Tri11, tri3, and tri4 genes are required for trichodermin biosynthesis of Trichoderma brevicompactum.

Trichoderma brevicompactum and T. arundinaceum both can synthesize trichodermin with strong antifungal activity and high biotechnological value. The two Trichoderma species have a tri cluster, which includes seven genes (tri14, tri12, tri11, tri10, tri3, tri4, and tri6) that encode transport and regulatory enzymes required for the biosynthesis of trichodermin. Here, we isolated T. brevicompactum 0248 transformants with disrupted tri11, tri4, or tri3 gene. We also described the effect of tri11, tri3, or tri4 deletion on the expression of other genes in the tri cluster. Targeted Δtri3 knockout mutant exhibited a sharp decline in the production of trichodermin, and trichodermol, which is a substrate for trichodermin production, accumulated. Thus, the results demonstrated that tri3 was responsible for the biosynthesis of trichodermin, and the tri3 gene-encoded enzyme catalyzed the acetylation reaction of the hydroxy group at C-4 of the trichodermin skeleton. In addition, tri4 and tri11 deletion mutants were generated to evaluate the roles of tri4 and tri11 in trichodermin biosynthesis, respectively. Deletion mutant strain Δtri4 or Δtri11 did not produce trichodermin in T. brevicompactum, indicating that tri4 and tri11 are essential for trichodermin biosynthesis. This is the first to report the function of tri3, tri4 and tri11 in T. brevicompactum, although the role of tri4 and tri11 has already been described for T. arundinaceum by Cardoza et al. (Appl Environ Microbiol 77:4867-4877, 2011).