccine 116E has beendeveloped in India and phase-II human clinical trialssuccessfully completed. The vaccine induced immuneresponse was reported in ~90% of the recipients.The Society for Applied Studies (SAS), New Delhihas conducted a clinical trial with financial supportfrom DBT, NIH & Programme for AppropriateTechnology in Health (PATH) to evaluate the safetyand immunogenicity of a live attenuated rotavirusvaccine (116E), manufactured by M/s BharatBiotech International Limited, Hyderabad underGMP conditions. No adverse events have beenreported in the trial till date. The investigators arenow doing the disease burden study and sitepreparation to conduct large phase-III clinical trialsat three sites i.e. SAS, New Delhi; CMC, Velloreand KEM Hospital, Pune.MalariaContinued support was provided for research onmalaria vaccine at ICGEB, New Delhi and NationalInstitute of Malaria Research, New Delhi. A fieldtrial site has been identified at Malaria ResearchCentre, Rourkela, to evaluate the candidate malariavaccines.The Indian Malaria Vaccine Programme (IMVP)was established after discussions among DBT, MVI,EMVI and ICGEB. The overall objective of IMVPis to promote the development of vaccines againstP. falciparum and P. vivax malaria in infants indeveloping countries by leveraging the work doneat ICGEB and available competencies in productdevelopment in India and clinical development inIndia and outside. The Bill & Melinda GatesFoundation have provided $2.95 million formanagement of IMVP. The MOU has been signedbetween DBT, ICGEB, and MVI. A ProgrammeDirector was appointed to coordinate the activitiesof IMVP.Dengue VaccineUnder the project on experimental tetravalent denguevaccine project implemented at ICGEB, New Delhi,a novel tetravalent chimeric protein was underdevelopment. Efforts were on for animal studies inAGK0129 mice strain for the development oftetravalent dengue vaccine and start preclinical andclinical studies on dengue with the help ofinternational experts. An Indo-US consultationmeeting on Dengue vaccine development wasorganized in July, 2009. The experts observed thatthe dengue vaccine work carried out at ICGEB wasinnovative and the only subunit vaccine approachin the world today that had the potential to be ofrelevance to the pediatric population. It was alsoobserved that the ICGEB’s tetravalent vaccine designwas elegant and the neutralization titers elicited wererespectable by world standards.Vaccine Grand Challenge ProgrammeVaccine Grand Challenge Programme has been underAnnual Report 2009-2010 87

Research and Developmentimplementation since 2008-09 with an overallobjective to accelerate development of candidatevaccines for Rotavirus, Cholera, Typhoid, Rabies,Malaria, Dengue and Tuberculosis. Proposals wereinvited through open advertisement in leadingnewspapers for support in the identified areas underthe programme. Altogether 23 proposals werereceived and these were evaluated through twomeetings of Apex Committee of the programme.Efforts have been made to establish a separate unitto handle the programme through a programmemanagement team.Indo-US meeting on Low Cost Diagnostics andTherapeutic TechnologiesA new initiative was taken during the year withNational Institute of Biomedical Imaging andBioengineering (NIBIB), NIH, USA. The groupreviewed. The programme implementation andfinalized the Request for Application (RFA) to inviteproposals in the area of low cost glucose monitoring,low cost platform technologies for multiple(multivalent) diagnostics tests, low cost ultrasoundimaging, low cost ultrasound therapy, innovativeimaging technology etc.Joint Working Group meeting of Indo-USVaccine Action ProgrammeThe 23 rd meeting of the Joint Working Group(JWG) of Indo-US Vaccine Action Programme(VAP) was held in November, 2009 at NIH,Washington DC, USA. The JWG reviewed theprogress made under ongoing, completed and newprojects, especially development of rotaviral diarrhoeavaccine dose escalation studies and initiation ofphase-III clinical trials in India, projects on malaria,dengue and tuberculosis, alongwith therecommendations of the TB and dengue consultationmeetings held in July, 2009 in India. The JWGalso discussed technical support to TranslationalHealth Science & Technology Institute of DBTand providing pre-clinical services by NIAID/NIH.Vaccine & Infectious Disease Research Centre(VIDRC)Vaccine & Infectious Disease Research Centre(VIDRC) has been established as one of the nichecentres of the newly established Translational HealthScience & Technology Institute, with an overallobjective to conduct multi disciplinary basic andclinical research on vaccines and biologics and takeup some of the available vaccine candidates and assistdevelopment activities in a mission mode as well asdevelop newer vaccines relevant to India with a broadfocus on the progression of vaccine development.INFECTIOUS DISEASESTuberculosisThe department has identified tuberculosis as afrontline area of research since the last two decadesand has been funding basic, applied and translationalprojects. Currently funding has been granted toabout 120 groups; and about Rs 100 crores ofinvestment has been made in the last three years.Major virtual Centres of Excellence with hubs atICGEB, New Delhi; University of Hyderabad,Hyderabad; & IISc, bangalore have been created.The major areas of research include: - diagnostics,biomarker research, immuno-modulation, systemsbiology, basic biology, crystallography and chemicalgenomics88Annual Report 2009-2010

Research and DevelopmentBesides funding R & D projects, DBT alsoestablished exclusive facilities such as a MycobacterialRepository at JALMA, Agra, Crystallography Unitsat IIT-Kanpur & Kharagpur and an AerosolChallenge Facility each at JALMA, Agra and ICGEB,New Delhi. The facility conforms to the guidelineslaid down by WHO on Biosafety in handling M.tuberculosis.DBT’s Tuberculosis Aerosol Challenge Facility (DBT-TACF) atICGEB, New DelhiUnder a project at the UDSC, New Delhi, anELISA based test system for confirming MTBcomplex for treatment decision using MPT64 alongwith another antigen (patented) has been developedin association with the M/S Span Diagnostics, Surat.Initial results are encouraging and a multi-centricvalidation trial is underway.In an effort to develop novel anti-mycobacterialagents against latent and MDR tuberculosis, researchwas pursued at M/s Ranbaxy Research Foundation,Gurgaon to design and synthesize novel analoguesof J & J compound-TMC-207 & screening ofNCEs from proprietary library against Mycobacteriausing enzyme-based and whole-cell based assays.Several hits with MIC against H 37Rv were identified.In a study conducted at NII, New Delhi, resultsshow that MIP immunization provides significantprotection against challenge with M. tuberculosis inmice as well as guinea pig model. Also,immunization with MIP by aerosol route inducesearly local lung responses as well as systemicresponse. The studies showed that MIP inducedprotective immune response against M. tuberculosisby activating macrophage and lymphocytes asindicated by IFN-α secretion and cytolytic activity.Activated macrophages were found to inhibit theintracellular growth and multiplication of M.tuberculosis.Cholera Vaccine DevelopmentThe department continued its support for furtherdevelopment of the live oral cholera vaccine. Thecandidate vaccine strain VA1.3 developed byIMTECH, Chandigarh had undergone Phase I/IIstudies at SAS, Kolkata; SGPGIMS, Lucknow andCMC, Vellore and it was found to be safe andsignificantly immunogenic. The vaccine was notshed and hence, did not have any environmentalconnotation. A new strain VA1.4 was made withthe removal of ampicillin marker. It is proposed toclinically test the new strain for safety andimmunogenicity for which M/s Shantha BiotechnicsLimited has been engaged as a contract manufacturerAnnual Report 2009-2010 89

Research and Developmentof the cGMP material. A distinct formulation thathas the desired stability till six months has beenprepared. The vaccine batch prepared under GMPconditions would now undergo regulatory toxicologyat INTOX, Pune, the vendor chosen by DBT foroutsourcing regulatory toxicology. Phase I/II trialsare underwayChronic Disease BiologyChronic Diseases Biology (CDB) has emerged as animportant area of research over the years. Significantfunding was provided to projects covering cancer,neurosciences, cardiac diseases, reproductive biology,diabetes etc. A special impetus was given to generatenationally relevant projects through three ExpertGroups on :1. Cancer biology, biomarkers & immunology2. Neurosciences and neuro-engineering3. Endocrine, cardiac & renal, autoimmunediseases & reproductive biologyCancerCancer continues to attract most researchers andthere were about 130 projects in cancer area coveringbiomarkers, drug design, genetic polymorphism,immunotherapy etc. The department is funding anumber of trials using curcumin in different cancerssuch as oral premalignant lesions, head & neckcancer and cervix cancer. While the former is aprophylactic trial, the latter is investigating the radiosensitizingand chemo-sensitizing ability of curcuminin different formulations. Curcumin is known tohave little bio-availability. Hence, a special effortwas made to create a “Curcumin Clinical Assay’sLab” at ACTREC, Mumbai. Samples from all siteswould be transported to the lab for evaluation forthe presence of curcumin or its metabolites inplasma. The results would then be correlated withthe clinical & biological biomarkers using validatedinstruments and protocols. A number of trials areunderway involving institutions across the country.NeurosciencesDBT constituted a new Expert Group onNeurosciences. The group discussed new ideas/novelapproaches to address various issues concerningneurosciences i.e. dementia (neuro-degeneration),stroke, spinal cord injury, neuro-inflammation,epilepsy and imaging, as also mentorship programmefor enlisted areas. The department also sponsoredan “Idea Generation Session” for discussing researchareas and funding opportunities in neurosciences.The efforts are designed to develop research projectsand establish long-term cohort for neurosciencerelated areas.Infectious Disease BiologyProgrammes were supported to address importantissues relating to pathogen biology and relevantaspects of infectious diseases like HIV/AIDS,Hepatitis, Chikungunya, Tuberculosis, Malaria,Leishmaniasis, Rotaviral infection, Cholera etc. Thelong term goal is to develop products and processesto prevent, detect and treat these diseases. Theendeavor of the department was to create an efficientpathway to bring scientific innovation from the labto the translational phase and leading to developmentof vaccines, diagnostics, and therapeutics. Duringthe year a total of 58 new R&D projects wereconsidered for financial support, out of which 23were supported this, the Besides, progress of 5990Annual Report 2009-2010

Research and Developmentongoing and 26 completed projects was alsoreviewed. Significant achievements of some of theR&D programmes are summarized below:Viral DiseasesTowards understanding the potential role of HIV-1 subtype-C Vif gene variations among Indianisolates in differential neutralization of APOBEC3G/3F and its possible role in diversification of HIV-1, mammalian expression vectors for HIV-1 vifvariants from infected samples were amplified andconstructed. Also, sub-type B/C chimeric vif clonewas generated using pNL4.3 and Indian C isolate.A study on the virological and immunologicalaspects associated with opportunistic viral infectionsin HIV-1 infected individuals in South India,indicated that individuals with < 100 CD4 + T cellcounts had comparatively more opportunistic viraltypes detectable. Studies towards understanding therole of HIV-1 trans-activating protein Tat on theexpression profile of inflammatory cytokines fromthe human fetal CNS progenitor cells and well asthe differentiation indicated that Tat modulateshNPCs proliferation by altering the phosphorylationstatus of MAKP/ERK-1/2. Tat was also found todown-regulate the expression of cyclin-D1, whichcritical for cell proliferation.Studies involving viral proteins and other molecularmarkers in hepatitis E virus (HEV) infected cellsshowed that the fate of specific RNA can befollowed experimentally during synthesis. This alsoindicates that premature termination duringreplication cycle resulted in detection of prematurefallout RNA species. Viral genome synthesis showedan inverse relation between the two strands in atime kinetic study with synthetic bursts in a timepoint periodicity. Inhibition studies of HCV RNAtranslation and replication using small RNAs showedthat appropriate targeting of the viral genome usingshRNA can significantly inhibit viral replication. Invivo liver targeted delivery of such interfering shRNAthrough Sendai virosome can be considered animportant therapeutic strategy. In future, appropriatesmall animal models with replicating HCV will bestudied to demonstrate efficacy of the virosomesystem in vivo.Role of Vaccinia virus complement control proteinin viral pathogenesis revealed that virus complementcontrol protein (VCP) played an important role invirulence of vaccinia virus. The virulence activity ofVCP, however, was much more pronounced inrabbits in comparison to mice.In silico identification of Chikungunya virus(CHIKV) inhibitors from natural product librariesbased on E1 envelope protein structure was carriedout. Immunological tests for early detection ofChikungunya viral infection are being developed.PCR assays for detection of viral RNA was doneon nearly 20% of specimens. A 40-kDa proteinwas found to be present in most of the specimenstesting positive for CHIKV RNA. Towardsdevelopment of Chikungunya vaccine candidates,various CHIKV isolates were tested for growth inVero cells. One isolate that showed titer of 108pfu/ml in Vero cells has been found. This isolatehas purified and visualized by electron microscopy.Bacterial DiseasesStudies on quorum sensing in immune modulationand virulence of Pseudomonas aeruginosa inpathogenesis of urinary tract infections showed thatthe levels of macrophage inflammatory protein-2,Annual Report 2009-2010 91

Research and Developmenttumour necrosis factor- alpha and IL-6 increased inmice with the standard strain PAO1. In an effortto assess zinc as an immunomodulator in thetreatment of possible serious bacterial infections ininfants more than 7 days and up to 4 months ofage, a double-blind randomized controlled trial wasconducted Seven hundred children were recruitedand the analysis has been made by groups withoutunblinding. The results have the potential toinfluence the management of childhood bacterialinfections which canses considerable morbidity andmortality. Another study has also been undertakento investigate the effects of administering 10 mgzinc orally to young infants aged 7 days and up to4 months with bacterial infection, on the survivaland function of peripheral blood mononuclear cells.The preliminary results in children with sepsis whencompared with patients without sepsis demonstratedlower total T lymphocytes, higher CD4:CD8 ratio,lower serum zinc, a positive correlation betweenCD4:CD8 ratio and serum zinc, no correlationbetween serum zinc and total leukocyte counts andno difference in apoptotic cells.Parasitic DiseasesStudies on the effect of gene silencing on themodulation of cell signaling mechanisms duringthe early phase of Leishmania donovani infectionshowed that exogenous chemokines like IP-10 andMIP-1alpha could act directly on parasites andreduce the survival of Leishmania promastigotes.Leishmania target antigens from promastigotes andamastigotes have been identified on the basis ofth1 stimulatory proteins for immunoprophylacticstudies against experimental visceral leishmaniasis.Toward cloning, expression and characterization offilarial Acetylcholinesterase (AchE), two isozymesof filarial AchE were found to be biochemicallyand immunochemically similar but different fromthe host enzyme. One of the monoclonal antibodiesproduced against the parasite AchE specificallyinhibited the enzyme activity of both the isoformsof parasite AchE. These findings suggested thatthe same antifilarial agent/compound can be usedto target both the isoenzymic forms of parasiteAchE.DBT-ICMR Collaborative EffortThe DBT-ICMR collaborative effort in the HIV/AIDS and Microbicides area has laid emphasis ondeveloping HIV viral load assays, CD4 assays;designing new vectors & adjuvant formulations;understanding mucosal immunity, developingcombinational microbicide formulations; preclinicaland clinical testing of microbicides etc. Currently,19 R&D projects are operational. The salientachievements were:HIV/AIDSIn order to design stable folded fragments of HIVenv for use as immunogens, CD4Bs a gp120 basedfragment was designed and it was found to elicitsignificantly better neutralization than gp120.Attempts are being made towards patenting theCD4Bs immunogen. Indian clade C envelope clonesisolated from recently transmitted HIV-1 showedmoderate sensitivities to a variety of antibodiesincluding broadly neutralizing monoclonalantibodies. They are being further characterized tofind domains conferring neutralization sensitivity. Anew class of specific and tight binders weresynthesized which inhibit Rev-RRE (Rev ResponsiveElement) interactions in HIV that can lead to92Annual Report 2009-2010

Research and Developmentdevelopment of inhibitors of HIV replication. Avirtual Knowledge and Resource Centre is functionalat NARI, Pune. Fifty eight participants registered atthe website will provide a list of reagents andtechnologies available with them. Various institutesprovided protocols, assays, cell lines for HIVisolation from peripheral blood mononuclear cells,Hetroduplex Mobility Analysis, biological isolationof HIV from semen from HIV-1 infected individualsand purification of Tat protein and its functionalanalysis etc.MicrobicidesVarious chemically synthesized reverse transcriptaseinhibitors showed potential to be developed as anti-HIV molecules. Several test preparations fromdifferent plant sources showed promising anti-HIVactivity. ‘BASANT’, a polyherbal gel as vaginalmicrobicide in terms of safety and acceptability hasshown promising results among women tested in aphase I clinical trial. At National facility (BSL-3Level), NII, New Delhi, anti-HIV activity has beenshown in various plant extracts using variousstandardized assays.Stem Cell ResearchOver the past few years, human stem cell researchhas emerged as a new and exciting field of lifescience in view of its potential therapeuticapplications. Being a highly integrative field oflife sciences, stem cell research requires closeinteraction of basic researchers, clinicians and theindustry. Programmes were initiated on differentaspects of embryonic and adult stem cells. Priorityareas were identified and categorized into basicresearch, pre-clinical & clinical research, institutionaldevelopment, creation of facilities and humanresource development.Currently, the challenges in this area includeavailability of expertise, adequate infrastructure,interdisciplinary network of researchers and cliniciansfor theme-based research, well defined basic researchleading to clinical/translational research andappropriate regulatory mechanisms. Efforts weremade to create a critical mass in the country. Duringthe period, about 30 projects were received and 20sanctioned on various aspects of basic andtranslational research.Adult Stem CellsIn the area of adult stem cells, programmes wereimplemented on in-vitro differentiation ofmesenchymal stem cells (MSCs) of adult and fetalorigin into neuron progenitor cells, role ofmicroRNAs in self renewal and tumorigenicity ofbrain tumor stem cells, identification of themolecular mechanisms involved in the induction ofproliferative response in primitive hematopoieticstem cells (HSCs) and biology of cancer stem cells.The ex-vivo expansion of HSCs, generation of stemcell lines and knockdown mice that lack specificgene products, the identification of novel therapeuticstrategies, cancer stem cells in squamous cellcarcinoma of buccal cavity and tongue, pluripotencyand differentiation of stem cells using the mouseESC line, MSCs construct on osteogenesis forrepairing large bone defects in animal models etc.were taken up for study.Embryonic Stem CellsIn the area of embryonic stem cells, programmeshave been supported for the generation of humanAnnual Report 2009-2010 93

Research and Developmentautologous embryonic stem cell lines by somaticcell nuclear transfer and parthenogenesis, assaysfor toxicity testing and drug discovery usingcardiomyocytes differentiated from hESCs,technologies of parthenogenesis and somatic cellnuclear transfer using sheep eggs, generation ofinduced pluripotent stem cells (iPSCs) from humansomatic cells, in-vitro differentiation of hESC linesto hepatocytes and assessing the potential role ofseroronin in its expansion and/or differentiation,etc. The hESC lines deposited in UK Stem CellBank were distributed to the scientists in the countryfor research purposes.Clinical ResearchPatients were recruited under multi-centric phase-IIIclinical trials on acute myocardial infarction usingautologous bone marrow mononuclear cellsimplemented at six hospitals in the country. About80% recruitment was completed by all the centres.A CRO was engaged for monitoring this study .Similarly, all the five sites were taken up for therecruitment of patients under phase-II clinical studyon acute ischemic stroke. The patients are also beingrecruited under critical limb ischemia after obtainingethical clearances at four centres.Guidelines for Stem Cell ResearchGuidelines for stem cell research in the countryfinalized jointly by the Department and IndianCouncil for Medical Research (ICMR) has beensubmitted to the Ministry of Health and FamilyWelfare for further processing for legislation. TheNational Apex Committee for Stem Cell Researchand Therapy (NAC-SCRT) is being constituted asper these guidelines.BioengineeringBioengineering integrates physical, chemical,mathematical, computational sciences with principlesof engineering to study biology, medicine, behaviour,and health. It advances fundamental concepts, createsknowledge from the molecular to the organ systemlevels and helps develop innovative, processes,implants, devices, and informatics for the prevention,diagnosis, and treatment of disease. The key aim ofthe bioengineering programme is to facilitate thedevelopment of novel devices and drugs. Thesignificant achievements of some of the ongoingprojects are presented.Tissue EngineeringResearch done at IIT, Kanpur has shown that hybridpolymer-ceramic biocomposites can be developedusing tailored processing conditions on a compressionmolding machine. In vitro biocompatibility testswith mouse fibroblast cells revealed good celladhesion, cell-cell interaction and cellular bridgeformation among the adhered cells on HDPE-20%HAp-20% alumina biocomposites. Silk proteinsfrom domesticated Bombyx mori and wild nonmulberry;Antheraea mylitta silkworms were foundpromising as natural biopolymer in tissue engineeringand other applications. Multilayer 2D films basedon silk fibroin protein and gelatin was fabricated inaqueous solution for controlled drug release. A silkfibroin-HPMC-PEG blended film was developedas a vehicle for transmucosal delivery by virtue ofits good mechanical strength, water stability, ex-vivobioadhesive strength and ideal swellability.Studies conducted at KEM, Mumbai resulted inisolating endothelial cells from human saphenous94Annual Report 2009-2010

Research and Developmentvein (HSVEC), human umbilical cord veinendothelial cells (HUVEC) and goat jugular vein(GJVEC). Characterization of HSVEC, HUVEC,& GJVEC was done by using Von Willebrandfactor as a molecular marker specific for endothelialcells. Glutraldehyde treated material showed arobust and rubbery nature with much lessdecellularisation. Decellularised bovine carotid arteryprovided evidence of maximum decellularisationin various regions.Research work done at IIT, Kanpur focused on thedevelopment of a dual drug delivery system for thetreatment of lung cancer. The system developedcould be extrapolated as a therapeutic option forother solid tumors as well. Chitosan micro/nanoparticles were synthesized by the process ofelectro spraying. A model drug, ampicillin wasincorporated in these micro nanoparticles, and itsrelease from the polymeric nanoparticles studied.Biomedical SensorThrough a project implemented at IIT, Mumbaidissolved core alginate microsphere biosensors weredeveloped by using the FRET transductiontechnique. The biocompatibility of the alginatemicrosphere glucose sensor was examined using invitro cytotoxicity assays performed with L929 mousefibroblast cell lines. Results provide clear leads tothe development of an implantable sensor withincreased functional longetivity for continuousglucose monitoring. A biosensor for medicaldiagnostics based on the detection of diseased ssDNA sequence was developed at UDSC, NewDelhi. The sensitivity and selectivity of thisbiosensor was improved and it was useful in detectingmutations in DNA sequence.Implants and BiomaterialsResearchers at IIT, Kharagpur investigated themechanical causes of failure of the implant-bonestructure, using finite element modeling based onCT-scan data. A hip/knee joint simulator wasdeveloped for friction and wear testing for accuratemeasurement of wear characteristics of newlydeveloped polymer composites. Studies wereundertaken to develop new biomaterials forhealthcare. Research at CMC, Vellore hassuccessfully established an animal model of physealinjuries in goat and standardized treatment withautologous chondrocytes expanded in the culture.Further studies are underway to obtain improvedyield of chondrocyte. A novel carbon nanotube(CNT) reinforced hydroxyapatite (HAp)polyetheretherketone (PEEK) nanocompositefor biomedical applications was under developmentat IIT - Kanpur. The CNFs-PEEK-HApnanocomposite showed much higher mechanicalstrength and modulus than the PEEK-HApnanocomposite (Fig).SEM images of nanocompositesAt NIPER, Mohali, studies indicated that the cellsmigrate into the wound bed from cell-scaffoldconstruct in in vivo conditions. Dense sheets ofcells covering the scaffolds were seen in SEM imagesof cell-scaffold constructs in case of cytomodulinmodified particles. These experiments indicate theAnnual Report 2009-2010 95

Research and Developmentpotential of cytomodulin modified particles tosupport quality growth and proliferation of dermalfibroblasts.Bio-InstrumentsWork done at IISc, Bangalore resulted in designand development of a measurement system suitablefor body pressure distribution by using MEMSbased pressure sensors. The sensing mat essentiallyconsists of a number of array of MEMS basedpressure sensors (Fig).Block diagram of the complete measurement system for pressure sensorsSince packaging plays a vital role, a special adapterhas been designed and fabricated which is weldedto the pressure sensor assembly. After filling theSilicone oil (used as the pressure transmittingmedium), the adapter is covered with elastic material(Fig.)Sixty sensors are arranged in an array on woodenmat. In order to study the response of themeasurement system subjected to differentialpressure/force, thin sheets covering the sensors areplaced to ensure smooth transfer of pressure. Thematrix of sensors subjected to pressure, show anoutput which is monitored using signal processingcircuit.Stanford – India BiodesignStanford-India Biodesign (SIB) was initiated by theDepartment in July 2007 in collaboration with theStanford University, USA and the Indo-US S&TForum. The programme was initiated jointly at AllIndia Institute of Medical Sciences (AIIMS) andIndian Institute of Technology (IIT) New Delhi.SIB offers a multidisciplinary, team-based, one totwo- year fellowship program that will train thefuture leaders of the Indian medical technologyindustry to identify clinical needs in India anddevelop solutions that are cost-effective. SIBFellows commit to remain in India at thecompletion of the program, with the intent ofreturning to academia, creating entrepreneurialventures or joining medical technology companies.In addition to the fellowship, this programme ispromoting medical technology innovation among abroader audience, including engineering students andphysicians. A key component of this is an intenseinternship program that is conducted by facultyand fellows. By imparting training to a large numberof India’s finest engineers and physicians, SIB willhelp to fuel industry growth.The fellows of SIB have developed three novelmedical devices that are in the process ofcommercialization using public and private96Annual Report 2009-2010

Research and Developmentinvestments. These devices include an automateddevice for cardiac-pulmonary resuscitation, a novelway to deliver fluids to critically ill patients inemergency situation and a critical care product tomanage and contain bodily secretions in nonambulatorypatients in ICU’s and other acute caresettings.In addition to fellowships, SIB also initiated aninternship program to attract a larger audience topotential careers in India’s growing medicaltechnology industry. The three-month internshipuse unmet clinical needs (problems that need to besolved) identified by fellows and teach the biodesignmethodologies to interns using real world casestudies. The prototype intensive training resulted inmultiple patents and prototypes.The interns developed multiple prototypes,completed clinical validation, developed a businessmodel and conducted live demonstrations to showthe value of their novel device. Their achievementswere also recognized in an all India level businessplan competition (ABLE BEST 2009), where theSIB intern’s team was judged the best team out ofover 170 applicants. Furthermore, the fellows wonat the Intel’s India Innovation Pioneers Challenge2009 and represented India at a global businessplan competition at University of California atBerkeley. In addition, to the fellowships andinternships, SIB also decided to host an annualMedTech Summit each year in India.Human Genetics & Genome AnalysisThe programme was one of the earliest onesinitiated, keeping in view the national priorities andinternational efforts to identify, map and characterizegenes associated with genetic disorders and alsoexploit the knowledge created to develop molecularmedicine tools for evolving better healthmanagement strategies. Major projects were initiatedinvolving clinicians, molecular geneticists andanthropologists, utilizing India’s rich genetic resourceto develop baseline data on the various ethnic groupsfor disease susceptibility. Genetic services and prenataldiagnosis were provided to a large number of familiesthe affected families were counseled to increaseawareness among public regarding prevalence ofcommon genetic disorders. Major infrastructurefacilities were established at various institutions acrossthe country to undertake genome research includingsequencing. The existing Bioinformatics networkwas strengthened by creating Genome Mirror Sites.The major achievements are as follows.A project implemented on Sickle Cell Disease (SCD)in Chhattisgarh with special reference to morbiditypattern at Pt. J.N.M. Medical College, Raipur,collected SCD samples and provided diagnostic andcounseling services to the affected families.Counseling was done among the tribal communityfor prevention of the disease, and enhancingawareness. The information will be utilized fordevelopment of novel treatment strategies.Under a project on polymorphisms in thephysiological anti - hypertensive peptide catestatinin Indian population implemented at IIT, Chennai,600 subjects were recruited. A total of 159 subjectswere sequenced and showed naturally occurringgenetic variations. Two amino acid substitutionvariants: Gly364Ser and Gly367Val were discovered.The Gly364Ser variant was also reported in aEuropean population. It was further observed thatthe Gly367Val is a novel variant in this endogenousAnnual Report 2009-2010 97

Research and Developmentanti-hypertensive peptide. Notably, the allelefrequency of 364Ser variant was about 3-fold higherin the Indian population as compared to theEuropean population.A project was implemented at JNU New Delhi;Jammu University; Jammu and GNDU, Amritsaron analysis of genetic determinants of T2DM andunderstanding their role in type2 diabetes in someNorth West Indian population. More than 1200blood samples were collected and anthropometricmeasurements were taken. Information on castes,sub castes ethnicity and genetic heritage werecollected. Further, PCR primers were designed andstandardized for use by the investigators to maintainuniformity in protocol.A project was undertaken to study unexplainedmental retardation in children at CREMERE,Mumbai. No significant subtelomeric rearrangementwas detected in the cases showing mentalretardation. More cases were being studied with theobjective of estimating the frequency of thesubtelomeric abnormalities in mental retardationcases in Indian population. Genetic counseling wasgiven to the affected families and further testingand therapy management were recommended forthe 44 mentally retarded children.Under a project on pharmacogenetics at SGPGIMS,Lucknow, 155 renal transplant patients werecategorized based on SNPs in drug metabolisingand drug transporter gene. The results suggested asignificant association of CYP3A5*3 with CsAabsorption at 1 month post transplantation. Patientswith CYP3A5*3/*3 genotype showed lower meanC 2level and were therefore, at a higher risk ofrejection due to under immunosuppression. Thus,pharmacogenetic identification of CYP3A5 andABCB1 in combination with therapeutic drugmonitoring could contribute to the individualizedcyclosporine therapy and enhance safety and efficacyof the drug.Cancer Genome ConsortiumThe International Cancer Genome Consortium(ICGC) programme was launched in November,2008 with the commitment of eight countries i.e.India, Australia, Canada, China, France, Japan, Spainand UK alongwith eleven funding organizations togenerate comprehensive, high-resolution analyses ofgenomic changes for 8 forms of cancer based onthe International Cancer Genome Atlas programmeimplemented by National Cancer Institute ofNational Institute of Health (NIH), USA. TheICGC is one of the most ambitious biomedicalresearch efforts since the Human Genome Project.The Consortium will help to coordinate currentand future large-scale projects to understand thegenomic changes involved in cancer. This willaccelerate efforts to develop better ways ofdiagnosing, treating and preventing cancer. The DBTimplemented India specific ICGC project on oralcancer at Indian Statistical Institute, Kolkata andACTREC, Mumbai with the setting up of ICGClab. at Kalyani, West Bengal. Support was providedto implement a pilot study by collecting samplesfrom 100 oral cancer patients as per internationalstandards. A facility was established at the NationalInstitute of Bio-medical Genomics (NIBMG),Kolkata for the project.Environmental BiotechnologyThe overall well being of the environment is thekey essentiality that determines the health of various98Annual Report 2009-2010

Research and Developmentecosystems and the species there in. The need fordevelopment of ecofriendly techniques for sustainabledevelopment and the use of biotechnological toolsfor amelioration of environmental perturbations cannot be over emphasized.BioremediationAt IITR, Lucknow, a hybrid 60-mer microarraywas developed and its utility evaluated for thepetroleum refinery contaminated soil samplesobtained from 5 different sites. The results suggestthat the system has great potential for ‘fingerprinting’microbial communities and for biodegradationcapabilities of various hydrocarbons contaminatedhabitats.(Fig1)In a project implemented in the contaminated soilof oil fields of upper Assam, an efficient bacterialstrain that can reduce the surface tension significantlyin three days was isolated. Keeping this in view,the researchers isolated a bacterial strain Pseudomonassp C8B using enrichment techniques and utilizedits potential for bioremediation of endosulfan atpesticide contaminated sites. Pseudomonas sp C8Bdegraded endosulfan sulphate along with othermetabolites. Four bacterial strains degradingendosulfan were isolated. The strain namelyOchrobactrum intermedium with nar G genedegraded á – and â – endosulfan 82 % and 66 %respectively after 45 days. This bacterial strain tookpart in nitrogen metabolism along withbiodegradation of endosulfan and can be a choicein development of bioremediation technology forendosulfan. (Fig2)Experimental field was set up using PVC pipes open at both end andtreated with endosulfan and our bacterial strains. (Fig2)Hybrid 60-mer microarray: Hierarchical gene tree of functional(biodegradation) genes from five contaminated sites based on signalintensity values obtained upon hybridization Cy-3 labeled DNA ontoBiodegPhyloChip. (Fig1)Environmental MetagenomicsA study was conducted on soil sample collectedfrom the vicinity of an industry involved inAnnual Report 2009-2010 99

