Comparative Proteomic Analysis of Botrytis cinerea Secretome

Abstract

Botrytis cinerea (B. cinerea) is a filamentous fungus infecting more than 200 plant species, causing significant economic losses worldwide. Secreted proteins are released as an initial response of the fungus to its plant host. We report the use of a high-throughput LC-MS/MS approach to analyze B. cinerea BO5.10 secreted proteins. Secretions were collected from fungus grown on a solid substrate of cellophane membrane while mock infecting media supplemented with the extract of full red tomato, ripened strawberry or Arabidopsis leaf extract. Overall, 89 B. cinerea proteins were identified from all growth conditions. Sixty proteins were predicted to contain a SignalP motif indicating the extracellular location of the proteins. Seven proteins were observed in all the growth conditions implying a constitutive nature of their secretion. Identified in the secretions were transport proteins, proteins well-characterized for carbohydrate metabolism, peptidases, oxidation/reduction, and pathogenicity factors that provide important insights into how B. cinerea may use secreted proteins for plant infection and colonization.

Secretome of the Coprophilous Fungus Doratomyces stemonitis C8, Isolated from Koala Feces

“…The aminopeptidases, which bear homology to the vacuolar protease aminopeptidase Y of Saccharomyces cerevisiae (aminopeptidase Y; PA_ScAPY-like; Table 1), are exoproteases that cleave at the amino termini of proteins or peptides (27). Proteases have been found in the secretomes of most filamentous fungi explored to date, including pathogenic (27,28,35,43) and saprophytic (13,31,42,47) species. The proteases in D. stemonitis could be predominantly involved in the degradation of microbial bodies or sloughed epithelial cells in the koala feces (32) but may also play a role in increasing the efficiency of the degradation of the plant cell wall matrix (4).…”

“…Integral to their survival is the ability to secrete enzymes to break down complex materials in the environment into small molecules that can be absorbed into the hyphae and used for nutrition (53). In the past decade, advances in protein identification techniques and genome sequencing have enabled detailed investigation of the secretomes of saprophytic (31,42,47,50,51), pathogenic (28,35,43), and symbiotic fungal species (29), revealing rich and diverse enzyme arrays. The fungal secretomes have been explored to find enzymes and enzyme combinations for various industrial applications, such as paper, textile, and food manufacture (7,33) and economically and industrially sustainable hydrolysis of plant biomass to fermentable sugars for biofuel production (1,13,46,51).…”

The Penicillium Chrysogenum Extracellular Proteome. Conversion from a Food-rotting Strain to a Versatile Cell Factory for White Biotechnology

“…The recent advances in the Proteomics tools and the availability of genome sequences, has allowed an analysis of the secretomes of a few filamentous fungi, but the available information is still scarce (17)(18)(19). However, because of the availability of several fungal genomes and diverse prediction programs for secretory proteins, an integrated platform for annotation of fungal secretomes (Fungal Secretome Database) has been established and implemented in a web-based database (20).…”

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

“…Thus, the knowledge of the protein abundance at the intracellular proteome helps to clarify possible cell lysis events. As an example, malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase in B. cinerea or flavohemoglobin and manganese superoxide dismutase in P. chrysogenum should be highly detected in the case of lysis, since these are the most abundant intracellular proteins (Shah et al, 2009;Jami et al, 2010a). However, sometimes it is difficult to ascertain whether a protein is truly secreted or it is present in the culture medium as a consequence of cell lysis.…”

“…It was called SecretomeP (http://www.cbs.dtu.dk/services/SecretomeP/). Even when this program was developed for mammalian applications and later improved for bacteria, it has shown good results in the fungal protein analysis (Shah et al, 2009;Jami et al, 2010b). The availability of several fungal genomes in databases together with the use of those prediction programs for secretory proteins have allowed the recent development of platforms for the annotation of fungal secretomes, such as the Fungal Secretome Database [FSD (Choi et al, 2010)] or the Fungal Secretome KnowledgeBase [FunSecKB (Lum & Min, 2011)], which allow the proper identification of secreted proteins by the compilation of different software.…”