In vitro assessment of the biocontrol activities of some Trichoderma species in suppressing Fusarium oxysporum f. sp. lycopersici

Abstract

Tomato Fusarium wilt is one of the crucial diseases threatening tomato production worldwide. In this study, the in vitro antagonistic potential of four Trichoderma species (Trichoderma koningii K-19, T. harzianum K-20, T. brevicompactum K-26, and T. atroviride TR-1) was evaluated using dual culture assays, volatile organic compound (VOC) bioassays, and cell-free culture filtrate (CFCF) tests. All tested isolates significantly inhibited pathogen mycelial growth in different bioassays. Among them, T. koningii K-19 consistently exhibited the strongest antagonistic activity, achieving inhibition rates of 70.37% in dual culture assays, 45.48% in sealed plate assays, and 43.10% in culture filtrate assays. In comparison, T. harzianum K-20, T. atroviride TR-1, and T. brevicompactum K-26 inhibited pathogen growth by 65.31%, 60.68%, and 66.40% in dual culture assays; 24.29%, 32.38%, and 31.43% in sealed plate assays; and 34.52%, 28.81%, and 35.95% in culture filtrate assays, respectively. These findings indicate that T. koningii K-19 suppresses the Fusarium wilt pathogen through multiple antagonistic mechanisms, such as competition, antibiosis, and volatile antifungal compounds. These findings demonstrate the potential of T. koningii K-19 as a promising biocontrol agent against tomato Fusarium wilt, requiring evaluation of its effectiveness in field trials in further studies.

Keywords

• Biological control
• Fusarium wilt
• multiple control effect
• Solanum lycopersicum
• Trichoderma spp.

References

1. Abdelmoteleb, A., Gonzalez-Mendoza, D., & Zayed, O. (2023). Cell-free culture filtrate of Trichoderma longibrachiatum AD-1 as alternative approach to control Fusarium solani and induce defense response Phaseolus vulgaris L. plants. Rhizosphere, 25, 100648.
2. Abdelrhim, A. S., Hemeda, N. F., Mwaheb, M. A., Omar, M. O., & Dawood, M. F. (2024). The role of Trichoderma koningii and Trichoderma harzianum in mitigating the combined stresses motivated by Sclerotinia sclerotiorum and salinity in common bean (Phaseolus vulgaris). Plant Stress, 11, 100370.
3. Algifari, M. H., Prismantoro, D., Hafsari, A. R., Miranti, M., Wan-Mohtar, W. A. A. Q. I., Ozturk, A. B., & Doni, F. (2025). Performance of hydrolytic enzymes produced by Trichoderma in sustainable crop production: Current insights and future perspectives. Sustainable Environment, 11(1), 2593746.
4. Alori, E. T., Onaolapo, A. O., & Ibaba, A. L. (2025). Cell free supernatant for sustainable crop production. Frontiers in Sustainable Food Systems, 9, 1549048.
5. Alqahtani, F. S. (2025). The utilization of microorganisms for biological control of soil-borne plant pathogens: a sustainable strategy for managing plant diseases-a comprehensive review. Journal of Plant Pathology, 107, 1815–1839.
6. Ateş, M., & Karaca, G. H. (2024). Antagonistic activities of mycoparasitic Pythium species against Fusarium oxysporum f. sp. lycopersici and Botrytis cinerea on tomatoes. International Journal of Agriculture Environment and Food Sciences, 8(1), 176-185.
7. Athinuwat, D., Ruangwong, O. U., Harishchandra, D. L., Pitija, K., & Sunpapao, A. (2024). Biological control activities of rhizosphere fungus Trichoderma virens T1-02 in suppressing flower blight of flamingo flower (Anthurium andraeanum Lind.). Journal of Fungi, 10(1), 66.
8. Aydın, M. H. (2015). Bitki fungal hastalıklarıyla biyolojik savaşta Trichoderma’lar. Türkiye Tarımsal Araştırmalar Derg., 2(2), 135-148.