Research and Developmentmanufacturing chlorinated pesticides and solventsfor more than 20 years. Upon enrichment of thesoil in minimal medium using 1, 2, 4-trichlorobenzene as sole carbon source, a pure culturewas obtained. The 16S rDNA sequencing showedthe bacterium to be Bordetella sp. The strain wasdesignated as IITR-02 (GenBank EU752498).Molecular characterization of genes for 1, 2, 4-TCB biodegradation from IITR-02 was done. Alarge sized plasmid of approximately 45 kb wasisolated from IITR-02. The plasmid was stable evenupon growth on nutrient rich medium (Luria-Broth).Metagenomic DNA obtained from samplescontaminated with chlorobenzenes were cloned intodifferent cosmid vectors, cloned into E. coli hostsand were being screened for the recombinant cloneexhibiting activity on varieties of substitutedchlorobenzenes including 1,2,4-trichlorobenzene.More than 30 anoxic consortia and 10 aerobicconsortia were developed for dye degradation.Stabilized consortia were selected for isolation ofmetagenomic DNA. Amplified products were ligatedinto pGEM-T vector and transformed into E. coliDH5á competent cells and subjected to blue-whitescreening. Plasmids of positive clones were isolatedand subjected to gene sequencing using M13 primers.All the clones had similar sequences. Hence byusing this set of primers only one azoreductase genecould be amplified. Azoreductase gene amplified bydegenerate primers was of 537bp and contained anORF of 178 amino acids.Water was collected from a depth of 700-800 meterdepth of a coalbed in Jharia for extraction of themetagenomic DNA from the formation. Themetagenomic DNA was used as a template to PCRamplify 16S rRNA gene. The amplicon (mix) wascloned into pGEMT easy vector to make a clonelibrary of 119 clones. PCR-RFLP of the clonesrevealed 4-5 groups of bacteria. A total of 15 clonesrepresenting each group were subjected to sequencingand the information was used to search forhomology with the 16S rRNA sequences in thedatabase. The bacteria present in the coalbed wereshown to be Dechloromonas, Azonexus, Azospira,Azoarcus and Thiobacillus. The metagenomic DNAwas also used for making a partial plasmid(pBluescript) library of about 100 clones.In a project at AIIMS, a multiplex PCR wasstandardized for detection of hepatitis-A & E,Salmonella spp., E. histolytica & G. lamblia. About65 % of the drinking water samples werecontaminated. Among the samples, 17.31% werepositive for Salmonella spp., 21.73% for E. histolyticaand 25.21% were positive for G. lamblia. Ametagenomic library consisting of 70,000recombinants clones with average genome coverageof 240 MB was constructed.In a project at MKU, Madurai, microbial diversityon the goat skin surface under fresh and putrefyingcondition was analyzed. Phylogenetic analysis of16S r-RNA gene sequences in the metagenomerevealed that 78% of culturable and 12 % ofunculturable bacteria were present on the fresh skinsurface and 67% of unculturable and 33% ofculturable bacteria were present under putrefyingconditions. By functional based screening of themetagenomic library, a novel protease coding genethat has 98% sequence similarity with peptidase S8and S53 subtilisin kexin sedolisin sequence ofShewanella baltica was identified. By PCR basedscreening method, two antibacterial protein codinggenes tmp1 and tpm3 were identified. These100Annual Report 2009-2010

Research and Developmentproteins exhibited antibacterial activity against anumber of gram-positive and gram-negative bacteria.Industrial Waste TreatmentIn this study, researchers isolated fungal and bacterialstrains capable of decolourization and detoxificationof highly colored and toxic effluents from pulp andpaper mills. Emericella nidulans var. nidulans andCryptococcus albidus fungal species were foundeffective for decolourization and detoxification ofthe effluents. Among the three bacterial strains, themost potent one was LP1, Bacillus sp., having highdecolourization and COD reduction potency. A threestep sequential bioreactor consisting of aerobicCryptococcus albidus (stage I), aerobic Emericellanidulans var. nidulans (stage II) and aerobic Bacillissp. (stage III) were evaluated for the treatment ofhighly coloured and toxic effluent released at thepulping stage from pulp and paper mills. After thethree step treatment there was 87% reduction incolour, 62% reduction in lignin content and 78%reduction in chemical oxygen demand (COD). Thegenotoxicity of the effluent was reduced by 80%.Thus the sequential treatment by fungi and bacterialed to successful decolourization and detoxificationof pulp and paper mill effluent.Scientists at NEERI developed a two-stage PANAMprocess. This was based on the principle of PartialNitritation and Anammox (PANAM). The processcomprised two units of anoxic fixed film bioreactor(FFB) followed by an anaerobic, completely stirredtank reactor (CSTR). The process could also operateat very low DO concentrations of

Research and Developmentdetermined by Cryptococcus albidus. The sugar canebagasse treated initially by fungus and subsequentlytreated by Saccharomyces cerevisiae indicated theproduction of ethanol (38.4gl -1 ). However, anincrease in 1.82 fold (70.0gl -1 ) ethanol was measured,indicating the potential use of sugarcane bagasse ofpulp and paper mill effluent for production ofbioethanol (Fig.).Climate Change/Carbon SequestrationPreparation of biopulp by Cryptococcus albidus and chlorine bleaching:1, indicates bio-chemo pulp where treatment by 25% chemical andfungus was used; 2, by 50% chemical and fungus; 3 by 100%chemicals and no fungus, and 4 where pulp is treated by Cryptococcusalbidus and chemical was not used.Another study aimed production of ethanol fromsugar cane bagasse used primarily for the preparationof pulp and paper. One fungal strain isolated frompulp and paper mill effluent, identified asCryptococcus albidus together with Saccharomycescerevisiae was used for the production of ethanol.Lignocellulolytic enzymes, CMCase (34 Uml -1 ),FPase (3 Uml -1 ), â-glucosidase (2.3 Uml -1 ), laccase(32 Uml -1 ), and xylanase (12 Uml -1 ) wereSemiarid tropics are likely to undergo changes infuture due to the critical environmental conditionsof changing temperature and precipitation patterninduced by global climate change. A study wascarried out at Maduari Kamaraj University forcontinuous measurement of CO 2and H 2O fluxesand energy fluxes on Cenchrus ciliaris L. grasslandin semi-arid region using Eddy Covariance Systemwith energy balance components. The study showedthat the Net Ecosystem Exchange of CO 2(NEE)was strongly controlled by soil moisture. High CO 2sequestration was observed during the wet seasonwhich lasted for 7 months and the grasslandProduction of bioethanol from sugar cane bagasse residue in pulp and paper mill effluent by Cryptococcus albidus and Saccharomyces cerevisiae102Annual Report 2009-2010

Research and Developmentecosystem was a carbon source during the dry season.Seasonal changes in ecosystem respiration (Reco)also showed a similar pattern with changes in soilmoisture while a negative correlation was observedbetween Reco and soil temperature. The phase andamplitude of NEE was a strongly regulated rainfallpattern that induced dynamics in soil moistureavailability.At University of Hyderabad, a study was carriedout to dissect out the CO 2fertilization effects onphotosynthetic gas exchange characteristics, keyresponses of photosynthetic enzymes along withoverall plant growth performance in a fast growingtree species, Gmelina arborea Roxb (Verbenaceae).They were grown under ambient (360 µmolmol -1 ) and CO 2-enriched conditions (460 µmolmol -1 ) in open top chambers (OTCs). Growthcharacteristics along with biochemical measurementswere carried out at regular intervals. PhotosyntheticMorphological characteristics of Gmelina grown in ambient and elevatedCO 2atmosphere showing the increased growth enhancement in shootand roots (C and D) in elevated CO 2- grown Gmelina (A and B).rates (P n) were correlated with other leaf gasexchange and biochemical variables to localizebottlenecks for acclamatory responses throughoutthe growing season. Gmelina significantly respondedto CO 2enrichment with periodic and potentialincrease in biomass which was also seen with almost32 % higher P nand a radical increase in CA (~60.82%). CA was believed to act as a keydeterminant of enhanced rates of CO 2fixation,preventing photosynthetic acclamatory responses inthe tree species. Fast growing tree species likeGmelina exhibiting CA - mediated photosyntheticup-regulation can be used as potential tree speciesfor carbon sequestration (Fig).Biodiversity ConservationA study was conducted in a 200-acre plot oflimestone mined out area consisting of overburdendumps (OBDs), and a 40 m deep water bodyextending over 150 acres for ecological rehabilitationthrough restoration technologies. The inputs forthese technologies were native grasses, bamboos,legumes and woody forest species, and microbesassociated with them. Aquatic flora and fauna werethe inputs for restoration of water bodies. Socialbenefits for local communities formed the basis ofselection of species and plant communities.Consequently, a bamboo thicket with associatedspecies, grassland, an orchard, blocks of tassar andlac culture plantations, a native mixed moistdeciduous forest community and a sal dominatedforest were developed. The barren OBD wassuccessfully transformed into a bamboo thicket with95% grass cover and associated woody species withintwo years of restoration. The overburden dumpsand cut benches were developed into luxuriantgrasslands with rich insect and bird fauna. A totalAnnual Report 2009-2010 103

Research and Developmentof 35 forest and 10 horticultural species wereintroduced and these were inoculated with forestsoil containing spores and microbial inoculants. Theestablishment rate of saplings of different speciesvaried from 80 to 90% (Fig).of sympatric carnivores. Microsatellite markers weredeveloped for the identification of individual tigerswith a sibling identity probability of 0.005 whichcould distinguish even closely related individualswith a 99.9% certainty. Results demonstrated thefeasibility of using tiger faecal matter as a source ofDNA for population estimation.Biodegradable Plastics/BiopolymerOverview of limestone mined out area at Purnapani site showingOBDs, void with water, and cut benches and their slopes.The Indian Black Buck is considered to be anendangered species. Scientists at NaFCONES,CCMB developed a technique to successfully inducepregnancy in the species by artificial insemination.Scientists induced estrus in adult female black bucksusing prostaglandin F2-alpha and then inseminatedthem non-surgically using an endoscope. Ultrasoundanalysis detected a fetus after 55 days in one of theanimals, and after 180 days the animal delivered alive fawn. This was the first successful artificialinsemination in an estrus induced black buck bynon-surgical insemination.A pilot study using DNA extracted from fecalsamples of Bengal tiger, Panthera tigris tigris for thepurpose of population estimation was initiated. PCRprimers were developed based on tiger-specificvariations in the mitochondrial cytochrome b forreliably identifying tiger fecal samples from thosePolyhydroxy butyrate (PHB) is biodegradablemacromolecule polyester accumulated by manymicroorganisms as an intracellular carbon andenergy storage compound, under unfavorablegrowth conditions. The biodiesel industry generatesabout 10 % (w/w) waste glycerol as the main byproduct.A study was undertaken to develop aprocess for the production of PHB from glycerolgenerated from biodiesel industry. Bacterial strainssuch as, Alcaligenes latus, Bacillus firmus, B.sphaericus and Pseudmonas putida were screenedfor PHB production. Bacillus firmus appeared tobe the best producer of PHB (31.23% of biomass)when using crude glycerol from biodiesel industry.When this strain was grown on commercialglycerol, it showed almost the same PHB yield(33.83 % of biomass) indicating that there wasno need for pretreatment of crude glycerolobtained from biodiesel industry.In an earlier study, the moderately haloalkali tolerntbacterium Halomonas sp. was shown to producepolyhydroxyalkanoates (PHA) and the copolymerwas found to be biodegradable. To generate sufficientquantities of polymer for preparation of blends, ascale up production was carried out. Optimizationof PHA production was carried out using differentNaCl concentrations, temperatures of incubation,104Annual Report 2009-2010

Research and Developmentwith addition of antifoam agent and differentvolumes of production media. To extract themaximum amount of PHA from dry cell mass ofHalomonas obtained from 10 L fermentation inminimum time, the rotary film evaporatorextraction was consideration as the best methodand the polymer obtained was found to bebiodegradable.Nano Science and NanobiotechnologyThe potential of nanotechnology to revolutionisethe health care, textile, materials, information andcommunication technology, and energy sectors iswell understood. Several products enabled bynanotechnology are in the market. Keeping these inview the department is encouraging the developmentof products employing nanotechnology. So far,ninety four projects have been sanctioned, in thelast two years and some of the research highlightsare as follows:Nano medicineNanobiotechnology approaches are being activelyinvestigated in the designing of functionalizedbiodegradable nanoparticulate drug delivery system(NPDDS) of Doxorubicin hydrochloride (DOX)using an asialoglycoprotein receptor (ASGPR)mediated targeting agent Pullulan, á-1, 4; 1, 6-glucan for application in hepatic cancer therapy.The NPDDS with a high entrapment efficiency(~70%), high drug loading (20%) and low size(~100 nm) of highly water soluble drug DOX in ahydrophobic polymer PES, was successfully designed.The in vitro cytotoxicity of gold nanorods(AuNRs) and gold nanoparticles (AuNPs) inEGFR over-expressing skin carcinoma cell line,A431 was evaluated by MTT assay and NeutralRed Uptake assay and results compared with astandard anticancer drug – Cisplatin. This revealedthe non toxic nature of AuNRs and AuNPs.Inflammatory response of AuNRs in monocyteswas determined by Lipoxygenase (LOX-12) andLDH release assay. These elicited no inflammation.Cell morphology was also observed by phasecontrast microscopy and found to be intact evenafter 48h of treatment.The work on the nanoparticle-aided delivery ofbioactive molecules is in progress. Considerableefforts were made by various research groupsincluding pharmaceutical companies indeveloping smarter non-viral vectors capable ofdelivering therapeutic agents safely, efficientlyand in a specific manner. Among various cationicpolymers, branched PEI was the most widelystudied polymer for gene delivery purposesbecause of its unique properties. Studies onalternative systems to deliver antianaplasmosisagents in therapy are in progress wherenanoparticles were successfully prepared by thenanoprecipitation method using poly (methylmethacrylate) polymer and with or withoutoxytetracycline antibiotic, purified by cross flowfiltration and evaporated to dryness. Thenanoparticles prepared were assessed by routinephysicochemical characterization studies toqualify them for future therapeutic studies.Work was in progress at B.V.Patel PharmaceuticalEducation & Research Development Centre(PERD), to develop two types of nanoparticles fordrug delivery i.e. SPION based and liposome based.In both the cases, targeting of cancer was achievedAnnual Report 2009-2010 105

Research and Developmentusing folate-folate receptors which are present onlyon cancer cells (Fig).To develop nanotherapeutics with ipidicnanoparticles, a study was undertaken at IICT,Hyderabad to develop colloidal nanocarriers suchas Solid Lipid Nanoparticles (SLN),Nanostructured Lipid Carriers (NLC), Nanoemulsionsand Nanosuspensions for antimalarialdrugs such as Primaquine, Atovaquone,Halofantrine and formulate them suitably for thetreatment of malaria.At Amrita Institute of Medical Sciences, Kochi, aproject on Novel Nanodelivery Carrier for SystemicControl and Release of Parathyroid Hormone forthe treatment of bone diseases resulted in a novelconcept for chemically modifying chitosan-derivativebased nanoparticles for use as a vehicle or carrier todeliver the PTH 1-34 peptide. This was possiblewith a controlled release rate in pH-selective fashionso as to bypass acidic environments wherein thedrug may be metabolized. Work was undertaken atBHU on the elucidation of the role of nanosilverin thrombosis biology where significant antiplateleteffects of silver nanoparticles (10-15 nm size) bothfrom in vitro and ex vivo were studied. Nano silverwas found to inhibit all the platelet functions.At the Institute of Pharmaceutical Sciences, PunjabUniversity, studies are under progress in thedevelopment of drug loaded solid lipid nanoparticlesfor treating cerebral tuberculosis where accurate andprecise analytical methods for analysis of isoniazid,pyrazinamide, streptomycin and rifampicin usingUV-Visible spectrophotometry were established.Standard plots in organic solvents have also beenprepared. Optimization studies are under progress.Work was undertaken at MKU Madurai on proteinengineering of self-assembly systems for variousapplications. Polyclonal antibodies which were raisedagainst CdMV-CP-N-E1 and ELISA showed goodtitre of the antiserum. In addition, the epitope E1was expressed at the C-terminus and also at both Nand C termini after engineering suitable restrictionsites in the coat protein gene by site-directedmutagenesis. An oligo nucleotide corresponding totwo other major epitopes of gp41, 2F5 & 4E10,was engineered at the N-terminus of CdMV-CP.Development of nano particles using the chimericproteins is in progress at Anna University, Chennai.Nano-AgricultureSilica nanoparticles were synthesized from biogenicsilica rich resources (O. sativa, P. purpureum,B. bambos, S. officinarum L., Z.mays). X-raydiffraction revealed that husk of paddy andleaf of elephant grass had a pre-dominant crystallinephase of SiO2 in the form of quartz andcristobalite. Translocation of silica nanoparticle inthe plant system was studied for C. sativus, L.acutangula and O.sativa. Impact of synthesizedSi nanoparticle on fresh water edible snail andtilapia fish was studied. Phytotoxic studies ofsynthesized Si nanoparticle (15nm) with other106Annual Report 2009-2010

Research and Developmentnanoparticles like ZnO (50nm), Al 2O 3(40nm.),TiO 2(50nm) were carried out. TiO 2(50nm) washighly toxic to O.sativa, and Z.mays. Optimizationof process parameters for the merging ofSi nanoparticle with HA (binder in thenanocomposites) from low rank coal (Neyvelilignite-supplied by NLC, Neyveli) was completed.Insecticidal properties of synthesized Si nanoparticleswas fungal mediated.Nanopesticidenon hazardous to human beings. ISI, Kolkata,developed a spherical, amorphous, silicananoparticles based pesticide named as Dipentoxand it is being patented (Figure A & B). Due tothe lipophilic nature of the product, Dipentox isalso effective in highly humid conditions.Nanocides of this category are needed in smallquantities when compared to the conventionalpesticides. Moreover, they were found to be costeffective. Bioassay studies with various group ofinsects established their biosafety (Fig).Amorphous or non crystalline silica naoparticle isFigure (A) Control S. oryzae, (B) lipophilic silica nanopowder treated S. oryzaeFigure: XRD data shows amorphous nature of lipiophilic nonosilica.Annual Report 2009-2010 107

Research and DevelopmentThe majority of the methodologies established foraflatoxin detection are sensitive, but they requireskilled operators and extensive sample pretreatment.New integrated magnetometric nanosensors with aminimal size, weight and power consumption willsignificantly impact the practice of precisionagriculture. An attempt in this direction was madeusing immunomagentic separation for detection ofaflatoxin. An other ongoing investigation is aimedat the development of nanosilver-nanohydrogel(nSnHG) for use as an antimicrobial agent. ThenSnHG may be used as the coating material forimplants, as additives in wound dressing materialsand as an additive in membranes for disinfectingwater. The approach for the development of thesenanomaterials has been based on nanoemulsionpolymerization.RNAi Technology PlatformsThe discoveries of RNA interference and RNAmediatedpost transcriptional gene silencing haveopened unexpected avenues for the regulation ofgene expression and knocking down geneexpression in organisms and cells. These are likelyto be exploited for human therapeutics. Globally,about 20 RNAi based candidate molecules are invarious stages of clinical trials. Keeping in viewthe importance of RNAi technology thedepartment started a programme in this area inthe year 2007. Fifty five projects are beingimplemented and the progress made in some ofthem is as follows:RNAi Technology in MedicineStudies at IISc, Bangalore investigated the role ofmicroRNA trafficking with reference to humancancer. Significant expression of miRNAs todifferentiate (i) malignant astrocytoma and normalbrain sample, (ii) glioblastoma and anaplasticastrocytoma, (iii) secondary glioblastoma and primaryglioblastoma, (iv) progressive pathway and de novopathway were validated by the grade specificexpression of miRNAs, by measuring the relativelevels of miRNA transcripts by quantitative realtimePCR.Given the prevailing epidemic of Type 2 diabetesin India, the preliminary microarray studiesconducted implied that altered miRNAscontribute to the disease mechanism(s) underlyingnot only Type 2 diabetes but also prediabetes.This was the first report in India on thedysregulated microRNAs in Type 2 diabetes andthe lead gives directionality for the identificationof novel drug targets. A sub-study also revealed arole for novel miRNA (miR-146a) that linksalterations in transcription factor(s), inflammationand insulin resistance.Use of chemically modified siRNAs seems to bethe most promising approach for development ofRNA Interference (RNAi) based therapeutics. Withthe specific aim to target drug resistant malarialparasite (Plasmodium falcifarum) genes dhfr and dhpsstudies were envisaged for the synthesis andevaluation of novel chemically modified siRNAs.The modifications are expected to enhance thespecificity, stability and efficacy of anti-malarialsiRNAs. It is expected that the outcome from thesewill lead to the development of siRNAs withdrug like properties and those siRNAs canbe used to target various therapeutically relevantgenes.108Annual Report 2009-2010

Research and DevelopmentRNAi Technology in AgricultureStudies were taken up for the identification of thefunctional homologue(s) of Arabidopsis MYC2 liketranscription factor from rice (OsMYC2) throughRNAi mediated gene silencing. The bioinformaticsanalysis of MYC2 transcription factor family inrice, identification of two potential candidate genes,OsMYC2-1 and OsMYC2-2 and generation ofRNAi construct with siRNA targets for both thecandidate genes was completed. In a significantfinding, blue light specific response in diseaseresistance in Arabidopsis was observed. MYC2 wasrequired for blue light specific development inArabidopsis. MYC2 negatively regulated salicylic acidsignaling. It was also seen that atmyc2 mutantswere more resistant to bacterial pathogen.In a project undertaken for reduction of coldinduced sweetening in potato the notableachievement was that a total number of 600 putativetransformants were obtained, which were screenedfor NPTII expression by RT-PCR. A total of 141NPTII positive amiRNA transgenic lines, 31 forUGPase amiRNA, 87 for INV amiRNA and 23 forboth UGPase and INV combined amiRNAs, werealso obtained. These putative transformants werescreened for NPTII expression by RT-PCR, and 38UGPase-TGS transgenic lines were obtained. Allthe 179 transgenic lines were multiplied in vitroand about 3000 plantlets were planted in the soilin the glasshouse of Central Potato Research Station,Jalandhar for evaluation of cold-chipping attributes.Research on development of transgenic tomato plantsresistant to a wilt causing fungal pathogen, Fusariumoxysporum by plant RNAi- mediated silencing ofimportant fungal genes involved in growth,development and pathogenecity resulted in (i) clonedornithine decarboxylase (ODC) gene from Fusariumoxysporum, (ii) RNAi construct with ODC gene ofFusarium oxysporum. (iii) transgenic tomato plantswith RNAi construct harboring ODC gene ofFusarium oxysporu, and (iv) protocol for transformationof Fusarium oxysporum using glass beads.Transgenic male sterile tobacco plants were developedusing the gene construct, TCON (TA29 promoterfrom tobacco-CoxIV presequence from yeast –orfH522 gene from sunflower) at DOR Hyderabad.To restore fertility in these transgenic plants,orfH522 gene was to be silenced using differentPTGS approaches: ihp-RNAi (intron-hairpin RNAinterference) and SHUTR (silencing by heterologous3’-UTR). Suitable gene constructs were developedand tobacco plants were transformed with these(Fig).A(A) Transgenic tomato plants (raised with ODC-RNAi construct)showing enhanced resistance to wilt disease caused by Fusariumoxysporum (right) as compared to wild-type plants (left) after 30 daysof infection; (B), Disease severity index values against Fusarium wilt ofODC RNAi transgenic lines and untransformed control.Annual Report 2009-2010 109

Research and DevelopmentVirus Induced Gene Silencing (VIGS) is based ona silencing mechanism that regulates gene expressionby the specific degradation of RNA. Gemini viruseswith their small DNA genomes and ease ofinoculation through agrobacterium, are excellentcandidates for VIGS vector development. The resultsobtained at MKU, Madurai show that theDelbC1bPTR carrying the PDS gene acts as VIGSvector and silences the endogenous PDS, showingthe bleached phenotypes. The VIGS vector wasfurther tested by cloning the gfp gene and gfpexpressingNicotiana benthamiana plants (16c) wereagroinfiltrated. Confocal microscopy of theagroinfiltrated plants show that the gfp transgenewas silenced by the gfp present in the VIGS vector.Thus an efficient VIGS vector based on the bDNA of Bhendi yellow vein mosaic virus has beendeveloped (Fig).Agroinfiltrstion of N. benthamiana with BYVMVAPTR and delâC1âPTR PDS (A) Buffer alone, (B) PTR alone, (C and D) APTR+ del âC1âPTR PDSEnergy Bioscience ProgrammeThe Department has been supportingprogrammes on biofuels and bioenergy with afocus on using different feed stocks as a rawmaterial for ethanol and biodiesel production.Salient achievements of the ongoing programmesare as follows:Energy Bioscience CentreThe DBT–ICT (Institute of Chemical Technology)Centre for Energy Biosciences (CEB) is the firstEnergy Biosciences Centre in India with integratedbasic and translational science capability. Theprimary focus of the centre is the developmentof biotechnologies for deriving energy fromrenewable resources. The major research areasidentified are bioethanol, biodiesel, biohydrogenand biomethane.The centre has developed an industry-readytechnology for producing lignocellulosic ethanol.The technology employs a combination of pretreatmentmethods and use of depolymerasesfor cost-effective breakdown of celluloses andhemicelluloses to sugars and their conversion toalcohol. The yield of bioethanol per ton of drybiomass is ~300L and lignin is produced as adry solid up to 150-200kg/ton biomassdepending on biomass variety. The technologydeveloped was being validated at pilot scale withan industrial partner – Indian Glycols Ltd. Studieswere undertaken on synthetic biology, with helpfrom collaborating partners, to design enzymeslike lipases and cellulases that will be morerobust and safe against product/substrateinhibitors.110Annual Report 2009-2010

Research and DevelopmentJATROPHA R&D, DEMONSTRATIONJatropha MicromissionA network of partner institutions was employed forJatropha improvement in order to enhanceproductivity and yield. These institutions made morethan 1000 collections and these were accessioned atNBPGR, New Delhi. The accessions were furtherevaluated at 12 institutes in the country for oilyield, productivity and performance.A total of 216 accessions were found promising onthe basis of percentage oil and tolerance to localconditions. These accessions were further bulked upunder controlled nursery conditions at four centres(TERI, New Delhi; Biotech Park, Lucknow;Nandan Biomatrix, Hyderabad; and Ruchi Biofuels,Indore). A total of 17 lakh clonally raised saplingswere produced for distribution to states andinstitutions.Multilocational TrialsEarlier studies undertaken at 12 institutions onevaluation of nearly 1000 accessions from theJatropha Field Demonstrationcountry revealed that Jatropha curcas was prone tofrost damage in some parts of Himachal Pradesh,Hissar and Uttrakhand. The 100 most promisingaccessions were evaluated under different agroclimatic regions of the country. Experiments with20 accessions under multilocational trials werepromising with respect to performance in differentagro climatic conditions.R&D in JatrophaStudies were supported on various R&D aspects inJatropha curcas. Using tissue culture protocols, fiveelite selections of high yielding Jatropha curcas plantswere multiplied at LabLand, Mysore. About 11,475tissue culture derived Jatropha curcas plants wereproduced and 2265 plants transferred to the field.RAPD analysis revealed that there were no geneticvariations among tissue culture regenerants and theirrespective mother plants. The tissue cultureregenerants exhibited flower induction within 6months of planting.Candidate marker genes for selection of high oilcontent genotypes of Jatropha were identified at JPUniversity, Solan. One marker was from 5’UTR ofoleoyl desaturase, two from desaturase and one fromintron of KAS II. These markers are expected to beof value in marker–assisted breeding for high oilcontent in Jatropha. A total of 130 microsatellitemarkers were developed through enrichment ofgenomic libraries and sequencing. Studies were takenup to generate EST’s for identification of genes foroil biosynthesis pathway at NBRI, Lucknow,MSSRF, Chennai, and SRM University, Chennai.So far 6000 odd EST’s were generated and nearly3000 EST’s were sequenced.Annual Report 2009-2010 111

Research and DevelopmentSeed oil analysis of 14 genotypes from Southernregion was completed at Vittal Mallya ResearchFoundation, Bangalore. The mutagen assembly isready in binary vector pBIN. Two genes (GPAT &LPAT) identified for oil enhancement were clonedinto plant expression vectors of Jatropha with thedual gene constructs at Avasthagen, Bangalore.Transformation was completed & transformed plantswere confirmed by PCR.BioethanolIn order to enhance the availability of cellulosicfraction for enzymatic hydrolysis, different chemicalpretreatments were used, out of which chloritetreatment, alkaline peroxide treatment, and alkalitreatment resulted in > 80 % delignification. Thechemical pretreatments were scaled up to 1.0 Kg.In order to secure efficient fermentation ofhemicellulosic hydrolysate (pentose sugars), 20 yeaststrains were screened. Out of these, 10 were foundto be ethanol producing and among these, Pichiastipitis NCIM 3499 was found to be the mostefficient.At Osmania University, Hyderabad, thermotolerantyeasts were isolated from 250 samples collectedfrom various high temperature regions of India,and 95 yeasts were selected based on their growthbetween 40°C – 50°C and characterized. Amongthe four Saccharomyces cerevisiae strains, OC21showed the maximum fermentation efficiency upto 25% sugar concentration. Sub cloning of threeengineered genes in yeast surface display vector andtransformation in E.coli was completed. Studies wereinitiated at Vallabh Vidyanagar, Gujarat to studydelignification of various lignocellulosic feed stockslike wheat straw, and banana stems using white rotfungi etc. Maximum delignification of 16% wasobtained on the 32 nd day by Pleurotus ostreatusHP -1 with wheat straw. Further, work is in progressto increase the rate of delignification.Biodiesel from AlgaeAlgae, was identified as a research material forbiodiesel production. An algal network programmewas launched with a focus on (i) collection andcharacterisation of algal strains from ecological niches.(ii) establishment of 3 repositories. (iii) developmentof production system. Studies were initiated tocollect, identify and characterize algal strains thathave higher oil/lipid content. The identified strainswill be taken up further for improvement to producebiodiesel from algae.112Annual Report 2009-2010

CHAPTER 5BIOTECHNOLOGY PROGRAMMESFOR SOCIETAL DEVELOPMENTThe department continued the implementationof biotechnology programmes for societaldevelopment; to improve the livelihood conditionsof SC and ST populations and also for the welfareof women. The success stories of some of theseare given below.Programmes for SC and ST PopulationSericultureSilkworm chawki rearing centers were establishedby the self help groups of SC/ST and weakersections for employment generation andimprovement of cocoon yield in selected villages ofMysore district in Karnataka. Two sericulture growergroups involving 15 farmers each were formed. Theknowledge and skills were upgraded through trainingprogrammes.VermicompostingSocio-economic development of rural SC/ST/weakersections was initiated in Bijapur district of Karnataka.Earthworm hatchery and vermicompost productionunits were established. Around 600 SC/ST/weakersections farmers attended the training programmesfrom different villages. One hundred vermicompostdemonstration units were constructed in individualbeneficiary’s fields in different villages. After theproject activity their income levels had gone up toRs. 2000/month.Integrated Pest ManagementPromotion of bio-control based IPM for majorinsect pests of cruciferous vegetables was taken upamong the ST populations of Arunachal Pradesh.The natural enemies viz. Steinernema carpocapsaeand Hetrorhabditis indica were procured from theProject Directorate of Biological Control, ICAR,Bangalore. Coccinella septempunctata and Menochilussexmaculatus were collected from the field. Thebio-control agents were maintained in the laboratoryin their host insects viz. wax moth, Galleriamellonella for S. carpocapsae & H. indica and Aphiscraccivora for C. septempunctata and M.sexmaculatus. Awareness campaigns were carried out.Entrepreneurship training was given to fifteenunemployed persons. They were trained on the massproduction of important natural enemies of pests.Mushroom CultivationThrough participatory research by utilizing theabundantly and locally available agricultural wastes,SC/ST women in Bangalore district were benefited.The project empowered 80 SC/ST resource poorwomen from four self help groups in addition to240 rural women. Agricultural wastes like coir pith,sugarcane trash, areca nut husk and coffee pulpwere used as substrates.AquacultureScientific fish farming technology was disseminatedAnnual Report 2009-2010 113

