Plant Protect. Sci., X:X | DOI: 10.17221/41/2025-PPS

Characterisation of Colletotrichum species associated with anthracnose disease in red chilli pepper (Capsicum annuum L.) in the South of Vietnam and the effectiveness of the consortium Bacillus amyloliquefaciens and Pseudomonas fluorescensOriginal Paper

Vo Thi Ngoc Ha ORCID...1, Huynh Thuong Vuong1, Tran Bao Thang1, Huynh Thanh Hung1
1 Plant Protection Department, Faculty of Agronomy, Nong Lam University Ho Chi Minh City, Ho Chi Minh City, Vietnam


Anthracnose is one of the most destructive diseases that limits pepper production and quality worldwide. In this study, the causal agent of anthracnose in red chilli pepper in Southern Vietnam was collected and identified based on morphological characteristics and multilocus sequence regions (ITS, β-tubulin, GPDH, ACT). The antifungal activity of Bacillus amyloliquefaciens and Pseudomonas fluorescens was evaluated in vitro and in vivo under greenhouse conditions. The results revealed that the morphological analysis categorised the Colletotrichum isolates into three species: C. acutatum, C. gloeosporioides, and C. scovillei. Sequence analysis of the four genes confirmed that C. scovillei was the causal agent of anthracnose in chilli pepper in Southern Vietnam. B. amyloliquefaciens and P. fluorescens bacteria demonstrated antifungal activity against C. scovillei in vitro, with mycelial growth inhibition rates ranging from 20.79% to 78.69%. The consortium of B. amyloliquefaciens CC-LD2.4, P. fluorescens CC-FN1.1, and P. fluorescens O-BT1.2 achieved 84.4% control efficacy at 7 days after inoculation (DAI), which declined to 68.5% at 14 DAI and 41.7% at 21 DAI, at the flowering stage, and achieved 100% control efficacy at the fruiting stage. B. amyloliquefaciens CC-LD2.4 showed very high chitinase, protease, and cellulase activities (halo diameter of 26.7 mm, 22.7 mm, 21.5 mm), whereas P. fluorescens CC-FN1.1 was very high in protease and cellulase (14.3 mm, 12.4 mm) but weak in chitinase (5.1 mm), and P. fluorescens O-BT1.2 exhibited overall lower activities (3.4–9.9 mm). There is still considerable room to optimise bacterial consortia to develop bio-fungicides that meet the requirements for an alternative or advanced solution for controlling anthracnose in red chilli peppers in sustainable agriculture.

Keywords: anthracnose; biological control; bacterial consortium; sustainable agriculture

Received: March 14, 2025; Revised: December 8, 2025; Accepted: December 10, 2025; Prepublished online: May 6, 2026 