9. Bailey, B. A., Bae, H., Strem, M. D., Crozier, J., Thomas, S. E., Samuels, G. J., & Holmes, K. A. (2008). Antibiosis, mycoparasitism, and colonization success for endophytic Trichoderma isolates with biological control potential in Theobroma cacao. Biological Control, 46(1), 24-35.
10. Baiyee, B., Pornsuriya, C., Ito, S. I., & Sunpapao, A. (2019). Trichoderma spirale T76-1 displays biocontrol activity against leaf spot on lettuce (Lactuca sativa L.) caused by Corynespora cassiicola or Curvularia aeria. Biological Control, 129, 195-200.
11. Behiry, S., Soliman, S. A., Massoud, M. A., Abdelbary, M., Kordy, A. M., Abdelkhalek, A., & Heflish, A. (2023). Trichoderma pubescens elicit induced systemic resistance in tomato challenged by Rhizoctonia solani. Journal of Fungi, 9(2), 167.
12. Benítez, T., Rincón, A. M., Limón, M. C., & Codón, A. C. (2004). Mecanismos de biocontrol de cepas de Trichoderma. International Microbiology, 7(4), 249-260.
13. Hernandez Castillo, F. D., Berlanga Padilla, A. M., Gallegos Morales, G., Cepeda Siller, M., Rodriguez Herrera, R., Aguilar Gonzales, C. N., & Castillo Reyes, F. (2011). In vitro antagonist action of Trichoderma strains against Sclerotinia sclerotiorum and Sclerotium cepivorum. American Journal of Agriculture Biology Science, 6(3), 410-417.
14. Datta, P., Dasgupta, B., & Sengupta, D. K. (2011). Efficacy of Trichoderma spp. against Phytophthora parasitica and Pythium spp. causing foot rot and leaf rot of betelvine (Piper betle L.). J.of Crop Weed, 7, 202-209.
15. De Angelis, G., Simonetti, G., Chronopoulou, L., Orekhova, A., Badiali, C., Petruccelli, V., & Palocci, C. (2022). A novel approach to control Botrytis cinerea fungal infections: uptake and biological activity of antifungals encapsulated in nanoparticle based vectors. Scientific Reports, 12(1), 7989.
16. Dennis, C., & Webster, J. (1971). Antagonistic properties of species-groups of Trichoderma: I. Production of non-volatile antibiotics. Transactions of the British Mycological Society, 57(1), 25-39.
17. dos Santos, J. M. R. (2025). Underground Warfare: Mechanisms of Trichoderma in the Suppression of the March of Fusarium in Banana Plants—Advances, Limitations and Perspectives. Journal of Phytopathology, 173(6), e70205.
18. D'souza, A., Roy, J. K., Bibekananda Mahanty, B. M., & Dasgupta, B. (2001). Screening of Trichoderma harzianum against major fungal pathogens of betelvine. Indian Phytopathology, 54(3), 340-345.
19. Elmeihy, R. M., Hewedy, O. A., Alhumaidi, M. S., Altammar, K. A., Hassan, E. O., & El-Debaiky, S. A. (2025). Co-inoculation of Trichoderma viride with Azospirillum brasilense could suppress the development of Harpophora maydis-infected maize in Egypt. Frontiers in Plant Science, 15, 1486607.
20. Fenta, L., & Mekonnen, H. (2024). Microbial biofungicides as a substitute for chemical fungicides in the control of phytopathogens: Current perspectives and research directions. Scientifica, 2024(1), 5322696.
21. Fontana, D. C., de Paula, S., Torres, A. G., de Souza, V. H. M., Pascholati, S. F., Schmidt, D., & Dourado Neto, D. (2021). Endophytic fungi: Biological control and induced resistance to phytopathogens and abiotic stresses. Pathogens, 10(5), 570.
22. Hajji-Hedfi, L., Rhouma, A., Wannassi, T., Utkina, A. O., & Rebouh, N. Y. (2025). Biocontrol assessment of Trichoderma species on tomato crops infested by Curvularia spicifera: Towards sustainable farming systems. Frontiers in Sustainable Food Systems, 9, 1627903.
23. Harman, G. E. (2024). Integrated benefits to agriculture with Trichoderma and other endophytic or root-associated microbes. Microorganisms, 12(7), 1409.