Biotechnology Programmes for Societal Developmentto fish farmers belonging to SC/ST and weakersections of Champawat district of Uttarakhand.Demonstrations showed a production of 94.43 kg/100m 2 fish in composite carp farming and 90 kg/100m 2 in poultry cum fish farming with 22kg poultrymeat and about 200 eggs. In a project implementedat Keonjhar and Kendrapara Districts of Orissa, therewere 163 beneficiaries (113 ST and 50 SC fishfarmers including three women self-help groups)covering a water area of 9.49 and 9.88 ha respectively.The average production levels were increased by 2.13and 1.43 times in Keonjhar and Kendrapararespectively involving 27 beneficiaries. A total of 642farmers including 150 women were trained.Bee-keepingPersons from the SC/ST population of Hasanganjtehsil of Unnao District (UP) were trained in beekeeping.Altogether, 40 demonstration units wereestablished in 5 villages. A total to 678 farmerswere trained in bee-keeping and managementtechnology. One technical bulletin “Madhumakhhipalan-mahatva, upyogita evam prabandhan” and threeextension pamphlets were published. The programswere covered by Doordarshan.Post Harvest ManagementEntrepreneurship development was initiated for thebackward population of ginger growing areas ofGolaghat district in Assam through post harvestmanagement of ginger. Eight eco-friendly, low costfresh ginger storage structures were constructed intwo districts. Demonstration and training on valueaddition of ginger was taken up. More than 90women from the backward sections were trained invalue addition of ginger.In a project implemented in Kanyakumari districtof Tamil Nadu, standardization was undertaken onthe production of low-cost vitamin rich HUFA forhuman consumption and feed for poultry frommarine fish, Odonus niger, for income generationby SC/ST population. The liver mass was asepticallycollected and the oil extracted. The percentage oilyield was found to be the maximum (70.3%) withthe Soxhlet extraction method. The de-livered fishwere dried, powdered and used as a major ingredientfor making poultry feed. Training programmes wereconducted for the SC/ST population.Bio-control ManagementAbout 250 small and marginal farmers wereidentified from different self help groups inPeermade and Udumbanchola taluks of IdukkiDistrict. They were provided training on thescientific organic management of cardamom rootgrubs. In addition, detailed information was alsoprovided on the control of root grubs with the useof entomo pathogenic nematodes (EPN).In a project implemented at Hingoli district ofMaharashtra, 71 farmers had the benefit of releasesof Trychocards and spraying of HaNPV on cotton.The redgram crop and bengalgram crop were sprayedtwice with HaNPV at the 50% flowering stage andthe pod formation stage in the fields of 164 farmers.The increase in cotton yields varied from 7.3 to13.25%. In redgram, the increase in yield rangedfrom 3.13 to 23.05% and in bengalgram it was 7.5to 31.7%. About 1000 farmers and rural youth weretrained on the production and uses of bio-pesticides.Rural Bio-resource ManagementAn integrated rural bio-resource complex was114Annual Report 2009-2010

Biotechnology Programmes for Societal Developmentestablished in the SC/ST villages of Thoothukudidistrict of Tamil Nadu. A model fish farm complexwas constructed in a 1600 sq. m. area for integratedfish farming and ornamental fish breeding. Trainingmanuals, leaflets, and extension publications relatedto fisheries were made available. A total of 760SC women from SHGs were trained and 3groups started practicing fish culture.PiggerySocio-economic development of resource poor tribalfamilies of Kangpokpi block of Manipur wasinitiated through piggery. The project covered eightvillages under the block. Ten SHGs were formed.Two (1+1) weaned piglets, cost of pigsties, feedsand veterinary medicines were provided to thebeneficiaries. Five demonstration cum trainingprogrammes were conducted.In a project implemented in Pantnagar,Uttarakhand, back yard rearing of pigs using probioticsfor effective utilization of non-conventionalfeed for the benefit of SC/ST population wasundertaken. A nucleus herd of large white Yorkshirepigs was established. A total of 196 pigs weredistributed among 72 beneficiaries both at Kotdwarand Pantnagar. Training programmes were organizedand relevant literature made available to the targetgroup.Goat RearingA profitable entrepreneurship for weaker sections offarm women and rural masses through goat farmingwas initiated in Amravati district of Maharashtra.About 300 persons were trained in goat rearing.Clinics and disease diagnostic centres were established.Booklets on goat farming were published anddistributed to the beneficiaries. Proven buck ofOsmanabadi breed were supplied to about 100beneficiaries for the upgradation of non- descriptgoats.In a project implemented at Bidar district ofKarnataka, employment of SC/ST youth throughgoat rearing under stall feeding was taken up. Theproject was implemented in 20 villages covering200 beneficiaries. Eighteen LIGs were promoted byinvolving 200 unemployed SC/ST youth.Beneficiaries were trained on goat rearing byorganizing 37 ‘on campus’ and ‘35 on farm’ trainingprogrammes, 16 demonstrations and 2 ‘on farm’trials. The project activities created an asset of 4200goats and 200 managers among the beneficiaries ofthe lower socio-economic strata. Further, goat rearingalso contributed to enhanced nutrition of the ruralpoor.Empowerment of Tribal CommunityThe tribal communities in Kanyakumari district ofTamil Nadu were empowered through sustainablebiotechnological options. Training programmes anddemonstrations were conducted on farm wasterecycling, composting, organic farming, bee keeping,mushroom production, multiple cropping system,medicinal plant cultivation, etc. Construction ofvermi-compost units and installation of bee hivesfor honey production were completed. Apart fromthese, multiple cropping system models wereintroduced for coconut, spices, banana, medicinalplants and bamboo involving beneficiaries.PoultryBackyard poultry was used as a tool for sustainablelivelihoods of rural SC/ST population inAnnual Report 2009-2010 115

Biotechnology Programmes for Societal DevelopmentNalgonda district of Andhra Pradesh. A total of1890 sexed dual purpose chicks (Rajasree) suitablefor back yard rearing were purchased from AICRPon poultry breeding. They were reared up to 9weeks of age in a deep litter system at farm leveland data was collected every week on bodyweights, feed consumption, mortality, etc. Aftercompletion of 9 weeks the grower birds andnight shelters were distributed to the selectedbeneficiaries. Training programmes were conductedfor back yard poultry rearing. A booklet on thesubject was published for distribution amongbeneficiaries.Medicinal and Aromatic PlantsStrengthening of traditional health care practicesand conservation of medicinal and nutritious plantsneeded for primary health care & nutrition wastaken up in 100 villages of two mandals of AndhraPradesh. About one lakh seedlings of various specieswere raised. Workshops were conducted oncultivation of the identified species. More than 60SHG women groups participated in kitchen gardenlaying, identification and usage of MAPs. About 50healers from the tribal areas participated in thetraining in the preparation of herbal medicines andnutritive foods.Livestock DevelopmentLive stock development was taken up for thebenefit of families in the desert areas of Rajasthan.Five Tharparkar breeding bull calves wereprocured. Three of them were introduced fordonating semen. A one hectare silvi-pasturedemonstration model was developed inconsultation with the villagers as well as GramPanchayat members to demonstrate the conceptof silvi-pasture. Training on new technologies andgeneral management practices for animals wasarranged for about 250 participants. A total 510artificial inseminations were done and 224pregnancies were recorded.Entrepreneurship DevelopmentEntrepreneurship development was initiated in therural areas of Ganjam district of Orissa through theproduction of biofertilisers. A laboratory formaintenance of microbial culture, massmultiplication, quality control and packing wasestablished. A series of training programmes wereconducted on biofertilizer production, quality controland use on different crops, covering 722beneficiaries. Thirty two persons started their ownbiofertilizer production centres. They earnedapproximately Rs. 60,000/- per annum. Field trialswere conducted regularly for various crops usingCyanobacteria, Rhizobium, and Azatobatcer.BiopesticidesParticipatory research for empowerment of SCand ST rural women through production of neembased biopesticide and Trichoderma by utilizinglocally available raw materials was taken up inTumkur district of Karnataka. Two Trichodermaproduction units were established by involving40 SC/ST women SHGs members. To improvethe knowledge and skills about the use ofTrichoderma, ‘hands on’ training anddemonstrations were conducted involving 350farmers. The production units generated nearly730 labour days and, produced 983 Kgs ofTrichoderma which were sold to local farmers,116Annual Report 2009-2010

Biotechnology Programmes for Societal Developmentresearch stations, KVKs and NGOs, and thesegenerated an income of Rs. 78,640/-. Five newTrichoderma production units were established byother farmers and NGOs after being trained underthis project.Integrated Crop ManagementIntegrated Crop Management (ICM) technologyon pigeon pea was demonstrated for SC/ST andeconomically weaker sections of NorthernKarnataka. Altogether, 200 farmers were trainedon INM (Integrated Nutrient Management), IPM(Integrated Pest Management) and IWM (IntegratedWeed Management). The beneficiaries wereprovided with seeds of improved varieties ofpigeonpea, fertilizers, herbicides and pesticides tothe extent of 50 percent of the total cost on theinputs. Income level increase was to the tune ofRs. 11000 to 15000 per hectare over that of theprevious year.Nutritional InterventionEffect of nutritional intervention on mother andchild health in the tribal populations of the NorthEast was studied. Screening camps were organizedamong tribal people in Assam, Meghalaya, Tripuraand Arunachal Pradesh. It was found that theincidence of anemia was 76.30% among females ofAssam, whereas the percentage of anemia was lessthan 50% in Tripura, Shillong and Eastern region.The incidence of anemia was 42.5% in Shillongcases, 36.09% in Tripura cases, 63.78% in Itanagarcases and 38.20% in Eastern region cases. Out of248 non pregnant cases, a one year follow up wasdone on 113 cases and in 106 cases, the conditionimproved.Animal HusbandryUpliftment of the economic conditions of SC/STpeople was attempted in Kota district of Rajasthanthrough training in animal husbandry. Theinformation related to socio-economic conditions,type & breeds of animals, types of vegetationavailable, crops grown, and common diseases werecollected. Programmes were organized for awareness.More than 675 villagers (men & women)participated in these programmes. A total of 120beneficiaries (20 from each village) were selected fortraining. Altogether 120 goats & 12 bucks ofBarbari breeds distributed with 50% of the costborne by the beneficiaries. Beneficiaries had startedearning around Rs. 350/- per month through thesale of milk.Quail FarmingThe potential of quail farming in Mizoram wasstudied. More than 100 beneficiaries were trainedin quail farming on the aspects of reproductiveperformance of quails, housing & sanitation of shed,management of quail chicks, nutrition and diseases.Awareness campaigns for popularization of quailfarming were organized in seven villages of Aizawl,and incubation and brooding facilities were providedto the trained farmers.Programmes for Rural DevelopmentApicultureRural populations of Jhalawar district of Rajasthanwere trained in bee-keeping. Training anddemonstration activities were conducted in 14villages of Jhalrapatan Panchayat Samiti. Twotrainings in apiculture were held for the benefit ofAnnual Report 2009-2010 117

Biotechnology Programmes for Societal Development42 rural youth. Honey production during the yearwas 836 kg. The number of bee colonies increasedfrom 77 to 205 during this period. Two trainingprogrammes were held on organic cultivation ofspices for 52 farmers/farm women.Mushroom CultivationThe rural population of the Jammu region wastrained in mushroom cultivation. Training wasconducted for 75 farmers from 22 villages. Inanother project implemented in Greater Noida,Uttar Pradesh, a centralized facility was establishedfor spawn production, and a demonstration unitestablished for technology transfer. In a projectimplemented at Bangalore, transfer of appropriatetechnologies for entrepreneurial cultivation ofmushroom for income generation was taken up. Atotal of 14 on–campus training programmes wereconducted involving 524 women, on spawnproduction, cultivation, preparation of value addedproducts and marketing of mushroom/products. Inaddition, 21 off campus training programmes wereconducted for 736 trainees.SericultureThe chawki rearing centre (CRC) model developedby CSRTI, Mysore was popularized in identifiedclusters of Karnataka, Andhra Pradesh and TamilNadu, for bivoltine sericulture. Six farmers wereselected, two from each state and they catered tothe needs of 100-120 farmers in the vicinity. Atraining programme on chawki rearing managementwas organized at CSRTI, Mysore for a period ofone week in different batches for the farmers.About 150 farmers were benefited by harvestingan average cocoon yield of 58-65kg/100dfls (diseasefree laying). Similarly, two CRCs brushed a totalof 90,000 dfls of hybrids and 250 farmers werebenefited. The average cocoon yield was 55-65kg/100dfls.Bio-control ManagementTraining and demonstration, activities were undertaken in Jabalpur district of Madhya Pradesh.Awareness campaigns on the use of biopesticideswere conducted for using simple, low cost,sustainable biopesticide production technology. Inanother project implemented at Noida, UttarPradesh, training and demonstrations were conductedon ecofriendly management of soil borne fungaland root knot nematode diseases infecting vegetables.For production of fungal bioagents, pressure cookers,polypropylene bags and mother cultures of weredistributed free of cost. For the application of IPMtechnology package on directly seeded crops (okra,cowpea), seed treatment was done with fungalbioagents. For the application IPM technologypackage on transplantable crops (tomato, brinjal,chili etc.) fungal bioagents were grown on sorghumgrains.Banana CultivationAgri-Entrepreneurship was promoted among small& marginal farmers through tissue culture basedbanana cultivation in Bhubaneswar, Orissa. About80 beneficiaries from four Gram Panchayats weretrained in banana cultivation. One demonstrationfarm was set up in 1.5 acres. Free supply of plantletsto the farmers was undertaken. A nursery cum lowcosttissue culture laboratory was established.Linkages were developed for finance to theentrepreneurs and marketing of the product.118Annual Report 2009-2010

Biotechnology Programmes for Societal DevelopmentTurkey FarmingPoverty alleviation through scientific farming ofturkey was initiated in selected rural areas ofKancheepuram district of Tamil Nadu. A total of13 villages with 1172 self help group women werecovered to create motivation and awareness on thepotential of turkey farming as an entrepreneurialactivity. The selected target groups were imparted‘hands on’ training on commercial turkey farming,feed mixing and hatchery operation. A total of 500self help group women were trained. The criticalinputs viz., 477 kg turkey feed, 575 kg feedingredients and 318 hatching eggs were supplied tothe target groups along with pulverisers andincubators.Medicinal and Aromatic PlantsCultivation of medicinal plants through organicfarming was taken up in the Mathura district ofUttar Pradesh. The project was implemented in10 villages and five medicinal plants viz tulsi,kalmegh, sannai, satavar and safed musli weretaken up for commercial cultivation. Six farmers’training programmes were organized in whichthe specialists trained 300 farmers. Four trainingcum-demonstrationunits of 0.2 hectares eachof tulsi, kalmegh and sannai were established,while two training-cum-demonstration units of0.2 hectares each of satavar and safed Musliwere established. The farmers got benefitsranging from 25% to 80% in comparison totraditional crops like sorgham, bajra, green gram,arhar, mustard, gram, barley, wheat etc. Thesehave encouraged the farmers to take upcultivation of medicinal plants and organicfarming.AquacultureMagur culture was demonstrated in farm pondsfor the upliftment of small and marginal farmersin Washim district of Maharashtra. A number oftraining programmes and demonstrations wereorganized on brood stock maintenance, breeding,rearing of seed, feeding, harvesting and marketingtechnology for 262 beneficiaries of the project.Fish seed were reared in nursery ponds up tofingerling stage and made available to 79 trainedbeneficiaries for culture.Rural Livelihood EnhancementRural livelihood enhancement through communityparticipation was undertaken in the Ernakulumdistrict of Kerala. A total of 120 beneficiaries wereselected for practicing interventions suitable for theagro-ecosystems of Moothakunnam viz. mushroomcultivation, vermi-compost and nutrition garden,and integrated fish cum poultry farming. Thebeneficiaries were divided into 4 groups of 30members each for practicing the interventions.Altogether 17 training programmes includingexposure visits and demonstration classes wereorganized. Extension leaflets in the vernacular onthe oyster mushroom cultivation, mushroom recipe,vermitechnology, and integrated fish-cum-poultryfarming were brought out. Four mushroomdemonstration units, 4 integrated fish-cum-poultryfarms and 40 vermicompost units & nutritiongarden were established.Agri Waste ManagementFiber extraction from agri-waste was successfullydemonstrated in the Ernakulum district of Kerala.Training manuals, in both English and MalayalamAnnual Report 2009-2010 119

Biotechnology Programmes for Societal Developmentwere distributed to 224 beneficiaries. In the secondyear, 204 beneficiaries from self help groups weretrained. Several production units were set up andtwo of them made their first sales of handmadepaper and products. The microbial strains isolatedwere purified and enzyme assays carried out toselect strains producing high yields of pectinase,hemicellulases and ligninase. The Post Master Generalof Kerala expressed an interest to procure paper ofarchival quality from the SHGs for printing ofpostal life insurance certificates.BiopesticidesBiopesticides as a cottage industry andentrepreneurship development among rural youthswas successfully demonstrated in Pantnagar area ofUttarakhand. Mass production of native strains ofTrichogramma was made in the biocontrol laboratoryand Trichocards were developed. Studies wereconducted on the efficacy of Trichogramma specieson different crops. A low cost technology wasdeveloped for mass production of alternate hostand production of Trichogramma. About 300 ruralyouth were trained to produce biopesticides at thecottage industry level.Tribal Health ManagementThe prevalence and awareness among Thalassaemicpopulation of their condition was studied in thevarious tribes belonging to different districts of WestBengal. Twenty seven camps were organized in ruralas well as urban areas for screening for Thalassemiacarriers. A total of 3210 individuals (Male:1858,Female:1352 ) were screened which included Totos,Rabhas, Mundas, Baraik, Kerketta, Ekka, Lohar,Gowala, Kheria, Mohali, Nagbanshi, Murmu andKera. About 35.3% of Totos and 36.0% of Rabhaswere carriers and 13.7% among Totos and 33% ofRabhas were homozygous for the mutation whichgives rise to the hemoglobin variant E (HbE).Mobile Veterinary ClinicsMobile clinic services were provided for the benefitof dairy farmers of rural western Vidarbha regionof Maharashtra. Survey of selected villages wascarried out to evaluate the health status of animals/livestock. Training programs were conducted to helpadoption of scientific animal husbandry practices.Over 1900 animals belonging to 1200 farmers weretreated for various diseases. Technical guidance wasextended to various Govt. agencies and institutionsfor better management of their projects. Medicines,surgical equipment, office materials and audio visualaids were provided.Rural Bio Resource ComplexA multicentric approach on integratedbiotechnological interventions for societaldevelopment was initiated in Pondicherry. Theproject developed four different interventions viz.vermicomposting, clonal propagation, biomassbriquetting and mushroom cultivation. Presently664 rural beneficiaries are utilizing the infrastructurecreated in the villages and practicing incomegenerating technologies. Apart from producingcharcoal briquettes, the beneficiaries were trained toproduce “incense briquettes” as a part of valueaddition. Moulds designed to produce these weredistributed. Beneficiaries were engaged in theproduction of amla seedling and flowers likekanakambaram, malli and vegetables namelycapsicum, tomato, leafy greens etc. So far, 16 activity120Annual Report 2009-2010

Biotechnology Programmes for Societal Developmentbased societies/ help groups were formed in differentoperational areas. The products generated by thelike mushrooms, briquettes, vermicompost andvegetables were sold in the nearby markets and alsothrough societies.Groundnut Seed ProductionDevelopment of knowledge based farmers ongroundnut quality seed production was taken up inthe Bijapur district of Karnataka. Seed input kitswere supplied to 10 beneficiary farmers. Regularfield visits were made during the crop period androuging of off-types was done at regular intervals.Training was given to 70 farmers. Beneficiary farmersgot additional income by selling their produce asseed. The technology had become self-sustainable.Vegetable Seeds ProductionProgrammes were taken up for multiplication,demonstration and training on vegetable hybrid seedproduction for generating additional income in ruralareas of Udham Singh Nagar and Dehradun districtsof Uttarakhand. Farmer’s families were selected ateach block level for hybrid seeds productiontechnology demonstration in which 50% were ST/SC farmers. The seeds of parental lines of bottlegourd, bitter gourd, cucumber and brinjal weredistributed to the farmers. The knowledge and skillsof farmers were enhanced through trainingprogrammes.Rabbit RearingImprovement of the economic and health statusof rural people, through backyard rabbit rearing,was undertaken in Bangalore, Kolar and Tumkurdistricts of Karnataka. The beneficiaries earned anincome of Rs.10,000/- to 12,000/- per year.Further, rabbit rearing activity also contributed tobalanced nutrition of rural poor people throughmeat and wool. In addition, the beneficiariesacquired skills with respect to marketing, feedpreparation and purchase etc, which had a positiveimpact on their livelihoods.Post Harvest TechnologyEmpowerment of coastal rural folk through trainingand demonstration on fishery products developmentwas done successfully. Fishery products such as fishmacaroni and fish chips were developed and theirstorage characteristics studied. Preparation of fisheryproducts like fish wafer, fish noodles, fish burger,fish puff, fish biscuits were standardized and thesetechniques were transferred to 250 coastal rural folk.A model unit was established at Iddinthakarai fishingvillage in Tirunelveli district for the production offish pickle, prawn pickle, crab pickle, masminpowder and masala flavored fish. Another fisheryproduct unit was established in Nazarath village ofThoothukudi district, for producing surimi basedproducts, fish pickle and spiced fish. Three largescale entrepreneurs established processing units.FloricultureFloriculture was popularized for economicupliftment of marginal farmers in Shimla, Kulluand Mandi districts of Himachal Pradesh. Theproject on lilium cut flower cultivation wasimplemented successfully. Extensive communitymotivation and awareness was promoted with theorganization of 28 camps and participation of 421farmers. A total of 36 farmers were organized inthree clusters, one in each district. Sixteen farmersAnnual Report 2009-2010 121

Biotechnology Programmes for Societal Developmentwho adopted the technology in the first year earnedmore than 100 days employment per annum withlilium cultivation on 100 sq m land. Thirteenpolyhouses of 1364 sq m floor area were fabricatedwith bank loan and were linked to the NationalHorticulture Technology Mission for partial financialassistance. Artisans were trained in low cost polyhousefabrication and 16 farmers were provided with shadenets to establish low cost cultivation and seedmultiplication plots.Programmes for WomenDuring the year, a total of 65 ongoing and newprojects were supported which benefited over 12,000women. The projects covered training,demonstration, extension activities, enterprisedevelopment, marketing linkages etc. in areas suchas floriculture, horticulture, mushroom cultivation,biofertilisers, biopesticides, organic farming,vermicomposting, aquaculture, animal husbandry,poultry farming, production of value addedmarketable items etc.Tamil Nadu, ‘hands-on’ training and demonstrationwere organized in sericulture, vermicomposting andorganic farming which benefited 360 farm women.In Thiruvallur district, 300 women farmers weretrained and given a demonstration onvermicomposting and biopesticide preparation. Amodel vermicomposting unit was set in the LoyolaCollege campus, Chennai.Mushroom cultivationForty unemployed women were given training oncomposting and cultivation of mushrooms at Urpashand Darind districts of J&K. The trainees were alsogiven a demonstration on preparation of mushroomrecipes. They were also imparted skill on marketingof their produces.Vermicomposting & Organic FarmingIn a vermitechnology project, 80 vermicompostproduction units with a capacity of 5-6 metrictonnes were set up in Aligarh and Hathras districtsof Western UP, which benefited 266 rural women.There was a production of 2560 quintals ofvermicompost generating an income of Rs. 6396for each beneficiary. Annamalai University trained330 marginal farm women from coastal Tamilnaduon vermitechnology and set up 42 vermicultureunits. The surplus vermicompost that was soldsupplemented their monthly income by Rs.1100-1500 per month per beneficiary. At Dindigul inThe SHG members and farmers were taught about vermicompostingmethod and biopesticides at Thiruvallar district.FloricultureAt Madakkathara in Kerala, training on cultivationof selected ornamental crops (gladiolus, gerbera,heliconia, gomphera etc) was imparted to farmwomen. Five such trainings on commercial122Annual Report 2009-2010

Biotechnology Programmes for Societal Developmentfloriculture and flower arrangements were given to62 beneficiaries. Demonstration plots wereestablished. Sale of flowers & flower arrangementsgave employment to 10 beneficiaries, and 33 of thebeneficiaries established flower production units,which resulted in substantial income.AquacultureA project on economic upliftment of rural womenthrough integrated fish farming was operated inPuri and Cuttack districts of Orissa and covered atotal of 300 beneficiaries. Out of these beneficiaries,282 belonged to different SHGs and had taken thecommunity ponds on lease. At Thoothkudi, TamilNadu, 40 women were trained in freshwaterornamental fish culture, and a few of them adoptedcottage level breeding activities of ornamental fishes.Under a project on lobster fattening, 410 womenbeneficiaries were trained. One lobster fattening unitwas established at Thoothukudi, Tamil Nadu.Undersized lobsters were purchased from localfisherman and again sold to local seafood processorsafter fattening for 3-4 months. At Thootikudi, TamilNadu, in a freshwater ornamental fish cultureproject, 186 women beneficiaries were trained inornamental fish reasing. Techniques for artificialhatching and live feed preparation were disseminatedto SHG women through training & fielddemonstrations.At Pantnagar, in a project on goat rearing, 968women got benefited through rearing 5562 goatsin 29 villages and through demonstration-cumtrainingprogrammes. This helped the beneficiariesto obtain enhanced growth of animals and also asaving of Rs.1.50 lakhs. At Anand, around 1100farm women were trained on scientific livestockmanagement through on-campus and off-campustraining programmes. In addition, around 700animals (cattle and buffalo) were given treatmentfor infertility.In Dharmapuri, Tamil Nadu 180 women weretrained on backyard poultry farming, 54 womenon clean milk production, and 27 on ornamentalfish production which gave them some additionalmonthly income. Each of the households wasgiven 9+1 improved poultry strains. About 23-25breeding stocks of ornamental fishes were developed.In a project implemented at Kolkata, forconservation & improvement of local white ducksto support shuttle cock feather industry, two trainingprograms were organized on modern duck farmingfor 175 women. It was observed that a white duckcould produce 96 eggs/year/bird and 27 good qualityfeathers. A pure line of white duck was establishedand distributed to women farmers. In a project forimproving socio-economic status of the ruralwomen, local desi birds were replaced with highyielding birds procured from the Project Directorateof Poultry, Hyderabad. The birds added to therevenue of the self help groups.Under a project at KVK, Madurai, training wasimparted on rearing of honey bees, and informationwas provided on site selection & bee foraging crops.About 100 women were benefited through theproject. The SHGs had a tie up with the Khadi &Village Industries Commission for marketing thehoney extracted by individuals.Annual Report 2009-2010 123

Biotechnology Programmes for Societal DevelopmentDistribution of beehives and accessories to the traineesHealth Awareness Campaign for Rural Women with special emphasison genetic disorders at Jaunpur District:A study was conducted at Tirupati on serum vitaminD and related hormonal and biochemical indices tosee the efficacy of vitamin D supplementation duringthe third trimester. The affected families wereprovided with calcium and folic acid supplementsin consultation with the hospitals and around 500women were benefited. Another study was inprogress in large slums of Hyderabad where localwomen were provided calcium rich recipes (tilladdus) twice a week i.e; for a period of one yearto lactating women after delivery and their bonedensity assessed every six months. Over 700 familieswere benefited through this project. The beneficiarieswere trained in the preparation of various valueadded food products using local fruits supplementedwith calcium rich local bioresources like ragi,amaranthus, til etc. At Jaunpur, UP, in ademographic study of maternal A66G and C667Talleles as risk factors for Down Syndrome andmental disorder, blood samples from 1000 womenand some patients were analyzed. Awarenesscampaigns for health & genetic disorders were held.At Thiruchirapalli, 50 farm women were trained inhybrid maize seed production and value addition ofmaize grains. By producing and selling hybrid seed,they were able to get a 30% increase in incomecompared to with the conventional grain. AtCoimbatore, 154 women from 4 villages weretrained in hybrid maize production. Eleven farmwomen were provided with inputs for conductingdemonstrations programme. Four of themsuccessfully maintained the crop as per thecertification standards and sold the seed to theAgriculture Department @ Rs 40 per kg. Fivewomen registered with the department and the foursuccessfully produced marketable seed according toseed certification norms.At Unnao in UP, 165 women were trained forpreparing value added products from mango and11 demonstration cum production units were setup in 5 villages. The project also facilitated the124Annual Report 2009-2010

Biotechnology Programmes for Societal DevelopmentKVK Unnao to start functioning as a IGNOUstudy centre for offering diploma in value addedproducts from fruits & vegetables.In a project at Kanyakumari, training on valueadded fish products was imparted to tsunami affectedfisherwomen. Four units were set up for productionof value added fishery products. Through improvedquality standards & marketing technologies, theincome of beneficiaries were improved to Rs. 3000per month /beneficiary.Under a project taken up at the DBT-ICARComplex at Meghalaya, a total of 740 beneficiariesfrom East Khasi Hills, Jaintia Hills and Ri-Bhoiwere given ‘hands-on’ training on post harvest lossesand management of horticultural crops like gingerand turmeric. Through a project on agri-hortimediculture approach for the benefit of womenfarmers in Uttarakhand, around 900 householdswere directly benefited in 7 villages. About 150nalis of fallow/under utilized land were broughtunder cultivation. A saving of water by 50% couldbe achieved through cultivation of aromatic herbsvis-à-vis potato. Garlic and turmeric showed goodacceptance by farmers.Initiatives were taken to promote the use of localbioresources in the preparation of commerciallyimportant items for additional income. Consideringthe availability of Bhimal fiber (Grewia optiva) andits application in preparation of various marketableproducts, a project was implemented at Uttarkashi,under which 470 women from 24 villages weretrained in the processing of Bhimal fibre and itsutilization. This resulted in an income of Rs. 1000-1500 per month per beneficiary. Women fromSrinagar area (Jammu & Kashmir) were trained incultivation of selected varieties of salix. They hadbeen trained in rearing the plant in their nurseriesand now the plants were ready for harvesting. Whilethe Salix plants were raised in nurseries, thebeneficiaries were trained in the preparation of variousitems like baskets, cricket bats etc. from the bioresources made available to them by the Universityof Kashmir.PrasadUnder the network, 13 projects components wereimplemented for creating employmentopportunities for women residing around majorshrines in Uttarakhand, Karnataka, Kerala, UttarPradesh, Himachal Pradesh, Arunachal Pradeshand Assam. Women were trained through localNGOs/ trusts of temples, churches, dargah, etc.in the preparation and marketing of various itemsfor offerings to respective shrines in accordancewith their tradition, rituals and belief. Emphasiswas on the utilization of locally available andnutritionally rich raw material for preparation ofedible items used as prasad. Other optionsadopted were the preparation of incense anddhoop-bati from local wild plants, floricultureand banana cultivation. The beneficiaries wereencouraged to prepare suitable souvenirs fortourists and generated additional income. Over4000 families were benefited through theseinnovative projects.Annual Report 2009-2010 125

CHAPTER 6BIOTECH PRODUCT ANDPROCESS DEVELOPMENTINDUSTRY SCHEMESSmall Business Innovation ResearchIntiative (SBIRI)The “Small Business Innovation Research Initiative(SBIRI)” scheme was initiated by the departmentto encourage small and medium enterprises to takeinnovative, high risk R&D projects for establishingproof of concept as well as for development andcommercialization of research leads. Some of theachievements under the completed and on-goingprojects are :HealthcareA platform for production of complex peptidesand proteins has been successfully developed andthe company is scaling up the processes to producethese peptides and proteins on a commercial scale.The company has applied for 2 patents coveringthe platform. It has identified a few therapeuticproteins of high value that could be produced in acost effective manner using this platform.A recombinant follicle stimulating hormone (r-FSH)that has been developed & commercialized underthe brand name “Foligraf” has a market share ofabout 30% in the Indian recombinant FSH market.The product is useful in the treatment of fertilitydisorders in women. The product has been shownto effectively stimulate follicular development insubjects whose endogenous gonadotrophin secretionis suppressed.FOLIGRAF : Recombinant human follicle stimulating hormonedeveloped by Bharat SerumsA silk protein blend film was developed for burnwound management. This has been demonstratedto be hypo-allergenic and non-toxic in rat models.Clinical trials are being undertaken.Commercially viable MAB’s for RBC phenotypingwere developed for the domestic market. Theproducts developed were standardized to producebulk quantities under GMP conditions. Theproduction standards are as per the standards ofcGMP with quality management system of ISO9001:2008 and with the regulatory approvals. Inanother project an alternative technology to replacethe polyclonal anti snake venom serum (ASVS)with a monoclonal cocktail has been developed.The achievements in other completed and on-goingprojects are the development of a homologousnatural bio-material for treating cancer lesions, burnwounds etc.; production of humanized monoclonalantibodies against epidermal growth factor receptorand development of Mycobacterium W. as an adjuvantfor anti-rabies vaccine.Annual Report 2009-2010 127

Biotech Product and Process DevelopmentAgricultureThe projects here are largely focused on thedevelopment of improved strains and generation ofplanting material in large quantities. Efforts are underway to genetically modify vegetable crops tomato,okra & brinjal for insect, pest and disease resistance.A full-fledged tissue culture lab, plant hardeningfacilities and a highly advanced greenhouse has beenestablished for the mass multiplication of highquality Jatropha curcas plants to produce biodiesel.Projects have also been supported for thedevelopment of salinity tolerant transgenic ricehybrids; drought tolerant genotypes of rice; diseaseresistant varieties of cotton and stacking of genesfor stress resistance in maize (Zea mays).Industrial Products and ProcessesThe focus of this programme is on thedevelopment, production and scaling up ofindigenous, cost-effective industrially importantenzymes through up-scaling the downstream processand manufacture of value added products.A nitrifying bioreactor technology based packed bednitrifying bioreactor, useful for organic recirculationof prawn seed production system in hatcheriesespecially small ones for management ofwater quality was developed. This is ready forcommercialization.Bio-instrumentationAn auto-dispenser has been designed and developedindigenously for generic bench top uses andprotocol-specific systems for specific diagnosticapplications. The development of an automatedcell-counter is also underway. A large size costeffectiveindigenous multi deck incubator shaker isalso being manufactured.Rapidec: An auto-dispenser developed by M/s Customised Technologies,BangaloreBiotechnology Industry PartnershipProgramme (BIPP)The BIPP scheme was approved in November 2008for partnership with industries on a cost sharingbasis for:(i)(ii)Path-breaking research in frontier, futuristictechnology areas with major economicpotential and making Indian industry globallycompetitive, and focused on IP creation withownerships by Indian industry andcollaborating scientists.Development of appropriate technologies inthe context of recognized national prioritiesin the area of agriculture, health, bioenergy,green manufacturing, when the scale of the128Annual Report 2009-2010

