See discussions, stats, and author profiles for this publication at: http://www.researchgate.net/publication/279201259 INHIBITORY EFFECT OF LEAF EXTRACTS ON SPORE GERMINATION OF PUCCINIA ARACHIDIS SPEG ARTICLE · JANUARY 2015 DOWNLOADS VIEWS 3 3 2 AUTHORS, INCLUDING: Suhas Kamble Shivaji University, Kolhapur 6 PUBLICATIONS 0 CITATIONS SEE PROFILE Available from: Suhas Kamble Retrieved on: 16 July 2015 Vyas Sonali and Yardi Veena / 35 J. Microb. World, Vol. 17(1-2), PP: (35-39) 2015 Microbiologists Society ISSN 0972 - 1487 INHIBITORY EFFECT OF LEAF EXTRACTS ON SPORE GERMINATION OF PUCCINIA ARACHIDIS SPEG. Bhimrao J. Patil and S. K. Kamble Department of Botany and Plant Protection, Sadguru Gadage Maharaj College, Karad, Dist.- Satara. Pin Code- 415124, Maharashtra, India. ABSTRACT Rust caused by puccinia arachidis Speg. is a destructive disease of groundnut on a world scale. An ecofriendly approach and alternative to chemical control measures of this disease, various plant extracts and are being involved to control the disease. The present investigation deals with the study of inhibitory effects of some plant extracts against spore germination of Puccinia arachidis Speg. causing groundnut rust. Leaf extracts of Eupatorium odoratum L., Hyptis suaveolens (L.) Poit., Cassia uniflora Mill., Vitex nigundo L., Blumea lacera (Burm. f.) DC. and Xantium indicum Koen. were used as phytofungicides against urediospore germination of Puccinia arachidis Speg. A chemical fungicide Propiconazole (0.05%) used as standard check and distilled water as a control. All plants showed the antifungal activity against spore germination over control. An aqueous leaf extract of Eupatorium odoratum L. showed the highest fungitoxicity followed by Hyptis suaveolens (L.) Poit. against spore germination. Key words:Leaf extracts, urediospores, Puccinia arachidis. INTRODUCTION Rust disease caused by Puccinia arachidis Speg. is prevalent throughout the groundnut growing areas. Losses in yield due to rust disease alone have been reported in the range of 1052% depending upon the variety. Rust disease also lowers the seed quality by reducing seed size and oil content (Lukose, C.M. et. al., 2008). There is reduction in pod yield due to this disease may be as high as 50-60%. As per Subrahmanyam et al., (1983) suggestion although fungicides are effective against rust disease, their mammalian toxicity and possible phytotoxic nature may restrict their use to control groundnut rust. Research activities proved that Neem leaves and seeds have the ability to kill some disease causing fungi, viruses and parasites (Basak and Chakraborty., 1968 ;Thind and Dahiya., 1977). A Spraying of neem leaf extract (2%) in combination with recommended fungicides recorded numerical superiority in reducing leaf spot and rust incidence in groundnut (Shivashankar and Kadam., 1993).The main objective of this study is to evaluate the fungicides and plant extracts for their efficacy against inhibition of urediospores germination under laboratory condition. 36 / Journal of Microbial World, Vol. 17, 2015 MATERIALS AND METHODS Fresh and Healthy selected plants like Hyptis suaveolens, Eupatorium odoratum, Cassia uniflora, Vitex nigundo, Blumea lacera and Xanthium strumariumwere collected from agricultural fields. Five grams of leaves were weighed, washed with tap water and wiped with blotting paper. They were crushed in a mortar and pestle with 5 ml with distilled water. The extract was filtered through four layered muslin cloth. The filtrate was centrifuged at 1600 rpm for 5 minutes and considered as stock solution and the concentrations of 1%, 2% and 3% were made by adding distilled water. The fungicide Propiconazole (0.05%) was used as standard check for their efficiency against spore germination. The distilled water was used as control. The fungal spores (urediospores) of Puccinia arachidis were isolated from diseased groundnut leaves and used as test organism. All experiments were carried out under laboratory condi- B A C D Photographs: (A) Groundnut rust infected leaf, (B) Rust pustules, (C) Spore germination in distilled water & (D) Spore germination in plant extract. Bhimrao J. Patil and S. K. Kamble / 37 tions. The effect of plant extracts on fungal spore germination was studied on slide. A drop of different concentrations of plant extract was taken on different slides taken. Fresh spores collected from single pustule were placed in the thin film of plant extract. In such a way that 30- 40 spores per microscopic field could be easily observed. All the slides were kept on moist blotting paper in petriplates to maintain humidity for 6, 12, 18 and 24 hours respectively. Counted the total number of spores and germinated spores under the single field of microscope (10 x 45). The mean of three observations (number of germinated spores) from different microscopic fields were calculated for replication I. In this way calculated replication II & III. Percentage of spore germination in three replications was calculated by the formula given by Raghavendra(2005).Percent inhibition of urediospores over control was calculated by Vincent’s formula (1947). Mean of three observations were considered as replication I. Likewise replication II and III considered. RESULT AND DISCUSSION It was revealed from the results (observation table) that different concentrations of plant extracts caused significant inhibition in the spore germination.However the maximum inhibition in the spore germination