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Biocontrol of Phyllosticta citricarpa by Bacillus spp.: biological and chemical aspects of the microbial interaction

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Abstract

Citrus fruits are the most produced fruits in the world, but they are threatened by several pathogens, including the fungus Phyllosticta citricarpa, the causal agent of citrus black spot (CBS). The fungus affects most citrus species and the infection results in economic losses in citrus-producing areas. This disease causes the aesthetic depreciation of fresh fruit, impairing its commercialization. As an alternative to the use of synthetic fungicides to control the pathogen, the biological control, using bacteria of the genus Bacillus, is highlighted. Such microorganisms enable biocontrol by the production of volatile organic compounds (VOC) or non-volatile. Therefore, this work aimed to evaluate the production of VOC by isolates of Bacillus spp. grown in different culture media; to evaluate the effects of these compounds on the evolution of CBS lesions in orange fruits; to study the effects of VOC on resistance induction in orange fruits; to evaluate the effects of VOC on P. citricarpa morphology in CBS lesions, and to identify the produced VOC. Tryptone soya agar (TSA) and tryptone soya broth (TSB) media used to culture the bacterium resulted in up to 73% pathogen inhibition by VOC. Volatile compounds from Bacillus spp. ACB-65 and Bacillus spp. ACB-73 when cultured in TSB culture medium provided 86% inhibition of freckles that evolved to hard spots. The volatile fractions produced by the bacteria were identified as alcohols, ketones, amines, ethers, aldehydes and carboxylic acids that can serve as arsenal against the phytopathogen. The present work demonstrated the potential of VOC produced by Bacillus spp. in the control of P. citricarpa.

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References

  • Adaskaveg JE, Förster H, Sommer NF (2002) Principles of postharvest pathology and management of decays of edible horticultural crops. In: Kader A (ed) Postharvest technology of horticultural crops, 4th edn. UC DANR Publ 3311. Oakland CA 163–195

  • Alamri SA (2015) Enhancing the efficiency of the bioagent Bacillus subtilis JF419701 against soil-borne phytopathogens by increasing the productivity of fungal cell wall degrading enzymes. Archives of Phytopathol and Plant Prot 48:159–170

    Article  CAS  Google Scholar 

  • Allain CC, Poon LS, Chan CSG, Richmond W, Fu PC (1974) Enzymatic determination of total serum colesterol. Clin Chem 120:470–475

    Article  Google Scholar 

  • Araujo FF, Menezes D (2009) Indução de resistência a doenças foliares em tomateiro por indutores biótico (Bacillus subtilis) e abiótico (Acibenzolar-S-Metil). Summa Phytopathol 35:169–172

    Article  Google Scholar 

  • Arrarte E, Garmendia G, Rossini C, Wisniewski M, Vero S (2017) Volatile organic compounds produced by Antarctic strains of Candida sake play a role in the control of postharvest pathogens of apples. Biol Control 109:14–20

    Article  CAS  Google Scholar 

  • Asari S, Matzén S, Petersen MA, Bejai S, Meijer J (2016) Multiple effects of Bacillus amyloliquefaciens volatile compounds: plant growth promotion and growth inhibition of phytopathogens. FEMS Microbiol. Ecology 92:fiw070

    Article  Google Scholar 

  • Ashwini N, Srividya S (2014) Potentiality of Bacillus subtilis as biocontrol agent for management of anthracnose disease of chilli caused by Colletotrichum gloeosporioides OGC1. 3 Biotech 4:127–136

    Article  CAS  PubMed  Google Scholar 

  • Bautista-Baños S (2014) Postharvest decay. Control Strategies. Elsevier, London, UK. Cano MP, Ancos B de, Matallana MC, Cámara M, Reglero G, Tabera J (1997) Differences among Spanish and Latin-American banana cultivars: morphological, chemical and sensory characteristics. Food Chem 59:411–419

    Google Scholar 

  • Cano MP, Ancos B, Matallana MC, Cámara M, Reglero G, Tabera J (1997) Differences among Spanish and Latin-American banana cultivars: morphological, chemical and sensory characteristics. Food Chem 59:411–419

    Article  CAS  Google Scholar 

  • Capdeville GD, Souza Júnior MT, Santos JRP, Miranda SP, Caetano AR, Falcão R, Gomes ACMM (2007) Scanning electron microscopy of the interaction between Cryptococcus magnus and Colletotrichum gloeosporioides on papaya fruit. Pesq Agropec Bras 42:1537–1544

