title: Secretome Analysis of Metarhizium anisopliae Under Submerged Conditions Using Bombyx mori Chrysalis to Induce Expression of Virulence-Related Proteins.
abstract: The entomopathogenic fungus Metarhizium anisopliae is used to control insect pests. This species is specialized for the secretion of an enzymatic complex consisting of proteases, lipases, and chitinases related to pathogenicity and virulence. In this context, the secretomes of strains IBCB 167 and IBCB 384 of M. anisopliae var. anisopliae, grown under submerged fermentation in the presence of chrysalis as an inducer, were analyzed. Analysis of two-dimensional gels showed qualitative and quantitative differences between secreted proteins in both isolates. Around 102 protein spots were analyzed, and 76 % of the corresponding proteins identified by mass spectrometry were grouped into different classes (hydrolases, oxidases, reductases, isomerases, kinases, WSC domains, and hypothetical proteins). Thirty-three per cent of all the proteins analyzed were found to be common in both strains. Several virulence-related proteins were identified as proteases and mannosidases. Endo-N-acetyl-β-D-glucosaminidase expression was observed to be 10.14-fold higher for strain IBCB 384 than for strain IBCB 167, which may be an important contributor to the high virulence of IBCB 384 in Diatraea ssaccharalis. These results are important for elucidation of the host-pathogen relationship and the differences in virulence observed between the two strains.
authors.affiliation: Department of Biology, Faculty of Philosophy, Science and Literature of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, São Paulo, 14040-901, Brazil.
authors.forename: Cynthia Barbosa
authors.initials: CB
authors.lastname: Rustiguel
authors.affiliation: Department of Molecular and Cell Biology, and Pathogenic Bioagents, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, São Paulo, 14040-901, Brazil.
authors.forename: José Cesar
authors.initials: JC
authors.lastname: Rosa
authors.affiliation: Department of Biology, Faculty of Philosophy, Science and Literature of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, São Paulo, 14040-901, Brazil.
authors.forename: João Atílio
authors.initials: JA
authors.lastname: Jorge
authors.affiliation: Department of Chemistry, Faculty of Philosophy, Science and Literature of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, São Paulo, 10040-901, Brazil.
authors.forename: Arthur Henrique Cavalcanti
authors.initials: AHC
authors.lastname: de Oliveira
authors.affiliation: Department of Biology, Faculty of Philosophy, Science and Literature of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirão Preto, São Paulo, 14040-901, Brazil. lhguimaraes@ffclrp.usp.br.
authors.forename: Luis Henrique Souza
authors.initials: LHS
authors.lastname: Guimarães
mesh_terms: D000818:Animals; D012831:Bombyx / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004798:Enzymes / Q000378:metabolism; D005285:Fermentation; D005656:Fungal Proteins / Q000378:metabolism*; D013058:Mass Spectrometry; D052981:Metarhizium / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D037521:Virulence Factors / Q000032:analysis*
chemical_list: D004798:Enzymes; D005656:Fungal Proteins; D020543:Proteome; D037521:Virulence Factors