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Investigation on the infection mechanism of the fungus Clonostachys rosea against nematodes using the green fluorescent protein

Abstract

The fungus Clonostachys rosea (syn. Gliocladium roseum) is a potential biocontrol agent. It can suppress the sporulation of the plant pathogenic fungus Botrytis cinerea and kill pathogenic nematodes, but the process of nematode pathogenesis is poorly understood. To help understand the underlying mechanism, we constructed recombinant strains containing a plasmid with both the enhanced green fluorescent protein gene egfp and the hygromycin resistance gene hph. Expression of the green fluorescent protein (GFP) was monitored using fluorescence microscopy. Our observations reveal that the pathogenesis started from the adherence of conidia to nematode cuticle for germination, followed by the penetration of germ tubes into the nematode body and subsequent death and degradation of the nematodes. These are the first findings on the infection process of the fungal pathogen marked with GFP, and the developed method can become an important tool for studying the molecular mechanisms of nematode infection by C. rosea.

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Acknowledgments

We sincerely thank Dr. PJ Punt (TNO Nutrition and Food Research Institute, Netherlands) and Dr. Zhang YJ (Biotechnology Research Center of Southwest University, China) for providing plasmids pAN52 and pBS/Sk-hph. This work was funded by the National Basic Research Program of China (approved no. 2007CB411600), by projects from the National Natural Science Foundation of China (approved nos. 30630003, 30570059 and 30660107), and by the Department of Science and Technology of Yunnan Province (approval nos. 2004C0001Z, 2005NG05).

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Correspondence to Ke-Qin Zhang.

Additional information

Lin Zhang and Jinkui Yang contributed equally to this work.

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Zhang, L., Yang, J., Niu, Q. et al. Investigation on the infection mechanism of the fungus Clonostachys rosea against nematodes using the green fluorescent protein. Appl Microbiol Biotechnol 78, 983–990 (2008) doi:10.1007/s00253-008-1392-7

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Keywords

  • Clonostachys rosea
  • Green fluorescent protein (GFP)
  • Protoplast transformation
  • Nematode
  • Infection