Fungicide tolerance of Trichoderma asperelloides and T. harzianum strains


Tolerance in isolations of Trichoderma was developed by exposing two strains of T. harzianum and three of T. asperelloides to increasing concentrations of chemical fungicides. These isolation of Trichoderma were exposed to three fungicides: Captan, Thiabendazol and the mixture Captan-Carboxin. Some selected lines of these strains reached tolerance to Captan and partial tolerance to the mixture Captan-Carboxin. The biological and genetic changes in these tolerant lines were monitored by determining the relative growth rate of the fungus, inhibition of Fusarium and by analyzing the genomic changes through UP-PCR. The results show that the tolerance to fungicides can be developed without affecting the parameters of biological activity in these lines of Trichoderma (growth and parasitism against Fusarium). Chemical tolerance to the fungicide was verified by means of changes at the DNA level (UP-PCR), mainly in the lines tolerant to Captan. This suggests that Trichoderma survives in environments with remnants of fungicide molecules.

Share and Cite:

Chaparro, A. , Carvajal, L. and Orduz, S. (2011) Fungicide tolerance of Trichoderma asperelloides and T. harzianum strains. Agricultural Sciences, 2, 301-307. doi: 10.4236/as.2011.23040.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Brunner, K., Zeilinger, S., Ciliento, R., Woo, S.L., Lorito, M., Kubicek, C.P. and Mach, R. L. (2005) Improvement of the fungal biocontrol agent Trichoderma atroviride to enhance both antagonism and induction of plant systemic disease resistance. Applied Environmental Microbiology, 71, 3959–3965.
[2] Hanson, L.E. and Howell, C.R. (2004) Elicitors of plant defense responses from biocontrol strains of Trichoderma virens. Phytopathology, 94, 171–176.
[3] Hoyos, L., Orduz, S. and Bissett, J. (2009b) Growth stimulation in bean (Phaseolus vulgaris L.) by Trichoderma. Biological Control, 51, 409–416.
[4] Sahebani, N. and Hadavi, N. (2008) Biological control of the root knot nematode Meloidogyne javanica by Trichoderma harzianum. Soil Biology and Biochemistry, 40, 2016–2020.
[5] Lenteren, V. and Woets, J. (1988) Biological and integrated pest control in greenhouses. Annual Review of Entomology, 33, 239-269.
[6] Ezzi, I.M. and Lynch, J.M. (2005) Biodegradation of cyanide by Trichoderma spp. and Fusarium spp. Enzyme Microbial Technology, 36, 849–954.
[7] Tang, J., Liu, L., Hua, S., Chen, Y. and Chen, J. (2009) Improved degradation of organophosphate dichlorvos by Trichoderma atroviride transformants generated by restriction enzyme-mediated integration (REMI). Bioresource Technology, 100, 480–483.
[8] Zhou, X., Xu, S., Liu, L. and Chen, J. (2007) Degradation of cyanide by Trichoderma mutants constructed by restriction enzyme mediated integration (REMI). Bioresource Technology, 98, 2958–2962.
[9] Goldman, G., Temmerman, W., Jacobs, D., Contreras, R., Van Montagu, M. and Herrera-Estrella, A. (1993) A nucleotide substitution in one of the beta-tubulin genes of Trichoderma viride confers resistance to the antimitotic drug methyl benzimidazole 2-yl-carbamate. Molecular and General Genetics, 240, 73-80.
[10] Mukherjee, P.K., Sherkhane, P.D. and Murthy, N.B. (1999) Induction of stable benomyl-tolerant phenotypic mutants of Trichoderma pseudokoningii MTCC 3011, and their evaluation for antagonistic and biocontrol potential. Indian Journal of Experimental Biology, 37, 710-2.
[11] Yan, K. and Dickman, M. (1996) Isolation of a β tubulin gene from Fusarium moniliforme that confers cold-sensitive benomyl resistance. Applied and Environment Microbiology, 62, 3053-3056.
[12] Deyle, C., Laigret, F. and Corio-Costet, M. (1997) A mutation in the 14 α demethylase gene of Uncicula necator that correlates with resistance to a sterol biosynthesis inhibitor. Applied and Environment Microbiology, 63, 2966-2970.
