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Research Articles

Vol. 12 No. sp3 (2025): Advances in Plant Health Improvement for Sustainable Agriculture

Bacillus subtilis (Bs-PLM), a bacterial bio-fungicide, effectively suppresses the chilli pathogen Colletotrichum truncatum by eliciting the host defense response

DOI
https://doi.org/10.14719/pst.8646
Submitted
3 April 2025
Published
08-12-2025

Abstract

The present study was conducted to evaluate the efficacy of the bacterial bio-fungicide Bacillus subtilis against Colletotrichum truncatum, the causal agent of anthracnose and fruit rot in chilli (Capsicum annuum L.). A total of 10 Bacillus spp. isolates were obtained from the rhizosphere soil of major chilli-growing regions in Tamil Nadu, India. Characterisation through various methods, including molecular analysis, confirmed that these isolates belonged to the Bacillus genus. Additionally, molecular and in vitro screening identified Bacillus subtilis (Accession No. MZ618269) and Bacillus cereus (Accession No. MZ618270) as highly effective in inhibiting C. truncatum. Furthermore, PCR analysis revealed that Bacillus spp. isolates Bs-PLM and Bc-ADP possessed the genes responsible for the synthesis of the antibiotics iturin and surfactin. GC–MS analysis indicated that these isolates produced more than 30 antimicrobial compounds at different retention times, with the most notable antifungal compounds being n-Nonadecanol-1, 1-Hexadecanol, Behenic alcohol and Dibutyl phthalate. The volatile compounds released by Bs-PLM and Bc-ADP exhibited the highest mycelial growth inhibition. Under in vivo conditions, fruit lesion areas were significantly reduced when pathogen-inoculated fruits were treated with volatiles produced by these Bacillus strains compared to the untreated control fruits. Therefore, the Bacillus spp. could be used for the effective management of anthracnose and fruit rot disease in chillies.

