Effects of culture conditions on spore types of Clonostachys rosea 67-1 in submerged fermentation.

UNLABELLED: The promising biocontrol isolate Clonostachys rosea 67-1 was investigated to clarify the effects of culture conditions on chlamydospore production in submerged fermentation. Culture conditions significantly affected both performance and types of C. rosea sporulation. C. rosea 67-1 was hard to generate chlamydospores under conventional conditions. However, the proportion of resistant spores increased to 17·4 and 15·5% in PD and rice meal media, respectively, in 8 days. Chlamydospore productivity was boosted (>threefold) with the addition of 50-200 mg l(-1) CuSO4 . The pH of the medium played a vital role in 67-1 sporulation. The percentage of chlamydospores decreased rapidly with increased pH (88·1% at pH 3·0 to 1·0% at pH 6·5). The optimal pH for conidia production was 6·0-6·5, at which chlamydospore forming was strongly inhibited. Regulating pH during fermentation contributed to improving output and proportion of resistant spores. When 67-1 was inoculated into broth with an initial pH of 6·5, followed by adjustment to pH 3·5 after 48 h, the number of chlamydospores reached 1·1 × 10(8) ml(-1). The impact of temperature and rotational speed was also analysed; an ultimate capacity of chlamydospores was achieved at 30°C and the speed above 120 rev min(-1) (P < 0·05). SIGNIFICANCE AND IMPACT OF THE STUDY: Clonostachys rosea is one of the most promising biocontrol agents in countering many plant fungal diseases. However, large-scale production and commercialization are hampered by the lack of understanding of the impacts of culture conditions on performance and types of C. rosea sporulation and subsequently inadequate research on the techniques for chlamydospore production. In addressing these concerns, this study provides a unique insight into the manipulation of C. rosea sporulation and chlamydospore fermentation of the biocontrol fungus.