[13] Yamamoto, E. and Baird, V. (1999) Molecular characterization of four beta-tubulin genes from dinitroaniline susceptible and resistant biotypes of Eleusine indica. Plant Molecular Biology, 39, 45-61.
[14] Hoyos, L., Orduz, S. and Bissett, J. (2009a) Genetic and metabolic biodiversity of Colombia and adjacent neotropic regions. Fungal Genetics and Biology, 46, 615-631.
[15] Bell, D., Wells, H. and Markham, C. (1982) In vitro antagonism of Trichoderma species against six fungal plant pathogens. Phytopathology, 72, 379-382.
[16] Bulat, S., Lubeck, M., Mironenko, N., Jensen, D. and Lubeck, P. (1998) UP-PCR analysis and ITS1 ribotyping of strains of Trichoderma and Gliocladium. Mycological Research, 102, 933-943.
[17] Lubeck, M., Alekhina, A., Lubeck, S., Jensen, F. and Bulat, A. (1999) Delineation of Trichoderma two different genotypic groups by a highly robust fingerprinting method, UP-PCR, and UP-PCR product cross-hybridization. Mycological Research, 103, 289-298.
[18] Raeder, U. and Broda, P. (1985) Rapid preparation of DNA from filamentous fungi. Letters in Applied Microbiology, 1, 17-20.
[19] Ca?as, G. (2004) Identificación de cepas de Mycosphaerella fijiensis resistentes al benomyl usando la reacción en cadena de la polimerasa PCR. Trabajo de grado Magíster of Science en Biotecnología. Universidad Nacional de Colombia, sede Medellín. Colombia.
[20] Harman, G.E. (2006) Overview of mechanisms and uses of Trichoderma spp. Phytopathology, 96, 190-194.
[21] Sanz, L., Montero, M., Redondo, J., Llobell, A. and Monte, E. (2005) Expression of an a-1,3-glucanase during mycoparasitic interaction of Trichoderma asperelloides. FEBS Journal, 272, 493–499.
[22] Viterbo, A. and Chet, I. (2006) TasHyd1, a new hydrophobin gene from the biocontrol agent Trichoderma asperelloides, is involved in plant root colonization. Molecular Plant Pathology, 7, 249–258. doi/10.1111/j.1364-3703.2006.00335.x
[23] Ma, Z. and Michailides, J. (2005) Advances in understanding molecular mechanisms of fungicide resistance and molecular detection of resistant genotypes in phytopathogenic fungi. Crop Protection, 24, 853–863.
[24] Brent, K. (1995) Monitoring fungicide resistance in crop pathogens: how can it be managed?. FRAC Monograph No. 1 GIFAP Brussels, 48 p.
[25] Ruocco, M., Lanzuise, S., Vinale, F., Marra, R., Turrà, D., Woo, S.L. and Lorito, M. (2009) Identification of a new biocontrol gene in Trichoderma atroviride: The role of an ABC transporter membrane pump in the interaction with different plant-pathogenic fungi. Molecular Plant-Microbe Interactions, 22, 291–301.
[26] De Waard, M.A. (1997) Significance of ABC transporters in fungicide sensitivity and resistance. Pesticide Science, 51, 271-275.
[27] Holmes, J.G. and Ecker, J.W. (1995) Relative fitness of Imazalil-resistant and sensitive biotypes of Penicillium digitatum. Plant Disease, 79, 1068-1073.
[28] Marra, R, Ambrosino, P., Carbone, V., Vinale, F., Woo, S.L., Ruocco, M., Ciliento, R., Lanzuise, S., Ferraioli, S., Soriente, I., Gigante, S., Turrà, D., Fogliano, V., Scala, F. and Lorito, M. (2006) Study of the three-way interaction between Trichoderma atroviride, plant and fungal pathogens by using a proteomic approach. Current Genetics, 50, 307-321.
[29] Mukherjee, M., Hadar, R., Mukherjee, P.K. and Horwitz, B.A. (2003) Homologous expression of a mutated beta-tubulin gene does not confer benomyl resistance on Trichoderma virens. Journal of Applied Microbiology, 95, 861-867.
[30] Kawchuk, L.M., Hutchison, L.J., Verhaeghe, CA., Lynch, D.R., Bains, P.S. and Holley, J.D. (2002) Isolation of the ?-tubulin gene and characterization of thiabendazole resistance in Gibberella pulicaris. Canadian Journal of Plant Pathology, 24, 233-238.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.