    Article  Google Scholar 

  • Chaves-López C, Serio A, Gianotti A, Sacchetti G, Ndagijimana M, Ciccarone C, Paparella A (2015) Diversity of food-borne Bacillus volatile compounds and influence on fungal growth. J Appl Microbiol 119:487–499

    Article  PubMed  Google Scholar 

  • Chowdappa P, Kumar SM, Lakshmi MJ, Upreti KK (2013) Growth stimulation and induction of systemic resistance in tomato against early and late blight by Bacillus subtilis OTPB1 or Trichoderma harzianum OTPB3. Biol Control 65:109–117

    Article  Google Scholar 

  • Cui W, He P, Munir S, He P, Li X, Li Y, Wu J, Wu Y, Yang L, He Y (2019) Efficacy of plant growth promoting bacteria Bacillus amyloliquefaciens B9601–Y2 for biocontrol of southern corn leaf blight. Biol Control 139:104080

    Article  CAS  Google Scholar 

  • Di Francesco A, Ugolini L, Lazzeri L, Mari M (2015) Production of volatile organic compounds by Aureobasidium pullulans as a potential mechanism of action against postharvest fruit pathogens. Biol Control 81:8–14

    Article  Google Scholar 

  • Donahaye EJ (2000) Current status of non-residual control methods against stored product pests. Crop Prot 19:571–576

    Article  CAS  Google Scholar 

  • Effmert U, Kalderás J, Warnke R, Piechulla B (2012) Volatile mediated interactions between bacteria and fungi in the soil. J Chem Ecol 38:665–703

    Article  CAS  PubMed  Google Scholar 

  • Ezra D, Strobel GA (2003) Effect of substrate on the bioactivity of volatile antimicrobials produced by Muscodor albus. Plant Sci 165:1229–1238

    Article  CAS  Google Scholar 

  • Ezra D, Hess WM, Strobel GA (2004) New endophytic isolates of Muscodor albus, a volatile-antibiotic-producing fungus. Microbiol 150:4023–4031

    Article  CAS  Google Scholar 

  • Feliziani E, Romanazzi G (2013) Preharvest application of synthetic fungicides and alternative treatments to control postharvest decay of fruit. Stewart Postharvest Rev 9:1–6

    Article  Google Scholar 

  • Fernando WD, Ramarathnam R, Krishnamoorthy AS, Savchuk SC (2005) Identification and use of potential bacterial organic antifungal volatiles in biocontrol. Soil Biol and Biochem 37:955–964

    Article  CAS  Google Scholar 

  • Fujimoto A, Kupper KC (2016) Production of Antifungal Compounds and Hydrolytic Enzymes by Bacillus spp. as Mechanisms of Action against Phyllosticta citricarpa. IOSR J Agric and Vet Sci 9:19–27

    Google Scholar 

  • Giassi V, Kiritani C, Kupper KC (2016) Bacteria as growth-promoting agents for citrus rootstocks. Microbiol Res 190:46–54

    Article  PubMed  Google Scholar 

  • Gotor-Vila A, Teixidó N, Di Francesco A, Usall J, Ugolini L, Torres R, Mari M (2017) Antifungal effect of volatile organic compounds produced by Bacillus amyloliquefaciens CPA-8 against fruit pathogen decays of cherry. Food Microbiol 64:219–225

    Article  CAS  PubMed  Google Scholar 

  • Graham JH, Gottwald TR, Timmer LW, Bergamin Filho A, Van Den Bosch F, Irey MS, Takeuchi Y (2014) Response to “Potential distribution of citrus black spot in the United States based on climatic conditions.” Eur J Plant Pathol 139:231–234

    Article  Google Scholar 

  • Herman A, Tambor K, Herman A (2016) Linalool affects the antimicrobial efficacy of essential oils. Curr Microbiol 72:165–172

    Article  CAS  PubMed  Google Scholar 

  • Hynes J, Müller CT, Jones TH, Boddy L (2007) Changes in volatile production during the course of fungal mycelial interactions between Hypholoma fasciculare and Resinicium bicolor. J Chem Ecol 33:43–57

    Article  CAS  PubMed  Google Scholar 

  • Kim PI, Ryu J, Kim YH, Chi YT (2010) Production of biosurfactant lipopeptides Iturin A, fengycin and surfactin A from Bacillus subtilis CMB32 for control of Colletotrichum gloeosporioides. J Microbio and Biotechnol 20:138–145

    Article  CAS  Google Scholar 

  • Klein MN, Silva AC, Lopes MR, Kupper KC (2013) Application of microorganisms, alone or in combination, to control postbloom fruit drop in citrus. Trop Plant Pathol 38:505–512