References

  1. 1. Darshan S, Seeja G, Manju RV, Priya RU, Kumar SM. Combining ability analysis in chilli (Capsicum annum L.) to identify suitable parents for hybrid production. Life Sci Intl Res J. 2017;4(1):15-8.
  2. 2. Olimpia P, Ion S, Gigel P, Eliza TS. Content of vitamin C, beta-carotene, potassium and phosphorus in the local pepper populations. https://www.cabidigitallibrary.org/doi/pdf/10.5555/20183050179
  3. 3. Sanati S, Razavi BM, Hosseinzadeh H. A review of the effects of Capsicum annuum L. and its constituent, capsaicin, in metabolic syndrome. Iran J Basic Med Sci. 2018;21(5):439. https://doi.org/10.22038/ijbms.2018.25200.6238
  4. 4. Sahitya UL, Sri Deepthi R, Krishna M. Anthracnose, a prevalent disease in capsicum. Res J Pharm Biol Chem Sci. 2014; 5(3):1583-94. http://rjpbcs.com/pdf/2014_5(3)/[163].pdf
  5. 5. Cannon PF, Damm U, Johnston PR, Weir BS. Colletotrichum: current status and future directions. Stud mycol. 2012;73(1):181-213. https://doi.org/10.3114/sim0014
  6. 6. Damm U, Woudenberg JH, Cannon PF, Crous PW. Colletotrichum species with curved conidia from herbaceous hosts. Fungal Diversity. 2009;39(1):45-87.
  7. 7. Bajpai VK, Kang SC. In vitro and in vivo inhibition of plant pathogenic fungi by essential oil and extracts of Magnolia liliflora Desr. J Agric Sci Technol. 2025;14(4):845-56.
  8. 8. Kohl J Kolnaar R, Ravensberg WJ. Mode of action of microbial biological control agents against plant diseases: Relevance beyond efficacy. Front Plant Sci. 2019. https://doi.org/10.3389/fpls.2019.00845
  9. 9. Fungicide Resistance Action Committee Fungicide Resistance Management. 2020. Available from: https://www.frac.info/fungicide-resistance-management/background
  10. 10. Hadi MR, Balali GR. The effect of salicylic acid on the reduction of Rhizoctonia solani damage in the tubers of marfona potato cultivar. Am Eurasian J Agric Environ Sci. 2010;16;7(4):492-96. https://www.cabdirect.org/cabdirect/abstract/20103179522
  11. 11. Sarwar N, Zahid MH, Ashfaq S, Jamil FF. Induced systemic resistance in chickpea against Ascochyta blight by safe chemicals. Pak J Bot. 2011;43(2):1381-7.
  12. 12. Jaiganesh V, Eswaran A. Biochemical changes in rice plants due to application of bioinoculant, organic product, plant activator and moculation of Pyricularia oryzae. Int J Plant Prot. 2012; 5(2): 405-12
  13. 13. Agrios GN. Plant pathology Fifth Edition Elsevier Academic Press, California, USA. 2005. p. 348-57.
  14. 14. Canton H. Food and Agriculture Organization of the United Nations — FAO. In: The Europa Directory of International Organizations 2021. 23rd ed. London: Routledge. 2021. p. 297–305. https://doi.org/10.4324/9781003179900-41
  15. 15. Rangaswami G, Mahadevan A. Diseases of crop plants in India. PHI Learning Pvt. Ltd. 1998.
  16. 16. Altschul SF, Carroll RJ, Lipman DJ. Weights for data related by a tree. J Mol Biol.1989;207(4):647-53. https://doi.org/10.1016/0022-2836(89)90205-9
  17. 17. He Y, Zhu M, Huang J, Hsiang T, Zheng L. Biocontrol potential of a Bacillus subtilis strain BJ-1 against the rice blast fungus Magnaporthe oryzae. Can J Plant Pathol. 2019;41(1):47-59. https://doi.org/10.1080/07060661.2018.1564792
  18. 18. Sambrook J. Molecular cloning: A laboratory manual. Cold Spring Habor Laboratory. 1989.
  19. 19. Amutha K, Kokila V. PCR amplification, sequencing of 16S rRNA genes with universal primers and phylogenetic analysis of Pseudomonas aeruginosa. Int J Sci Res. 2014;3(8):257-61.
  20. 20. Dennis C, Webster J. Antagonistic properties of species-groups of Trichoderma: I. Production of non-volatile antibiotics. Trans Br Mycol Soc. 1971;57(1):25-IN3. https://doi.org/10.1016/S0007-1536(71)80077-3
  21. 21. Vincent JM. Distortion of fungal hyphae in the presence of certain inhibitors. Nature. 1947;159(4051):850. https://doi.org/10.1038/159850b0
  22. 22. Prapagdee B, Tharasaithong L, Nanthaphot R, Paisitwiroj C. Efficacy of crude extract of antifungal compounds produced from Bacillus subtilis on prevention of anthracnose disease in Dendrobium orchid. 2012;5(1):32-38.
  23. 23. Zheng M, Shi J, Shi J, Wang Q, Li Y. Antimicrobial effects of volatiles produced by two antagonistic Bacillus strains on the anthracnose pathogen in postharvest mangos. Biol Control. 2013;65(2):200-6. https://doi.org/10.1016/j.biocontrol.2013.02.004