Biotech Product and Process Developmentproblem has serious consequences for socialand economic development.So far 5 rounds of call for proposals were announcedand more than 250 proposals received. Altogether,20 proposals were cleared and 11 projects supportedso far.Some major projects supported are:(i)(ii)(iii)(iv)(v)(vi)(vii)“Development and pilot scale production ofanti-TNF-a antibody scFv for treatment ofinflammatory diseases” – ARA Healthcare Pvt.Ltd., Gurgaon.“Development of self-glucogenic pearl milletadapted for marginal lands” -Avesthagen Ltd,Bangalore.“A multicenter randomized, double-blind,placebo control study of IN-105 tablets,Biocon Limited, Bangalore“Development of a cost effective prophylacticand therapeutic recombinant human Papillomavirus vaccine”- Gennova BiopharmaceuticalsLimited, Pune“Enhancement of ethanol yield from molassesfermentations by adding a specific enzyme toconvert an un-fermentable sugar to afermentable sugar” - Richcore Lifesciences Pvt.Ltd., Bangalore.“Process for asymmetric synthesis ofhexahydrobenzophenanthrenes, Dopamine D1agonists”- TCG Life Sciences Ltd., Kolkata.“A strategy for the development of alternativetreatment for heart failure complicated withdiabetes mellitus”- Torrent PharmaceuticalsLtd., Gandhinagar, Gujarat.(viii) “Development of a novel mucosal vaccine forHpv” - Virchow Biotech Pvt. Ltd.,Hyderabad.Biotechnology Industry ResearchAssistance Programme (BIRAP)A pilot BIRAP was established in November 2008with the mandate to promote and nurtureinnovation in the biotech industry especially SME’swith emphasis on:(i) promoting public private partnership programme(ii) facilitating IP management (iii) nurturing andsupporting innovation and research and (iv) capacitybuildingThe first programme which was operationalizedfrom the pilot BIRAP was the BIPP scheme.Besides BIPP, the pilot BIRAP in its first year ofoperation got equipped with the required staffwhich executed different programmes for capacitybuilding in partnership with ABLE and BCIL.Workshops on biotech licensing were organizedin New Delhi and Bengaluru. The pilot BIRAPalso initiated an electronic journal consortiumwith the help of DELCON to provide access toexperts and industry, to over 2000 journals andpublications. It also provided support for travelof experts to workshops, meetings andconferences.Biotechnology Industry ResearchAssistance CouncilIt is proposed to set up a Biotechnology IndustryResearch Assistance Council (BIRAC) as anindependent autonomous organization to deal withAnnual Report 2009-2010 129

Biotech Product and Process Developmentindustry related activities of DBT i.e. ignition grant,pre-SBIRI, SBIRI, BIPP, Incubation centre, BiotechPark, Research Resource Facilities, Industry Platformetc. This organization would be the interface agencyof DBT for industry-academia interaction and willalso help in promoting start-ups, SMEs and facilitateinnovation research in large industries. AManagement cum Advisory Board and an ExecutiveCommittee were constituted and national levelconsultations were held.R & D SCHEMESBiotech Product and Process DevelopmentThe emphasis of this theme was on developmentof research leads as well as products and processesfor utilization by various biotech industries.Significant achievements of some of the projectsare:In a project implemented at Punjab University,Chandigarh, the gene coding for chitinase was clonedin Escherichia coli. The recombinant chitinase waspurified to near homogeneity. Like native chitinase,the recombinant chitinase produced medicinallyimportant N-acetyl D-glucosamine and chitobiosefrom swollen chitin used as a substrate and it alsoinhibited the growth of many fungi.Work done at NIIST resulted in completesequencing of the gene encoding phytase gene(phyA), isolated from Aspergillus ficuum NRRL 3135and A. niger BTCF 5 using primer walking. Toexpress the phyA gene in a eukaryotic expressionsystem, Kluyveromyces lactis GG799, phyA maturegene sequence was ligated in to vector, pKLAC1and the recombinant plasmid was transformed into E. coli JM 107. The clones were confirmed bycolony PCR and double digestion. The confirmedclones were linearized using SacII restriction enzymefor integration in K. lactis genome.A team working at Shri AMM Murugappa ChettiarResearch Centre, Chennai, optimized the growth ofmicrofungi viz., Curvularia lunata, Alternariaalternata, Sclerotinia sp., Pestalotiopsis sp., and Phomafoveata for pigment production in a liquid nutrientmedia. Purple coloured intra- and extracellularpigments were produced by the fungi and pigmentproduction was enhanced under light. The pinkand beige shades were developed in cotton yarnswith Phoma foveata and Curvularia lunata pigmentsrespectively. Two different shades were developedfrom Pestalotia sp. and the wash fastness was tested.Sclerotinia sp. showed different shades of light green,green, brown, and pink in cotton yarn. The fungalpigments were found to be stable at different pH(2.0 – 14.0), different temperature (40 – 120C), inhot air oven (90’C) and steaming at 120 lbs for 20min.In a project on solid state fermentation of deoiledJatropha seed cake carried out at IIT, New Delhifor production of industrially importantextremophilic enzymes, a xylanase producerthermophilic Scttalidium thermophilum strain wasisolated The SSF conditions were optimized and aproduction of 4691 U/dry seed cake was achieved.The xylanase exhibited novel properties i.e., beingheat stable and free of cellulase. The detoxificationof phorbol ester was achieved by fermentationwith Enterobacter sp strain isolated by them. Thestrain was found to hydrolyze 60-70% toxicphorbol esters when grown on deoiled Jatrophaseed cake.130Annual Report 2009-2010

Biotech Product and Process DevelopmentIn a study undertaken at TERI, New Delhi, anefficient technology for production of bioethanolfrom the hemicellulose fraction of sweet sorghumbagasse was developed. Fermentation ofhemicellulose hydrolysate of sweet sorghumbagasse was achieved with Pichia stipitis NCIM3497 and Debaryomyces hansenii sp. Thefermentation of hemicellulose hydrolysate showeda higher ethanol yield than that from syntheticmedium containing only pure xylose as a carbonsource.In a study conducted at JNU, New Delhi, thetranscriptomic profiling and structured modeling ofrecombinant E. coli high cell density cultures forthe design of improved host strains was taken up.The different sets of genes were shown to get up/down regulated during induction, clearlydemonstrating that cellular stress is a function ofthe recombinant protein expressed, environmentalparameters, and the host cell background. Crosscomparison of the transcriptomic profiles helped inidentifying the key genes which need to be upregulatedfor enhancing the recombinant proteinyield.Biotechnological Approaches for Foodand Nutritional SecurityProbiotics for holistic healthThe use of probiotics for a holistic approach tohealth management is gaining increased acceptance.A number of research programmes were taken upin this regard. NDRI, Karnal, developed arepository of 102 probiotic lactobacilli, out ofwhich 68 were identified and characterized. AIIMS,New Delhi, studied the claims of 20 probioticpreparations available commercially and foundthat in all cases, the numbers and viability claimswere not substantiated and the identification ofthe microbes was inadequate. Assays for cytokinesin stool of 11 breast-fed and 21 non-breast-fedchildren showed high basal levels, not altered byprobiotics. A project at CMC, Vellore studied 36probiotic isolates from human and dairy sourcesand observed an intrinsic resistance to a widevariety of antibiotics in almost all strains. Assayswere developed for evaluation of probioticactivities using epithelial and monocytemacrophagecell lines and peripheral bloodmononuclear cells from health volunteers. Someprobiotics induced tumor necrosis factor secretionfrom macrophage cell lines, while they were ableto suppress IL-8 secretion in epithelial cell linesin response to appropriate stimuli. Through aproject at PGI, Chandigarh a trial was conductedon stool colonization in premature infants. Itwas observed that Lactobacilli with eitherBifidobacteria or Saccharomyces colonized the gutin over 90% of the infants using any of thecombinations. A study supported at NDRI, Karnalevaluated three probiotic dahi preparations in ratsfed a high cholesterol diet. All three dahis loweredthe serum cholesterol and triglyceride levels andraised the HDL cholesterol compared to buffalomilk or normal dahi. This lead is being taken upfor validation.Detection of Genetically Modified Foods(GMOs) and Food ProductsThe GMO detection methods are a pre-requisite toassessing the consequences of genetic modificationin terms of food security, environmental risk andconsumer preference. A brainstorming session onAnnual Report 2009-2010 131

Biotech Product and Process Development“GM chip technology was organized at NBPGR,New Delhi to develop a cost-effective micro-arraysystem for detection of GMOs. A networkprogramme involving, CDFD, Hyderabad; NBPGR,New Delhi; ITRC, Lucknow; NIN, Hyderabadand CFTRI, Mysore developed assays and kits forrapid detection of various transgenic traits. TheCDFD has also developed efficient duplex andmultiplex PCR methods for simultaneous detectionof two GM cotton events i.e. Mon 531 and Mon15985. At NIN, Hyderabad, methods foridentification, screening and detection of Bt-cotton,RR-soya and Bt-maize were standardized based ontheir endogenous reference genetic markers, universalGMO screening elements and the presence oftransgenes.Nutritional and nutraceutical potential of underutilized plant resourcesAt CFTRI, Mysore, studies were carried out onpigment rich extracts from Tinospora cordifolia(carotenoids), Santalum album (anthocyanins) andRivina humilis (betalains) fruits. Tocopherol showeda mild stabilizing effect on carotenoid rich extractof T. cordifolia fruits. All the fruits tested showed ahigh content of sugar, protein, phenolics etc.Methanol extracts of Rivina leaves, root, and stemshowed inhibition against B. subtilis whereas B. cereuswas sensitive to only leaf and stem extracts. Inanother study, it was observed that the Indian redseaweed, Acanthophora spicifera had conjugatedpolyenoic fatty acids viz., conjugated eicosapentaenoicacid and conjugated arachidonic acid. Severalseaweeds are also indicated as potential sources ofnatural antioxidant compounds. Brown seaweedsespecially Padina tetrastromatica and Sargassummarginatum containing considerable quantities offucoxanthin (FUCO), were found to amelioratethe oxidative stress induced by vitamin-A deficiency(VAD). In a potential commercial application, itwas shown that Indian brown seaweed (Sargassummarginatum) can be incorporated up to 2.5%(w/w) to obtain Pasta of better biofunctionality/quality.Strategy to increase omega-3 fatty acid contentin the foodThe major aim of this approach was to developrecombinant yeast strains capable of generating theessential omega–3 fatty acid and, alfa linoleic acidby desaturating the omega-6 fatty acid and linoleicacid. Work was carried out on cloning of FAD -3gene from yeast strains identified as expressingomega-3 desaturase, and characterization of FAD-3fatty acid products in additional strains identified aspositive in hybridization studies.Chitosan based films for food packagingapplicationsChitosan (CH) and synthetic biodegradablepolymer, poly-å-caprolactone (PCL) blend filmswere under development to reduce water vapourpermeability rate to minimize water loss frompackaged fruits/ vegetables, extending theirshelf life. Differential scanning calorimetrythermograms showed better miscibility forCH80/PCL20 blend ratio by lowering PCLmelting peak temperature. SEM studiessupported these results.Potential of black tea as bio-enhancer in cancerchemopreventionBlack tea is known to exhibit antioxidant,132Annual Report 2009-2010

Biotech Product and Process Developmentantiproliferative, anti-inflammatory and immunomodulatoryproperties. A study carried out at ITRC,Lucknow showed that black tea had drug resistancereversing potential against several cancer (breast,prostate, skin and liver) cell lines. Cancer chemopreventive bio enhancer aspect of tea was analyzedin animal models with cancers of different organs.Post translational modifications of hemoglobinin iron deficiency and iron overloadAt St. John’s National Academy of Health Sciences,Bangalore, studies were carried out onglutathionylated hemoglobin which is a novelbiomarker for iron deficiency anemia. Levels of thehemoglobin were significantly elevated whencompared to levels seen in age matched controls (p

Biotech Product and Process Developmentcommercialization. R&D activities were underwaywith various other crops like rice, okra, brinjal,mustard, tomato, cabbage, cauliflower, ground nut,pigeon pea, chick pea, cowpea, finger millets,sunflower, soybean, bamboo, castor, wheat, blackgram, black pepper, chillies, onion, papaya and rubber.Nine public/private institutions/seed companieswere permitted to conduct confined field trialsof six transgenic crops viz., cotton, corn, rice,brinjal, potato and sorghum containing new genes/events with the approval of RCGM and GEAC.The list of approvals accorded for conduct ofconfined field trials of biosafety research level-1(BRL-1)/ event selection on transgenic cropsdeveloped by public/private institutions/seedcompanies within the country is given inTable-1.Table-1Transgenic crops approved for conduct of confined limited fieldtrials by the Institutes/ Industries/Seed Companies during 2009-10.Sl. No. Crop Institute/Industry Transgene1. Brinjal Bejo Sheetal Seeds Pvt. Ltd., Jalna cry1Fa12. Cotton Dow AgroSciences India Pvt. Ltd. cry1Ac & cry1FJK Agri Genetics, Hyderabadcry1Ac (Event-1) & cry1ECMahyco, Mumbaicry1Ac,cry2Ab and CP4EPSPS3. Potato CPRI, Shimla RB gene derived fromSolanum bulbocastanumvacuolar acid and invertaseRNAi-transgenic eventsGA20 Oxidase 1 gene4. Rice Mahyco, Mumbai cry2AbBayer BioSciences, Hyderabadcry1Ab, cry1Ca and bar5. Corn Dow AgroSciences India Pvt. Ltd. cry1FPioneer Overseas Corporation, Hyderabad cry1F and CP4EPSPSMonsanto India Ltd., Mumbaicry2Ab2 and cry1A.105 &CP4EPSPS6. Sorghum National Research Centre for Sorghum cry 1B gene(NRCS), Hyderabad134Annual Report 2009-2010

Biotech Product and Process DevelopmentFourteen Central Compliance Committees (CCC)were constituted to interact with the SAUs formonitoring of the BRL-1 confined field trialsconducted on GM crops under the jurisdiction ofthe respective SAUs. Under the event based approvalmechanism for commercialization of Bt Cottonhybrids expressing approved events, the StandingCommittee constituted by the Ministry ofEnvironment & Forests was serviced by thedepartment since April 2009. Based on the approvalof the Standing Committee in its two meetings heldin the department, 398 Bt cotton hybrids expressingfour approved events were recommended to the StateAgriculture Departments for commercialization.In the pharmaceutical sector, 15 rDNA productswere approved for conducting pre-clinical toxicitystudies at 11 private/public/autonomous institutions& companies. Information on the pharmaceuticalproducts approved by RCGM for conduct of preclinicaltoxicity studies during 2009 is given inTable-2.Table-2List of r-DNA Pharmaceuticals approved for pre-clinical studies during 2009-10.S. No. Products Institute/Industry1. • Recombinant human growth hormone (rhGH) M/s. Reliance Life Sciences, Mumbai• Recombinant human Chorionic GonadotrophinHormone (rhCG)• Rituximab-monoclonal antibody• Recombinant chimeric monoclonal antibody(IgG1 k) Abciximab• Recombinant Palivizumab2. • Recombinant tick vaccine antigen (Bm95) M/s. Indian Immunological Ltd.,• Recombinant epsilon toxoidHyderabad3. • Recombinant pegylated G-CSF M/s. Biocon India Limited,• Bmab 200 (Trastuzumab) Monoclonal antibody• Putatively targeting epidermal growth factorreceptor (EGFR)• Bmab 100 Monoclonal antibody putatively targetingvascular endothelial growth factor (VEGF)Bangalore4. • Recombinant Human Chorionic Gonadotropin M/s. Bharat Serums & Vaccines Ltd.,(rHCG)MumbaiAnnual Report 2009-2010 135

Biotech Product and Process DevelopmentS. No. Products Institute/Industry5. • Filgrastim Injection (480mcg/vial) M/s. USV Limited, MumbaiPEG-Filgrastim Injection (6mg/vial)6. • Malaria vaccine formulations ICGEB, New Delhi7. • Recombinant Human Insulin API (r-DNA origin) M/s. SciGen BioPharma Pvt. Ltd.,Pune8. • Recombinant Parathyroid Hormone (rPTH) M/s Bharat Biotech InternationalLtd., Hyderabad9. • Recombinant human granulocyte colony M/s. Lupin Limited, Mumbaistimulating factor (rhGCSF)10. • Recombinant Anthrax vaccine M/s Panacea Biotec Ltd., New Delhi11. • Recombinant Granulocyte Colony Stimulating M/s. Shasun Chemicals and DrugsFactor (rGCSF) and recombinant Pegylated Granu- Ltd., Chennailocyte Colony Stimulating Factor (r Peg- GCSF)Based on the pre-clinical study reports, 8 rDNAproducts developed by 6 private/public institutions& companies were recommended by the RCGM tothe DCGI for clinical trials. Table 3 gives the detailsof rDNA products recommended by RCGM toDCGI for clinical trials.Table-3List of r-DNA Pharmaceuticals recommended by RCGM to DCGIfor appropriate phase of clinical trials during 2009-10.S. No. Products Institute/Industry1. • Recombinant human follicle stimulating M/s. Reliance Life Sciences, Mumbaihormone (rFSH)• Recombinant Human Interferon gamma-1b (IFNg)2. • Recombinant human Parathyroid Hormone Intas Biopharmaceuticals Ltd.,(rHu PTH)Ahmedabad136Annual Report 2009-2010

Biotech Product and Process DevelopmentS. No. Products Institute/Industry3. • Teriparatide (rDNA origin) Injection (750 mcg/vial) M/s. USV Ltd., Mumbai• Recombinant human Platelet Derived GrowthFactor-BB (PDGF-BB)4. • Combination malaria vaccine composed of ICGEB, New Delhirecombinant PfMSP-1 195. • Recombinant Darbepoetin-alpha M/s. Avesthagen Ltd., Bangalore6. • Recombinant Etanercept (anti TNFr-antibody M/s. Zenotech Laboratories Ltd.,fusion protein)HyderabadThe DBT received a number of applications frominstitutions working with GMOs for constitutionof IBSCs in their research institutions. Altogether,412 IBSCs were constituted at various public-fundedinstitutions, universities, private R&D institutionsand industries involved in rDNA research activities.Two dedicated dynamic & interactive web sites on“Biosafety” (http://dbtbiosafety.nic.in) and “IndianGMO Research Information System (IGMORIS)”(http://igmoris.nic.in) aimed to provide informationon research work going on in Indian laboratoriesand disseminate information on the statutoryrequirements to be adhered to by the researcherswere in operation. The details of projects weremade available on the website. All the relevantActs, Rules and Guidelines for processing applicationsfor biosafety clearances including new gene constructs,collaborations, genotypes, proforma, stepwiseprocedures to be adopted for transgenic crops andr-DNA therapeutics etc. were made available in thepublic domain through these websites. Thesewebsites also facilitated in tracking of regulatoryclearances on applications to RCGM and e-monitoring of IBSCs on personalized web features.Patent Facilitation CellThe patent facilitation cell of the DBT has beenworking towards the creation of awareness andunderstanding related to IPR, among students,scientists and faculty of colleges, universities andresearch institutes, through workshops, seminarsand conferences. The department providedadministrative and financial support for filing ofpatent applications and maintenance of patentsgranted in the areas of life sciences andbiotechnology from the publicly fundedinstitutions. The patent filing is done throughNational Research Development Corporation(NRDC). The number of patent applications filedby DBT is 5 and the number of patents granted is11 during 2009-10.Annual Report 2009-2010 137

CHAPTER 7BIOTECHNOLOGY INFORMATIONSYSTEM NETWORKAdvances in bioinformatics and computationalbiology are possible only with the activecollaboration of specialists from diverse fields suchas biology, computer science, mathematics, statistics,physics and related sciences. Bioinformatics hasclearly emerged as an important discipline forresearch and various industrial applications. Thelarge size of biological data sets, the inherentcomplexity of biological problems and the need tohandle data without errors call for large memoryspace and huge computation time. TheBiotechnology Information System Network(BTISnet) program of DBT promotes developmentof specialized manpower and undertaking ofchallenging research activities with experimentalbiologists. The system also promotes the creationof database, software tools, and infrastructurefacilities. The growth and the current status ofBTISnet are outlined here.Biotechnology Information System(BTISnet)BTISnet, functional since 1986-87, has emerged asa major distributed scientific network in the world.To day it spreads across the country and comprises155 Bioinformatics centers with varied subjectspecializations. More than 25 of these centers arelocated in the North Eastern states of India. ANorth East Bioinformatics network (NEBInet) hasbeen specially designed for these states to speed upgrowth in the region. Altogether, 80 majorinstitutions of the country have been included underthe BTISnet by creating Bioinformatics InfrastructureFacilities for Biology Teaching throughBioinformatics (BTBI). These are networkedthrough high speed internet connectivity.Centres of Excellence (COE) inBioinformaticsSignificant progress has been registered by thecenters in terms of research publications and humanresource development. These CoEs are located atthe (i) Bose Institute, Kolkatta, (ii) IISc. Bangalore,(iii) JNU, New Delhi, (iv) MKU, Madurai,(v) University of Pune, Pune, and (vi) IIT, Delhi.Bose Institute, KolkataThe centre undertakes research in modeling ofprotein structures; drug design; molecular interactionand docking; chromosomal dynamics; stress biologyof plants; host-pathogen interactions and functionalgenomics. A number of webservers such as ProFace,IntGeom, ConfPlot, ContPlot are available at: http://www.boseinst.ernet.in/resources/bioinfo/stag.html.Computer programs were developed for displayingevolutionary conservation of protein interfaces.Altogether, 13 peer reviewed research papers werepublished during the year.Indian Institute of Science, BangaloreThe key focus of the centre is on systems biology.Annual Report 2009-2010 139

Biotechnology Information System NetworkSystems–level modeling and simulation ofmycobacterial metabolism was carried out and thishas provided insights into key reactions and criticalproteins involved. It has provided leads in identifyingpoints for possible therapeutic intervention. Anotherarea of focus is the development of algorithms forsequence and structural bioinformatics. An algorithm(FAIR) has been developed to find all distant repeatsin a protein sequence that will be useful inestablishing correlations of the repeats with regardto their function and three-dimensional structureduring evolution. A geometric clustering algorithmhas also been developed using backbone phi, psiangles to group together conformationally similarpeptide fragments of any length. More than 35papers were published in high impact journals.Jawaharlal Nehru UniversityThe bioinformatics center (BIC) was established in1989 for providing information to the researchersin the Northern Region. The center is now focusingon the development of Chemoinformatics baseddrug designing tools. Research is targeted towardsnew molecules as potential drugs for Entamoebahistolytica, simulation of viral proteins and inhibitordesigning, prediction of miRNAs, dynamics ofintracellular oscillators etc. A high performancecomputing facility has been set up which will assistin large scale data analysis, systems biology researchand simulation of large systems. The centre haspublished 25 research papers during the year.Madurai Kamraj University, MaduraiThe centre at MKU, Madurai specializes in structuralbioinformatics and genetic engineering. Majorresearch activities pertain to structural bioinformaticsof membrane proteins, structural genomics ofprophage proteins, genome analysis for non codingRNAs, proteomics and biomarkers discovery,molecular modeling and drug design, and strainmanipulation of industrially useful microorganisms,The centre has been working in collaboration withWeizmann Institute, Israel; University of Bath, UK;University of Aalborg, Denmark; NCBS, Bangalore;IISc, Bangalore; AIIMS, New Delhi and Avesthagen,Bangalore.University of PuneThe bioinformatics centre at the university is activelyinvolved in human resource development inbioinformatics. It has expanded its portfolio ofresearch to include genome assembly andcomparative genomics, metabolic pathway analysis,chemoinformatics and target based drug design,organization and interaction of proteins in virus,epitope mapping, etc. The centre was awarded aUS and Vietnam patent for synthetic peptide vaccineagainst Japanese encephalitis virus jointly with theNational Institute of Virology, Pune. The centrehas published three important research papers in thearea of metabolic alphabets in bacterial systems andmolecular phylogeny analysis.Super Computing Facility for Bioinformatics(SCFBio), IIT, DelhiThe facility was established in 2002 and is currentlymanaged by 20 personnel comprising Ph.D.scholars, post graduate students and project staff.About 10, 000 hits are registered per day. Thecentre has developed indigenous software packagessuch as” Gene to Drug” for genome annotation,tertiary structure prediction of small proteins and140Annual Report 2009-2010

Biotechnology Information System Networkactive site directed lead molecule design. Thesepackages are accessible to researchers. The SCFBioprovides the computational resources, bothhardware and software to the newly created Schoolof Biological Sciences at IIT, New Delhi. TheSCFBio has been recently upgraded through DBT’sprogramme support as a multi-tera facility. Thisunique facility which includes Gene to Drugsoftware will be operational and freely accessibleover the net.A start-up company called ‘Leadinvent’ has beenunder incubation and working on converting Geneto Drug software into products (new molecules).The company has handled projects from SBIRI,Ranbaxy and Novartis. They are also working on acollaborative project with Genus Ontology, HarvardMedical School and TIFR, Mumbai.New Bioinformatics InfrastructureFacilities (BIF) for Biology Teachingthrough Bioinformatics (BTBI)The Bioinformatics Infrastructure Facilities (BIF)scheme was launched to promote innovation inBiology Teaching through Bioinformatics (BTBI)three years ago. The scheme is designed to exposeteachers and students to the real-world of scienceand the use of bioinformatics for solving hard corebiological problems. These centres use lecturematerials, video clippings, demonstrations, tutorialsand on-line lectures to help students and teachers tonavigate and gather data that are useful for researchin biological sciences. Several sites provide free accessto biological information resources to develophuman resource to undertake challenging researchin the field of modern biology. Eighty eighteducational institutions have so far been supportedwith BIF centers. Five new centers were establishedin the current year.North Eastern Bioinformatics Network(NEBInet)NEBInet which provides linkages to 25bioinformatics centers located in the North Easternregion of India is part of BTISnet. These centersare located at various universities, colleges andinstitutions to promote communication among thescientific community, sharing of information andhelp in initiating collaborative R&D activities invarious fields of life sciences. During the currentyear, two centers were established in the region.Human Resource DevelopmentBioinformatics is a key tool to handle biologicalresearch problems. It requires highly trainedmanpower to deal with molecular biology and theapplication of software tools. The department hadtherefore accorded this area high priority and hasintroduced several innovative educational activitiesto meet the requirements.M. Sc., M.Tech & Advanced P.G. DiplomaCourses in BioinformaticsThe MKU, Madurai, Pondicherry University,Pondicherry and Anna University, Chennai initiateda Network programme on higher education in“M.Sc. Computational Biology” on a consortiumbasis. The objective of this network programme isto share the expertise of teachers and the resourceswhich are created by these universities, through videoconferencing and virtual class room approaches. Theprogram envisages creation of a strong computationalAnnual Report 2009-2010 141

Biotechnology Information System Networkand experimental foundation to bioinformaticseducation at the post graduate level. The otheruniversities i.e. JNU, New Delhi; Pune University,Pune; and Calcutta University, Kolkatta arecontinuing the M.Tech., M.Sc. and Post Graduatediploma courses in bioinformatics respectively.Ph.D. Program in BioinformaticsThe COEs of BTISnet including the supercomputing facility at IIT, New Delhi are runningPh.D. programs in bioinformatics to meet the hugerequirement for high end human resource inbioinformatics and computational biology.Short Term Training ProgramsIn order to provide exposure and hands on experienceto researchers, and students in emerging areas, theBTISnet centres organized 139 short-term trainingprograms. Over 2000 researchers were trained duringthe year through these programs that were well received.Bioinformatics National CertificationExamination (BINC)The BINC examination was introduced in 2005 byDBT. It is designed for the purpose of certifyingprofessionals in the area of bioinformatics. Thisexam is unique and employs a three-tier system toevaluate the knowledge and skill of the candidates.All successful candidates are awarded certificates ofproficiency in bioinformatics. The certificate isgenerally considered equivalent to the AdvancedDiploma in Bioinformatics offered by severaluniversities and DBT-supported institutions. Theexamination was conducted at seven centers(Bangalore, Chennai, Delhi, Hyderabad, Kolkata,Pune & Shillong). Out of 752 registrants, 519appeared for the examination, and 30 candidateshave qualified the BINC 2009. The exam isconducted every year.BINC prospectus being released on October 27, 2009: L to R: Prof. A. S. Kolaskar (Chairman BINC Coordination Committee), Dr. M. K.Bhan (Secretary, Dept. of Biotechnology) & Dr. Urmila Kulkarni-Kale, Director, Bioinformatics Centre, University of Pune, Pune.142Annual Report 2009-2010

Biotechnology Information System NetworkR&D in Bioinformatics andComputational BiologyIn-silico analysis has become a part of almost everytopic in modern biology research. During this year,the department supported 8 new R&D projectsrelated to collagen, peptides, regulatory systemanalysis with respect to miRNAs, insulin sensitiveglucose transporters, electrostatic mechanism ofhistone tail modifications, computational methodfor riboswitch detection covariance matrix and itsapplications in bioinformatics.Consortium Projects on BioinformaticsFor the development of a database on repository ofall information resources, two consortium projectswere sanctioned. These are on (i) TB-bioinformaticsand (ii) rice bioinformatics. Seven institutions arepartners in this project. The rice bioinformaticsproject sanctioned during the year has the partnershipof eight institutions namely viz. (1) NBPGR, NewDelhi (2) NRCPB (IARI), New Delhi (3) CRRI,Cuttack (4) IGKV, Raipur (5) DRR, Hyderabad(6) CSSRI, Karnal (7) IAS (BHU), Varanasi and(8) UDSC, New Delhiprocessing. Proposals are also invited through thesystem for organizing exhibitions and popularlectures in India. The TERI has been identified asan associate partner to carry out the management ofthis program.DBT’s e-Library Consortium (DeLCON)A major initiative taken by the DBT was thelaunching of the e-Library Consortium (DeLCON)in January 2009, which provided access to 600journals, by the autonomous institutions of DBT.This facility was upgraded to include more than900 journals for online access. The portal of theconsortium can be accessed through www.nbrc.ac.in/delcon. The DeLCON facility has been extendedthis year to 15 institutions in the North Easternstates with full access to more than 900 importantbiology journals and biotechnology.Websites and PortalsOnline System for Financial Support toConference, Travel, Exhibition & PopularLecture (CTEP)The Department of Biotechnology, launched a newuser friendly interactive website URL http://www.dbtctep.gov.in on CTEP. This website enablesthe submission of proposals and expediting theirSnapshot of online system for DBT’s e-Library Consortium(DeLCON)BTISnet websiteThe BTISnet website with URL http://www.btisnet.gov.in provides a central resource to allAnnual Report 2009-2010 143

Biotechnology Information System NetworkAnnual Report 2009-2010 145

CHAPTER 8BIOTECHNOLOGY PARKS, INCUBATORSAND INDUSTRY PLATFORMSBiotechnology Parks and IncubatorsBiotechnology parks are created to facilitateinnovation through the development of abiotechnology industrial cluster and create a skillpool of biotechnologists and entrepreneurs who havea strong foundation in research and innovation.Incubators are a platform to encourage entrepreneurialspirit, by providing the basic minimum facilitieslike research laboratories for product and processdevelopment, quality control and validation oftechnologies. With a large number of multinationaland bio pharma companies initiating their R&Dand manufacturing operations in India, the demandof biotech incubators has increased. The incubatorshelp start up companies and young entrepreneurswho are not able to invest in setting up facilitiesbut have capabilities to develop and design newbiotech products and processes by using the facilitiesin incubators. The Central and State Governmentsare making earnest efforts to promote biotechnologyby setting up biotechnology parks, incubators aswell as pilot projects through public-privatepartnership.Golden Jubilee Biotechnology Park for Women,Chennai, Tamil NaduThe park was registered as a Society under theTamil Nadu Society's Registration Act 1975. It isa joint project of the Government of Tamil Naduand DBT. It is located in Siruseri, Chennai, and isfully functional. The production units that havebeen set up are for herbal cosmetics, bio-pesticides,bio-fertilizers, spice fortified with herbs, and essentialoil, ready to eat snacks etc. An amount of Rs 400lakhs has been provided by the Department ofBiotechnology.TIDCO Centre for Life Sciences (TICEL) BioPark, Chennai, Tamil NaduThe park was established by the Government ofTamil Nadu with technical collaboration from theCornell University, USA. The Park has 87 modulesof BSL-2/BSL-3 lab spaces, each of 1525 sq. ft.,which have been leased out to companies for theirR&D activities. The broad areas of focus includepharma R&D, medical biotechnology, andbioinformatics. The DBT has sanctioned Rs. 1420lakhs towards establishment of a Bio ResourceCentre (BRC) at TICEL Bio Park Ltd andRs. 186.5 lakhs has been released as the firstinstallment.Guwahati Biotech Park Technology IncubationCentre, GuwahatiThe DBT has sanctioned the setting up of GuwahatiBiotech Park Technology Incubation Centre(GBPIC) at a total cost of Rs. 2799 lakhs for aperiod of three years. An amount of Rs. 2437.9lakhs has been released as the first year grant. Theprogress of the park is regularly monitored by theManagement Committee of GBPIC.Annual Report 2009-2010 147

