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Developing Resources for Analysis of Secreted Proteins from Magnaporthe oryzae

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Advances in Genetics, Genomics and Control of Rice Blast Disease

Abstract

Extracellular proteins of fungal pathogens are candidate effector mole-cules. We have cloned a large number (∼300) of putative secreted proteins from Magnaporthe oryzae. The proteins were fused to a His(× 6) tag to allow for affinity purification of the proteins. The genes were transformed into M. oryzae to test for expression. Approximately one-third of the genes could be expressed and secreted to sufficient levels to allow detection by western blot analysis of culture filtrates. In most cases, constitutive expression of the proteins in the transformed strains did not markedly alter their interaction with rice. However, a number of transformed lines may have reduced aggressiveness as a result of protein expression. Proteins were purified from culture filtrates of M. oryzae and symptoms were observed on rice leaves in only very few cases. The results to date are summarized and the resources developed in the project are being used for functional analysis of secreted protein. The resources generated are being distributed to facilitate functional characterization of the secretome of M. oryzae

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References

  • Finn, R.D., Tate, J., Mistry, J., Coggill, P.C., Sammut, J.S., Hotz, H.R., Ceric, G., Forslund, K., Eddy, S.R., Sonnhammer, E.L. and Bateman., A. (2008). The PFAM protein families database. Nucleic Acids Res. Database Issue 36:D281–D288.

    Google Scholar 

  • Jeong, J.S., Mitchell, T.K., and Dean, R.A. (2007). The Magnaporthe grisea snodprot1 homolog, MSP1, is required for virulence. FEMS Microbiol. Lett. 273:157–165.

    Article  PubMed  CAS  Google Scholar 

  • Kamoun, S., Hamada, W., and Huitema, E. (2003). Agrosuppression: a bioassay for the hypersensitive response suited to high-throughput screening. Mol. Plant Microbe. Interact. 16:7–13.

    Article  PubMed  CAS  Google Scholar 

  • Laugé, R., Goodwin, P. H., de Wit, P. J. G. M., and Joosten, M. H. A. J. (2000). Specific HR-associated recognition of secreted proteins from Cladosporium fulvum occurs in both host and non-host plants. Plant J. 23:735–745.

    Article  PubMed  Google Scholar 

  • Lu, G., Filippi, C., Li, D., and Ebbole, D. J. 2004. Identification and characterization of secreted proteins in Magnaporthe grisea. In Rice Blast: Interaction with rice and control. Proceedings of the Third International Rice Blast Conference. S. Kawasaki, (Ed.) Kluwer Academic Publishers, The Netherlands, pp. 47–55.

    Google Scholar 

  • Nielsen, H., and Krogh, A. (1998). Prediction of signal peptides and signal anchors by a hidden Markov model., In Proceedings of the Sixth International Conference on Intelligent Systems for Molecular Biology (ISMB 6), AAAI Press, Menlo Park, California, pp. 122–130.

    Google Scholar 

  • Nielsen, H., Engelbrecht, J., Brunak, S., and von Heijne, G. (1997). Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng., 10:1–6.

    Article  PubMed  CAS  Google Scholar 

  • Soderlund, C., Haller, K., Pampanwar, V., Ebbole, D., Farman, M., Orbach, M.J., Wang, G.L., Wing, R., Xu, J.R., Brown, D., Mitchell, T., and Dean, R. (2006). MGOS: A resource for studying Magnaprothe grisea and Oryza sativa interactions. Mol. Plant-Microbe Interact. 19:980–986.

    Article  Google Scholar 

  • Takken, F.L., Luderer, R., Gabriels, S.H., Westerink, N., Lu, R., de Wit, P.J., and Joosten, M.H. (2000). A functional cloning strategy, based on a binary PVX-expression vector to isolate HR-inducing cDNAs of plant pathogens. Plant J. 24:275–283.

    Article  PubMed  CAS  Google Scholar 

  • Zhou, J., Zheng, X.Z., Lan, L., Lin, C.Z., Wu, Y.B., Lin, X.J., Ebbole, D., Lu, G.D., and Wang,. Z.H. (2008). Biochemical and molecular characterization of a putative endocglucanase in Magnaporthe grisea. Curr. Genet. 53:217–224.

    Article  PubMed  CAS  Google Scholar 

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Lu, Gd. et al. (2009). Developing Resources for Analysis of Secreted Proteins from Magnaporthe oryzae . In: Wang, GL., Valent, B. (eds) Advances in Genetics, Genomics and Control of Rice Blast Disease. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-9500-9_12

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