  24. 24. Chung S, Kong H, Buyer JS, Lakshman DK, Lydon J, Kim SD, et al. Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soilborne pathogens of cucumber and pepper. Appl Microbiol Biotechnol. 2008;80(1):115-23. https://doi.org/10.1007/s00253-008-1520-4
  25. 25. Senthil Kumar M, Vinothkumar D, Saravana Kumar A, Aslam A, Shajahan A. The phytochemical constituents of Withania somnifera and Withania obtusifolia by GCMS analysis. Int J Pharmacogn Phytochem Res. 2011;3:31-4.
  26. 26. Gomez KA, Gomez AA. Statistical procedures for agricultural research. John wiley & sons. 1984.
  27. 27. Bharathi NK, Revathy N, Ebenezar EG, Gnanamalar RP. In vitro antagonistic activity of fungal and bacterial bio control agents against chilli fruit rot incited by Colletotrichum capsici. Int J Curr Microbiol Appl Sci. 2019;8(5):190-8. https://doi.org/10.20546/ijcmas.2019.805.023
  28. 28. Dev D, Konda S, Puneeth ME, Tanuja N, Singh P, Narendrappa T. In vitro evaluation of bioagents and botanicals against Colletotrichum gloeosporioides (Penz.) Penz&Sacc. causing anthracnose of pomegranate. Int J Ecol Environ Conserv. 2016;22(3):1229-32.
  29. 29. Hernandez Montiel LG, Zulueta Rodriguez R, Angulo C, Rueda Puente EO, Quiñonez Aguilar EE, Galicia R. Marine yeasts and bacteria as biological control agents against anthracnose on mango. J Phytopathol. 2017;165(11-12):833-40. https://doi.org/10.1111/jph.12623
  30. 30. Prasanna Kumar MK, Amruta N, Manjula CP, Puneeth ME, Teli K. Characterisation, screening and selection of Bacillus subtilis isolates for its biocontrol efficiency against major rice diseases. Biocontrol Sci Technol. 2017;27(4):581-99. https://doi.org/10.1080/09583157.2017.1323323
  31. 31. Lincoln L, More SS. Comparative evaluation of extracellular β-d-fructofuranosidase in submerged and solid-state fermentation produced by newly identified Bacillus subtilis strain. J Appl Microbiol. 2018;125(2):441-56. https://doi.org/10.1111/jam.13881
  32. 32. Li XY, Mao ZC, Wu YX, Ho HH, He YQ. Comprehensive volatile organic compounds profiling of Bacillus species with biocontrol properties by head space solid phase microextraction with gas chromatography-mass spectrometry. Biocontrol Sci Technol. 2015;25(2):132-43. https://doi.org/10.1080/09583157.2014.960809
  33. 33. Gao H, Li P, Xu X, Zeng Q, Guan W. Research on volatile organic compounds from Bacillus subtilis CF-3: biocontrol effects on fruit fungal pathogens and dynamic changes during fermentation. Front Microbiol. 2018;9:456. https://doi.org/10.3389/fmicb.2018.00456
  34. 34. Lourenço A, Kamnetz MB, Gadotti C, Diez-Gonzalez F. Antimicrobial treatments to control Listeria monocytogenes in queso fresco. Food Microbiol. 2017;64:47-55. https://doi.org/10.1016/j.fm.2016.12.014
  35. 35. Lopez DC, Sword GA. The endophytic fungal entomopathogens Beauveria bassiana and Purpureocillium lilacinum enhance the growth of cultivated cotton (Gossypium hirsutum) and negatively affect survival of the cotton bollworm (Helicoverpa zea). Biol Control. 2015;89:53-60. https://doi.org/10.1016/j.biocontrol.2015.03.010
  36. 36. Pathak KV, Keharia H. Identification of surfactins and iturins produced by potent fungal antagonist, Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) tree using mass spectrometry. 3Biotech. 2014;4(3):283-95. https://doi.org/10.1007/s13205-013-0151-3
  37. 37. Chowdhury SP, Hartmann A, Gao X, Borriss R. Biocontrol mechanism by root-associated Bacillus amyloliquefaciens FZB42–a review. Front Microbiol. 2015;6:780. https://doi.org/10.3389/fmicb.2015.00780
  38. 38. Mihalache G, Balaes T, Gostin I, Stefan M, Coutte F, Krier F. Lipopeptides produced by Bacillus subtilis as new biocontrol products against fusariosis in ornamental plants. Environ Sci Pollut Res. 2018;25(30):29784-93. https://doi.org/10.1007/s11356-018-3119-8
  39. 39. Ongena M, Jacques P, Touré Y, Destain J, Jabrane A, Thonart P. Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis. Appl Microbiol Biotechnol. 2005;69(1):29-38. https://doi.org/10.1007/s00253-005-1940-3
  40. 40. Kumar A, Rabha J, Jha DK. Antagonistic activity of lipopeptide-biosurfactant producing Bacillus subtilis AKP, against Colletotrichum capsici, the causal organism of anthracnose disease of chilli. Biocatal Agric Biotechnol. 2021;36:102133. https://doi.org/10.1016/j.bcab.2021.102133

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