    Article  Google Scholar 

  • Klein MN, Silva AC, Kupper KC (2016) Bacillus subtilis based-formulation for the control of postbloom fruit drop of citrus. World J Microbiol and Biotechnol 32:205

    Article  Google Scholar 

  • Kupper KC, Gimenes-Fernandes N (2002) Isolamento e seleção de Bacillus spp. para o controle de Colletotrichum acutatum em flores destacadas de lima ácida ´Tahiti´. Summa Phytopathol 28:292–295

    Google Scholar 

  • Kupper KC, Correa FE, de Azevedo FA, Silva AC (2012) Bacillus subtilis to biological control of postbloom fruit drop caused by Colletotrichum acutatum under field conditions. Sci Hortic 134:139–143

    Article  Google Scholar 

  • Kupper KC, Moretto RK, Fujimoto A (2019) Production of antifungal compounds by Bacillus spp. isolates and its capacity for controlling citrus black spot under field conditions. World J Microbiol and Biotechnol 36:7

    Article  Google Scholar 

  • Lanza FE, Metzker TG, Vinhas T, Behlau F, Silva Junior GJ (2018) Critical fungicide spray period for citrus black spot control in São Paulo state, Brazil. Plant Dis 102:334–340

    Article  CAS  PubMed  Google Scholar 

  • Leelasuphakul W, Hemmanee P, Chuenchitt S (2008) Growth inhibitory properties of Bacillus subtilis strains and their metabolites against the green mold pathogen (Penicillium digitatum Sacc.) of citrus fruit. Postharvest Biol and Technol 48:113–121

    Article  CAS  Google Scholar 

  • Lisker N, Cohen L, Chalutz E, Fuchs Y (1983) Fungal infections suppress ethylene-induced phenylalanine ammonia-lyase activity in grapefruit. Physiol Plant Pathol 22:331–338

    Article  CAS  Google Scholar 

  • Liu W, Mu W, Zhu B, Liu F (2008) Antifungal activities and components of VOCs produced by Bacillus subtilis G8. Cur Res in Bacteriol 1:28–34

    Article  CAS  Google Scholar 

  • Lopes MR, Klein MN, Ferraz LP, Silva AC, Kupper KC (2015) Saccharomyces cerevisiae: a novel and efficient biological control agent for Colletotrichum acutatum during pre-harvest. Microbiol Res 175:93–99

    Article  PubMed  Google Scholar 

  • Lu L, Lu H, Wu C, Fang W, Yu C, Ye C, Shi Y, Yu T, Zheng X (2013) Rhodosporidium paludigenum induces resistance and defense-related responses against Penicillium digitatum in citrus fruit. Postharvest Biol and Technol 85:196–202

    Article  Google Scholar 

  • Mari M, Di Francesco A, Bertolini P (2014) Control of fruit postharvest diseases: old issues and innovative approaches. Stewart Postharvest Rev 10:1–4

    Article  Google Scholar 

  • Palou L, Valencia-Chamorro SA, Pérez-Gago MB (2015) Antifungal edible coatings for fresh citrus fruit: a review. Coatings 5:962–986

    Article  CAS  Google Scholar 

  • Parafati L, Vitale A, Restuccia C, Cirvilleri G (2017) Performance evaluation of volatile organic compounds by antagonistic yeasts immobilized on hydrogel spheres against gray, green and blue postharvest decays. Food Microbiol 63:191–198

    Article  CAS  PubMed  Google Scholar 

  • Paul I, Van Jaarsveld AS, Korsten L, Hattingh V (2005) The potential global geographical distribution of Citrus Black Spot caused by Guignardia citricarpa Kiely: The likelihood of disease establishment in the European Union. Crop Prot 24:297–308

    Article  Google Scholar 

  • Poveda J (2021) Beneficial effects of microbial volatile organic compounds (MVOCs) in plants. Appl Soil Ecol 168:104118

    Article  Google Scholar 

  • Poveda J, González-Andrés F (2021) Bacillus as a source of phytohormones for use in agriculture. Appl Microbiol Biotecnol 105:8629–8645

    Article  CAS  Google Scholar 

  • Queiroga CL, Duarte MCT, Ribeiro BB, Magalhães PM (2007) Linalool production from the leaves of Bursera aloexylon and its antimicrobial activity. Fitoterapia 78:327–328

    Article  CAS  PubMed  Google Scholar 

  • Shafi J, Tian H, Ji M (2017) Bacillus species as versatile weapons for plant pathogens: a review. Biotech & Biotech Equip 31:446–459