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References

  1. Anderson J.M., Coates L.M., Aitken E.A.B., Mitchell R.W., McTaggart A.R., Dann E.K. (2024): The pathogenic diversity and host range of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia. Plant Pathology, 73: 1334-1348. Go to original source...
  2. Benbrook C., Kegley S., Baker B. (2021): Organic Farming lessens reliance on pesticides and promotes public health by lowering dietary risks. Agronomy, 11: 1266. Go to original source...
  3. Cai L., Hyde K.D., Taylor P.W.J., Weir B., Waller J., Abang M.M., Zhang J.Z., Yang Y.L., et al. (2009): A polyphasic approach for studying Colletotrichum. Fungal Diversity, 39: 183-204.
  4. Carbone I., Kohn L.M. (1999): A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia, 91: 553-556. Go to original source...
  5. de Andrade L.A., Santos C.H.B., Frezarin E.T., Sales L.R., Rigobelo E.C. (2023): Plant growth-promoting Rhizobacteria for sustainable agricultural production. Microorganisms, 11: 1088. Go to original source... Go to PubMed...
  6. De Silva D.D., Groenewald J.Z., Crous P.W., Ades P.K., Nasruddin A., Mongkolporn O., Taylor P.W.J. (2019): Identification, prevalence and pathogenicity of Colletotrichum species causing anthracnose of Capsicum annuum in Asia. IMA Fungus, 10: 8. Go to original source... Go to PubMed...
  7. Diao Y.Z., Zhang C., Liu F., Wang W.Z., Liu L., Cai L., Liu X.L. (2017): Colletotrichum species causing anthracnose disease of chili in China. Persoonia, 38: 20-37. Go to original source...
  8. Dikbas N., Ucar S., Tozlu G., Özer T., Kotan R. (2023): The effect of immobilised chitinase enzyme on the biocontrol of Sitophilus zeamais. Turkish Journal of Agriculture and Forestry, 47: 170-177. Go to original source...
  9. Dimkić I., Janakiev T., Petrović M., Degrassi G., Fira D. (2022): Plant-associated Bacillus and Pseudomonas antimicrobial activities in plant disease suppression via biological control mechanisms - A review. Physiological and Molecular Plant Pathology, 117: 101754. Go to original source...
  10. FAOSTAT (2024): Food and agriculture data. Available at: https://www.fao.org/faostat/en/#data/QCL. (Accessed Dec 24, 2024)
  11. Glass N.L., Donaldson G.C. (1995): Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology, 61: 1323-1330. Go to original source... Go to PubMed...
  12. Guerber J.C., Liu B., Correll J.C., Johnston P.R. (2003): Characterisation of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs, and mating compatibility. Mycologia, 95: 872-895. Go to original source...
  13. Hegde G.M., Monisha L. (2024): Challenges and potentials of microbial consortia for plant disease management anand sustainable productivity. Asia-pacific Biofertilizers and Biopesticides Information Platform. Available at https://apbb.fftc.org.tw/article/611 (accessed Dec 24, 2024).
  14. Ishii H., Watanabe H., Yamaoka Y., Schnabel G. (2022): Sensitivity to fungicides in isolates of Colletotrichum gloeosporioides and C. acutatum species complexes and efficacy against anthracnose diseases. Pesticide Biochemistry and Physiology, 182: 105049. Go to original source...
  15. Kanto T., Seiji U., Toshihide T., Jouji M., Naho Y., Toshiyuki U., Toyozo S. (2013): Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan. Journal of General Plant Pathology, 80: 73-78. Go to original source...
  16. Khalimi K., Darmadi A.A.K., Suprapta D.N. (2019): First report on the prevalence of Colletotrichum scovillei associated with Anthracnose on chili pepper in Bali, Indonesia. International Journal of Agriculture & Biology, 22: 363-368. Go to original source...
  17. Khodadadi F., González J.B., Martin P.L., Giroux E., Bilodeau G.J., Peter K.A., Doyle V.P. Acimovic S.G. (2020): Identification and characterisation of Colletotrichum species causing apple bitter rot in New York and description of C. noveboracense sp. nov. Scientific Reports, 10: 11043. Go to original source... Go to PubMed...
  18. Kumar M., Chakdar H., Pandiyan K., Thapa S., Shahid M., Singh A., Srivastava A.K., Saxena A.K. (2022): Bacterial chitinases: genetics, engineering and applications. World Journal of Microbiology & Biotechnology, 38: 252. Go to original source...
  19. Liu F., Tang G., Zheng X., Li Y., Sun X., Qi X., Zhou Y., Xu J., et al. (2016): Molecular and phenotypic characterisation of Colletotrichum species associated with anthracnose disease in peppers from Sichuan Province, China. Scientific Reports, 6: 32761. Go to original source...
  20. Malik M.S., Rehman A., Khan I.U., Khan T.A., Jamil M., Rha E.S., Anees M. (2023): Thermo-neutrophilic cellulases and chitinases characterised from a novel putative antifungal biocontrol agent: Bacillus subtilis TD11. PloS One, 18: e0281102. Go to original source... Go to PubMed...
  21. Masi C., Tebiso A., Kumar S.K.V. (2023): Isolation and characterisation of potential multiple extracellular enzyme-producing bacteria from waste dumping area in Addis Ababa. Heliyon, 9: e12645. Go to original source... Go to PubMed...
