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Biochemical and molecular characterization of a putative endoglucanase in Magnaporthe grisea

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Abstract

Microbial pathogens secrete an array of cell wall-degrading enzymes to break down the structure of the host cell wall to facilitate colonization of the host tissue. To better understand their role in the pathogenesis, a putative endoglucanase from Magnaporthe grisea was characterized in this paper. SignalP-3.0 analysis indicates that the protein encoded by gene MGG_02532.5 in M. grisea (named MgEGL1 for M . g risea endoglucanase 1 ) contains a secretory signal peptide. Multiple alignment shows that MgEGL1 has high level of homology to endoglucanases (EC 3.1.1.4) from Aspergillus nidulans and Trichoderma reesei. The three proteins share a conserved catalytic domain, but only the one from T. reesei contains a cellulose binding module. MgEGL1 was constitutively expressed with the highest level in mycelia and the lowest in conidia. Interestingly, the MgEGL1 RNA could be alternatively processed when cultured in vitro and after infection of rice. Expression analysis confirmed that the MgEGL1 is a secreted protein. Its endoglucanase activity was assayed by Congo red plates, and further confirmed by the dinitrosalicylic acid method. The finding in this paper will provide the basis for further determination of the biochemical properties of the endoglucanase protein and its relevant function in fungal pathogenesis.

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Acknowledgments

This work was supported by the grant National Natural Science Funds of China (No. 30471132 and No. 30671348) and US National Science Foundation (grant DBI0115642). We thank Kiran Bhattarai, Hanno Wolfe, Cristina Fillipi, and Dan Li for technical assistance.

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Authors and Affiliations

  1. The Key Laboratory of Biopesticide and Chemistry Biology, Ministry of Education, Fujian Agriculture and Forestry University, 350002, Fuzhou, People’s Republic of China

    Jie Zhou, Xiong-jie Lin, Guo-dong Lu & Zong-hua Wang

  2. The School of Life Sciences, Fujian Agriculture and Forestry University, 350002, Fuzhou, People’s Republic of China

    Jie Zhou, Xiang-zi Zheng, Lan Lan, Chen-zeng Lin, Yu-bing Wu & Zong-hua Wang

  3. Departmant of Plant Pathology and Microbiology, Texas A & M University, College Station, TX, 77843-2132, USA

    Daniel Ebbole

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  1. Jie Zhou
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  2. Xiang-zi Zheng
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  3. Lan Lan
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  4. Chen-zeng Lin
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  5. Yu-bing Wu
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  7. Daniel Ebbole
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  8. Guo-dong Lu
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  9. Zong-hua Wang
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Correspondence to Guo-dong Lu or Zong-hua Wang.

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Communicated by U. Kück.

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Zhou, J., Zheng, Xz., Lan, L. et al. Biochemical and molecular characterization of a putative endoglucanase in Magnaporthe grisea . Curr Genet 53, 217–224 (2008). https://doi.org/10.1007/s00294-008-0179-9

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  • DOI: https://doi.org/10.1007/s00294-008-0179-9

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