    Article  CAS  Google Scholar 

  • Silva FDAS, Azevedo CAV (2016) The Assistat Software Version 7.7 and its use in the analysis of experimental data. Afr J Agric Res 11:3733–3740

    Article  Google Scholar 

  • Silva RF, Pascholati SF, Bedendo IP (2007) Indução de resistência em tomateiro por extratos aquosos de Lentinula edodes e Agaricus blazei contra Ralstonia solanacearum. Fitopatol Bras 32:189–196

    Article  Google Scholar 

  • Solanki MK, Singh RK, Srivastava S, Kumar S, Kashyap PL, Srivastava AK (2015) Characterization of antagonistic-potential of two Bacillus strains and their biocontrol activity against Rhizoctonia solani in tomato. J Basic Microbiol 55:82–90

    Article  CAS  PubMed  Google Scholar 

  • Thilagavathi R, Saravanakumar D, Ragupathi N, Samiyappan R (2007) A combination of biocontrol agents improves the management of dry root rot (Macrophomina phaseolina) in greengram. Phytopathol Mediterr 46:157–167

    CAS  Google Scholar 

  • Toffano L, Fialho MB, Pascholati SF (2017) Potential of fumigation of orange fruits with volatile organic compounds produced by Saccharomyces cerevisiae to control citrus black spot disease at postharvest. Biol Control 108:77–82

    Article  CAS  Google Scholar 

  • Torres MJ, Brandan CP, Petroselli G, Erra-Balsells R, Audisio MC (2016) Antagonistic effects of Bacillus subtilis subsp. subtilis and B. amyloliquefaciens against Macrophomina phaseolina: SEM study of fungal changes and UV-MALDI-TOF MS analysis of their bioactive compounds. Microbiol Res 182:31–39

    Article  CAS  PubMed  Google Scholar 

  • Wheatley RE (2002) The consequences of volatile organic compound mediated bacterial and fungal interactions. Ant Van Leeuwenhoek 81:357–364

    Article  CAS  Google Scholar 

  • Xu B, Zhang H, Chen K, Xu Q, Yao Y, Gao H (2013) Biocontrol of postharvest Rhizopus decay of peaches with Pichia caribbica. Cur Microbiol 67:255–261

    Article  CAS  Google Scholar 

  • Yamamoto S, Shiraishi S, Kawagoe Y, Mochizuki M, Suzuki S (2015) Impact of Bacillus amyloliquefaciens S13–3 on control of bacterial wilt and powdery mildew in tomato. Pest Man Sci 71:722–727

    Article  CAS  Google Scholar 

  • Zohora US, Ano T, Rahman MS (2016) Biocontrol of Rhizoctonia solani K1 by Iturin A Producer Bacillus subtilis RB14 Seed Treatment in Tomato Plants. Adv Microbiol 6:424

    Article  CAS  Google Scholar 

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Acknowledgements

To the financial support of CAPES, which provided a scholarship to Andréia Fujimoto; to the Agricultural Microbiology Program of Universidade Estadual Paulista “Júlio de Mesquita Filho”, Jaboticabal/SP, Brazil, is gratefully acknowledged and the financial support of the INCT Citrus (Proc. CNPQ465440/2014–2 and FAPESP 2014/50880–0).

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Authors and Affiliations

  1. Agricultural Microbiology Program of Universidade Estadual Paulista “Júlio de Mesquita Filho”, Jaboticabal, São Paulo, 14884-900, Brazil

    Andréia Fujimoto

  2. Institute of Chemistry, Universidade Estadual de Campinas, Campinas, SP, CP 6154, 13083-970, Brazil

    Fábio Augusto & Taicia Pacheco Fill

  3. Programa de Pós-Graduação em Produção Vegetal e Bioprocessos Associados/Universidade Federal de São Carlos, Araras/São Paulo, CEP 13600-970, Brazil

    Rafael Kupper Moretto

  4. Centro Avançado de Pesquisa de Citros “Sylvio Moreira”, Instituto Agronômico, Cordeirópolis/São Paulo, CEP 13490-970, Brazil

    Katia Cristina Kupper

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  1. Andréia Fujimoto
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  2. Fábio Augusto
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Correspondence to Katia Cristina Kupper.

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Fujimoto, A., Augusto, F., Fill, T.P. et al. Biocontrol of Phyllosticta citricarpa by Bacillus spp.: biological and chemical aspects of the microbial interaction. World J Microbiol Biotechnol 38, 53 (2022). https://doi.org/10.1007/s11274-021-03214-z

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