was found at highest concentration 3%. It was followed by 2% and 1% leaf extracts. The chemical fungicide (Propiconazole) showed the 100% inhibition of spore germination which is significantly superior over all plant extract treatments and control. The preliminary phytochemical analysis of Eupatorium odoratum showed the presence of alkaloids, saponins, tannins, flavonoids, phenolics, resins, glycosides and steroids (Umedum CU., 2013). The leaves of Blumea balsamifera afford the flavanoid luteolin (Ragasa et. al., 2005).The fungicidal activities of Cassia tora L. and its active principles are determined against Botrytis cinerea, Erysiphae graminis, Phytophthora infestans, Puccinia recondita, Pyricularia grisea and Rhizoctonia solani (Kim et. al., 2004). The treatment with Eupatorium odoratum and Hyptis suaveolensaresignificantly superior over the Blumea lacera, Cassia unifloraXanthium indicum and Vitex nigundo treatments. The treatments of all plant leaf extracts were not found to be significant but are superior over the control. Eupatorium odoratumshowed the higher inhibition of spore germination (68.93%) for 3% concentration at 6 hour incubation period followed by Hyptis suaveolens (65.34%) for the same concentration and incubation period. The disease is usually controlled by conventional fungicides applied as foliar spray however; high cost of fungicides and the problem of environmental pollution have stimulated investigation of alternatives strategies for the control of pest and pathogens (Lyon et al., 1995). Traditionally, the plant has been used for control of diseases of plant. Plants possess chemical compounds which are toxic to growth of diseases.Applications of plant extracts have shown significant reduction in the incidence of many diseases with corresponding increase in yield (Khan et al. 1996). According to Ghewande(1989),the leaf water extract of Neem was found very effective against the fungal disease leaf rust of groundnut plants caused by fungi Puccinia arachidis. Ganapathy and Narayanaswamy (1990) reported that neem oil from Azadirachta I ndica and leaf extract of Nerium odorum reduced the incidence of rust, late leaf spot and ring mosaic of groundnut. Therefore, the plant leaf extracts are the safe and ecofriendly option to control the groundnut rust. One can resourcefully use these extracts against the rust disease. 38 / Journal of Microbial World, Vol. 17, 2015 Table: Effect of leaf extracts on inhibition ofspore germination (%) over control of Pucciniaarachidis (Groundnut rust) after various incubation periods. Tr. No. Name Plant of the Concentrations of extract 1% 2% 3% Observations after various incubation periods (in hours). 6 12 18 24 12 18 24 6 12 18 54.83 52.65 51.13 61.10 58.72 56.19 53.78 65.34 62.18 60.52 57.91 60.56 58.86 56.35 54.41 65.58 62.05 59.86 57.63 68.93 66.08 64.54 61.20 49.39 47.24 45.58 43.76 53.01 49.73 49.44 46.60 54.22 52.29 53.22 49.19 43 41.93 40.55 38.01 45.85 43.67 42.13 39.38 46.52 44.53 43.09 40.44 46.71 47.01 44.37 53.37 48.97 46.22 43.38 54.96 52.21 51.46 48.55 49.32 47.64 46.84 44.14 51.96 53.05 51.63 47.73 51.04 54.94 54.17 50.43 Hyptissuaveolens 6 24 56.71 1 2 Eupatorium odoratum 3 Cassia uniflora 4 Vitexnigundo 5 Blumealacera 6 Xantiumindicum 7 Propiconazole (0.05%) 90 90 90 90 90 90 90 90 90 90 90 90 8 Control (Distilled Water) 00 (55.28) 00 (61.03) 00 (65.81) 00 (71.06) 00 (55.28) 00 (61.03) 00 (65.81) 00 (71.06) 00 (55.28) 00 (61.03) 00 (65.81) 00 (71.06) SE ± 1.57 0.92 1.04 1.38 1.61 0.81 0.62 0.66 1.51 1.17 0.57 1.11 C.D. (at 5%) 4.28 2.61 2.95 2.93 4.56 2.31 1.66 1.87 4.27 3.31 1.62 3.15 52.23 (Note: Figures in the parenthesis show germination percentage.) References: Basak, S. P., Chakraborty, D. P. (1968). Chemical investigation of Azadirachta indica leaf. Journal of the Indian Chem. Society, 45(5):466-467. Ganapathy, T. and Narayanaswamy, P. (1990). Effect of plant products on the incidence of major diseases of groundnut. Internation. Arachis Newslett., 7: 20-21. Ghewande, M. P. (1989). Management of foliar diseases of ground nut (Arachis hypogaea) using plant extracts. Indian J. of Agric. Sci., 59:133- 144. Khan, T. Z., Gill, M. A., Nasir M. A. and Bukhari S. A. (1996). Antifungal properties of some plant extracts. Pakistan Journal ofPhytopathology 10: 62-65. Kim, Y. M., Lee, C. H., H. G. Kim and Lee, H. S. (2004). Anthraquinones isolated from Cassia tora (Leguminosae) seed show an antifungal property against phytopathogenic fungi. J. Agric. Food Chem., 52: 6096-6100. Lukose, C.M., Moradia, A.M. and Kunadia, B.A. (2008). Diseases of groundnut in Gujarat and their management. Published research scientist (Groundnut). Main Oilseeds Research Station. Junagadh Agricultural University, Junagadh: 1-16. Bhimrao J. Patil and S. K. Kamble / 39 Lyon, G.D, Relinski, T. and Newton, A.C. (1995). Novel disease control compounds: Potential to immunize Plants against infection. Plant Pathology, 44:407-427. Ragasa C. Y., Co A. L. and Rideout. (2005). Antifungal metabolites from Blumea balsamifera. Nat. Prod. Res., 19: 231-237. Raghavendra, B. K.(2005). Epidemiology and management of chilli powdery mildew caused by Leveillula taurica (Lev). Arn., University of Agricultural Sciences, Dharwad: 20. Shivashankar, S. P. and Kadam, D. N.(1993). Efficacy of neem leaf extract against foliardiseases of groundnut Indian Phytopath., 45: 72 Subrahmanyam, P. and Mc Donald, D. (1983). Rust disease of groundnut. 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