Zhang Lin, Yang Jinkui, Niu Qiuhong, Zhao Xuna, Ye Fengping, Liang Lianming, Zhang Ke-Qin
Laboratory for Conservation and Utilization of Bio-resources, Yunnan University, Kunming, PR China.
Appl Microbiol Biotechnol. 2008 Apr;78(6):983-90. doi: 10.1007/s00253-008-1392-7. Epub 2008 Feb 22.
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.
粉红粘帚霉（同义词：粉红粘帚菌）是一种潜在的生物防治剂。它可以抑制植物病原真菌灰葡萄孢的孢子形成并杀死致病线虫，但对线虫致病过程的了解却很少。为了帮助理解其潜在机制，我们构建了含有增强型绿色荧光蛋白基因egfp和潮霉素抗性基因hph的质粒的重组菌株。使用荧光显微镜监测绿色荧光蛋白（GFP）的表达。我们的观察结果表明，致病过程始于分生孢子附着在线虫角质层上进行萌发，随后芽管穿透线虫体内，随后线虫死亡并降解。这些是关于用GFP标记的真菌病原体感染过程的首次发现，并且所开发的方法可以成为研究粉红粘帚霉对线虫感染分子机制的重要工具。