Biotechnology Parks, Incubators and Industry PlatformsBiotechnology Park, KarnatakaThe Department of Biotechnology has supportedthe establishment of an incubation centre andcommon instrumentation facility at the park inBangalore. The cost of the project was Rs. 3226lakhs and the DBT's share was Rs.1445.45 lakhs,out of which Rs.1113 lakhs has been released sofar. The Karnataka Biotechnology & InformationTechnology Services which is the implementingagency for development of the park is now inthe process of developing an incubation centre in5 acres out of 14 acres of land earmarked for theproject and also procuring the necessaryequipment.Biotechnology Park, KeralaThe proposal for setting up of a biotechnologyincubator in the Kerala Biotechnology Park wassanctioned in March 2005 at a total cost ofRs.2318.90 lakhs. The share of DBT is Rs.1100.00lakhs and the remaining is to be contributed by theState Government. An amount of Rs. 150 lakhshas been released so far by DBT.Biotechnology Park, OrissaThe DBT has released an amount of Rs.200 lakhsto the Society for Development of Biotechnology,Government of Orissa for setting up of theBiotechnology Incubator Centre in the Biotech-Pharma-IT Park in Bhubaneswar.Nano Biosym Technology Park, HimachalPradeshThe Government of Himachal Pradesh hadsubmitted a proposal in April 2007 for establishmentof Nano-biosym Park in Himachal Pradesh. TheGovernment of Himachal Pradesh were allowed toutilize the earlier released fund of Rs 50 lakhs forHP Biotechnology Park for engaging a consultantfor preparation of detailed feasibility report for thesetting up of Nano- Biosym Park.Biotechnology Park, LucknowThe park houses 16 entrepreneurs. Five of them areUS based. Four had signed agreements with thepark and have occupied nearly all of the 50,000square ft. of air-conditioned space available for R&Dand production facilities. The park providesknowledge based support to two companies, one ofwhich is located in Rajasthan. Two companies, i.e.,M/s Chandan Health Care and M/s LifecareInnovations Pvt. Ltd. have started construction oftheir facilities with an investment around Rs. 450lakhs. The facility of Lifecare Innovations would bethe first WHO compliance GMP facility in theState. The park has also received the QualityManagement System Certificate from NorskAkkreditering of Norway. The NS-EN ISO9001:2008/ISO 9001:2008 certificate has beengranted for running a biotechnology park withAnalytical Quality Control & Molecular BiologyLaboratories, Aqueous & Solvent Extraction Units,Biofertilizer & Tissue Culture units andBioinformatics Facilities. The tissue culture facilityat the park was recognized under the NationalCertification of Tissue Culture Raised Plants(NCSTCP). Production of banana and bambooplants were in full swing in the tissue culture unitbeing run in participation with M/s Sheel BiotechLtd. During the year, 6.75 lakhs tissue culturedplants were supplied. The biofertilizer unit in thepark produced 240 tonnes / annum of Azotobact,and 500 tonnes / annum Trichoderma apart from148Annual Report 2009-2010

Biotechnology Parks, Incubators and Industry Platformsphosphate solublizing bacteria (PSB). The produceis being utilized in military farms.M/S IQRA Biotech services that has set up the firstHuman DNA Bank in the area will be developingan umbilical cord blood bank that will be a resourcefor stem cells. The park signed agreements withMs/ Cognate Bioservices Inc., USA and M/s VivoBiosciences, USA who are planning to set up aGMP stem cell facility for cell based therapeutics.The park also signed an agreement with GenetechLaboratories.The Park continued to play an important rolein human resource development. It is developinga 30,000 square-ft commercial wet labbuilding to accommodate nearly 15 wet lab suites.The park also organized six training programmesfor farmers on agri-biotechnology incollaboration with the Department of RuralDevelopment, Government of U.P. andnearly 200 farmers were trained in these programmes.Biotechnology Incubation Centre at ShapoorjiPallonji Park, HyderabadThe Biotechnology Incubation Centre (BTIC) atHyderabad was set up with the support of IICT,Hyderabad, Govt. of Andhra Pradesh and theDepartment of Biotechnology. For this purpose,the DBT provided a financial assistance of Rs 1729lakhs. The project has since been completed andBTIC, Hyderabad is now in the process ofoperationalizing an agreement for outsourcing themanagement and operation of the incubation centre.Industry PlatformsR&D in biotechnology is becoming increasinglyexpensive owing to growing complexities, whichrequire integration of research efforts. Thisnecessitates joint efforts through public-privatepartnerships. Such partnerships would have a catalyticeffect on the innovative capability of the industryand the academia, especially in a multi-disciplinaryresearch setting through the establishment of aframework for national needs, infrastructuredevelopment and bringing together the industry andresearch institutions through industry platforms.These platforms are expected to promote sectoralresearch, and developmental programmes through along term vision and strategic research agenda.Two brain storming meetings were organized toassess the viability of the approach and thestakeholder’s response. On the basis of the responsefrom various stakeholders, it was agreed to launchIndustry Platforms in the area of agriculturebiotechnology, biopharmaceuticals and implants &medical devicesThe agriculture biotechnology industry platformconstituted four working groups on abiotic stress,biotic stress, infrastructure and seed production. Draftstrategic research planning papers were prepared,covering all aspects based on the following framework:(i)(ii)(iii)(iv)Existing technologies: Proven at bothnational and international levelPotential of the sector to include typeand extent of support requiredInformation exchange among themembers of the platformInfrastructure requirements includingtechnology validation mechanismAnnual Report 2009-2010 149

Biotechnology Parks, Incubators and Industry Platforms(v)Grand Mission with a view to bringlong term perspective for the sectorDBT organized a brain storming session on thestrategic research planning and development ofprospective project proposals along with experts fromthe relevant research institutions as identified by theindustry.A meeting of the Biopharmaceuticals IndustryPlatform was held on 4 th March, 2009 in NewDelhi. Six working groups were formed with thefollowing charter:• The working group on cell lines wouldwork with the objective of creating theavailability of WHO approved cell linesthrough specialized banks and also facilitatecell lines development and characterizationof master cell lines and working cell lines.• The working group on animal modelswould work towards developing theinfrastructure and for the procurementof animal model (efficacy andimmunogenicity model).• The working group on clinical trial sitedevelopment will endeavour to create anational clinical trials network.• Safety and pharmacology will be the focusarea for the working group on biologics/biogenerics• The working group on stem cells therapywould coordinate with NCBS, Bangaloreto plan how stem cells therapy can betaken forward in the country.• The working group new drug discoveryand development will address thedeliberated and related issues concerningNDDS based pharmaceuticals/biotherapeutics.A meeting of the industry platform on medicaldevices and implants was held on 16 th September,2009 in New Delhi. Working groups wereconstituted in the area of infrastructure, R&D andsupport services.150Annual Report 2009-2010

CHAPTER 9INTERNATIONAL COLLABORATIONThe DBT has endeavored to open up new avenuesin international collaboration for taking research leadsforward and developing products and processes. Thedepartment has established successful collaborationswith strategic partners like Australia, Canada,Denmark, Finland, Norway, Sweden, Switzerland,UK, EU and USA. It is also working very closelywith non-governmental funding organizations suchas Welcome Trust, UK. Bilateral collaborations werepursued and the achievements of some of theprogrammes are given below:Indo-CanadaDBT launched a call for proposals in the area ofConvergent Medical Technologies: Application toClean Technologies and Bio-Pharma and Health withInternational Science and Technology PartnershipsCanada Inc. (ISTP Canada).Four research projects were approved.(i)(ii)(iii)PET-MRI automatic tumor detection andrecognition, at Soft Team Solutions Pvt. Ltd.,Chennai; CMC, Vellore; Kamakshi MemorialHospital, Chennai and A.U.G Signals Ltd,Ontario, Canada.Translating head and neck cancer markers intodiagnostic arrays, at AIIMS, New Delhi;IMGENEX INDIA Pvt. Ltd., Orissa and YorkUniversity, Ontario, Canada.Cost effective multiplex assay for predictionand tracking of sexually transmitted infections,(iv)at St. Johns Research Institute, Karnataka;Kentropy Technologies Pvt., Ltd., Karnatakaand Centre for Global Health Research,Ontario, Canada.International Malaria Research Consortium forthe development of novel antimalarials, atICGEB, New Delhi; BITS, Pilani, Rajasthan,Life Care Innovations, Gurgaon and UniversityHealth Network, Toronto.As a follow up of the MoU signed between DBTand National Research Council, Canada a letter ofagreement was signed between the two parties toinvest up to 10 million Canadian dollars for fiveyears (50:50 basis) for collaborative research in plantbio-technology.Three projects were approved by both the fundingagencies:I. Identification and manipulation of geneticfactors involved in seed yield and oil contentin rapeseed and mustard, at UDSC, New Delhiand NRC-PBI, Saskatoon.II.III.Indo-Canadian Pulse Genomics Initiative(InCan-PGI), at NIPGR, New Delhi;CSKHPAU, Palampur; IARI, New Delhijointly with NRC-PBI, Saskatoon.Genetic improvement of flax for oil andagronomic traits, at NCL, Pune; IRSHA, Pune;College of Agriculture, Nagpur with NRC-PBI, SaskatoonAnnual Report 2009-2010 151

International CollaborationUnder the DBT – National Research Council (NRC)–Plant Biotechnology Institute (PBI), Saskatoonprogramme, efforts were initiated to locate majorloci and identify candidate genes responsible for theoil content and seed size in Brassica juncea at UDSC,New Delhi. Twenty lines from double haploid (DH)mapping population for the two traits were plantedto isolate RNA for microarray analysis at PBI,Saskatoon. Work was taken up on development ofnew genomic tools in chickpea and lentil, jointly atNIPGR, New Delhi and NRC-PBI, Saskatoon. Inanother joint project at NCL, Pune and NRC-PBI,Saskatoon, about 500 EST-SSR primers were designedin flax. Out of these, 100 primers were screened forpolymorphism using seven flax varieties and 29primers were found to be polymorphic.Indo-DenmarkThe MoU between DBT and the Danish Agency forScience, Technology and Innovation was extendedfor a period of five years. DBT and Denmark hada joint call for proposals in food, feed and bioenergywith a commitment of 1 million euros from boththe sides. One project was approved for funding.Altogether there were 7 ongoing projects resulting in8 publications and transfer of one technology.Significant results of ongoing projects are:In a project at Delhi University, an efficient enzymaticprocedure for the discrimination of locked nucleicacids (LNAs) has been developed. LNAs are knownto possess favorable features towards development ofantisense and / or antigene drug candidates. Efficientbiocatalytic methodology for the separation ofmixture of five- and six-membered nucleosides wasdeveloped. This highly selective and efficientbiocatalytic procedure has been patented.At the IIT, Kharagpur and Technical University ofDenmark, the feasibility of a two-stage process ofbiohydrogen production followed by biomethaneproduction was tested. The first stage hydrogenproduction was done using E. cloacae IIT-BT 08 ina dark fermentation process and the spent media ofthe first stage was used for biomethane productionusing aceticlastic methanogens enriched from cowdung samples. The study holds promise in themaximization of gaseous energy recovery from organicwastes. Studies on protein aggregation were carriedout at MKU, Madurai in collaboration with AalborgUniversity, Denmark. The aggregation of humanalpha synuclein, a major component of intracellularinclusions in several neurodegenerative diseases, wascharacterized.Indo-FinlandA successful trilateral call for proposal involving DBT,Academy of Finland (AoF) and TEKES in the areaof medical diagnostics was completed. An MoU withTEKES was signed for a one to one bilateralcooperation. Altogether there were 12 ongoingprojects and 16 publications. Achievements underthe ongoing projects are given below:Medical BiotechnologyThe goal of a project being carried out at ICGEB,New Delhi and Univ. of Turku, Finland was tounderstand the molecular basis of interaction betweenthe B cell antigen receptor (BCR) and IL-4 receptorin B cells, and also to resolve the mechanism of Th2differentiation. With an immature B cell line, it waspossible to successfully delineate the interactionsbetween BCR- and IL-4 receptor signaling. Thesestudies revealed new insights into the receptor-152Annual Report 2009-2010

International Collaborationdependent control of cellular responses. It has alsobeen seen that the canonical MAP kinase pathwayserved as a novel regulator of human Th2 celldifferentiation. The terminal MAP kinase, ERK,directly regulated transcription of the IL4 gene,thereby controlling Th2 differentiation.The ongoing project at NIPER, Punjab and AboAkademi University, Abo/Turku, Finland aims tointegrate in vitro and in silico analysis for designingdual binding site Acetylcholinesterase (AChE)inhibitors for the treatment of Alzheimer’s disease.High-throughput in vitro screening was performedon a structurally diverse set of compounds towardsAChE inhibition at Åbo Akademi University, Finland.Two highly selective AChE inhibitors were obtained.Pharmacophore models developed from theseinhibitors followed by sequential virtual screening ofdifferent small molecule databases provided potentialleads.Through a project at ICGEB, New Delhi andUniversity of Turku, Finland, a novel dual labeltime resolved fluoroimmunoassay was developedfor the simultaneous detection of HBsAg and anti-HIV-1 antibodies in human sera from the samewell. For the detection of HBsAg, two mousemonoclonal antibodies were used in a sandwichformat utilizing intrinsically fluorescent europium(III) nanoparticles as label. Intrinsically fluorescentterbium(III) chelates were used as label for thedetection of antibodies to HIV-1 in a doubleantigen sandwich format. The unique emissionwavelength of Eu(III) and Tb(III) chelates enabledthe time resolved measurement of fluorescencefrom the same well (Fig).(A) Assay format for HIV-HBV dual assay. (B) Histogram showing the Tb signals obtained from HIV negative and positive human sera. (C) Histogramshowing the Eu signals obtained from HBV negative and positive human sera.Annual Report 2009-2010 153

International CollaborationAgriculture BiotechnologyWork done in the ongoing project at UDSC, NewDelhi and University of Helsinki, Finland on wholegenome expression profiling showed that various heatshock transcription factor (HSF) genes were affecteddifferently in rice against varied abiotic stresses. In anongoing research project at G.B.Pant Univ, Pantnagarand MTT Agrifood Research, Finland, Jatrophacompost were screened against the Fusarium oxysporumf. sp. lycopersici for their disease suppressive nature underglass house conditions. Results clearly showed theimportance of rhizocompost in reducing abiotic andbiotic stresses.At ICGEB, New Delhi and University of Helsinki,Finland, RCD1 and RCD2 (Radical Induced CellDeath) full length cDNA were cloned from rice andexpressed using pET28ain E.coli to obtainrecombinant proteins. Using yeast two hybridanalysis and transactivation monohybrid assay, severalinteracting partners of RCD1 were identified and theRNA binding domain containing (RBD) protein wascloned and its interaction confirmed. RCD1 wasfound to be over expressed.Indo-GermanyEight new proposals were approved for funding.Altogether there were 17 ongoing projects. Therewere a total of 22 publications and two technologieswere transferred. Some highlights of the projects are:In an ongoing project, researchers from IIT,Kharagpur availed the automated high throughputcrystallization facility at EMBL Hamburg forcrystallizing the proteins: Rv0040c, Rv0398c andRv1284. Diffraction data of native and derivatizedRv0040c, Rv0398c, Xe and inhibitor soaked Rv1284were collected using the Synchrotron facility at EMBLHamburg (Fig).Crystals of (a) Rv0398c, (b) Rv1284, (c) Rv0040cAt NEERI, Nagpur and University of Leipzig,Germany four marker proteins involved in thesenescence mechanism of Her-2 induced MCF-7 cellswere identified using the proteomics approach. Keyregulators of major cellular processes were observedto be up regulated. HSP27 a major cellular chaperonewas found to be down regulated and its downregulation was reported to induce apoptosis orsencence by p53/P21 pathway activationWork done in an ongoing project at the NationalInstitute of Interdisciplinary Science and Technology,Kerala, and Westfalian Wilhelms-UniversityMuenster, Germany, showed the feasibility of biomassprocessing. Attempts were made to incorporate thepentose (xylose and arabinose) assimilating genes ofE. coli MG 1655 into C. glutamicum wild type ATCC13032 as well as into the lysine producing strainDM1729. The recombinants were characterized /analyzed in terms of growth and amino acidproduction from pure pentose sugars and in varioushemicellulosic hydrolysates. The lab demonstrated theproof-of- principle of both growth and amino acidproduction with pentose/xylose/arabinose as the solecarbon and energy source.At the ICAR Research Complex, Goa and Justus-Liebig University Frankfurter, more than 250 strainsof Listeria were serotyped and analyzed using154Annual Report 2009-2010

International Collaborationmultiplex PCR and pulsed field gel electrophoresis(PFGE). A PCR based diagnosis of listeriosis fromclinical samples was ready for commercialization. Anelectronic database for the characterized strains wascreated, indicating geographical distribution,phenotypic and genotypic characteristics, and DNAfingerprint. As part of a collaborative project at BharatBiotech Foundation, Hyderabad and University ofWurzburg, Germany, pre-clinical development ofrecombinant lysostaphin for the eradication ofsystemic S.aureus infection was completed. ThecGMP product was tested in vivo for the efficacy oflysostaphin in eradicating catheter infections mediatedby MRSA (Methicillin resistant S.aureus). New leadswere identified for developing a novel multi-epitopevaccine by improvising the existing S.aureus vaccinecandidate.Indo-SwedenThe DBT and Swedish Governmental Agency forInnovation Systems, Sweden agreed to supportresearch co-operation between Indian and Swedishscientists in the fields of biology, diagnosis andtreatment of tuberculosis. Four proposals wereselected for funding. These were: (i) Doctors officediagnostic instrument for detection of M. tuberculosisunder field conditions, for use by unskilled personnel,at University of Delhi, Delhi and Imego AB.(ii) Mechanisms of protein synthesis and ribosometargeting antibiotic drugs in Mycobacteria at IISc,Bangalore and Uppsala University. (iii) Biology ofgene-deleted M. tuberculosis strains - immunologicalmarker profiling at IISc, Bangalore and KarolinskaInstitute. (iv) Structure-guided design of newantibacterial agents against dormant Mycobacteriumtuberculosis at BITS, Hyderabad and KarolinskaInstitute.Indo-USUnder the Indo-US Contraceptive and ReproductiveHealth Research (CRHR) programme, threenew projects were approved. These were:(i) Randomized controlled trial to enhance dualprotection among people living with HIV (PLHIV)in India, at Indian Network for People Living withHIV/AIDS, Chennai and Yale University, USA.(ii) Gender, equity-focused, male-centered familyplanning for rural India, at NIRRH, Mumbai andBoston University, USA. (iii) Evaluation of aprogesterone vaginal ring as a new contraceptiveoption in India, at ICMR and R&D PopulationCouncil, USA. Altogether, there were 13 ongoingprojects. A total of 48 publications were made andtwo technologies were transferred. Some highlightsof the ongoing CRHR projects were:- Sperm-Associated Antigen 9 (SPAG9), a novelcancer testis antigen, was found to be a potentialtarget for immunotherapy in epithelial ovariancancer and the humoral immune responseagainst SPAG9 in early stages suggested a rolein early diagnosis. (NII, New Delhi)- A recombinant vaccine was developed basedon heat labile enterotoxin chain B of E.colifused to α-hCG at the C-terminal end. It wasexpressed in Pichia pastoris to obtain theproduct with glycosylation of the typeacceptable to Regulatory Authorities for humanuse. The work was carried out at TRF, NewDelhi & National Cancer Institute, USA.- In a PGF 2α-induced luteolysis model,microarray analysis was done to compare thePGF 2αresponsive transcriptome to that of LHAnnual Report 2009-2010 155

International Collaborationresponsive transcriptome and to examine theexpression of select genes as common targetsof LH and PGF 2αactions in CL duringdifferent stages. (IISc., Bangalore & MorehouseSchool of Medicine, USA)- In a study conducted during treatment forChlamydia trachomatis infection, the numbersof both myeloid DCs (mDCs) andplasmacytoid DCs (pDCs) were found to behigher in peripheral blood confirming theirrecruitment from the blood. In women withchlamydial MPC significantly lower numbersof pDCs were present after therapy suggestinga possible role of this subset in the pathogenesisof chlamydial infection (ICMR, New Delhi& Dartmouth Medical School, USA)Khorana ProgrammeDepartment of Biotechnology, Govt. of India andthe Indo-US Science and Technology Forum(IUSSTF) have partnered to support theprestigious Khorana Program for Scholars namedin honor of Dr. Har Gobind Khorana, who wonthe Nobel Prize in 1968 for his work at theinterface of chemistry and biology. The KhoranaProgram provides an opportunity to Indianstudents (enrolled in B.Tech. M.Tech. and M.Sc.programs in Biotechnology and allied areas) toundertake research internships at UW. A total of424 applications were received, and 30 wereshortlisted. Fifteen students were selected and 12availed the internship in 2009 in the areas ofgenetic recombination; biochemistry; bacteriology;cellular and molecular biology; geosciences;physiology, and chemical & biochemical engineering(Fig).The University of Wisconsin-Madison (UW), theThe selected Khorana Programme Scholars for the year 2009 alongwith representatives of DBT, Indo-US S&T Forum and WisconsinUniversity at a function held in New Delhi on 9 th January, 2010.156Annual Report 2009-2010

International CollaborationIndo-UKThe research collaboration between NII, New Delhiand Queens University, Belfast resulted in thefollowing significant findings:The overall approval was to advance cancer researchand improve patient outcomes by discoveringbiomarkers for multiple types of cancer. Cancer testis(CT) antigens represent a unique class of tumorantigens, which are potential targets for diagnosticbiomarkers, antigen-based vaccination and antigendirectedimmunotherapy. Studies showed that amajority of histotypes of breast cancer, cervical cancerand thyroid carcinoma tissues expressed CT antigenSPAG9 at both mRNA and protein levels. It wasalso seen that in early cancer, a substantial number ofpatients exhibited SPAG9 expression and generatedSPAG9 antibodies, suggesting its potential role inearly detection and cancer management. The findingsalso support a potential role for SPAG9 as a diagnosticbiomarker and a therapeutic target in cancertreatment.Wellcome Trust-DBT India AllianceThe Wellcome Trust/DBT India Alliance wasestablished to nurture talent of the highestinternational standards, in cutting-edge biomedicalresearch in India, leading to important scientificbreakthroughs and applications to society. TheWellcome Trust – DBT India Alliance has been setup in Hyderabad and the main function would be toaward fellowships to scientists. Early career fellowshipsare targeted at young scientists who have justcompleted their PhDs. They will also be funded tovisit any international lab for a short duration tofoster collaboration with leaders in the field. Theintermediate fellowships would be awarded toscientists with a post-doctoral experience of 3-6 yearswho wish to set up labs in India. The seniorfellowships are meant for scientists of proven trackrecord, who have 5-10 years of research after theirPhD. Three senior fellows, six intermediate fellowsand five early career fellows were selected.Indo-EUA joint call was made in the area of developmentof functional foods and ingredients and valorisationof by-products in food processing. The EC hascommitted Euros 1.5 million towards the call. Thetwo projects approved were: (i) Impact of agentswith potential use in functional foods on biomarkersfor induction of age related diseases, at AnnamalaiUniversity, Tamil Nadu and Stockholm University,Sweden. (ii) New advances in the integratedmanagement of food waste in India and Europe:Use of sustainable technologies for the exploitationof new foods and feed, at NEIST, Assam andBologna University, Italy.ERA-NETThe DBT collaborated with the first Era-netproject (European Research Area) named NEWINDIGO, aimed at fostering and coordinatingscientific cooperation between European ResearchArea and India. The project was launched withthe announcement of a joint call for proposals inthe area of biomarkers and diagnostics;bioinformatics for health; and structural biologyfor health. The other partners are FederalMinistry of Science and Research (BMWF) fromAustria; National Centre for Scientific ResearchAnnual Report 2009-2010 157

International Collaboration(CNRS) Ministry of Foreign and European Affairs(MAEE) and the National Institute for Healthand Medical Research (Inserm) from France;Federal Ministry of Education and Research(BMBF) from Germany; Netherlands Organisationfor Scientific Research (NWO) from TheNetherlands; Foundation for Science andTechnology (FCT) from Portugal and Scientific andTechnological Research Council of Turkey(TUBITAK) from Turkey.Indo-AustraliaAn MOU was signed between the DBT and theDepartment of Innovation, Industry, Science andResearch, Government of Australia at New Delhi on6 March 2006 for co-operation between India andAustralia in the field of biotechnology. The overallobjective of the Indo-Australian Biotechnology Fundis to develop and support collaborative researchactivities which draw upon strengths in India andAustralia. The fund supports Indian and Australianscientists, from both the public and private sectors,to collaborate in cutting edge areas of science andtechnology.Indo-NorwayA multicentric collaborative programme on aquahealth has been supported with the emphasis to studythe basic mechanisms of the pathogenesis of variousinfectious diseases in aquatic animals, with theintention of developing improved and new vaccinesand vaccination concepts. Besides this, advancedmolecular tools will be used to improve diseaseresistance in rohu carp and tiger shrimp farmed inIndia.Under the human health collaborative programme,projects on vaccine formulations, novel deliverysystems, immunomodulators & adjuvants are in theprocess of funding. Two collaborative proposals underreview are: (i) Development of biodegradablenanobeads as vaccines against tuberculosis. (ii) Effectof zinc and vitamin D supplementation on immuneresponses to vaccines in Indian infants. The 4thConference on Global Health and VaccinationResearch was held in Oslo, Norway in December,2009 involving the partners and stakeholders.Indo- JapanDBT and AIST, Japan have entered into an MOUto work in life sciences and biotechnology. Threebilateral meeting cum workshops were held so far.The first one was organized in 2008 at AIST, Tsukuba,Japan to discuss the schemes for Cooperative ResearchProjects (CRP) and fellowship for researchers in theidentified areas of bioinformatics, cell engineering andglycoscience. It was subsequently agreed to includethe study of biomarkers. The third bilateral meetingwas held in Tsukuba, Japan on 28th October, 2009.Four projects, in the areas of glycoscience and cellengineering were agreed to for collaboration betweenAIST, Japan and DBT institutions namely NII,NCCS and CDFD.India-CubaA delegation from DBT visited Havana, Cuba in2009, to explore the possibility of bilateralcooperation between the two countries under theIndo–Cuban Joint Commission. The delegation alsovisited some of the centers and institutes engaged inresearch and technology development. The delegation158Annual Report 2009-2010

International Collaborationrecommended the following lines of co-operationwith Cuba: (i) to invite a delegation from Cuba tovisit India and explore areas of collaboration, (ii) toset up a mechanism, with co funding by bothcountries, for joint R&D projects and (iii) to facilitatebiotech technology transfer from Cuba to Indianprivate companies by providing, incentives, andfunding under BIPP or SBIRI.DBT- IAVI CollaborationThe DBT and International AIDS Vaccine Initiative(IAVI) have forged a partnership to develop “NextGeneration” vaccine candidates. Indian MedicinalChemistry Programme towards advancement of novelneutralizing antibody approach has been operationalsince 2007. The major aim of the programme istowards the design of candidate vaccines to elicitneutralizing antibodies against HIV. A new IndianMedicinal Chemistry Programme (IMCP), cosponsoredand co-funded by IAVI and the Departmentof Biotechnology, comprising Indian and U.S.scientists was tasked with accelerating the pace ofAIDS vaccine discovery.The Indian Institute of Science component is focusedprimarily on polypeptide mimic immunogens ofthe b12 antibody epitope in gp120. They madesignificant progress towards generation and panningof phage libraries displaying derivatives of b121aand b122a, design of additional b12 binding siteconstructs and design of constructs to study thegp120:gp41 interface. The ICGEB, New Delhicomponent is focused on development ofconformationally preferred peptide immunogens asvaccine candidates against HIV infection. Six newpeptides were designed, synthesized, purified andcharacterized. Immunogenicity & conformationalstudies were done on 3 of the peptides. TranslationHealth Science & Technology Institute (THSTI) anautonomous institution of DBT has recently signedan agreement with IAVI for an Applied Science HIVdiscovery program. It will work towards discoveryand fill gaps to identify immunogens that elicitbNabs.Indo-SwissThe Indo-Swiss programme in Biotechnology (ISCB)jointly funded by Department of Biotechnology(DBT) and Swiss Agency for Development andCooperation (SDC) was launched in 1998. So far,the programme has completed two phases. The phaseIII programme (2007-2011) is basically aimed attechnology advancement and product development.The achievements under the phase-II of theprogramme in terms of prototype products are beingvalidated for technology transfer and co-developmentwith industrial partners. The phase III has 120researchers including the private sector. Somehighlights are:Technology transfer of transgenic chickpea with insectresistance was taken up. Regeneration andtransformation protocols of chickpea were established.Transgenic lines with cry genes and garlic lectin geneswere assessed for their potential commercial value. Atechnology show-casing workshop for private sectorseed companies was conducted. Technologieswere transferred on a non-exclusive basis toM/s Maharashtra Hybrid Company Ltd., Mumbai.A technology was transferred to UAS, Dharwad fordeveloping insect resistant varieties of chickpea. Theevents selected for technology licensing to privateseed companies and state agricultural universitiesinclude (i) insect pest resistance through the use ofAnnual Report 2009-2010 159

International CollaborationBt (cry1Ac and cry2Aa) technology for podborer (ii)insect pest resistance through the use of Lectin (ASAL– Garlic lectin) technology for sucking pests.Work was taken up for development of prototypebiosensor for detection of pesticides of barathion,attrazine, and 2,4-D. The specific antibodies detectingtrace levels were produced in combination with achemiluminescence-based detection system forquantitative analysis. An SBIRI scheme was recentlyapproved for development of commercial kits by aBangalore based company.A fungal biopesticide was developed against podborer,a major insect pest on pulses and other crops inIndia. Studies for large-scale production includingnegotiations for the transfer of the technology toindustry are underway. Trials to test the efficacy ofthe prototype mycoinsecticide against podborer inchickpea and pigeonpea were carried out. Field trialsshowed that the efficacy of the new biopesticide wascomparable to chemical control measures.Significant leads were obtained on the function ofmycorrhizal fungi, free-living and plant-associatedmicro-organisms, with regard to soil quality andproductivity. A large number of fungal and bacterialstrains were screened. Large scale field trials in variousagro-ecological regions with varying soils, wheatvarieties was carried out at five locations incollaboration with Agricultural Universities. Initialresults indicate a 30-40% yield increase in low fertilitysoils alongwith increased level of zinc in grains.160Annual Report 2009-2010

CHAPTER 10AUTONOMOUS INSTITUTIONSThe existing autonomous institutions and newgeneration institutions being established clearlydemonstrate the department’s increasing emphasison translational research and process-productdevelopment. These institutions are generating anew synergy for capturing research leads andconverting them into products. It is a boldexperiment that would be redefining the contoursof biotechnology and identifying new mechanismsthat will hasten product development.National Institute of Immunology (NII),New DelhiThe main thrust of the institute is to undertakeresearch in basic and applied immunology; carryout research for development of new vaccines,immunodiagnostic kits, immunological reagents;interact with industry for development of productsfrom research leads; organize post-graduate coursesleading to Ph.D. degree; organize scientificinteractions; serve as a national reference centre forimmunology and also provide consultancy services.It also endeavors to promote linkages amongresearch agencies in immunology, vaccinedevelopment and also collaborate with internationalagencies.HRD ProgrammesSome of the salient achievements of the HRDprogramme were:Around 180 students from all over the country andother south Asian countries received their PhDdegrees through the Jawaharlal Nehru University,New Delhi. The Ph.D. program was launched in1986-87. Admissions tests are conducted regularlyat different places in India.R&D ActivitiesThe possibility of apoptosis-based therapies forvarious tumours was investigated. Studies onestrogen-induced modulation of death pathways inhuman macrophages revealed the importance ofintegrative signaling from multiple estrogen receptorpools in modulating estrogen effects and openingnew approaches to explore the use of estrogenreceptor modulators in apoptosis-based therapies.Modulation of acetyltransferase activity in M.tuberculosis was attempted. Serine/threonine kinasePknB mediated phosphorylation of a novel substrate,N-acetylglucosamine-1-phosphate uridyltransferasewas found to modulate acetyltransferase activity.Protein kinase K was seen to regulate the activationof transcription using promoter of mycobacterialmonooxygenase operon through phosphorylation oftranscriptional regulator VirS.Presence of novel HIV-1 LTR and VPr sequencesin North Indian populations was investigated. Severalbiologically active catalytic RNAs (Ribozymes) andDNAs (DNA-enzymes) and siRNAs were designedagainst the X gene of Hepatitis B, the InternalAnnual Report 2009-2010 161

