Antagonistic Potential and Molecular Characterization of Trichoderma Harzianum Isolates against Sclerotium Rolfsii Infecting Tobacco


Ten isolates of Trichoderma harzianum isolated from rhizosphere of tobacco and other crops were evaluated for their genetic diversity and antagonistic potential against Sclerotium rolfsii, a fungal pathogen causing collar rot in tobacco. In dual culture, isolates WG 1, WG 3, GT 1 and DH were more aggressive in inhibiting the mycelial growth of the pathogen. The isolate WG 1 showed maximum production of HCN which reflected in the maximum inhibition of S. rolfsii growth by volatile compounds. Production potential of hydrogen cyanide, indole acetic acid and siderophore varied among the isolates. Siderophore production was not observed in the isolate KP. The level of polymorphism detected with different primers ranged from 70 to 100%. Out of 249 amplified fragments, 216 (86.74%) were polymorphic. The primers OPP 11, OPAB 1, OPL 4, PL 5 and OPL 6 were highly polymorphic (100%). Based on RA PD profiles, the pair-wise similarity coefficients among the isolates ranged from 0.46 to 0.85 with an average of 0.65. The minimum genetic similarity was observed between the isolates KP and BK (46%), whereas the highest (85%) was between the isolates GT 1 and GT 2. Cluster analysis by unweighted pair group method on arithmetic averages (UPGMA) grouped the isolates into two major clusters. The clustering pattern of the isolates was based on host and region with the exception of isolate KP. There was no correlation between the RA PD pattern and the antagonistic potential of T. harzianum isolates.


Antagonism, Genetic Diversity, Non-volatile And Volatile Compounds, Sclerotium Rolfsii, Siderophores, Tobacco and Trichoderma Harzianum

Subject Discipline

Agriculture Sciences


Adams, P. B. 1990. The potential of mycoparasites for biological control of plant diseases. Annual Review of Phytopathology, 28: 59-72.

Bonde, M. R., Peterson, G. L., Emmert, R. W and Menge, J. A. 1991. Isozyme comparisons of Septoria isolates associated with citrus in Australia and United States. Phytopathology, 81: 517-521.

Bunker, R. N. and Mathur, K. 2001. Antagonism of local biocontrol agents to Rhizoctonia solani, inciting dry root rot of chilli. Journal of Mycology and Plant Pathology, 31: 50-53.

Chen, W. A. and Zhang, T. Y. 1994. Analysis of isozymes by PAGE of Alternaria spp. from cruciferae. Acta Mycologica Sinica, 13: 295-302.

Chet, I., Viterbo, A., Brotman, Y. and Lousky, T. 2006. Enhancement of plant disease resistance by biocontrol agent Trichoderma. Life Science, Weizmann Institute of Science, p. 1-2.

Cook, R. J. 1993. Making greater use of introduced microorganisms for biological control of plant pathogens. Annual Review of Phytopathology, 31: 53-80.

Dennis, C. and Webster, J. 1971a. Antagonistic properties of species group of Trichoderma I. Production of non-volatile antibiotics. Transactions of British Mycological Society, 57: 25-39.

Dennis, C. and Webster, J. 1971b. Antagonistic properties of species group of Trichoderma II. Production of volatile antibiotics. Transactions of British Mycological Society, 57: 41-48.

Doyle, J. J. and Doyle, J. L. 1990. Isolation of plant DNA from fresh tissue. Focus, 12: 13-15.

El-Khawas, H. and Adachi, K. 1999. Identification and quantification of auxins in culture media of Azospirillum and Klebsiella and their effect on rice roots. Biology and Fertility of Soils, 28: 377-381.

Gams, W. and Bisset, J. 1998. Morphology and Identification of Trichodema, pp. 3-34. In: Kubicek, E. and Harman, G. E. (Eds.). Trichoderma and Gliocladium Vol. I: Basic Biology; taxonomy and genetics. Taylor and Francis, London, UK

Glick, B. R., Penrose, D. M. and Li, J. 1998. A model for lowering of plant ethylene concentration by plant growth promoting bacteria. Journal of Theoretical Biology, 190: 63-68.

Harman, G. E., Howell, C. R., Viterbo, A., Chet, I. and Lorito, M. 2004. Trichoderma speciesopportunistic, avirulent plant symbionts. Nature Microbiological Review, 2: 43-56.

Harman, G. E. 2000. Myths and dogmas of biocontrol. Changes in perceptions derived from research on Trichoderma harzianum T22. Plant Disease, 84: 377-393.

Hermosa, M. R., Grondona, I., Itturriaga, E. A., Diaz- Minguez, J. M., Castro, C., Monte, E. and Garcia-Acha, I. 2000. Molecular characterization and identification of biocontrol isolates of Trichoderma spp. Applied and Environmental Microbiology, 66: 1890-1898.

Kloepper, J. W., Leong, M., Teintze, M. and Schroth, M. N. 1980. Pseudomonas siderophores: A mechanism explaining disease suppressive soils. Current Microbiology, 4: 317-320.

Kumar, D. and Dubey, S. C. 2001. Management of collar rot of pea by the integration of biological and chemical methods. Indian phytopathology, 54: 62-66.

Larissa, B.G., Ana Bolena Lima da, C., Laurineide L. C. F., and Neiva Tinti, O. 2002. Randomly amplified polymorphic DNA of Trichoderma isolates and antagonism against Rhizoctonia solani. Brazilian Archives of Biology and Technology, 45: 151-160.

Lone Gram, 1996. The influence of substrate on siderophore production by fish spoilage bacteria. Journal of Microbiological methods, 25: 199-205.

Muthumeenakshi, S., Brown, A. E. and Mills, P. R. 1998. Genetic comparison of aggressive weed mould strains of Trichoderma harziamum from mushroom compost in North America and the British Isles. Mycological Research, 4: 385-390.

Ozbay, N., Newman, S. E. and Brown, W. M. 2004. The effect of Trichoderma harzianum strains in the growth of tomato seedlings. Proc. XXVI. IHC Manage, Acta Horticulurae, 635: 131-135.

Prokkola, S. 1992. Antagonistic properties of Trichoderma spp. against Mycocentrospora acerina. Bulletin of OILB SROP, 15: 76-78.

Rohlf, F. J. 1998. NTSYS-pc: Numerical taxonomy and multivariate analysis system. Version 2.02. Exter Software, Setauket, New York. Shanmugam, V., Vivek Sharma and Anantha Padmanabhan. 2008. Genetic relatedness of Trichoderma isolates antagonistic against Fusarium oxysporum f. sp. dianthi inflicting carnation wilt. Folia Microbiology, 53: 130-138.

Shishido, M., Miwa, C., Usami, T., Amemiya, Y. and Jonhson, K. B. 2005. Biological control efficiency of Fusarium wilt of tomato by nonpathogenic Fusarium oxysporum F0-B2 in different environments. Phytopathology, 95: 1072-1080.

Wei, G., Clopper, J. W. and Tuzun, S. 1991. Induction of systemic resistance of cucumber to Colletotrichum orbiculare by select strains of plant growth promoting rhizobacteria. Phytopathology, 81: 221-224.

Williams, J. G. K, Kubelik, A. R., Livak, K. J., Rafalski, J.A. and Tingey, S. V. 1990. DNA polymorphism amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research, 18: 6531-6535.


  • There are currently no refbacks.