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实验室保存菌Fh对番茄灰霉病有明显的抑制作用,通过形态特征、生理生化特征进行初步鉴定归属于芽孢杆菌属(Bacillus circulans)。从质粒pUC1965中得到含有几丁质酶基因的6.5kb DNA片段,将该基因片段与大肠杆菌-枯草芽孢杆菌穿梭质粒pBE2连接,获得重组质粒pBE2-chib。将重组质粒转入芽孢杆菌Fh中获得工程菌株Fh-chib。几丁质酶基因的PCR检测和几丁质平板实验表明几丁质酶基因被成功转入,工程菌株Fh-chib的原始粗酶液几丁质酶活为4.06U/ml。与野生菌相比,Fh-chib工程菌株对番茄灰霉病(Botrytis cinerea)抑菌效果提高34.46%。
Fh had a significant inhibitory effect on Botrytis cinerea, which was initially identified as Bacillus circulans by morphological and physiological and biochemical characteristics. The 6.5kb DNA fragment containing the chitinase gene was obtained from the plasmid pUC1965, and the gene fragment was ligated with the E. coli-Bacillus subtilis shuttle plasmid pBE2 to obtain the recombinant plasmid pBE2-chib. The recombinant plasmid was transformed into Bacillus Fh to obtain engineering strain Fh-chib. Chitinase gene PCR test and chitin plate experiments showed that the chitinase gene was successfully transferred, the original crude enzyme solution Fh-chib chitinase activity was 4.06U / ml. Compared with the wild-type strain, Fh-chib strain increased the inhibitory effect on Botrytis cinerea by 34.46%.