2‐DE proteomic approach to the Botrytis cinerea secretome induced with different carbon sources and plant‐based elicitors

Abstract

Botrytis cinerea is a phytopathogenic fungus infecting a number of crops (tomatoes, grapes and strawberries), which has been adopted as a model system in molecular phytopathology. B. cinerea uses a wide variety of infection strategies, which are mediated by a set of genes/proteins called pathogenicity/virulence factors. Many of these factors have been described as secreted proteins, and thus the study of this sub-proteome, the secretome, under changing circumstances can help us to understand the roles of these factors, possibly revealing new loci for the fight against the pathogen. A 2-DE, MALDI TOF/TOF-based approach has been developed to establish the proteins secreted to culture media supplemented with different carbon sources and plant-based elicitors (in this study: glucose, cellulose, starch, pectin and tomato cell walls). Secreted proteins were obtained from the culture media by deoxycholate-trichloroacetic acid/phenol extraction, and 76 spots were identified, yielding 95 positive hits that correspond to 56 unique proteins, including several known virulence factors (i.e. pectin methyl esterases, xylanases and proteases). The observed increases in secretion of proteins with established virulence-related functions indicate that this in vitro-induction/proteome-mining approach is a promising strategy for discovering new pathogenicity factors and dissecting infection mechanisms in a discrete fashion.

LongSAGE gene-expression profiling of Botrytis cinerea germination suppressed by resveratrol, the major grapevine phytoalexin

“…We further checked whether these four putative secreted virulence effectors could be found in four recently published secretomes of B. cinerea (Shah et al 2009a(Shah et al , 2009bEspino et al 2010;Fernandez-Acero et al 2010). The putative xyloglucanase (BC1G_00594.1/BofuT4_P021270.1), the putative pectin lyase (BC1G_12017.1/BofuT4_P032630.1) and the putative aspartic protease (BC1G_08393.1/BofuT4_P010710.1) were found in one or several of the published secretomes.…”

Proteomic analysis of mycelial proteins from Magnaporthe oryzae under nitrogen starvation

“…Recently, a variety of plant pathogenic fungi has been investigated through proteomics (Garg et al, 2002). The pathogenesis of most of these fungi has been explored based on secreted proteins (Fernández-Acero et al, 2010), mycelia (Bregar et al, 2012), and conidia (Noir et al, 2009).…”

Proteomics Methodology Applied to the Analysis of Filamentous Fungi - New Trends for an Impressive Diverse Group of Organisms

“…This fungus is able to infect more than 200 different plant species, at any plant stage or tissue, including fruits during storage and distribution. As a result, it inflicts high economic damage at various levels in the food industry (Fernández-Acero et al, 2010). The ability for B. cinerea to infect is due to its wide variety of infection strategies, which are facilitated by a set of pathogenicity/virulence factors.…”

“…For example, the discovery of novel sources of host resistance, the evolution of strains due to the industrial improvement or the effects of carbon sources, antifungal drugs and gene deletion have been analysed in detail (Fernández-Acero et al, 2010;Jami et al, 2010a;Cagas et al, 2011b). One of the earliest intracellular filamentous fungal proteomic studies was performed on the wood-degrading fungi Phanerochaete chrysosporium and Lentinula edodes (Hernández-Macedo et al, 2002).…”

“…The ability for B. cinerea to infect is due to its wide variety of infection strategies, which are facilitated by a set of pathogenicity/virulence factors. The study of the secreted proteins has been faced up by means of different carbon sources from fruit (tomato, strawberry, kiwifruit, glucose, carboxymethyl cellulose, starch, pectin or tomato cell walls) and using solid and liquid cultures (Shah et al, 2008;Espino et al, 2010;Fernández-Acero et al, 2010 The ultrafast genome sequencing is a boosted fashion trend, which is generating thousands of gene sequences every day. In spite of this fact, several fungi which are producers of severe infections in fruit trees (fruit blight) and grapevines, like Diplodia seriata, keep fully unsequenced.…”