24. Harries, E., Berruezo, L., Mercado Cárdenas, G., Barrera, V., & Rajal, V. (2025). Trichoderma species from northwestern Argentina suppress Rhizoctonia solani infection in tobacco plants. International Journal of Pest Management, 1-12.
25. Jemil, N., Besbes, I., Gharbi, Y., Triki, M. A., Cheffi, M., Manresa, A., & Hmidet, N. (2024). Bacillus methylotrophicus DCS1: production of different lipopeptide families, in vitro antifungal activity and suppression of Fusarium wilt in tomato plants. Current Microbiology, 81(6), 142.
26. John, E., Singh, K. B., Oliver, R. P., & Tan, K. C. (2021). Transcription factor control of virulence in phytopathogenic fungi. Molecular Plant Pathology, 22(7), 858-881.
27. Joo, J. H., & Hussein, K. A. (2022). Biological control and plant growth promotion properties of volatile organic compound-producing antagonistic Trichoderma spp. Frontiers in Plant Science, 13, 897668.
28. Katyayani, K. K. S., Bindal, S., Singh, J. P., Rana, M., & Srivastava, S. (2020). In vitro evaluation of Trichoderma spp. against chickpea wilt. Int.Archive of Applied Sciences and Technology, 11(3), 1-4.
29. Loc, N. H., Huy, N. D., Quang, H. T., Lan, T. T., & Thu Ha, T. T. (2020). Characterisation and antifungal activity of extracellular chitinase from a biocontrol fungus, T. asperellum PQ34. Mycology, 11(1), 38-48.
30. Manzar, N., Kashyap, A. S., Goutam, R. S., Rajawat, M. V. S., Sharma, P. K., Sharma, S. K., & Singh, H. V. (2022). Trichoderma: advent of versatile biocontrol agent, its secrets and insights into mechanism of biocontrol potential. Sustainability, 14(19), 12786.
31. Mao, L., Yin, B., Ye, Z., Kang, J., Sun, R., Wu, Z., & Ping, W. (2024). Plant growth-promoting microorganisms drive K strategists through deterministic processes to alleviate biological stress caused by Fusarium oxysporum. Microbiological Research, 289, 127911.
32. Mishu, N. J., Hasan, M. R., Islam, S. M. N., Nayeema, J., & Hossain, M. M. (2025). Additive effects of Trichoderma isolates for enhancing growth, suppressing southern blight and modulating plant defense enzymes in tomato. Plos One, 20(7), e0329368.
33. Mukherjee, P. K., Mendoza-Mendoza, A., Zeilinger, S., & Horwitz, B. A. (2022). Mycoparasitism as a mechanism of Trichoderma-mediated suppression of plant diseases. Fungal Biology Reviews, 39, 15-33.
34. Rahman, M. A., Begum, M. F., & Alam, M. F. (2009). Screening of Trichoderma isolates as a biological control agent against C. paradoxa causing pineapple disease of sugarcane. Mycobiology, 37(4), 277-285.
35. Rajani, P., Rajasekaran, C., Vasanthakumari, M. M., Olsson, S. B., Ravikanth, G., & Shaanker, R. U. (2021). Inhibition of plant pathogenic fungi by endophytic Trichoderma spp. through mycoparasitism and volatile organic compounds. Microbiological Research, 242, 126595.
36. Raynaldo, F. A., Xu, Y., Wang, Q., Wu, B., & Li, D. (2024). Biological control and other alternatives to chemical fungicides in controlling postharvest disease of fruits caused by Alternaria alternata and Botrytis cinerea. Food Innovation and Advances, 3(2), 135-143.
37. Rizk, M. N., Ketta, H. A., & Shabana, Y. M. (2025). Trichoderma Culture Filtrates: Promising Biocontrol Agents for Potato Virus Y Management. Potato Research, 68(4), 4467-4492.
38. Rossi-Rodrigues, B. C., Brochetto-Braga, M. R., Tauk-Tornisielo, S. M., Carmona, E. C., Arruda, V. M., & Chaud Netto, J. (2009). Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system. Brazilian Journal of Microbiology, 40, 404-410.
39. Ruangwong, O. U., Wonglom, P., Suwannarach, N., Kumla, J., Thaochan, N., Chomnunti, P., & Sunpapao, A. (2021). Volatile organic compound from Trichoderma asperelloides TSU1: Impact on plant pathogenic fungi. Journal of Fungi, 7(3), 187.
