Applied Microbiology and Biotechnology

, Volume 78, Issue 6, pp 983–990 | Cite as

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

  • Lin Zhang
  • Jinkui Yang
  • Qiuhong Niu
  • Xuna Zhao
  • Fengping Ye
  • Lianming Liang
  • Ke-Qin ZhangEmail author
Applied Genetics and Molecular Biotechnology


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.


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



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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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Lin Zhang
    • 1
    • 2
  • Jinkui Yang
    • 1
    • 2
  • Qiuhong Niu
    • 1
    • 2
  • Xuna Zhao
    • 1
    • 2
  • Fengping Ye
    • 1
    • 2
  • Lianming Liang
    • 1
    • 2
  • Ke-Qin Zhang
    • 1
    • 2
    Email author
  1. 1.Laboratory for Conservation and Utilization of Bio-resourcesYunnan UniversityKunmingPeople’s Republic of China
  2. 2.Key Laboratory for Microbial Resources of the Ministry of EducationYunnan UniversityKunmingPeople’s Republic of China

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