Autonomous InstitutionsRibosome Entry Site (IRES) of Hepatitis C, andagainst HIV-1 genes. Molecular determinants forHIV-1 gene (Tat, Vif) mediated pathogenesissuggested the presence of novel HIV-1 LTR andVPr sequences suggested co-circulation of HIV-1subtypes B and C in North India.Studies demonstrated that SiRNA-mediatedknockdown of SPAG9 expression significantlyreduced cancer cell growth. The expression ofSPAG9 was shown to be associated with the knownrisk of breast cancer recurrence over 10 years. Theinvolvement of SPAG9 in renal cell carcinoma,which accounts for 2 to 3% of all malignancies,was further investigated. It was seen that siRNAmediatedknockdown of SPAG9 expressionsignificantly reduced cancer cell growth, colonyformation, promoted wound healing and reducedtumour growth in vivo in nude mice.Plasticity of adult and fetal haematopoietic cellswas pursued as a line of investigation. Both adultand fetal haematopoietic cells were found topossess inherent plasticity, as they were observedto directly differentiate into haepatocytes in thepresence of suitable micro-environmental cues.Preliminary results showed that a purifiedpopulation of bone marrow cells can correct thebleeding phenotype in haemophilic mice,suggesting a potential therapy for haemophiliaand related diseases.Sustained insulin release formulation was founduseful as a therapeutic for metabolic disordersparticularly diabetes. This has been demonstratedby studies in diabetic models of mice, rats andrabbit. The formulation shows potential for a longlasting treatment of diabetes and halting the advanceof secondary diabetic complications.National Centre for Cell Sciences(NCCS), PuneNCCS is the only National Cell Bank for animalcell lines in India catering to the needs of academiaand industry. NCCS provided 1980 cell lines ofdifferent types to scientific institutions in thecountry. In addition to serving as the National CellRepository, NCCS focuses on HRD and R & D inthe areas of cell biology, signal transduction, cancerbiology, diabetes, infection & immunity, generegulation, stem cells and regenerative biology.NCCS conducted training programmes on basictechniques in modern biology for researchers fromvarious institutes in the country and also participatedin the post graduate teaching and trainingprogrammes at Pune University, Devi AhilyaUniversity (Indore), Nagpur University and GoaUniversity. NCCS is currently training 89 Ph.D.students.NCCS developed novel methods to preserve andmaintain viable stem cells and has succeeded indifferentiating these into specific lineages, forpotential applications in cell/tissue replacementtherapy. Cell-cell signalling is fundamental to thegrowth and development of multi-cellular organisms,mis-regulation of which causes various diseasesincluding cancers. The centre is studying theregulation of critical players, such as p53, eNOS,Osteopontin, TNF-alpha, involved in these cell-cellcommunication pathways that go awry in cancers.The centre has demonstrated that deficiency ofosteopontin effectively curbs melanoma metastasisand angiogenesis, thus providing a new target forfuture cancer therapies. The team at NCCS identifieda specific non-coding RNA that is important for162Annual Report 2009-2010

Autonomous Institutionstumorigenesis. In recent years, cancer has beenillustrated as a stem cell disease. Studies have defineda subset of cancer stem cells that contribute totumour dormancy and resistance to chemotherapy.Understanding the molecular intricacy of infectionand immunity is of great importance, particularlydue to the emergence of new infectious agents.Studies carried out at NCCS suggest that successfulinfection by at least a group of bacteria involvesalteration of the host cell membrane and caveolaedynamics by the toxins they produce. Establishmentof an infection depends on how effectively thepathogens can counteract the adaptive immuneresponses of the host. Studies were done on howcertain viruses target the host complement systemin order to overcome the innate immune response.Furthermore, a role for CD40-CD40L interactionin regulating the activation of T cells in response toLeishmania and HIV infections was underinvestigation.The culture collection at NCCS greatly aids theconservation and exploitation of biodiversity ofIndia. Furthermore, the microbial distribution invarious habitats was studied using the metagenomicapproach. NCCS published 60 papers in peerreviewed national and international journals. Inaddition, NCCS has filed one Indian and oneinternational patent.National Brain Researesearch Centre(NBRC), GurgaonThe NBRC was established as a “Centre ofExcellence in Brain Research”. The major areasidentified for research include computationalneuroscience, system and cognitive neuroscience, stemcell research, developmental neurobiology and basicresearch aimed towards the understanding of thecomplexities of neurological and psychiatricdisorders. Translational and clinical neurosciences arealso research priorities. The salient achievements are:Human Resource DevelopmentNBRC is an unique institution in the country thatpursues an integrated approach to educate researchscholars from various backgrounds such as science,engineering and medicine, by pursuing a one-yeartransdisciplinary curriculum that encompasses subjectsfrom the various fields from neuroanatomy,neurochemistry, embryology, to cognitive studies,image processing and computer science. Thereafter,the scholars pursue their research topics. Anincreasing number of students with a backgroundin biological sciences are pursuing research work incomputational neuroscience areas, while a goodnumber of them coming from the physical orengineering sciences are working on problems inneurobiology.Networking ActivitiesNBRC and Salk Institute, San Diego organised anInternational Workshop on Cognitive Neuroscienceat NBRC with the sponsorship of InternationalBrain Research Organization (IBRO). TheDELCON centre at NBRC functioned as the digitaldocumentation resource centre for all the researchinstitutions of the DBT. Virtually, most of theinformation on biological sciences was made availablethrough over a thousand journals. NBRC has alsoactively collaborated with neuroscientists throughits extramural network centers. A collaborative effortwas explored by NBRC to track the ageing humanAnnual Report 2009-2010 163

Autonomous Institutionsbrain from 50 years of age onwards. This endeavourwould involve several centers across the India, withthe objectives of identifying biomarkers and imagingmarkers of Alzheimer’s dementia and vasculardementia.Molecular & Cellular NeuroscienceIt has been found that the expression of mutantHuntington’s disease results in massive transcriptionalinduction of several chemokines as MCP-1 andKC. In another study the methodology andmechanism of restoration of blood-brain barrierintegrity by minocycline-based treatment of Japaneseencephalitis was elucidated. In the Neuro-AIDSresearch it was observed that the HIV transactivatingprotein causes a marked deficit in the proliferationand differentiation of human neural progenitor cellor neural stem cells. This has implications fortherapy. Investigations showed that plant derivativesas curcumin metabolite have neuroprotective effectson Alzheimer’s disease, by reducing amyloidbetaoligomer-induced toxicity in hippocampalneurons.Systems & Cognitive NeuroscienceMarked reorganization of higher somatosensory areasafter spinal cord injuries were accurately mappedusing cytological, electrophysiological and behaviouralmethods. Investigations carried out on retinalprosthesis showed that photoreceptor degenerationleads to rise in synaptic proteins in amacrine cells,suggesting an enhanced inhibitory neurotransmission.Harnessing of adult neural stem cell/progenitor cellby modulating the opioid system by naloxone wasdemonstrated through studies in animal systemsand zebra finches.Computational Neuroscience & NeuroimagingNeurospectroscopic investigation and mapping ofthe biochemical dynamics of neurodegenerativediseases were undertaken, using proton andphosphorous 31 signals. Experiments were conductedfor locating the different cortical pathways inbilinguals i.e. English vis-à-vis Hindi. Importantinsights, with clinical applicability, were obtainedon the two paths which differ depending onwhether the languages were learnt simultaneously orsequentially. MRI-based structural and tensor imagingof normal brain ageing across the life span (from20-95 years age) was done, and the elastic phasemap of the brain developed, from which theprediction and risk of dementia was elucidated.Translational & Clinical NeuroscienceA unit has been set up in the Gurgaon GovernmentHospital that provides neurology, psychiatry andclinical psychology services to patients. Data fromsome of these patients are studied by NBRC.About 65% of the cases suffered from seizuredisorder or migraine and 5% had neuroses/psychoses.The remaining belonged to a miscellaneous groupcomprising cerebrovascular disease, neuropathies,palsy, trigeminal neuralgia, and brain tumours.Centre for DNA Fingerprinting andDiagnostics (CDFD), HyderabadThis centre combines two kinds of activities, i.e.basic sciences and R&D. DNA studies areundertaken for law-enforcement agencies.Diagnostic tests are being developed for geneticdisorders. There is also a good portfolio of researchin molecular biology.164Annual Report 2009-2010

Autonomous Institutions(A) ServicesDNA FingerprintingA total of 91 cases were received for DNAfingerprinting examination. Out of these, 49 caseswere related to paternity/maternity disputes, andthe others were mainly related to identificationof deceased. Eighteen states of India availedDNA fingerprinting services of CDFD. Rajasthanforwarded 27 cases, followed by Maharashtra andKarnataka (8 each).Agricultural and Processed Food Product ExportDevelopment Authority (APEDA) – CDFDCentre for Basmati DNA AnalysisAt the instance of APEDA, the APEDA-CDFDcentre was established for quantification ofadulteration of Basmati rice. Based on the noveltechnology developed by CDFD, DNA basedmethods were developed for the quantification. Atotal of 146 samples were analyzed.During the year, DNA experts defended theirreports in about a dozen cases in various Hon’bleCourts throughout the country. Training on DNAfingerprinting was provided to the scientists fromthe State Forensic Science Laboratory, Rajasthanand Jaipur. Lectures were delivered for thebenefit of foreign delegates from 22 countries,investigating officers from Central DetectiveTraining School, Hyderabad, IPS officers fromSardar Vallabhabhai Patel National PoliceAcademy, Hyderabad and Air Force Traineeofficers form Pune.DiagnosticsA total of 1664 patients attended the clinic forgenetic evaluation and counselling. These consistedof patients with chromosomal disorders, monogenicdisorders, mental retardation, congenitalmalformations, multiple malformation syndromes,haemolytic anaemia, short stature, skeletaldysplasias, myopathies, neurodegenerative disorders,ataxia, hypogonadism (male/female), ambiguousgenitalia, inborn errors of metabolism, and familialdisorders.Distribution of QTLs for Basmati grain quality traits in the molecularlinkage map of chromosome 5. The QTLs are indicated on the rightside of the linkage group. Numbers in parenthesis are genetic distanceof markers in centimorgans (cM).Annual Report 2009-2010 165

Autonomous Institutions(A) Research:Cell growth, cancer and inflammation were studied.GATA6 was identified as a candidate oncogene inpancreatobiliary carcinoma; an intronic DNAsequence within the neuronatin gene wascharacterized; and pharmacological modulation ofNF-kappaB as a means of controlling inflammationand proliferation investigated. The mechanism ofaction of the bacterial transcription terminationproteins Rho and NusG was studied. The Z sexchromosome of the silkmoth Bombyx mori wasshown to harbour a disproportionately large numberof testis-specific genes, which is likely to haveevolutionary significance. Development of improvedsilkworm strains and characterization of transgenicstrains rendered resistant by RNAi strategy tobaculovirus infection was undertaken.National Institute of Plant GenomeResearch (NIPGR), New DelhiNIPGR has made considerable progress in nurturingplant genomics and various aspects of plant molecularbiology. Genomics research is providing the bridgebetween different branches of natural sciences.Highlights of some of its scientific achievementsand new initiatives are:Computational BiologyComputational analyses were carried out to identifyprospective candidates as inhibitors to selectedenzymes and to decipher the molecular mechanismsby which they function. This has led to the discoveryof a novel mechanism of clustered gene biosynthesis,specifically for polyketide synthases from microbesand chalcone synthases in plants. Work on bacterialhyaluronidases led to the elucidation of a novelmechanism of anti-bacterial defence that may occurin all mammals.White arrow: Non-Transgenic eyesMFF 154C RedEyesThe red fluorescent eyes are seen in the transgenic silkmoths that areabsent in non-transgenic moths.Genome Analysis and Molecular MappingIsolation of genomic SSRs led to the developmentof about 300 novel STMS markers in chickpea.cDNA libraries were constructed from differentstages of chickpea seeds that yielded more than1500 unigenes. Two full-length cDNA libraries ofchickpea were generated. A total of 14,250 partialsubtracted cDNA clones were generated and aboutten thousand chickpea ESTs sequenced. In order toconstruct the framework linkage map of C. roseus,more than 400 markers targeting genes of TIApathway were developed. A F2 mapping populationwas generated and genotyped and a linkage mapdefining the position of 113 markers was generated.166Annual Report 2009-2010

Autonomous InstitutionsIn wheat, three phenologically plastic lines wereinter-crossed to pyramid the genes favourable fortandem cropping. The F 2generations were developedfrom these crosses for testing. A phenologically plasticline was crossed with two aplastic lines for thepurpose of mapping the loci responsible fortemperature tolerant seed formation and photoperiodinsensitive early flowering traits.Molecular Mechanisms of Abiotic StressResponseTranscriptome analysis during water-deficit stressgiven to IR64 cultivar of indica rice seedlingsunravelled more than 2500 up- and down-regulatedgenes, respectively. Co-regulation of water-deficitstress responsive genes with cold and salt stressshowed greater overlap with salt-induced changes ascompared to cold stress. The entire gene family ofmitogen activated protein kinase and MAPK kinase(MAPKK/MKK) was cloned from Oryza sativaindica cultivar group, var. Pusa Basmati-1.Transgenic rice plants over expressing constitutivelyactive forms of OsMKK6 ΕΕ showed tolerancetowards salt stress. Silencing OsMKK6 by RNAitechnology was found to be lethal to rice plants.Nutritional GenomicsTwo ripening specific N-glycoprotein modifyingenzymes, α–mannosidase and β-D-N-acetylhexosaminidasewere used to enhance fruit shelf lifeby reducing the rate of softening in transgenic tomatoRNA interference lines. Shelf life of the transgenictomatoes was found to be enhanced upto 30 days.Stable transgenic lines of AmA1-potato withenhanced protein and amino acids were developed.Food safety and allergenicity experiments indicatedtheir safety. Further, using the constitutive andseed specific AmA1 gene constructs, stable transgeniclines of three commercial indica rice cultivars wereobtained. Stable transgenic lines of sweet potatousing the constitutive AmA1 expression plasmidwere also developed.Plant Development and ArchitectureThe molecular basis of light mediated seedlingdevelopment in higher plants was investigated bycloning and functional characterization of severaltranscription factors and regulatory proteins (ZBF1/MYC2, ZBF2/GBF1, ZBF3/CAM7 and SHW1).They played important roles in lateral rootdevelopment and flowering time in adult plants.Further, the potential of ZBF1 in crop improvementwas investigated by generating transgenic tomatoplants that display early flowering and fruit setting.Analysis of molecular interrelations of GBF1 withtwo negative regulators of light signaling, COP1and SPA1, revealed that GBF1 protein is lessabundant in the dark grown seedlings, and COP1physically interacts with GBF1 and is required forthe optimum accumulation of GBF1 protein inlight grown seedlings.Variant genotypes of the pea plant were constructedby combining mutations in seven of the genes thatregulate various traits. Comparison of organ-wisebiomass partitioning in the mutant genotypes andwild type showed that the tendril-less genotype outyieldedall others in vegetative growth and grainyield.Plant ImmunityAt least 128 Ascochyta genes induced in response tooxidative stress were identified. The full lengthAnnual Report 2009-2010 167

Autonomous Institutionssequences of gene coding Old Yellow Enzyme(OYE), Alternate Oxidase (AOX), and SuperoxideDismutase (SOD) were determined. An efficienttransformation protocol for Agrobacterium-mediatedtransformation of Ascochyta was established. Usingchickpea- Fusarium systems, 6272 gene sequencesof immune-response pathway were identified andassembled into a CaUnigene set of 2013 transcriptscomprising of 973 contigs and 1040 singletons.A chickpea BAC library was developed and closelylinked molecular markers for wilt locus wereidentified.The model vegetable tomato was transformed withOXDC gene under constitutive, fruit specific andwound inducible promoters. Molecular and fieldevaluation upto T3/T2 progeny of the promisinglines showed tolerance to the fungal pathogenS. rolfsii. The food value, toxicity, allergenicityanalysis of the transgenic tomatoes suggest that theyare safe.Days1045Control fruitsá-mannosidase RNAi â-hexosaminidase RNAiFruits from transgenic (T1) lines and wild type plant were harvestedat pink stage and stored at room temperature (22-24oC in 55-60%relative humidity). The progression of fruit deterioration was recordedby time-lapse photography. Time after harvest is specified by days.Institute of Life Sciences, BhubaneswarThe key areas of research of the ILS pertained toinfectious diseases biology, various aspects of generegulation and an emphasis on translational researchand product development. Some of the achievementsare highlighted.Infectious Disease BiologyThe research on metazoan pathogens focused onapoptosis inducing factors in embryogic stagesresulting in imparting anti-fecundity immunityagainst such pathogens and induction of hostmonocytes/macrophage apoptosis by parasite factors.Genetic polymorphism of host genes in humansthat predispose them to infectious diseases likemalaria and metazoan pathogens were studied. Moreparticularly genes associated with immune-regulationwere addressed. A direct association of mannosebinding lectin (MBL) mutations was shown withplasma MBL levels. Two of the mutations wereassociated with increased susceptibility to humanfilariasis. Mutations in Endothelin-1 and TNFreceptor type 2 were correlated with developmentof two diverse forms of the disease viz., lymphedemaand hydrocele.Relationships among non-toxigenic V. cholerae O1biotype El Tor strains were studied. Housekeepinggenes, mdh, dnaE and recA sequences and ribotypepatterns indicated a phylogenetic relationship amongnon-toxigenic V. cholerae O1 biotype El Tor strainsisolated from India, Guam, and Brazil. Theoccurrence of SXT ICEs harboring sulII, strAB,dfr18 and/or dfrA1 genes in environmental andclinical V. cholerae O1 isolated prior to 1992 inIndia was reported.Gene Regulation and FunctionStudies on identification, cloning and characterizationof age-associated genes continued. Cloning of a1009-bp iPPase promoter fragment in the pGL3168Annual Report 2009-2010

Autonomous Institutionsbasic reporter vector followed by promoter analysisidentified two transcription factor binding sites.Analysis of the transcriptional regulation of SMP30gene suggested that the TATA, AP2 and CAATbox located between -128bp and +157bp plays animportant role in determining the promoter activity,sufficient to drive SMP30 gene expression.Biochemical characterization of recombinant riceCatalase A protein expressed in E. coli reportedearlier was completed. Thermostability, lightinsensitivity and NADPH binding characteristicswere studied. Significant progress was made onmolecular and biochemical characterization of athermo stable Superoxide dismutase of plant originthat was recombinantly expressed in E. coli.In depth screening of bio-molecules from diversemicroorganism collected from different ecologicalniches was undertaken. A moderately thermophilicalphaproteobacterium was isolated from a hot springsediment namely was reported. Exopolysaccharides(EPS) produced by the bacterium isolated from hotspring was characterized. A bacterial strain, designatedHT27 T , was isolated from sulfur spring samplescollected from Orissa, as and characterized. Thestrain HT27 T is to be assigned as the type strain ofa novel species of the genus Gulbenkiania,Gulbenkiania indiensis sp. nov (Fig).Two novel promoters FMV-Sgt-En-2X and FMV-Sgt3-En-3X (by inserting double and triple enhancerdomains into Figwort Mosaic Virus sub-genomictranscript promoter, FMV-Sgt) possessing 12-15times stronger promoter activity than CaMV 35Spromoter were successfully developed. In addition,several modified promoters were also developedthough single and multiple shuffling of MMV(Mirabilis Mosaic Virus) and FMV (Figwort MosaicVirus) promoter DNA. The activities of thesepromoters were checked both in transient andtransgenic system using tobacco protoplasts (XanthiElectron micrograph of Gulbenkiania indiensis (strain HT27 T ), showinga single polar flagellum.Assay of promoter constructs in tobaco protoplast using GFP reporter genesAnnual Report 2009-2010 169

Autonomous InstitutionsBrad) and transgenic tobacco (Sumsun NN) plants(Fig). Molecular mining of genes potentially involvedin disease transformation of Chronic MyeloidLeukemia to CML-blast crisis was studied. Resultsdemonstrated that DEFA4 status along with theBCR-ABL transcript could be used as a biomarkerfor identifying CML and CML disease progressionsince none of the patients in early CP or the nascentcases showed any mutation. The studies detectedsingle mutation only in six cases out of the 69 casesof CML.Translational Research and TechnologyDevelopmentIdentification and characterization of differentiallyexpressed genes under abiotic stress by southernhybridization showed seven copies of the PM-H + ATPase gene that exist in at least two isoforms.Gene mining for salt-stress tolerance provided aclear indication of possible vital role of glycinebetaineand proline in salt tolerance process in S. maritima.Studies on targeted EGFR- nanoparticlebioconjugates for breast cancer therapy wereundertaken. Results indicated that nuclear proteinp70 S6K1 may induce apoptosis in a breast cancercell line - MCF 7. Validation using rapamycin,rapamycin loaded nanoparticles and EGFRconjugated nanoparticles suggested that use of EGFRconjugated immune-particle can be a bettertherapeutic approach for treating breast cancer.Institute of Bioresources and SustainableDevelopment (IBSD), ImphalIn its ongoing endeavour to tap the rich bioresourcesof North East Region for developing processes andproducts, the institute strengthened its R &Dfacilities. The institute has also become a partnerin the DBT & Electronic journal Consortia(DELCON) and now has access to more than 800international research journals. Video conferencingfacilities have been installed and the communicationnetwork of the institute strengthened. During theyear, new staff were recruited. Two extramuralnetwork projects were sanctioned. The institute haspublished seven research papers in internationaljournals and 11 papers in national reference journals.The significant achievements of the differentprogramme during the year are summarized below:Bioresource Database UnitTransmission Electron micrograph of rapamycin loadednanoparticlesA digitized database of bioresources of North Eastwas developed with nearly 5500 records fromsecondary sources, inclusive of 1598 floral, 500ethno-botanical plants, 3296 faunal, 95 fungal and472 freshwater cyanobacterial resources. A databaseon Zingiberaceae and bamboo was developed.Efforts were initiated for analysis of microbial data170Annual Report 2009-2010

Autonomous Institutionsusing online/offline bioinformatics tools.Bioinformatics software was installed to facilitateanalysis of molecular data.Medicinal, Aromatic and Horticultural PlantsResourcesA germplasm repository of species belonging tofamily Zeingiberaceae was strengthened with theaddition of new collections including Etlingeralinguiformis, raising the tally to a total of 46 species.Seed germination method of Kaempferia galangalwas standardized. Two Zingiberals – Zingibermontanum and Z. zerumbet were found to containessential oil at 0.8% and 0.41% respectively. Theessential oil content in the Cinnamomum tamalaclones collected from different parts of Manipurwas found to very between 1.3 to 2.1%. Seeds ofpassion fruit (Passiflora edulis) were found to be arich source of unsaturated fatty acids with linoleicacid (69.33%) and oleic acid (14.38%).Microbial ResourcesVermicompost fortified with agriculturallyimportant microorganisms (AIMs) was found toincrease yield of pea, potato and onion in smallplots in farmers’ field. Saccharomyces cervisiae strainsof ‘Hamei’, a traditional starter for rice wineproduction from Manipur and Sikkim werecharacterized. Microorganisms associated with atraditional liquid starter used for ‘Soidon’ (fermentedbamboo shoot product) were identified. ISBDparticipated in a network programme on screeningof bacteria from different ecological niches foridentifying biomolecules of therapeutic potentialinvolving Piramal Life Sciences, Mumbai and nineresearch institutions. Several bacterial isolates thatwere screened showed promising anticancer,antidiabetes and anti-inflammatory activity.Aquatic BioresourcesInduced breeding technique for Bengana devdevi(Khabak) was standardized and the technologytransferred to fish farmers / entrepreneurs for massseed production. Demonstration of captive breedingtechniques of an ornamental fish, Gagata dolichonemawas in progress. Twenty species of edible insectsand fifteen species of medicinal insects werecollected, identified and documented based ontraditional knowledge. Nutritional and therapeuticalevaluations of these were progress.Rajiv Gandhi Centre for Biotechnology(RGCB), ThiruvananthapuramWith the RGCB becoming as a national instituteunder DBT, a redefining of its research anddevelopment programmes was done. The focus ofthe disease biology programme is to understand thebasis of human diseases & promote collaborationbetween clinical and basic scientists. The diseasesstudied include cancer, atherosclerosis heart disease,tuberculosis, viral infections, cholera, neurologicaldisorders and reproductive problems. Brief highlightson the results/achievements of programmes are asfollows:Cancer researchA new web resource, TarMir1.0 that was createdwill serve as a meta-server for detailed miRNAtarget prediction. The web resource now holds datafrom all the 10 publicly available, major targetprediction servers currently in use. It enables multipleAnnual Report 2009-2010 171

Autonomous Institutionsqueries to be submitted, based on miRNA andreverse queries based on a given list of targets.Analysis of differentially expressed miRNAs in oralcancer was done. A list of miRNAs was predictedbased on their mRNA target/anti-target status inoral epithelium. Currently, probe based validationof the expression levels of a selected set of thesemiRNAs was carried out in young versus old oralcancers.Studies have shown that nicotine inducedupregulation of survival signals in lung cancer cellsare dependent on the p53 status of the cells andare downregulated by curcumin irrespective of theirp53 status. Diaminothiazoles were seen to bepotential anticancer agents targeting microtubules.The apoptotic pathway analysis of diaminothiazolesshowed that they induced apoptosis in a novelway compared to other antimitotic agents so farreported.Infectious diseasesField strains of M. tuberculosis strains from Keralawere found to be different from that of other partsof the country on the basis of mycobacterialinterspersed repeat units (MIRU). Screening revealeda battery of promoters that are active on interactionwith macrophages. Ethyl p-methoxycinnamate(EPMC) isolated from Kaempferia galanga wasshown to inhibit drug susceptible and MDR clinicalisolates of M. tuberculosis. Three enzymes involvedin the mycolic acid biosynthetic pathway wereidentified as probable drug targets by in silico analysis.Extensive screening of the field isolates of theChikungunya virus was carried out. In silico dockingstudies by virtual screening of 1, 00,000 structuresfrom natural product libraries targeting viral envelopeproteins identified three potential molecules asCHIKV inhibitors.Chemical biologyDrug entrapped polyester based nanoparticles andfilm implants were developed as drug delivery systemsto treat cancer. Polyesters with different monomercombination based curcumin, paclitaxel and 5-fluorouracil entrapped polymer nanoparticles andpolymer films were synthesized and characterized.For neuronal transplantation, polyester basedscaffolds with interconnecting pores were developedby chemical free ice porogen method. Several novelbiomolecules from skin secretion of frog Ranacurtipes/ R. temporalis were isolated. Theirantimicrobial activity revealed that they may serveas promising candidates for peptide antibiotics.Plant molecular biologyTwenty two species of family Zingiberaceae werebarcoded using the chloroplast target mat K and114 collections of the medicinal rice Njavara werefingerprinted. A miRNA library was generated fromPiper nigrum for elucidating the functionalsignificance of microsatellite associated miRNA.Cloning and characterization of osmotin isoformsfrom resistant wild pepper was done. Theidentification of two forms with distinct sequencecharacters could have functional implications.Agreements with other agencies:RGCB will transfer its Rt-PCR diagnostic systemfor dengue and chickungunya to Hindustan Life-Care Limited (HLL) for the diagnostic facilities itplans to set up in all cities in the country. RGCBand HLL have jointly developed a drug transport172Annual Report 2009-2010

Autonomous Institutionssystem for blood and body fluids which allowsDNA and RNA extraction and transport of thesample without a cold chain. The RGCB haspartnered with Madras Medical Mission, Chennaion cardiovascular diseases.UPCOMING INSTITUTIONS/CENTRESInstitute for Stem Cell Biology andRegenerative Medicine, BangaloreAn institute for “Stem Cell Science andRegenerative Medicine” (InStem) is beingestablished at Bangalore in partnership with itstranslational unit at CMC, Vellore. TheGovernment of Karnataka has allotted 10 acres ofland for the Institute. A total of 20 acres of landwould be available for this institute. The interimlaboratory facilities including clean room facilitieswere constructed in NCBS, Bangalore to initiatethe activities of the Institute. Deans have beenappointed. The Hon’ble Minister of State forScience & Technology, Shri Prithviraj Chavan hadunveiled the foundation stone for this institute on14.11.2009. The programmatic linkages withhospitals and academic institutions are beingdeveloped. The first brainstorming session wasorganized by the institute with a focus on cardiachypertrophy and stem cells. The aim was togenerate a roadmap to initiate translationalresearch and forge interactions between basic andclinical researchers. A collaboration has beenestablished among the Stanford University,InStem, NCBS and others with a focus oncardiac hypertrophy.This institute has established a major collaborativeplatform technology and industry-interaction unit,the Centre for Cellular and Molecular Platforms(C-CAMP) with NCBS. The stem cell institutewill be a part of a Bangalore Bio-cluster which isan innovative institutional model for cutting edgescientific research.Translational Health Science &Technology Institute (THSTI)The activities of the Translational Health Science &Technology Institute, an autonomous institution ofDBT, were initiated after the necessary approvalswere obtained. The THSTI was registered as asociety. Action was taken to appoint the ExecutiveDirector and Deans. Discussions were held withnational and international experts to conceptualizethe design of the NCR cluster at Faridabad andsteps were taken to initiate the activities of HealthScience Technology (HST) component with MIT.Activities of the institute were initiated through theinterim facility established at Gurgaon. The firstmeeting of the Society and Governing Body ofTHSTI were organized and the search cum selectionprocesses for faculty recruitment was underway. Thefollowing niche centres are being established underTHSTI.Vaccine & Infectious Disease Research CentreActivities of the Vaccine & Infectious DiseaseResearch Centre under THSTI as a multidisciplinaryresearch center were initiated for basicand clinical research on vaccines and biologics.The centre would take up some of the availablevaccine candidates and assist their development ina mission mode and also develop newer vaccinesrelevant to India.Annual Report 2009-2010 173

Autonomous InstitutionsPaediatrics Biology Centre (PBC)Necessary approvals were obtained for theestablishment of Paediatrics Biology Centre as apart of THSTI. In order to initiate the activities ofPBC, two senior scientists of NII were appointedas adjunct faculty. The short term objectives inaddition to infrastructure and human resourcedevelopment would be biology of vaccineimmunogenicity; characterization of the immunesystem in neonates and infants; and establishmentof service labs and networking.National Development (Clinical) Service Agency(NDSA)The establishment of the NDSA was taken up as apart of THSTI, to promote pre clinical and clinicaldevelopment of health technologies and productsgenerated, provide expertise for pre clinical services,clinical trials and regulatory issues. The emphasiswould be the development of health related productsand processes developed in India or obtained throughpartnership with global entities for SMEs, publicfunded institutions and not for profit organizations.The NDSA would endeavour to provide low costand quality services for evaluating vaccines biotechproducts in a clinical trial set up.National Institute of BiomedicalGenomics, (NIBMG) Kalyani, WestBengalThe National Institute of Biomedical Genomics(NIBMG) is being established as an autonomousinstitution under the DBT at Kalyani, West Bengal.The institute has been registered under the SocietiesRegistration Act 1860. The major objectives of theinstitute are to promote basic research throughbiomedical genomic approaches and establish stateof-the-artinfrastructure for genomic and proteomicanalyses. The Governing Body, Scientific AdvisoryCommittee, Finance Committee and BuildingCommittee have been constituted. Efforts areunderway to recruit scientific, technical,administrative and support staff. The Governmentof West Bengal provided 30 acres of land forbuilding the institute and 120,000 sq. ft. of qualityspace at Kalyani, as an interim facility. The activitieson the International Cancer Genome Consortium-India Project are being launched. In addition, aproject on genomics of susceptibility to tuberculosisinfection in human subjects is being implementedin collaboration with clinicians of the JawaharlalNehru Memorial Hospital, Kalyani, and scientistsof the ICGEB, New Delhi. NIBMG has alsoinstituted a fellowship programme to attract youngIndian scientists working abroad. The website ofNIBMG has been launched (www.nibmg.ac.in).Regional Centre for Biotechnology,Faridabad, HaryanaThe mission of the centre is to promote researchand create physical infrastructure in critical platformtechnologies supporting interdisciplinary education,training and research in biotechnology. It wouldalso be a place for designing of new and novelprogrammes for education & training which couldthen be assimilated by the existing universities inIndia and the region. It would also act as the focalpoint for cooperation amongst the member countriesof the Asian Region in Biotechnology and oversightwould be provided by a Board of Governors withrepresentatives from India, UNESCO and themember countries. Collaboration with UNESCO174Annual Report 2009-2010

