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Verticillium dahliae , a vascular-colonizing fungus, causes economically important wilt diseases in many crops, including olive trees.  Trichoderma  spp. have demonstrated an effective contribution as biocontrol agents against this pathogen through a variety of mechanisms that may involve direct mycoparasitism and antibiosis. However, molecular aspects underlaying  Trichoderma – V. dahliae  interactions are not well known yet due to the few studies in which this pathogen has been used as a target for  Trichoderma . In the present study,  Trichoderma atroviride  T11 overgrew colonies of  V. dahliae  on agar plates and inhibited growth of highly virulent defoliating (D)  V. dahliae  V-138I through diffusible molecules and volatile organic compounds produced before contact. A  Trichoderma  microarray approach of T11 growing alone (CON), and before contact (NV) or overgrowing (OV) colonies of V-138I, helped to identify 143 genes that differed significantly in their expression level by more than twofold between OV and CON or NV. Functional annotation of these genes indicated a marked up-regulation of hydrolytic, catalytic and transporter activities, and secondary metabolic processes when T11 overgrew V-138I. This transcriptomic analysis identified peptidases as enzymatic activity overrepresented in the OV condition, and the  cpa1  gene encoding a putative carboxypeptidase (ID number 301733) was selected to validate this study. The role of  cpa1  in strain T11 on antagonism of V-138I was analyzed by a  cpa1 -overexpression approach. The increased levels of  cpa1  expression and protease activity in the  cpa1 -overexpressed transformants compared to those in wild-type or transformation control strains were followed by significantly higher antifungal activity against V-138I in  in vitro  assays. The use of  Trichoderma  spp. for the integrated management of plant diseases caused by  V. dahliae  requires a better understanding of the molecular mechanisms underlying this interaction that might provide an increase on its efficiency.

frontiers in microbiology

Transcriptomic Analysis of Trichoderma atroviride Overgrowing Plant-Wilting Verticillium dahliae Reveals the Role of a New M14 Metallocarboxypeptidase CPA1 in Biocontrol