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Applied and Environmental Microbiology, May 2000, p. 1890-1898, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Characterization and Identification of Biocontrol Isolates of Trichoderma spp.

M. R. Hermosa, I. Grondona, E. A. Iturriaga, J. M. Diaz-Minguez, C. Castro, E. Monte,* and I. Garcia-Acha

Departamento de Microbiología y Genética, CSIC/Universidad de Salamanca, 37002 Salamanca, Spain

Received 20 September 1999/Accepted 7 February 2000

The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as "Trichoderma harzianum" Rifai and one biocontrol strain recognized as T. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains in T. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae, Rosellinia necatrix, Botrytis cinerea, and Fusarium oxysporum f. sp. dianthi in three different media.

* Corresponding author. Mailing address: Departamento de Microbiología y Genética, Universidad de Salamanca, Edificio Departamental, Lab. 208, Avenida del Campo Charro s/n, 37007 Salamanca, Spain. Phone: 34-23-294532. Fax: 34-23-224876. E-mail: emv{at}gugu.usal.es.

Applied and Environmental Microbiology, May 2000, p. 1890-1898, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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