Autonomous Institutionswill ensure gateways to international institutionssuch as European Molecular Biology Organization(EMBO), International Union of Biochemistry &Molecular Biology (IUBMB) etc. and usher in skillacquisition in the global context.Steps have been taken to enrol students fromneighbouring countries in the DBT-sponsored M.Sc.(Biotechnology) programme through placements.Certain training programmes like Winter SchoolTraining Programme in Biotechnology wereconducted in October 2009 in collaboration withICGEB, New Delhi. The research and developmentactivities have been initiated towards developmentof knowledge based drug discovery approaches; andanalysis of complex diseases for identification ofintervention points. The second meeting of theBoard of Governors was held in UNESCO, Parison 31 st August, 2009 along with an interactivesession with the permanent delegations fromBangladesh, China, Indonesia, Iran, Japan,Kazakhstan, Malaysia, Myanmar, Nepal, Philippines,New Zealand, Uzbekistan, Vietnam, Republic ofKorea and the Russian Federation.Recruitment of the faculty, technical andadministrative staff is in progress. The interim facilityof the centre is being set up in Gurgaon, Haryana.The plans of the permanent campus at Faridabadhave been prepared. The Phase–I of the campusdevelopment has been initiated along with HealthScience Biotech Cluster, Faridabad.National Agri-Food BiotechnologyInstitute (NABI)& Bioprocessing Unit(BPU), Mohali, PunjabNecessary approval of the Cabinet was accorded inAugust 2008 for setting up NABI & BPU asautonomous institutions both NABI and BPU areplanned to be co-located with an Agri-food BiotechPark designed to house start-up companies. All thesecomponents – NABI, BPU and the Park – willform the Agri-food Biotech Cluster at Mohali, Punjab.The cluster will be a unique facility with a stronginter-disciplinary approach that is strengthened bythe synergy and close proximity of variousinstitutions and linkages with prospectiveentrepreneurs. It will link biotechnology of cropswith that of food and nutrition while facilitatingbench to market progression of products and serviceswhile catalyzing innovation in Punjab as well as theentire region.NABI will be an apical centre for plant genomicscience based designer crop breeding programmeand will address challenging areas of stress tolerance,drought, salinity, heat, post harvest loss and nutritionenhancement. It will also seek lateral technologytransfer nationally and globally in these areas forfurther development. NABI will be a centre ofexcellence in translational science and productdevelopment, working towards development ofinnovative food processes and products keeping inview the current and future markets both withinthe country and abroad. With the focus on novelfood processes and products, it will take advantageof Punjab’s strength in agriculture. It will also beinvolved in training world class human resource infood science & technology as well as nutritionalscience. NABI will have an enduring partnershipwith the BPU and the Park, and thus promotestart up companies. The BPU is designed to linkthe R&D system with a production facility to serveas an incubator for start-ups. The BPU will havethe process capacity to support a number of start-Annual Report 2009-2010 175

Autonomous Institutionsup companies for up to 5 years till they get a feelof the market and attain in-house capacity for settingup of their larger units. The DBT has alreadyrecruited the Executive Director of NABI andselected candidates for various administrative andfinancial positions. An interim research facility hasbeen set up for both NABI &BPU in Mohali.NABI is currently in the process of identifyingcandidates for specific research positions.BIOCLUSTERSBangalore Bio-clusterThe idea of a Bio-cluster is unique to India. TheCentral and State Governments have joined handsto set up the Bangalore Bio-cluster in the GKVKcampus of the University of Agricultural Sciences,Bangalore. The cluster will comprise the NationalCentre for Biological Sciences (NCBS), the Institutefor Stem Cell Science and Regenerative Medicine(InStem) and the Centre for Cellular and MolecularPlatforms (C-CAMP). The institutions will activelycollaborate, through join programmer. Thefoundation for the Bangalore Bio-cluster was laidby the Hon’ble Minister of State for Science andTechnology, Shri Prithviraj Chavan, on November14, 2009. In contrast to the usual model whereindividual scientists create groups that work onspecific areas of research, InStem, adjacent to theNCBS, will encourage inter-disciplinary work inthe special emphasis on translational and clinicalstem cell research. It will build on the existing linkswith the CMC-DBT Centre for Stem Cell Researchat CMC, Vellore.The, C-CAMP will act as the provider and developerof technology and enabler of scientific activity andentrepreneurship. All the three units of the BangaloreBio-cluster will share space, manpower, facilities andideas. This is expected to be a model for cuttingedge scientific research.Health Biotech Science Cluster(HBSC), Faridabad, HaryanaThe Department of Biotechnology has identified200 acres to set up the Health Biotech ScienceCluster at Faridabad, 40 km from New Delhi in theNational Capital Region. The master plans havebeen finalized and detailed architectural planning ofthe cluster is currently underway. The sitedevelopment work has already commenced and theconstruction of the main buildings is expected tostart by March 2010.The cluster is expected tobecome operational in 24 months from the date ofstart of construction. The Faridabad cluster willhave multiple institutions like a Translational HealthScience and Technology Institute (THSTI),comprising of several niche centers like the Vaccineand Infectious Diseases Center, a Pediatric BiologyCenter, Platform Technology Center, National(Clinical) Development Service Agency, Centre forBiodesign etc. and the Regional Centre forBiotechnology Training & Education. Faculty fromthe NII, New Delhi and the Harvard MIT HealthScience and Technology Institute (HST),Cambridge, USA are closely involved in thedesigning and setting up the Translational instituteat the NCR Bio-cluster. The objective of thecluster is to create a total eco-system for innovationon one campus. It will provide a new model ofeducational institutions along with the advancedcentres for product development, clinicaldevelopment, pilot facility, animal house, technology176Annual Report 2009-2010

Autonomous Institutionsplatforms, including specialized centres for VaccineDevelopment, Molecular Diagnostics and Biodesignso that all services for taking a product idea fromthe bench to bed side can be offered at one location.The HBSC cluster will interact with the Bangalorecluster which is being set up around the Stem CellInstitute, C-CAMP and Plant Genome Centre atNCBS, GKVK Campus, Bangalore.Annual Report 2009-2010 177

CHAPTER 11PUBLIC SECTOR UNDERTAKINGSBharat Immunologicals and BiologicalsCorporation Limited, BulandshaharBharat Immunologicals and Biologicals CorporationLimited (BIBCOL), Bulandshahar is a public sectorundertaking under the administrative control of theDepartment of Biotechnology. The company wasestablished in 1989 under the Integrated Long TermProgramme (ILTP) of Cooperation in Science &Technology with Russia for indigenous productionof Oral Polio Vaccine (OPV) in India to facilitatethe universal immunization programme ofGovernment of India. BIBCOL is currently engagedin the business of indigenous production of OralPolio Vaccine (OPV) and zinc dispersible tablets.The company has set up its new production facilitiesfor manufacturing zinc dispersible tablets, employingthe technology transferred from M/s Nutiset, France.After obtaining the manufacturing license and GMPcertification, BIBCOL is engaged in themanufacturing and supplying of zinc dispersibletablets to the State Governments under a project“Large scale demonstrative studies of zinc in themanagement of diarrhea as an adjunct to oralrehydration solution in high burden Indian States”.The Government of India currently holds 59.25%of the equity capital of BIBCOL. The shares of thecompany are listed on the Bombay Stock Exchange.Indian Vaccines Corporation Limited,GurgaonThe Indian Vaccines Corporation Limited (IVCOL)was set up as a joint venture company in 1989 forundertaking R&D, and engageing in themanufacture of viral vaccines. Due to a change inthe product mix policy and non-availability of thevero cell technology from Pasteur Merieux Serum& Vaccines (PMSV), France IVCOL was put onhold in 1992. In pursuance of a Cabinet decisionin 1998, the restructuring process of the companywas undertaken to utilize the assets created.Accordingly, the National Brain Research Centre(NBRC) facility has been established in a portionof the IVCOL premises.Concerted efforts were made to finalize therestructuring process with a portfolio of newactivities. The company has signed a lease agreementwith IPCL/Reliance Life Sciences, one of thepromoters of the company, to initiate new activitiesincluding production of vaccines, biopharmaceuticalsand also set up a superspeciality hospital to initiateactivities in translational research in collaborationwith NBRC. The IVCOL board has also beenreconstituted.Annual Report 2009-2010 179

CHAPTER 12INTERNATIONAL CENTRE FOR GENETICENGINEERING AND BIOTECHNOLOGYInternational Centre for Genetic Engineering andBiotechnology continued its significant contributionin promoting biotechnology and genetic engineeringresearch in the member countries. The centretransferred a technology on ‘Dengue specific antigensand monoclonal antibodies’ to an industrial partnerin India and filed three new patent applicationsincluding PCT applications. Three training programson (i) applications of structural & computationalbiology in biomedical sciences (ii) biologics:discovery to development and (iii) gemini viruswere conducted for the participants from India andmember countries of the developing world.HUMAN HEALTHImmunologyThe group working on this theme is involved inunderstanding the regulatory parameters thatinfluence the outcome of an immune response. Themain focus was on understanding the mechanismscontrolling plasticity in receptor-initiated signalingand how this pathway directs specific expressionprograms in lymphocytes.Under a project supported in a public-privatepartnership mode by DBT and the Institute of LifeSciences (ILS), Hyderabad, a genome-wide siRNAscreening was done to identify host factors thatregulated pathogen load in human macrophagesinfected with a virulent strain of Mycobacteriumtuberculosis. About 275 such molecules were shownto be functionally associated with each other througha dense network of interactions.A DBT – ICGEB partnership programme wasinitiated for identifying biomarkers in samples frompatients infected with active tuberculosis. Theproteome composition of serum, saliva, and urinewere analyzed using 2D-LC-MS/MS massspectrometry, whereas volatile components of thebreath were examined by GC-MS. A combinationof six serum proteins of host origin showed asignificant alteration in the tuberculosis disease status.Out of the six marker proteins, two were found tobe relevant in checking diseased susceptibility.Tuberculosis patients were studied through breathanalysis using GC-MS. A total of 350-380 volatilemolecules were identified in tuberculosis patientgroups, 300-350 in normal and healthy contacts,310-360 from lung cancer patients and 230-380 inCOPD patients. The result showed a metaboliteassociation with health status. The breath biomarkersseem to be a potential non-invasive method fordisease diagnosis.Malaria ResearchMalaria immunity and vaccine researchUnder a program on development of a vaccine forPlasmodium vivax malaria based on PvRII, methodsto produce recombinant PvRII were developed.Annual Report 2009-2010 181

International Centre for Genetic Engineering and BiotechnologyClinical grade JAIVAC-1 was manufactured undercGMP by Bharat Biotech, Hyderabad. Toxicologystudies with JAIVAC-1 were completed. Theexpansion of infrastructure for clinical developmentof malaria vaccines at ICGEB was underway.Malaria Drug ResearchA proteomics based approach was used to investigatethe mechanism of action of the antimalarial drug,artemisnin. ClpQY protease system in P. falciparumwas identified as potential drug target. In silicoscreening libraries of small molecules were beingdeveloped to identify possible specific inhibitorsagainst PfClpQ proteases that may act as leadcompounds to design drugs against malaria.Recombinant Gene ProductsGenetically engineered biomolecules and viraldiagnosticsThe primary focus of this initiative was on thedevelopment of novel recombinant multiepitopeproteins (MEPs), as inexpensive, highly sensitiveand specific diagnostic intermediates for viralinfections, like HCV, HIV and Dengue. Incollaboration with the University of Turku, Finland,CE certified designer proteins generated wereexplored for developing unique 3-in-1 assays forsimultaneous detection of HIV, HCV and HBVinfections, in blood bank.Protein trafficking machinery in the parasiteStudies on the trafficking and processing of animportant food vacuole protease, falcipain-2, usingthe GFP targeting approach were done and effortscontinued to identify and characterize novel parasitespecific protein-transporters that may be developedas novel targets against the parasite.Receptor-ligand interactions involved inerythrocyte invasion & cyto-adherenceWork continued on trying to understand theinteraction of a family of erythrocyte bindingproteins from P. vivax, P. knowlesi and P. falciparumEBA175, with host receptors during invasion. Anovel cyto-adherence receptor, gC1qR, was identifiedwhich binds complement component C1q and isexpressed both on endothelial cells as well as platelets.It has been demonstrated that P. falciparum isolatesuse gC1qR as a receptor to bind endothelial cells aswell as for platelet-mediated clumping, which isassociated with severe malaria.Flavicheck : Rapid Immunochromatographic test for HCV antibodiesRecombinant dengue vaccine developmentEfforts continued on the development of anexperimental sub-unit vaccine, based on a discretedomain of the major DENV E protein. With thepediatric dengue vaccine initiative in Seoul, a projectwas undertaken to examine the feasibility of linkingthe EDIII-based antigen to HBsAg and make viruslikeparticles (VLPs).Synthesis and selection of de novo antibacterialproteinsEfforts were directed towards ‘codon-shuffling to182Annual Report 2009-2010

International Centre for Genetic Engineering and Biotechnologycreate similar antibacterial proteins against pathogenicorganisms like Mycobacterium tuberculosis. Potentbinders against an essential protein of M. tuberculosis,called hupB were identified. Expression of this‘codon-shuffled’ protein in Mycobacteria causedgrowth retardation, and changes related with theinhibition of hupB inside Mycobacteria.Structural and Computational BiologyProtein crystallographic studies of malariaparasite proteinsWork was carried out on two proteins which areknown to perform DNA compaction and calledNucleosome Assembly Proteins (Pf-NAPz). Theseproteins interact with other essential parasite proteinslike histones to assemble and disassemblenucleosomes. Work was in progress to identify therole of PfNAPz in histone transport and nucleocytoplasmicshuttling.Biology of Mycobacterial membrane proteinsA strategy was devised for parallel cloning andexpression to facilitate faster and efficient screeningof as many transporters as possible fromM. tuberculosis in a short time to find candidatesamenable to structural and functionalcharacterization. One of the transporters wasexpressed using this strategy and work was in progressto adopt the method for other transporters in thesame family.Computational biology studiesSeveral protein sequence databases includingProtRepeatsDB: a database of different types ofprotein repeat sequences in genomes (http://bioinfo.icgeb.res.in/repeats), ProtvirDB: a databaseof protozoan virulent protein sequences (http://bioinfo.icgeb.res.in/protvirdb) were developed.Artificial intelligence based web servers for methodsto solve bioinformatics problems such as predictionof cyclin sequences virulence proteins and lipocalinswere also developed.VirologyHepatitis E virus (HEV)The main focus was on the role of the HEVORF3 protein in viral pathogenesis. Recent resultsshowed that the ORF3 protein protected cells frommitochondrial depolarization, and death. The bindingand endocytosis of capsid (ORF2) protein viruslikeparticles to cultured cells was studied as amodel to understand virus attachment and entry.Human immunodeficiency virusEfforts were made to study the HIV-1 accessoryproteins Nef and Vpu for their roles in pathogenesis.The immune responses and the dynamics of viralreceptors/coreceptors in HCV-HIV co-infectedpersons were investigated. A biomarker discoveryprogramme based on genomic, proteomic andmetabolomic analyses was initiated in various groupsof HIV-infected persons.Influenza A (H5N1/H1N1) virusUnder this research program, it was discovered thatthe nucleocapsid protein (NP) of H5N1 interactedwith heat shock protein (hsp) 40. The levels ofpPKR and phospho-eIF2alpha (the downstreamtarget of PKR) were found to be down regulated inNP transfected cells.Annual Report 2009-2010 183

International Centre for Genetic Engineering and BiotechnologyAGRICULTURAL BIOTECHNOLOGYPlant Molecular BiologyBiotic & Abiotic Stress regulated genesCharacterization of candidate genes that contributepositively to stress tolerance, and understanding theirexpression mechanisms in response to various stresses,was taken up as a priority. About 2000 differentiallyexpressed ESTs were obtained from Pokkali, a sensitiveand a saline tolerant rice cultivar. Transcript analysiswas done for 10 HSPs and 26 members of a CBSdomain gene family. One of the CBS domainscontaining proteins was functionally validated intobacco and it was shown to confer improvedtolerance to different stresses. A cold stress regulatedbi-polar DNA helicase (PDH47) and translationalinitiation factor 4A (eIF4A/PDH45) from pea andPennisetum were characterized. Results linked the G-proteins to high salinity and heat stress tolerance.Developing stress tolerant transgenic plantsAn antibiotic marker free system based on theglyoxalase system for the selection of transgenicswas developed. These efforts resulted in ‘engineered’rice over-expressing glyoxalase I and II, vacuolarantiporter and PDH45 genes and works onpyramiding genes, making mega vectors for geneencoding ROS pathway enzymes and Glyoxalase Iand II and NHX together. A few transgenic eventswere transferred to seed companies for validation.Plant TransformationBt. Cotton and RiceAn efficient Agrobacterium mediated transformationprotocol for cotton and rice was developed andfour Bt genes were introduced. Transgenic linesshowing stable expression of Bt. proteins showedprotection against various pests. A highly efficientregeneration and transformation system based onsomatic embryogenesis was developed for an elitecultivar Narsimha and used in cotton genome tovalidate candidate gene functions.Cotton improvementProtocols were developed to reduce the time involvedin, and improve the frequency of regeneration, andhastened transgenic plant development by directlytransforming the embryogenic callus with the requiredgenes.BiofuelsIn collaboration with the University of Pavia, Italy,a research program was initiated to express variouscell wall degrading enzymes in chloroplasts oftobacco with a view to utilizing these in degradingcarbohydrate polymers present in plant waste/biomass, and convert them simple carbohydratesand further into ethanol.184Annual Report 2009-2010

CHAPTER 13PROMOTION OF BIOTECHNOLOGY INNORTH EASTERN STATES OF INDIAThe North East Region (NER) of India is endowedwith exceptional natural beauty, floral and faunalbiodiversity and abundant mineral, water and forestresources. Although it occupies only 7.8% of thetotal geographical area of the Indian subcontinent, itharbors about 50% of the floristic wealth of thecountry. Moreover, the forest cover in the regionconstitutes 52% of the geographical area. Hence, ithas been identified as one of the world’s biodiversityhotspots. The Government of India has laid specialemphasis on the all round development of the regionfor ensuring economic progress and prosperity. TheNational Biotechnology Development Strategy of theDBT seeks to address a number of challenges relatingto the biotech sector in terms of R&D, creation ofinvestment capital, technology transfer and diffusion,IPR, regulatory issues, building public confidence,and tailor made human capital for these aspects. Thekey elements of the strategy relevant to NER are:(i)Promotion of biotech industry through publicprivate partnership programmes,(ii) Biotechnology Industry PartnershipProgramme for advanced technologies,(iii)Supporting small and medium sectorinnovation,2009-10 to promote these activities in theNorth Eastern Region.Major Activities of DBT in theNorth - East RegionEstablishment of Regional Research Centre ofIBSD in SikkimDBT sanctioned the setting up of a regional centreof IBSD at a total cost of Rs 9.61 crores. Thecentre will the have the ‘state-of-the-art’ facilitiesfor biotechnology research on the local bioresources,for their conservation, management and sustainableutilization, towards developing technology packages,for entrepreneurship development and employmentgeneration.Biotechnology Incubator in AssamThe Biotech Park located at Guwahati will have aBiotech Incubation Centre to develop and scale upbio-processes and technologies and provide researchlaboratories knowledge based service centers andsophisticated facilities. The incubation centre willhave facilities for medicinal plants extraction, micropropagation, drug discovery, bioremediation, biofuels etc.(iv)(v)Promotion of industrial R&D andBuilding expertise and infrastructure.Programme based support to BioresourcesDevelopment Centre, Shillong, MeghalayaDBT has earmarked 10% of its total budgetamounting to Rs.100 crores for the yearA programme-based support was provided forconservation and sustainable utilization of Bio-Annual Report 2009-2010 185

Promotion of Biotechnology in North Eastern States of Indiaresources of Meghalaya at Bioresources DevelopmentCentre (BRDC), Shillong, Meghalaya. Theprogramme aims at conservation, in vitropropagation and improvement of horticultural,ornamental, medicinal and aromatic plants of highcommercial important. Human resourcedevelopment forms an integral component of theproject.Programme support for Biotechnology, at IIT,GuwahatiThe DBT program support on “FundamentalMolecular Investigation in Biotechnology” is aimedat creating not only a dedicated infrastructure andstrengthening research activities of the investigatorsbut also develop a system that would help researchin frontline areas of proteomics and genomics.Establishment of North Eastern BioinformaticsNetwork (NEBINet)Twenty five bioinformatics centers established in allthe 8 states of the NER were networked as theNorth Eastern Bioinformatics Network (NEBINet).The North East Hill University, Shillong wasdesignated as the nodal center for NEBINet.DelCON E-Journals access to NER institutionsDBT’s e-library consortium (DeLCON) whichprovides access to 899 important journals in lifesciences was extended to cover 15 institutes in theNER which met certain laid down criteria by DBT,including presence of bioinformatics facility,institutes/universities offering Post Graduate levelto Ph.D. programmes, adequate expertise, computerhardware and internet connectivity.Establishment of Biotech Hubs for the NorthEastern StatesThis initiative was taken to promote education andresearch in life sciences and biotechnology, and toattract brilliant students to build their career indifferent fields of biological sciences/biotechnology.At least one nodal hub will be established to providebiotechnology facilities to the institutions in thestate. The following will be the key elements ineach biotech hub.• A biotechnology lab with a basic set ofequipment• A bioinformatics centre with at least 5computers and internet connectivity• Access to e-journals in biotechnology• Recurring budget for chemicals, glassware andinternet connectivity• Support for training and workshops forteachers• Travel support for conferences and training inIndia.2nd NEBinet Coordinators’ Meeting• Student fellowships etc.186Annual Report 2009-2010

Promotion of Biotechnology in North Eastern States of IndiaDevelopment of infrastructure in MedicalColleges in North-Eastern States of IndiaDBT took the initiative to strengthen biotechnologyin medical colleges in the North-Eastern India. Toinitiate the activity, an assessment of the state ofacademics especially in UG & PG medical education,existing facilities in various departments and stateof research in the Pathology/Microbiology/Biochemistry departments was undertaken. The selectGovernment Medical Colleges/Hospitals thereaftersubmitted proposals for infrastructure and manpowerdevelopment in various departments i.e.Microbiology, Pathology, Biochemistry, Anatomy,Forensic Medicine and Pharmacology. DBTorganized a brainstorming session on 20 th July 2009through the Department of Molecular Biology andBiotechnology, Tezpur University to facilitate theinitiative effectively. DBT also established a nodalcentre at Tezpur University. A Site-Visit Committeewas set-up for making recommendations on thevarious proposals. An Apex Committee was alsoformed to allocate funds for the proposal submittedby the medical colleges.A summary of initiatives/facilities taken so far• Establishment of Regional Research Centre ofIBSD in Sikkim State• Establishment of Biotechnology Incubator inAssam• Establishment of Women’s Biotechnology Park• Programme Support at IIT, Guwahati• Programme Support for Clinical ResearchNetwork in North East Region• Bioresources Centre at NEHU, Shillong• Network Programme on Developing Drugsfrom Medicinal Plants• Establishment of North EasternBioinformatics Network (NEBINet)• On-line Information Access Facility forSchools• Promotion of Star Colleges in Life Sciencesand Biotechnology• Network on Management of Classical SwineFever• Establishment of state and institutionalBiotech Hubs• Launching of Special Fellowship/Associateship• DNA Clubs (DBT’s Natural ResourcesAwareness Clubs)• Establishment of Rural Bioresource Complex• Biotechnology Infrastructure Development inMedical CollegesAnnual Report 2009-2010 187

Promotion of Biotechnology in North Eastern States of India• Biotechnology Overseas Associateships forNER Scientists• NER-DeLCON- E-Journals accessSetting Up of North Eastern RegionBiotechnology Programme Management Cell(NER-BPMC)There is a clearly felt need to launch newprogrammes/projects/policies for supporting thedevelopment of biotechnology in the NER. Theseare necessary in a time bound manner with regularmonitoring to ensure successful implementation.To facilitate effective coordination, theestablishment of the BPMC was taken up withBCIL. The action plan was drawn up with thenecessary technical, financial and administrativesupport from the DBT which will work in closecooperation with various Government Ministries/Departments/Agencies at the Central and Statelevels along with universities, research institutionsand NGOs located in the NER.188Annual Report 2009-2010

CHAPTER 14NEW LEGISLATIONS AND REGULATIONSDNA Profiling BillDNA based technology has been found to be veryeffective in enabling Court(s) of Law to decide ,for example, on the parentage of a child, or whethera particular individual has been the source of abiological specimen obtained from a scene of crime.The concerns regarding appropriate use of DNAtechnology by the Courts of Law and other agencieshas made it necessary for the DNA testingcommunity to establish effective guidelines andstandards. Therefore, there is a need to regulate theuse of DNA profiles through the enactment ofLaw by the Parliament, so that such use is restrictedto lawful purposes of establishing identity in acriminal or civil proceedings.Recognizing the need to create an appropriatelegislation to regulate various issues relating to DNAprofiling and establishment of DNA data banks,quality control, obligations and accreditation ofDNA laboratories, the DBT set up a DNA ProfilingAdvisory Committee (D-PAC) in December 2003to make recommendations for enacting a suitablelegislation. The D-PAC comprising experts fromthe relevant fields submitted its report and a draftHuman DNA Profiling Bill in 2006. One of theimportant provisions of the Bill is the establishmentof DNA data banks, that would help the criminaljustice system in solving cases and also in crimeprevention. It is known that a number of crimesare committed by repeat offenders, who could beapprehended and convicted by comparison ofevidence at the scene of crime with information aDNA data bank.The draft Bill was circulated to the relevantMinistries and Departments of the CentralGovernment to obtain their comments. It wasrevised keeping those comments in view. The draftBill will be sent to the Department of legal Affairs,Ministry of Law & Justice for their vetting.The Protection and Utilization ofPublic Funded Intellectual PropertyBill, 2008Public funded research in universities, academic andresearch institutions have often produced innovationsthat hold potential for public good. However, alarge number of innovations have not been reachingthe industry or the public due to:(i)(ii)(iii)Possession of the IP generated remaining withthe funding agency;Non-existence of an appropriate legalframework in universities, academic andresearch institutions for utilization, andfinancial incentives to the researchers;Procedural complexities in technology transfer,its absorption and utilization.It is envisaged to create a legal framework in theform of The Protection and Utilization of PublicAnnual Report 2009-2010 189

New Legislations and RegulationsFunded Intellectual Property Bill, 2008 to ensurethe seamless transfer of innovation from lab toplace of utilization. The expected benefits of theBill are:i) Uniform legal framework for protection andutilization of the IP generated out of publicfunded R&D;ii)iii)iv)Innovations in private sector as well as inuniversities, academic and research institutionsthat receive grants from Government will getencouraged;Collaboration between Government, non-Governmental organizations and private sectorwill get promoted;The legislation will facilitate commercializationof IP generated out of Government fundedR&D and also promote innovation culturewithin the country.v) Employment generation in niche areas, withunique skills required for IP management,development and marketing, will getencouraged;vi)Transfer of technologies from researchlaboratories to market place (nationally andinternationally) will spur technological andfinancial growth of the country.The Protection and Utilization of Public FundedIntellectual Property Bill, 2008 approved by theCabinet on October 30, 2008, was introduced inthe Rajya Sabha on 15 th December, 2008. TheBill is now under consideration of the Departmentrelated Parliamentary Standing Committee onScience & Technology and Environment &Forests. The Committee had its first meeting on27.1.2009 in which the Secretary, DBT tenderedoral evidence before the Committee. Subsequentmeetings were held on 17 th December, 2009 and24 th December, 2009 in which Secretary, DBTand other scientists/academicians tendered oralevidence before the Department-relatedParliamentary Committee on Science &Technology, Environment & Forests.Biotechnology Regulatory Authorityof India (BRAI)The Government of India directed the DBT to bethe nodal agency for establishment of theBiotechnology Regulatory Authority of India(BRAI). The DBT has prepared a draft establishmentplan for setting up the BRAI based on the reviewof structure and governance of other autonomousagencies in India, international models for theregulation of biotechnology and India’s internationalobligations. The BRAI is proposed to be anindependent, autonomous and professionally ledbody that will provide a single window mechanismfor the safety assessment of genetically engineeredproducts and processes. Establishing the BRAI willrequire the promulgation of new legislation, namelythe “Biotechnology Regulatory Authority of IndiaAct” or the BRAI Act, the draft bill of which hasbeen prepared by DBT through a consultative groupof experts.In order to obtain views of various stakeholders,DBT initiated an extensive consultative process toseek inputs on the draft establishment plan and thelegislation through countrywide consultations as wellas by placing both the documents on its websites.About 400 responses were received through emails.Seven stakeholder meetings were held involving190Annual Report 2009-2010

New Legislations and Regulationsmore than 700 participants from industry, NGOs,academia, farmers, consumer organizations, legalexperts and media representatives. More than 28State Governments responded with their writtenconsent. Based on the feed-back and commentsfrom consultations, the final draft for establishmentof BRAI, and the BRAI Bill were prepared andcirculated for comments from Ministries/Departments concerned. Taking into considerationthese comments, the final proposal of the bill, withclearance by form the Ministry of Law and Justice,Government of India has been submitted to theCabinet for approval to introduce the Bill in theParliament.Annual Report 2009-2010 191

CHAPTER 15ADMINISTRATION AND FINANCEAdministrationThe Administration Section provided support forthe implementation of various scientific programmesof the department. Logistical support was providedfor the meetings of the various Task Forces/ExpertCommittees that were organized. Office equipmentsuch as laptops, PCs, multimedia projectors,photocopying machines etc. were provided to variousdivisions and sections in the department. The officespace available in the department was utilized in anoptimal manner keeping in view the space constraintthat the department has.EstablishmentThe Establishment Section dealt with the personnelmatters of the department.(i)Recruitment and promotion to variousposts:- During the year, action was initiatedto fill up the 3 posts of Scientist ‘H’, 4 postsof Scientist ‘D’ and 21 posts of Scientist ‘C’.One post of Scientist ‘H’ which was notfilled was advertised again. Offers ofappointment have been sent to the selectedcandidates for the posts of Scientists ‘C’ &‘D’. Further, offers of appointment were sentto the 8 candidates selected candidates for theposts of Junior Technical Assistants. A proposalwas sent to the UPSC for filling up of 8posts of JA (Tech.) on a direct recruitment(ii)(iii)(iv)basis. Apart from the above, one JAO ondeputation in the department has beenabsorbed, and one Staff Car Driver (OrdinaryGrade) was appointed.Flexible Complementing Scheme:- Underthe Flexible Complementing Scheme (FCS),two Scientists of the department werepromoted w.e.f. 1.1.2009 i.e. one from thelevel of Scientist ‘D’ to Scientist ‘E’ and theother from Scientist ‘C’ to Scientist ‘D’.Training:- The officers and staff were deputedfor the training programmes conducted bythe Institute of Secretariat Training &Management to enhance their skills in relevantfields.Personal Information System:- Personalrecords relating to officers and staff wereupdated regularly. The department alsoinitiated the work relating to maintenance ofe-service books of its employees.The category wise position of posts sanctioned andfilled as on 31.12.2009 was as under:-Category of posts Posts sanctioned Posts filledGroup ‘A’ 73 39Group ‘B’ 79 57Group ‘C’ 65* 46Group ‘D’ 40 38Total 257 180Annual Report 2009-2010 193

Administration and Finance* This includes 10 posts of LDC, one post each of UDC andStenographer proposed for abolition in lieu of creation ofadditional scientific and technical posts in the department.Representation of SC/ST/OBC/PH:- The numberof SC/ST/OBC/PH employees in Grade ‘A’,’B’,’C’and ‘D’ categories as on 31-12-2009 was as under:-Group/ Group Group Group Group TotalCategory of ‘A’ ‘B’ ‘C’ ‘D’Post(i) Sanctioned 73 79 65 40 257strength(ii) Total 39 57 46 38 180Posts filledScheduled 02 08 05 19 34CastesScheduled 01 03 06 03 13TribesOther - 01 08 - 09BackwardClassesPhysically 01 01 03 - 05HandicappedSports and Cultural ActivitiesA number of DBT officials participated for thefirst time in the inter-ministry athletics andbadminton tournament 2009-10 and distinguishedthemselves with excellent performance. Ms. AlkaMankad and Ms. Rajani Kushwaha represented DBTin the inter-ministry badminton tournament 2009-10 held at AIIMS, New Delhi and Ms. AlkaMankad won the silver medal in the women singles(Veteran) event. Ms. Alka Mankad won bronzemedals in 100 mtrs and 200 mtrs race and Ms.Rajani Kushwaha won bronze medal in 400 mtrsrace in the inter-ministry athletic meet 2009-10held in New Delhi. The staff members of thedepartment participated in the inter-ministry music& dance and short play competition, 2009-10, inthe events of instrumental music, folk dance andfolk music.97 th Indian Science CongressThe Department participated in the 97 th IndianScience Congress held on 3-7 th January 2010 atThiruvanthpuram, Kerala. The focal theme was“Science & Technology Challenges of 21 st Century– National Perspective”. The science congress wasinaugurated by Dr. Manmohan Singh, Hon’blePrime Minister of India. It is the largest nationalscience event in the country. Indian Science Congressserves to inspire the students, academicians and thecommon man to appreciate the value of scienceand inculcate a scientific temper. The Departmentdeputed two officers Dr. Gulshan Wadhwa and Dr.Onkar Tiwari to the congress and participate in theScience Exhibition. The Science Exhibition was oneof the major events of the 97 th Indian ScienceCongress. This exhibition focused on India’sdevelopment and intended for common people,The stall set up by the DBT at the 97 th Indian Science Congress.194Annual Report 2009-2010

