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Secretome Analysis of Metarhizium anisopliae Under Submerged Conditions Using Bombyx mori Chrysalis to Induce Expression of Virulence-Related Proteins

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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.

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Acknowledgments

We thank Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP – Process No. 2011/50880-1) and CAPES (Coordenação de Aperfeiçoamento de Pessoal do Ensino Superior) for financial support, and Maurício de Oliveira for technical assistance. This manuscript is part of the CBR Ph.D. thesis presented to the Department of Biology, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto -USP, São Paulo, Brazil.

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Authors and Affiliations

  1. 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

    Cynthia Barbosa Rustiguel, João Atílio Jorge & Luis Henrique Souza Guimarães

  2. 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

    Arthur Henrique Cavalcanti de Oliveira

  3. 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

    José Cesar Rosa

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  1. Cynthia Barbosa Rustiguel
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  2. José Cesar Rosa
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  3. João Atílio Jorge
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  4. Arthur Henrique Cavalcanti de Oliveira
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  5. Luis Henrique Souza Guimarães
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Correspondence to Luis Henrique Souza Guimarães.

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Rustiguel, C.B., Rosa, J.C., Jorge, J.A. et al. Secretome Analysis of Metarhizium anisopliae Under Submerged Conditions Using Bombyx mori Chrysalis to Induce Expression of Virulence-Related Proteins. Curr Microbiol 72, 220–227 (2016). https://doi.org/10.1007/s00284-015-0943-2

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  • DOI: https://doi.org/10.1007/s00284-015-0943-2

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