Abstract
Verticillium wilt, caused by the Verticillium dahliae phytopathogen, is a devastating disease affecting many economically important crops. Previous studies have shown that the exoproteome of V. dahliae plays a significant role in this pathogenic process, but the components and mechanisms that underlie this remain unclear. In this study, the exoproteome of V. dahliae was induced in a cotton-containing C’zapek-Dox (CCD) medium and quantified using the high-throughput isobaric tag technique for relative and absolute quantification (iTRAQ). Results showed that the abundance of 271 secreted proteins was affected by the CCD medium, of which 172 contain typical signal peptides generally produced by the Golgi/endoplasmic reticulum (ER). These enhanced abundance proteins were predominantly enriched in carbohydrate hydrolases; 126 were classified as carbohydrate-active (CAZymes) and almost all were significantly up-regulated in the CCD medium. Results showed that CAZymes proteins 30 and 22 participate in pectin and cellulose degradation pathways, corresponding with the transcription levels of several genes encoded plant cell wall degradation enzyme activated significantly during cotton infection. In addition, targeted deletion of two pectin lyase genes (VdPL3.1 and VdPL3.3) impaired wilt virulence to cotton. This study demonstrates that the V. dahliae exoproteome plays a crucial role in the development of symptoms of wilting and necrosis, predominantly via the pathogenic mechanisms of plant cell wall degradation as part of host plant infection.
Highlights
Verticillium wilt, caused by the phytopathogen Verticillium dahliae, is an extremely destructive disease affecting hundreds of dicotyledonous plant species
The induced exoproteome of V. dahliae purified from modified medium containing the containing C’zapek-Dox (CCD) medium caused serious chlorosis and necrosis of cotton leaves (Figure 1B)
Abundances of CAZymes involved in pectin and cellulose degradation were up-regulated in the CCD medium; proteins 26 and 18, respectively, were upregulated by more than fivefold (Supplementary Figure S2). These results strongly suggest that CAZymes associated with the degradation of pectin and cellulose play a crucial role in V. dahliae germination and proliferation in the CCD medium
Summary
Verticillium wilt, caused by the phytopathogen Verticillium dahliae, is an extremely destructive disease affecting hundreds of dicotyledonous plant species. The pathogen produces toxic or elicitor-like substances as well as hydrolytic enzymes that facilitate its colonization and proliferation in the unique plant vascular system environment (Fradin and Thomma, 2006; Klosterman et al, 2009). Given this background, the V. dahliae exoproteome, including secreted proteins, is thought to play an important role in determining pathogenicity (Fradin and Thomma, 2006)
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