Armillaria mellea Induces a Set of Defense Genes in Grapevine Roots and One of Them Codifies a Protein with Antifungal Activity

Abstract

Grapevine root rot, caused by Armillaria mellea, is a serious disease in some grape-growing regions. Young grapevines start to show symptoms of Armillaria root rot from the second year after inoculation, suggesting a certain degree of resistance in young roots. We used a suppression subtractive hybridization approach to study grapevine's reactions to the first stages of A. mellea infection. We identified 24 genes that were upregulated in the roots of the rootstock Kober 5BB 24 h after A. mellea challenge. Real-time reverse-transcriptase polymerase chain reaction analysis confirmed the induction of genes encoding protease inhibitors, thaumatins, glutathione S-transferase, and aminocyclopropane carboxylate oxidase, as well as phase-change related, tumor-related, and proline-rich proteins, and gene markers of the ethylene and jasmonate signaling pathway. Gene modulation was generally stronger in Kober 5BB than in Pinot Noir plants, and in vitro inoculation induced higher modulation than in greenhouse Armillaria spp. treatments. The full-length coding sequences of seven of these genes were obtained and expressed as recombinant proteins. The grapevine homologue of the Quercus spp. phase-change-related protein inhibited the growth of A. mellea mycelia in vitro, suggesting that this protein may play an important role in the defense response against A. mellea.