Administration and Financeespecially the large student community, who visitedthe exhibition in large numbers. The DBT’sexhibition stall has highlighted the activities of theDepartment towards promotion of biotechnology,support for various R&D programmes itsautonomous institutions, and interaction withindustry.Progressive use of Hindi in the DepartmentThe Hindi Branch ensured progressive use of Hindiand implementation of the Government policy onOfficial Language in the department. An OfficialLanguage Implementation Committee, constitutedunder the chairmanship of the Joint Secretary(Admn.) in the department, reviewed theprogressive use of Hindi in every quarter andsuggested corrective measures for promoting useof Hindi. Under section 3(3) of the OfficialLanguages Act, 1963, all documents were issuedin a bilingual form and the letters received inHindi were replied to, only in Hindi. In order tostrengthen the monitoring system, the officers whosigned the papers have been made the check-points.Hindi fortnight was organized in the departmentduring 14 - 30 th September, 2009, in which 7different competitive events namely essay, writingdebate, scientific article writing, noting & drafting,hindi quiz, hindi typing and antakshari were heldto promote the use of Hindi in official work.Seven officers and twenty nine other employeeswere given certificates and cash prizes in a ceremonyorganized under the chairmanship of Secretary(DBT) on 30 Nov, 2009. Under Rule 8(4) ofthe Official Language Rules, 1976, Establishment,Administration, PPVC, Cash and Library sectionsin the Department have been notified to do100% their work, in Hindi.With a view to encourage publication of originalbooks on biotechnology related subjects in Hindi,the department has been carrying out an awardscheme namely “Dr. Jagdish Chandra Bose HindiGranth Lekhan Puraskar Yojna”. Under this scheme,for the year 2008, two cash awards of Rs. 10,000/-each were given to the authors of the books namely(1) JAIV PRODYOGIKI CHIKITSA EVAM ROGNIDAAN and (2) JAIVIK SANSADHAN.During the year, 60 officers/employees were impartedtraining in four Hindi workshops organized in thedepartment during every quarter of the year. TwoHindi computer training programmes were alsoorganized to impart training to the employees inorder to facilitate their working in Hindi oncomputers.Grievance RedressalThe Grievance Redressal Machinery dealt withthe public as well as the staff grievance petitions.Eight public grievances were received during theperiod from April, 2009 to December, 2009. Allthese were disposed off in a time bound manner.The Department of Biotechnology regularlyupdated the progress and pendency of publicgrievances.Vigilance UnitA Vigilance Cell continued functioning in theDepartment of Biotechnology. During the periodApril, 2009 to December, 2009, no vigilance casewas registered. The Vigilance Awareness Week wasobserved in the department from November 3-7,2009 in pursuance of the instructions of the CentralVigilance Commission. Complaints received fromvarious sources were processed in a timely mannerAnnual Report 2009-2010 195

Administration and Financeand the Commission was apprised of the progressand the final outcome of these complaints.FinanceThe Department of Biotechnology was allocated anamount of Rs.1000.00 crores (Plan) and Rs. 24.00crores (Non-Plan) for the year 2009-10. This wasrevised to Rs.902.00 crores (Plan) and Rs.23.90crore (Non-Plan) at R.E. stage. The budgetallocation for 2010-11 is Rs.1200.00 crores (Plan)and Rs.22.00 crores (Non-Plan). The financialstatement showing the details of actual expenditureduring 2008-09, B.E. and R.E. 2009-10 and B.E.2010-11 in respect of various programmes/schemesis at Annexure II.196Annual Report 2009-2010

ANNEXURESAnnexure-IGender BudgetingIn pursuance to the recommendations of the Inter-Departmental Committee set up under theChairmanship of Secretary (Expenditure) to considerissues regarding Gender Budgeting, the DBT hasestablished a “Gender Budgeting Cell” with thefollowing Terms of Reference:• To make an assessment of the benefitsreaching women through the existingprogrammes and schemes of the department.The assessment is to be reflected in the AnnualReport of the department.• To come up with specific schemes targetedtowards women.• To clearly bring out scheme-wise provisionsand physical targets for benefiting women inthe Annual Plan and Performance Budget ofthe department.• To provide inputs for the detailed Demandsfor Grant.The Gender Budgeting Cell has taken necessaryaction as per the guidelines circular of Ministry ofFinance for identifying specific schemes for whichbudget needs to be earmarked for the benefit ofwomen. A special programme on Biotechnologyfor Women targets the women beneficiaries in termsof employment and income generation. There isalso a Special Biotechnology Award Scheme forencouraging and giving due credit to outstandingWomen Scientists. In addition, quantificationregarding number of women scientists benefitedwas done in respect of various programmesimplemented. The details of the “Gender Budgeting”are as follows:Annual Report 2009-2010 197

AnnexuresAnnexure-IGender BudgetingScheme-wise provisions and physical targets benefiting womenSl. Details of Schemes : HRD, Total Budget Expenditure No. of women BudgetNo. Bioinfor-matics, Biotech allocated for incurred to benefited through estimates forfacilities, R&D, Societal the year support various activities Financial yearDevelopment, Bioprocess 2008-09 women during during 08-09 2010-11 to& Product Development, the Financial support womenInternational Cooperation year 2008-091. Human Resource 3500 550 1000 500Development2. Bioinformatics 250 530 3003. Biotech facilities Centres of 6500 270 120 300Excellence and Programmesupport4. Research & Development 34100 4435.00 515 100005. Biotechnology for Societal 1000 280 1350 300Development6. International co-operation 1500 535 17 100(Rs. in lakh)Benefits earmarkedfor women during theFinancial year 2009-10.The specific programmeshave been supported forcapacity building and toprovide training underbioinformatics. Also toprovide R&D support andinfrastructure strengtheningincluding InternationalCollaborations.In addition, special awardschemes are initiated toencourage women scientists/students.* In terms of R&D projects supported.198Annual Report 2009-2010

AnnexuresAnnexure-IIFinancial Statement for Annual Report : 2009-10(Rs. in crores)S. Name of the Programme/Schemes/Projects Expeniture during B. E. 2009-10 R.E. 2009-10 B.E. 2010-11No 2008-2009Plan Non- Plan Non- Plan Non- Plan Non-Plan Plan Plan Plan1 Secretariat Economic Services1.01 Secretariat 0.00 9.37 0.00 12.00 0.00 11.90 0.00 0.00Total = Secretariat Economic Service 0.00 9.37 0.00 12.00 0.00 11.90 0.00 10.002 Autonomous R&D Institutions2.01 National Institute of Immunology 57.31 1.50 53.00 1.60 53.00 1.60 53.00 1.602.02 National Centre for Cell Science 43.96 0.36 34.00 0.40 34.00 0.40 34.00 0.402.03 Centre for DNA Finger Printing & Diagnostics 27.27 0.00 24.00 0.00 24.00 0.00 24.00 0.002.04 National Brain Research Centre 29.07 0.00 24.00 0.00 24.00 0.00 24.00 0.002.05 National Institute for Plant Genome Research 26.48 0.00 29.00 0.00 29.00 0.00 29.00 0.002.06 Institute of Bioresources & Sustainable Development 12.12 0.00 0.10 0.00 0.10 0.00 0.10 0.002.07 Institute of Life Sciences 12.40 0.00 13.00 0.00 13.00 0.00 15.00 0.002.08 Rajiv Gandhi Research Centre for Biotechnology 26.25 0.00 24.00 0.00 24.00 0.00 24.00 0.002.09 Translational Health Science and Technology Institute 20.24 0.00 17.00 0.00 0.00 0.00 25.00 0.002.10 UNESCO Regional Centre for Education and Training 0.00 0.00 13.00 0.00 1.00 0.00 22.00 0.002.11 National Agri-Food Biotechnology Institute and Bioprocessing Unit 0.00 0.00 20.00 0.00 4.00 0.00 30.00 0.002.12 Institute for Stem Cell Research and Regenerative Medicine 0.00 0.00 14.00 0.00 14.00 0.00 18.00 0.002.13 Institute for Biomedical Genomics 0.00 0.00 14.00 0.00 7.00 0.00 25.00 0.002.14 National Institute of Animal Biotechnology 0.00 0.00 0.00 0.00 0.00 0.00 5.00 0.00Total = Autonomous R&D Institutions 255.1 1.86 279.10 2.00 227.10 2.00 328.10 2.00Annual Report 2009-2010 199

Annexures(Rs. in crores)S. Name of the Programme/Schemes/Projects Expeniture during B. E. 2009-10 R.E. 2009-10 B.E. 2010-11No 2008-2009Plan Non- Plan Non- Plan Non- Plan Non-Plan Plan Plan Plan3 Assistance to other Scientific Bodies3.01 Human Resource Development 29.93 0.00 35.00 0.00 36.00 0.00 54.00 0.003.02 Bioinformatics 21.91 0.00 20.00 0.00 22.00 0.00 19.00 0.003.03 Research and Development 303.08 0.00 340.90 0.00 308.90 0.00 394.40 0.003.04 Biotechnology for Societal Development 9.88 0.00 10.00 0.00 9.40 0.00 13.00 0.003.05 Grand Challenge Programmes 51.68 0.00 40.00 0.00 38.00 0.00 54.00 0.003.06 Programme for Promotion of Excellence and Inovation 66.16 0.00 45.00 0.00 42.40 0.00 42.50 0.003.07 Biotech Facilities 25.00 0.00 20.00 0.00 20.00 0.00 27.00 0.00Total = Assistance to other Scientific Bodies 507.64 0.00 510.90 0.00 476.70 0.00 603.90 0.004.01 I & M Sector - Assistance for Technology Incubators, Pilot Projects, 31.99 0.00 5.00 0.00 3.00 0.00 5.00 0.00Biotechnology Parks and Biotech Development Fund4.02 I & M Sector - Public Private Partnership 57.70 0.00 90.00 0.00 90.00 0.00 118.00 0.00Total - I&M Sector 89.69 0.00 95.00 0.00 93.00 0.00 123.00 0.005 International Cooperation 12.60 0.00 15.00 0.00 15.00 0.00 25.00 0.006 I.C.G.E.B 0.00 10.00 0.00 10.00 0.00 10.00 0.00 10.007 Provision for projects/schemes for the benefit of North Eastern 0.00 0.00 100.00 0.00 90.20 0.00 120.00 0.00Region and SikkimTotal = Revenue Section 865.03 21.23 1,000.00 24.00 902.00 23.90 1,200.00 22.00200Annual Report 2009-2010

AnnexuresAnnexure-IIIList of patents filed/applied by DBT institutions: 2009-2010National Institute of Immunology, New Delhi• Composition useful for treatment of diabetes & chronic disorder: (a) 914/DEL/2008 on 13.10.2008; (b)PCT/IN2008/000664 on 13.10.2008; (c) AUS-2009200906 on 6.3.2009; (d) EPO-09155775.1-2107 on 20.3.2009; (e) JP-2009-90219 on 7.4.2009; (f) US-12/419,669 on 6.4.2009; (g) SA-2009/2374• Mechanism-Based Inhibitors of Transthyretin Amylodosis Studies with Bephenyl Ethers and new StructuralTemplates. 2345/DEL/2008 0n 13.10.2008.• Pharmaceutical composition for therapeutic or prophylactic treatment of bacterial infections and associateddiseases: 2345/DEL/2008 on 13.10.2008• Process for detection of inclusion bodies and process thereof. PCT/IN2008/000297. 13.5.2008• The therapeutic effect of bone marrow-derived liver cells in the treatment of Haemophilia. US PatentApplication # 61/280,188; 30.10.2009National Centre for Cell Sciences, Pune• Synthesis of new fluconazole analogues containing 1,2,3-triazole moiety and having better antifungalspectrum. Aher, N.G., Pore, V.S., Mishra, N.N., Kumar, A., Shukla, P.K., Sharma, A., and Bhat, M.K.Indian patent application IPO/ 054/NF/2008 Jointly with NCL, Pune, 2008• “Tumor suppressor activation polypeptide and uses thereof” Indian patent file number: IPR/4.19.20/06083/2006. International patent is filed for USA and European countries. Ref: 06083 (PCT).Centre for DNA Fingerprinting and Diagnosis, Hyderabad• Chandrashekharaiah, Rao CG, Sivaprasad V, Babu M R, Goel AK, Prasad J and Nagaraju J . Superior,Novel quality mulberry silkworm hybrids and a method thereof. Indian Patent 218895, granted on 16April, 2008.• Nagaraju J. Novel FISSR-PCR primers and methods of identifying genotyping diverse genomes of plantand animal systems including rice varieties, a kit thereof. Indian Patent 219765, granted on 13 May, 2008.• Gowrishankar J and Harinarayanan R. A method of altering levels of plasmids. ; Japanese Patent 4142649,granted on 20 Jun, 2008.; German Patent DE 60224457 T2, granted on 5 Feb, 2009.Annual Report 2009-2010 201

Annexures• Hasnain SE and Prachee C. Peptide antigens which elicit high humoral Immune response and T-cell response.Indian Patent 227179, granted on 5 Jan, 2009.• Gowrishankar J and Nandineni M R. A microbial process for arginine production. Indian Patent 230540,granted on 27 Feb, 2009.National Institute of Plant Genome Research, New Delhi• Chattopadhyay D, Yadav R K, Shukla R K. Chimeric Construct of Mungbean Yellow Mosaic India Virus(MYMIV) and its uses thereof (IPA- 2571/DEL/2007 and PCT/IN/2008/000819)• Datta A, Chakraborty S, Chakraborty N, Ghosh S, Meli S V. Polynucleotide sequence of fruit softeningassociated α-mannosidase and its uses for enhancing fruit shelf life (IPA-1647/DEL/2008)• Datta A, Chakraborty S, Chakraborty N, Meli S V, Ghosh S. Polynucleotide sequence of fruit softeningassociated α-D-N-acetyhexosaminidase and its uses for enhancing fruit shelf life (IPA-1648/DEL/2008)• Datta A, Bhattacharya S, Pal Bikas C, Sen J, Dasgupta S, Biswas A and Batra J. Process for production ofanti-diabetic compound in root culture of Catharanthus roseus (IPA-1649/DEL/2008)• Kumar S. A method for tandem cropping for increased production of food grain crops (applied:0033NF2008)International Centre for Genetic Engineering and Biotechnology, New Delhi• 94/DEL/2009. Kanury V. S. Rao, Dhiraj Kumar. Identification of human proteins as targets for the developmentof drugs against tuberculosis. 19 th January, 2009.• PCT/IN2009/000685. Dr. V.S.Reddy, Sadhu Leelavati and Amit Bhardwaj. A method of expressingforeign protein in plastids. 26 th November, 2009• PCT/1N2009/000487. Shashank Gupta, Rupak Singla, Digamber, Behera, Khalid Khyyam, ReshmaKorde, Pawan Malhotra, K.Natarajan & Varsha Srivastava. Pharmaceutical compositions containing CalciumChannel Blokers for treatment of Tuberculosis. 4 th September, 2009.202Annual Report 2009-2010

AnnexuresAnnexure IVList of patent applications filed/granted by Department of Biotechnology during theyear 2009-10 (upto 12-2-2010)The patent filing is done through National Research Development Corporation (NRDC). The number of patentapplications filed by DBT is 5 and number of Patent Granted is 11 during 2009-10 (till 12.2.2010). The detailsare as under:List of patent applications filed by during the year 2009-10 (upto 12-2-2010)S.No. Title of Patent Filed in1 A process for isolation of secondary metabolite from haloarchaeon strain IAH-I India2 Bioprocess for organic pollution and other biological alterations on stone surface India3 Islet neogenic potential of methanolic extract of Enicostemma littorate herb India4 Chemical cloak of human red blood cell surface antigenic determinants by sodium Indiaperiodate and choloro-acetic acid5 Method for processing and preparation of a cellular dermal matrix of rabbit origin IndiaList of patents granted during 2009-10 (up to 12-2-2010)S. No. Title Country1 An improved process for Oak Tasar (Antheraea proylei ) coocon cooking using Indiapineapple extract2 A process for producing reusable granular formulation of Trichorderma India3 A process for preparing high yields of Xylanase protein from Bacillus species India4 Primers for developing genotype specific marker in Piper nigrum India5 Chimeric helper B cell peptide vaccine for Japanese Encephalitis virus USAAnnual Report 2009-2010 203

AnnexuresS. No. Title Country6 A novel DNA marker for drought tolerance in plants Sri Lanka7 A method for diagnosis of tuberculosis by smear microscopy, culture and EPpolymerase chain reaction using processed clinical samples & kit thereof8 A polyherbal preparation useful for prevention of anthrosclerosis and ChinaHyperlipidermia9 A salt tolerant L myo-inositol 1 phosphate synthase and a process for obtaining Japanthe same10 A microcrystalline DNA marker used for identifying disease resistant population Singaporeof Penaeus monodon11 Gene OSISAPI of Rice confers tolerence to stress and a method thereof USA204Annual Report 2009-2010

AnnexuresAnnexure-VList of technologies transferred/developed: 2009-2010List of technologies transferred/readyAquaculture and marine biotechnology• “Molecular signal intervention technology” was ready for transfer to shrimp aqua culture farms (M/s.VaishaliAqua farm; Best India Marine harvest farm) for commercial application after validation of results.National Institute of Immunology, New Delhi• Slow Insulin Release Formulation (SIRF) for Treatment of Diabetes: A slow insulin release formulation hasbeen developed and licensed to a US based company for up-scaling. An initial amount of US$ 50,000/-has been received. The terms and conditions of the agreement restrict the revealing of certain details till thetechnologies go to Phase-I clinical trials. Use of SIRF technology is being exploited for other diseases likeosteoporosis and arthritis.International Centre for Genetic Engineering and Biotechnology, New Delhi• Dengue specific antigens and monoclonal antibodies, technology was transferred to Diagnostics Enterprises,26, Sector-1, Industrial Estate, Parwanoo – 173220, Himachal Pradesh, IndiaList of technologies/products developedEnvironmental Biotechnology• Technology for pulp and paper mill effluent for removal of colour by fungus (Cryptococcus albidus), anddegradation of chlorinated phenols by bacterial consortium.• Technology for preparation of biopulp and paper from sugar cane bagasse by lignin-degrading fungus-Cryptococcus albidus.• Technology for preparation of bioethanol production from lignocellulosic waste of bagasse in pulp andpaper mill effluent by lignin-degrading fungus- Cryptococcus albidus.National Institute of Plant Genome Research, New Delhi• A highly virulent construct of Mungbean Yellow Mosaic India Virus (MYMIV) for screening ofMYMIV-resistant soybean cultivar by agroinoculation.Annual Report 2009-2010 205

AnnexuresAnnexure VIImportant Bioinformatics Trainings/Workshops organized during 2009-10Aligarh Muslim University, AligarhTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedExploring Proteins with Bioinformatics 15 -16 January, DISC, Aligarh Muslim2010 University, AligarhAnna University, ChennaiTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedWorkshop on Sequence Analysis and 2 - 5 December, Distributed InformationMolecular Simulation 2009 Centre, Centre forBiotechnology, AnnaUniversity, ChennaiCIMAP, LucknowTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedCIMAP Summer Training on Tools 23 June -22 July, Central Institute ofand Techniques of Biotechnology and 2009 Medicinal and AromaticBioinformatics (CST-2009)Plants, Lucknow206Annual Report 2009-2010

AnnexuresCSRTI, MysoreTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedWorkshop on Bioinformatics and its 9-10, November, SubDIC, CSRTI, MysoreApplications 2009G. B. Pant University of Agriculture & Technology, PantnagarTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedBiological Sequence Analysis and 2 - 4 September, Bioinformatics Centre,applications in Agriculture & Veterinary 2009 MBGE, G. B. PantSciencesUniversity of Agriculture &Technology, PantnagarHimachal Pradesh University, ShimlaTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedFunctional & Structural Proteomics: 26 - 30 October, Bioinformatics Center,An In Silico Approach 2009 Himachal PradeshUniversity, ShimlaIIT, MadrasTitle of the Workshop Date of the Event Name of the CentreWorkshop on Techniques in 5 - 7 March, 2009 BIF, Department ofBioinformaticsBiotechnology, MadrasAnnual Report 2009-2010 207

AnnexuresIIT, KharagpurTitle of the Workshop Date of the Event Name of the CentreWorkshop on Bioinformatics in 9 - 10 October, Bioinformatics Sub-DIC,Genomics and Proteomics 2009 IIT KharagpurIISR, CalicutTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedRecent advances in EST analysis and 20-23 October. Bioinformatics centre,their annotation 2009 Indian Institute of SpicesResearchKalyani University, KalyaniTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedCurrent trends in Evolutionary 17 - 20 February, BIF, Kalyani UniversityBiology 2010Kerala Agricultural University, ThrissurTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedTraining on Bioinformatics in genome 18 May to Bioinformatics Centre,Keralaand proteome analysis 18 June, 2009 Agricultural UniversityWinter School on Bioinformatics and 1-21 December, Bioinformatics Centre, Keralaits Applications in Agriculture 2009 Agricultural University208Annual Report 2009-2010

AnnexuresMadras Veterinary College, MadrasTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedAn ICAR Summer School on 02-22 Sep., 2009 Bioinformatics CentreBioinformatics and its applicationsMadras Veterinary Collegein Animal Health and VeterinaryResearchMaharani Science College for Women, BangaloreTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedTwo month courses (2) in Basic March to April, DBT-BIF, Maharani’s ScienceBioinformatics program for B.Sc. 2009 & August to College for Women,students Oct., 2009 Bangalore.Two months certificate course on 7th March - 2nd DBT-BIF, Maharani’s ScienceBioinformatics program for M.Sc., May, 2009 College for Women,students of Maharani’s ScienceBangalore, in collaborationCollege for Women, Bangalorewith Institute ofand Government Science College,Computational BiologyBangaloreMahatma Gandhi Institute of Medical Sciences, SevagramTitle of the Workshop/ Date of the Event Name of the CentreTraining Organized12 th Workshop on Biomedical 20-21 November, Bioinformatics Centre JBTDInformatics & Communication 2009 Research Centre, MahatmaGandhi Institute of MedicalSciences, Sevagram (Wardha)Online Health Informatics Certificate July – December,Course (6 months duration) 2010Annual Report 2009-2010 209

AnnexuresMizoram University, MizoramTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedTraining Course On Bioinformatics 27 November, BIF, Mizoram University– Current Approaches and Applications 2009Training Course On Basics of 29 - 30 March, BIF, Mizoram UniversityBioinformatics – An Overview 2010(for Undergraduate/School students)Shri Mata Vaishno Devi University, KatraTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedTrends in Bioinformatics 16 -19 March 2009 BIF Centre, Shri MataVaishno Devi University, KatraVidyasagar University, West BengalTitle of the Workshop/ Date of the Event Name of the CentreTraining OrganizedComputational and Structural 08.09.09 – BIF Centre, Department ofBiology - 10th Workshop 10.09.09 Microbiology, VidyasagarUniversity, West Bengal210Annual Report 2009-2010

AbbreviationsAbbreviations2D-LC-MSAAI-DUAAUAAUACTRECAICRAPAIDSAIIMSAMFANGRAUARIB.E.B.Tech.BACBCBCILBEBHUBLBBODBSIBTCARICARICBSCCMBCCSUCDFD2 Dimensional Liquid chromatography - Mass SpectrometryAllahabad Agricultural Institute - Deemed UniversityAssam Agricultural University, JorhatAnand Agricultural University, AnandAdvanced Centre for Treatment, Research & Education in CancerAll India Coordinated Research projectAcquired Immunodeficiency SyndromeAll India Institute of Medical SciencesArbuscular Mycorhigal FungiAcharya NG Ranga Agricultural UniversityAgharkar Research InstituteBachelor of EngineeringBachelor of TechnologyBacterial Artificial ChromosomeBack CrossBiotech Consortium India LimitedBudgeted ExpenditureBanaras Hindu UniversityBacterial Leaf BlightBiochemical Oxygen DemandBotanical Survey of IndiaBacillus thuringiensisCentral Avian Research Institute, IzatnagarCentral Agricultural Research Institute, Port BlairCystathionine beta - synthaseCentre for Cellular and Molecular BiologyChaudhary Charan Singh UniversityCenter for DNA Fingerprinting and DiagnosticsAnnual Report 2009-2010 211

AbbreviationscDNAcDNA- AFLPCDPKCECLRICMCCMERICMICOPDCRCCROCRRICSIRCSKHPCSMCRICSRTICSSRICTR&TIDBTDELCONDNADORDRRDWRECMEDELISAEPNESTComplimentary Deoxyribo Nucleic AcidcDNA - Amplified Fragment Length PolymorphismCalcium Dependent Protein KinasesCombinatorial ExtensionCentral Leather Research InstituteChristian Medical CollegeCentral Mechanical Engineering Research InstituteCell-mediated immune responseChronic Obstructive Pulmonary DiseaseChawki Rearing CentreClinical Research OrganizationCentral Rice Research InstituteCouncil of Scientific and Industrial ResearchCSK HP Agricultural UniversityCentral Salt and Marine Chemicals Research InstituteCentral Sericulture Research and Training Institute, MysoreCentral Soil Salinity Research Institute, KarnalCentral Tassar Research and Training InstituteDepartment of BiotechnologyDBT Electronic Library ConsortiumDeoxyribo Nucleic AcidDirectorate of Oilseeds ResearchDirectorate of Rice ResearchDirectorate of Wheat ResearchEctomycorrhizal FungiEnvelope DomainEnzyme linked immuno sorbant assayEntomopathogenic NematodesExpressed Sequence Tag212Annual Report 2009-2010

AbbreviationsEST-SSRExpressed Sequence Tags - Single Sequence Repeat MarkersFMDFoot and Mouth DiseaseFRETForster Resonance Energy TransferFTIRFourier Transform Infrared SpectroscopyGBPUATGobind Ballabh Pant University of Agriculture and Technology, PantnagarGC-MSGas Chromatography - Mass SpectrometryGEACGenetic Engineering Approval CommitteeGFPGreen Fluorescent ProteinGMGenetically ModifiedGMOGenetically Modified OrganismGMPGood Manufacturing PracticesGNDUGuru Nanak Dev UniversityGSTGlutathione S-TransferaseHAPHydroxyapatiteHCVHepatitis - C VirusHDLHigh Density LipoproteinHDPEHighdensity PolyethyleneHIVHuman Immunodeficiency VirusHPLCHigh Performance Liquid ChromatographyHPMCHydroxypropyl Methyl CelluloseHSPHeat Shock ProteinHsp70 Heat Shock Protein 70HUFAHighly Unsaturated Fatty AcidHUVECHuman Unbiblical cord Vein Endothelial CellHVAHordeum Vulgare AleunncheIARIIndian Agricultural Research Institute, New DelhiIBRCInstitute of Biotechnology & Research CentreIBSDInstitute of Bioresource and Sustainable DevelopmentICARIndian Council Agricultural of ResearchAnnual Report 2009-2010 213

AbbreviationsICGEBICMRICRISATICTICUIFNIGFRIIgGIGKVVIHBTIICBIICTIIHRIILIIPRIIRIIIScIISERIITIITRILSIMTECHIPCLIPMIPTGISSRITS regionIVRIJAInternational Centre for Genetic Engineering and BiotechnologyIndian Council of Medical ResearchInternational Crop Research Institute for Semi Arid TropicsInstitute of Chemical TechnologyIntensive Care UnitInterferonIndian Grassland and Fodder Research InstituteImmunoglobulin - GIndira Gandhi Krishi Vishva VidyalayaInstitute of Himalayan Bioresource TechnologyIndian Institute of Chemical BiologyIndian Institute of Chemical TechnologyIndian Institute of Horticultural ResearchIndian ImmunologicalsIndian Institute of Pulses ResearchInternational Rice Research InstituteIndian Institute of Science, BangaloreIndian Institute of Science Education & ResearchIndian Institute of TechnologyIndian Institute of Toxicology ResearchInstitute of Life SciencesInstitute of Microbial TechnologyIndian Petro - Chemical LimitedIntegrated Pest ManagementIsopropyl β-D-1-thiogalactopyranosideInter Simple Sequence RepeatsInternal Transcribed SpacerIndian Veterinary Research InstituteJunior Analyst214Annual Report 2009-2010

AbbreviationsJAI VACJAOJNUKSSR&DIKVKLC-MSLDCLIGsLMOLSMABMAPMAPMAPKMASMDCK CellsMEMSMIRSMKUMOUMPUATmRNAMSSRFMVScNARINBAGRNBFGRNBRCNCCSJai Vigyan Mission VaccineJunior Accounts OfficerJawaharlal Nehru UniversityKarnataka State Sericulture Research and Development InstituteKrishi Vigyan KendraLiquid Chromatography - Mass SpectrometryLower Division ClerkLaboratory Implementation GuidanceLiving Modified OrganismLinalool SynthaseMolecular AntibodiesMitogen-Activated ProteinMedicinal and Aromatic PlantsMitogen-activated Protein KinaseMarker Assisted SelectionMadin-Darby Canine Kidney CellsMicro - Electro Mechanical SystemMultiple Insecticide Tolerant (resistant) StrainMadurai Kamaraj UniversityMemorandum of UnderstandingMaharana Pratap University of Agriculture & TechnologyMessenger Ribonucleic AcidM.S. Swaminathan Research FoundationMasters of Veterinary ScienceNational Aids Research InstituteNational Bureau of Animal Genetics ResourcesNational Bureau of Fish Genetics ResourcesNational Brain Research CentreNational Centre for Cell ScienceAnnual Report 2009-2010 215

AbbreviationsNCLNDRINENGONIANPNIHNIINIISTNIPERNIPGRNIRRHNMUNPLNRCNRCPBNRSCOMPORFPAUPCPCRPCTPDBCPEGPGPRPLGAPPVCPSBPVR IINational Chemical LaboratoryNational Dairy Research InstituteNorth EastNon Governmental OrganizationNational Institute of Animal Nutrition and PhysiologyNational Institute of HealthNational Institute of ImmunologyNational Institute for Interdisciplinary Science and TechnologyNational Institute of Pharmaceutical Education and ResearchNational Institute of Plant Genome ResearchNational Institute of Research on Reproductive HealthNorth Maharashtra UniversityNational Physical LaboratoryNational Research CentreNational Research Centre on Plant BiotechnologyNational Remote Sensing CentreOuter Membrane ProteinsOpen Reading FramePunjab Agricultural UniversityPersonal ComputerPolymerase Chain ReactionPatent Cooperation TreatyProject Directorate of Biological ControlPolyethylene GlycolPlant Growth Promoting RhizobacteriaPoly (lactic-co-glycolic acid)Parliament, Protocol, Vigilance and CoordinationPhosphate Solubilizing BacteriaPlasmodium vivax Recombinant II216Annual Report 2009-2010

AbbreviationsQPMQTLRAPDRBCRERGCBRILRNA.irRNART- PCRS&TSASSCSCARSCTIMSTSDS PAGESEMSGPGIMSSHGSiRNASMESNPSNP- CAPSSPMESRMCRI, ChennaiSSFSSRSTSVVUQuality Protein MaizeQuantitative Trait LociRandom Amplified Polymorphic DNARed Blood CellRevised ExpenditureRajiv Gandhi Centre for BiotechnologyRecombinant Inbred LinesRNA InterferenceRibosomal Ribonucleic AcidReal Time Polymerase Chain ReactionScience and TechnologySociety for Applied StudiesScheduled CasteSequence Characterized Amplified RegionSree Chitra Thirunal Institute for Medical Sciences & Technology,Sodium Dodicyl Sulphate Polymerase Acryl Gel ElectrophoresisScanning Electron MicroscopeSanjay Gandhi Postgraduate Institute of Medical SciencesSelf Help GroupsSmall Interfering Ribonucleic AcidSmall and Medium EnterprisesSingle Nucleotide PolymorphismSingle Nucleotide Polymorphism - Cleaved Amplified Polymorphic Sequence markersSolid Phase Micro ExtractionSri Ramachandra Medical College & Research Institute, ChennaiSolid State FermentationSimple Sequence RepeatsScheduled TribeSri Venkateswara Veterinary UniversityAnnual Report 2009-2010 217

AbbreviationsTaVAPTDSTERITGMSTNTNAUToCLVUASUDCUDSCUGCUKUPUPSCURLUSUSAUVVMSRFVNTRWHOTranslation Initiation Vesicle -Associated Membrane ProteinTotal Dissolved SolidsThe Energy and Resources InstituteThermo Genetic Male SterilityTamil NaduTamil Nadu Agricultural UniversityTomato Leaf Curl VirusUniversity of Agricultural SciencesUpper Division ClerkUniversity of Delhi South Campus, New DelhiUniversity Grants CommissionUnited KingdomUttar PradeshUnion Public Service CommissionUniform Resource LocatorUnited StatesUnited States of AmericaUltra VioletVittal Mallya Scientific Research FoundationVariable Number Tandem RepeatWorld Health Organization218Annual Report 2009-2010

Annual Report 2009-2010 - Department of Biotechnology

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