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Overexpression of a cuticle-degrading protease Ver112 increases the nematicidal activity of Paecilomyces lilacinus

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Abstract

Due to their ability to degrade the proteins in nematode cuticle, serine proteases play an important role in the pathogenicity of nematophagous fungi against nematodes. The serine protease Ver112 was identified from the nematophagous fungus Lecanicillium psalliotae capable of degrading the nematode cuticle and killing nematodes effectively. In this study, the gene ver112 was introduced into the commercial biocontrol fungal agent Paecilomyces lilacinus by the restriction enzyme-mediated integration transformation. Compared to the wild strain, the transformant P. lilacinus 112 showed significantly greater protease activity, with nematicidal activities increased by 79% and 96% to Panagrellus redivivus and Caenorhabditis elegans at the second day, respectively. The crude protein extract isolated from the culture filtrate of P. lilacinus 112 also showed 20–25% higher nematicidal activity than that of the wild-type strain. Reverse transcription PCR results showed that the expression of gene ver112 in P. lilacinus 112 was correlated to protease activity of the culture filtrate. Our results demonstrated the first successful transfer of a virulence gene from one nematophagous fungus to another nematophagous fungus, and improved the pathogenicity of the recipient fungus against pest nematodes.

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Acknowledgements

We are grateful to Prof. Jian** Xu of the Department of Biology, McMaster University, for the valuable comments and critical discussions. This work was supported by National Basic Research Program of China (approved no. 2007CB411600), by projects from the National Natural Science Foundation of China (approved nos. 30630003, 30960229, and 30660107), by project from the Major State Basic Research Development Program (2009CB125905), the Department of Science and Technology of Yunnan Province (approved nos. 2007C007Z and 2009CI052), and the Yunnan Branch of China Tobacco Industrial Corporation (grant no. 2010yn17).

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  1. Laboratory for Conservation and Utilization of Bio-Resources, and Key Laboratory for Microbial Resources of the Ministry of Education, Yunnan University, Kunming, 650091, People’s Republic of China

    **kui Yang, Xuna Zhao, Lianming Liang, Xuemei Niu, Chenggang Zou & Ke-Qin Zhang

  2. Tobacco Agricultural Science Institute of Yunnan Province, Yuxi, 653100, People’s Republic of China

    Zhenyuan ** Lei

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  1. **kui Yang
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Correspondence to Ke-Qin Zhang.

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**kui Yang and Xuna Zhao contributed equally to this work.

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Yang, J., Zhao, X., Liang, L. et al. Overexpression of a cuticle-degrading protease Ver112 increases the nematicidal activity of Paecilomyces lilacinus . Appl Microbiol Biotechnol 89, 1895–1903 (2011). https://doi.org/10.1007/s00253-010-3012-6

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  • DOI: https://doi.org/10.1007/s00253-010-3012-6

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