40. Salwan, R., Rialch, N., & Sharma, V. (2019). Bioactive volatile metabolites of Trichoderma: An overview. Secondary Metabolites of Plant Growth Promoting Rhizomicroorganisms: Discovery and Applications, 87-111.
41. Sharfuddin, C., & Mohanka, R. E. E. N. A. (2012). In vitro antagonism of indigenous Trichoderma isolates against phytopathogen causing wilt of lentil. International Journal of Life Science and Pharma Research, 2(3), 195-202.
42. Shoaib, A., Munir, M., Javaid, A., Awan, Z. A., & Rafiq, M. (2018). Anti-mycotic potential of Trichoderma spp. and leaf biomass of Azadirachta indica against the charcoal rot pathogen, Macrophomina phaseolina (Tassi) Goid in cowpea. Egyptian Journal of Biological Pest Control, 28(1), 26.
43. Shoresh, M., Harman, G. E., & Mastouri, F. (2010). Induced systemic resistance and plant responses to fungal biocontrol agents. Annual Review of Phytopathology, 48(1), 21-43.
44. Singh, J., Kumar, V., Srivastava, S., Kumar, A., & Singh, V. P. (2018). In vitro evaluation of Trichoderma species against Fusarium oxysporum f. sp. lycopersici causing tomato wilt. Plant Pathology J., 17, 59-64.
45. Sunpapao, A., Chairin, T., & Ito, S. I. (2018). The biocontrol by Streptomyces and Trichoderma of leaf spot disease caused by Curvularia oryzae in oil palm seedlings. BioControl, 123, 36-42.
46. Tomah, A. A., Abd Alamer, I. S., Li, B., & Zhang, J. Z. (2020). A new species of Trichoderma and gliotoxin role: A new observation in enhancing biocontrol potential of T. virens against Phytophthora capsici on chili pepper. Biological Control, 145, 104261.
47. Villavicencio-Vásquez, M., Espinoza-Lozano, F., Espinoza-Lozano, L., & Coronel-León, J. (2025). Biological control agents: mechanisms of action, selection, formulation and challenges in agriculture. Frontiers in Agronomy, 7, 1578915.
48. Vinale, F., Sivasithamparam, K., Ghisalberti, E. L., Marra, R., Woo, S. L., & Lorito, M. (2008). Trichoderma–plant-pathogen interactions. Soil Biology and Biochemistry, 40(1), 1-10.
49. Yao, X., Guo, H., Zhang, K., Zhao, M., Ruan, J., & Chen, J. (2023). Trichoderma and its role in biological control of plant fungal and nematode disease. Frontiers in Microbiology, 14, 1160551.
50. You, J., Zhang, J., Wu, M., Yang, L., Chen, W., & Li, G. (2016). Multiple criteria-based screening of Trichoderma isolates for biological control of Botrytis cinerea on tomato. Biological Control, 101, 31-38.
51. Zehra, R., Moin, S., Enamullah, S. M., & Ehteshamul-Haque, S. (2022). Detection and identification of quarantine bacteria and fungi associated with imported and local potato seed tubers. Pakistan Journal of Botany, 54(3), 1157-1161.
52. Zhang, F., Huo, Y., Cobb, A. B., Luo, G., Zhou, J., Yang, G., & Zhang, Y. (2018). Trichoderma biofertilizer links to altered soil chemistry, altered microbial communities, and improved grassland biomass. Frontiers in Microbiology, 9, 848.
53. Zhang, Q., Zhang, J., Yang, L., Zhang, L., Jiang, D., Chen, W., & Li, G. (2014). Diversity and biocontrol potential of endophytic fungi in Brassica napus. Biological Control, 72, 98-108.
54. Zhang, Y., Lu, Y., Jin, Z., Li, B., Wu, L., & He, Y. (2024). Antifungal mechanism of cell-free supernatant produced by Trichoderma virens and its efficacy for the control of pear Valsa canker. Frontiers in Microbiology, 15, 1377683.
55. Zhu, N., Zhou, J. J., Zhang, S. W., & Xu, B. L. (2022). Mechanisms of Trichoderma longibrachiatum T6 fermentation against Valsa mali through inhibiting its growth and reproduction, pathogenicity and gene expression. Journal of Fungi, 8(2), 113.