Multiplex PCR for detection and differentiation of diverse Trichoderma species

Abstract

Trichoderma species are among the most common fungi frequently isolated as saprotrophs from free soil, soil litter, dead wood, and the rhizosphere of different crops. Four sets of species-specific primers were designed from the tef1 and rpb2 genes, in order to identify Trichoderma asperellum (tef1 gene), T. longibrachiatum (tef1 gene), T. virens (tef1 gene), and T. harzianum (rpb2 gene). Here, we report the development of a multiplex PCR assay to detect and distinguish each of these four most common Trichoderma species—viz., T. asperellum, T. harzianum, T. longibrachiatum, and T. virens—simultaneously in a single reaction through their distinct amplicons of 507, 824, 452, and 330 bp, respectively. The developed multiplex PCR technique will provide a rapid, simple, and reliable alternative to conventional methods and a new site for identification of different species of Trichoderma in a single reaction.