Two-dimensional electrophoresis protein profile of the phytopathogenic fungus Botrytis cinerea

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Abstract
Botrytis cinerea is a phytopathogenic fungi causing disease in a number of important crops. It is considered a very complex species in which different populations seem to be adapted to different hosts. In order to characterize fungal virulence factors, a proteomic research was started. A protocol for protein extraction from mycelium tissue, with protein separation by 2‐DE and MS analysis, was optimised as a first approach to defining the B. cinerea proteome. Around 400 spots were detected in 2‐DE CBB‐stained gels, covering the 5.4–7.7 pH and 14–85 kDa ranges. The averages of analytical and biological coefficients of variance for 64 independent spots were 16.1% and 37.5%, respectively. Twenty‐two protein spots were identified by MALDI‐TOF or ESI IT MS/MS, with some of them corresponding to forms of malate dehydrogenase and glyceraldehyde‐3‐phosphate dehydrogenase. Two more spots matched a cyclophilin and a protein with an unknown function.