Applied Genetics and Molecular Biotechnology

Applied Microbiology and Biotechnology

, Volume 78, Issue 6, pp 983-990

First online:

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

  • Lin ZhangAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Jinkui YangAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Qiuhong NiuAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Xuna ZhaoAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Fengping YeAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Lianming LiangAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University
  • , Ke-Qin ZhangAffiliated withLaboratory for Conservation and Utilization of Bio-resources, Yunnan UniversityKey Laboratory for Microbial Resources of the Ministry of Education, Yunnan University Email author 

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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.

Keywords

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