  22. Minchev Z., Kostenko O., Soler R., Pozo M.J. (2021): Microbial consortia for effective biocontrol of root and foliar diseases in tomato. Frontiers in Plant Science, 12:2021. Go to original source... Go to PubMed...
  23. Mongkolporn O., Taylor P.W.J. (2018): Chili anthracnose: Colletotrichum taxonomy and pathogenicity. Plant Pathology, 67: 1255-1263. Go to original source...
  24. Moreno-Velandia C.A., Izquierdo-García L.F., García-Arias F.L., Patiño-Moscoso M.A. (2024): Bacterial consortium based on Pseudomonas fluorescens, Lysinibacillus xylanilyticus and Bacillus velezensis reduces clubroot disease in broccoli. European Journal of Plant Pathology, 169: 171-186. Go to original source...
  25. Nguyen V.Q.H., Tran T.T.N., Tran L.T., Nguyen T.T.T., Pham T.T.T., Hoang Q.T., Pham T.T.D. (2023): Identification of fungal species associated with chilli fruit disease in north-central Vietnam. Journal of Plant Pathology, 106: 507-526. Go to original source...
  26. Niu B., Wang W., Yuan Z., Sederoff R.R., Sederoff H., Chiang V.L., Borriss R. (2020): Microbial interactions within multiple-strain biological control agents impact soil-borne plant disease. Frontiers in Microbiology, 11: 2020. Go to original source... Go to PubMed...
  27. Noor N.M., Zakaria L. (2018): Identification and characterisation of Colletotrichum spp. associated with chili anthracnose in peninsular Malaysia. European Journal of Plant Pathology, 151: 961-973. Go to original source...
  28. O'Donnell K., Cigelnik E. (1997): Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution, 7: 103-116. Go to original source... Go to PubMed...
  29. Peres N.R., Souza N.L., Peever T.L., Timmer L.W. (2004): Benomyl sensitivity of isolates of Colletotrichum acutatum and C. gloeosporioides from citrus. Plant Disease, 88: 125-130. Go to original source...
  30. Ray A.K., Roy T., Mondal S., Ringa E. (2010): Identification of gut-associated amylase, cellulase and protease-producing bacteria in three species of Indian major carps. Aquaculture Research, 41: 1462-1469. Go to original source...
  31. Raymaekers K., Ponet L., Holtappels D., Berckmans B., Cammue B.P.A. (2020): Screening for novel biocontrol agents applicable in plant disease management - A review. Biological Control, 144: 104240. Go to original source...
  32. Ro N.Y., Sebastin R., Hur O.S., Cho G.T., Geum B., Lee Y.J., Kang B.C. (2021): Evaluation of Anthracnose resistance in pepper (Capsicum spp.) genetic resources. Horticulturae (ISHS), 7: 460. Go to original source...
  33. Sarfraz S., Ali F., Hameed A., Ahmad Z., Riaz K. (2023): Sustainable agriculture through technological innovations. In: Hameed A. (Ed.): Sustainable Agriculture in the Era of the OMICs Revolution, Springer, Singapore. Go to original source...
  34. Shi X.C., Wang S.Y., Duan X.C., Wang Y.Z., Liu F.Q., Laborda P. (2021): Biocontrol strategies for the management of Colletotrichum species in postharvest fruits. Crop Protection, 141: 105454. Go to original source...
  35. Silva D.D.D., Ades P.K., Crous P.W., Taylor P.W.J. (2017): Colletotrichum species associated with chili anthracnose in Australia. Plant Pathology, 66: 254-267. Go to original source...
  36. Suprapta D.N., Darmadi A.A.K., Khalimi K. (2020): Potential antagonistic rhizobacteria to control Colletotrichum scovillei, the cause of anthracnose disease in chili pepper. Biodiversitas Journal of Biological Diversity, 21: 2727-2734. Go to original source...
  37. Sutton B.C. (1980): The Coelomycetes. Fungi Imperfecti with Pycnidia, Acervuli and Stromata. Commonwealth Mycological Institute, Kew, Surrey.
  38. Tamura K., Stecher G., Kumar S. (2021): MEGA11: Molecular evolutionary genetics analysis version 11. Molecular Biology and Evolution, 38: 3022-3027. Go to original source... Go to PubMed...
  39. Than P.P., Jeewon R., Hyde K.D., Pongsupasamit S., Mongkolporn O., Taylor P.W.J. (2008): Characterisation and pathogenicity of Colletotrichum species associated with anthracnose on chilli (Capsicum spp.) in Thailand. Plant Pathology, 57: 562-572. Go to original source...
  40. Wang L., Zhang X., Lu J., Huang L. (2025): Microbial diversity and interactions: Synergistic effects and potential applications of Pseudomonas and Bacillus consortia. Microbiological Research, 293: 128054. Go to original source...
  41. White T.J., Innis M.A., Gelfand D.H., Sninsky J.J. (2012): PCR Protocols: A Guide to Methods and Applications. Academic Press, London.
  42. Yanti Y., Hamid H., Reflin, Warnita, Habazar T. (2020): The ability of indigenous Bacillus spp. consortia to control the anthracnose disease (Colletrotricum capsici) and increase the growth of chili plants. Biodiversitas Journal of Biological Diversity, 21: 197-186. Go to original source...

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