Down-regulation of Sclerotinia sclerotiorum gene expression in response to infection with Sclerotinia sclerotiorum debilitation-associated RNA virus

We have previously presented convincing evidence in support of a viral etiology for the debilitation phenotype exhibited by strain Ep-1PN of Sclerotinia sclerotiorum. To explore the possible mechanisms underlying fungal pathogenicity and hyphal growth, potential genes whose expression was down-regulated in Ep-1PN were identified from a cDNA library of the virus-free strain Ep-1PNAa, which is a single ascospore derivative of strain Ep-1PN, using reverse northern blot analysis. A total of 1116 cDNA clones were targeted and, following PCR re-amplification, 210 cDNA clones were selected as candidates, of which 16 cDNA clones were subjected to northern blot analysis for further confirmation. The results showed that 12 clones represented genes that were differentially expressed in the virus-free strain compared to the virus-infected one. Of the 210 clones that were sequenced, 150 had non-redundant sequences and of these 92% (138 clones) had significant homology to fungal genes in the databases examined. The remaining 12 clones did not have any matches. The differentially expressed genes represented a broad spectrum of biological functions including carbon and energy metabolism, protein synthesis and transport, signal transduction and stress response. This study provides the first insight into genes differentially expressed between the virus-free strain Ep-1PNAa and the virus-infected strain Ep-1PN. The possible relationships between mycovirus-mediated changes in cellular gene expression and observed phenotypes are discussed.