| Summary: | <i>Clonostachys rosea</i> is an important mycoparasite, with great potential for controlling numerous plant fungal diseases. Understanding the mechanisms and modes of action will assist the development and application of this biocontrol fungus. In this study, the highly efficient <i>C. rosea</i> 67-1 strain was marked with the green fluorescent protein (GFP), and the transformant possessed the same biological characteristics as the wild-type strain. Fungal interactions with <i>Botrytis cinerea</i> during co-culture and encounter on tomato leaves were assessed by fluorescence confocal and electron microscopy. The results indicated that once the two fungi met, the hyphae of <i>C. rosea</i> grew alongside those of <i>B. cinerea</i>, then attached tightly to the host and developed special structures, via which the biocontrol fungus penetrated the host and absorbed nutrients, eventually disintegrating the cells of the pathogen. Mycoparasitism to <i>B. cinerea</i> was also observed on tomato leaves, suggesting that <i>C. rosea</i> can colonize on plants and act following the invasion of the pathogenic fungus.
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