番茄灰霉病高效拮抗菌株的鉴定及通过导入β-1,3-葡聚糖酶基因提高其生防效果

从淄博市温室土壤中分离到蜡样芽孢杆菌B-04菌株，对灰霉病菌表现较高的拮抗作用。本研究从质粒pUC1940得到4.1kb的β-1,3-葡聚糖酶基因片断，将该基因与大肠杆菌-芽孢杆菌穿梭质粒pBE2和pHY300PLK连接，获得重组质粒PBE2-glu和pHY300PLK-glu，转入蜡样芽孢杆菌（Bacillus cereus）B-04菌株，获得工程菌株B-04-glu。限制酶切分析、ABP平板、PCR实验证实B-04成功转入β-1,3-葡聚糖酶基因。与野生菌株相比，平板拮抗试验表明工程菌株较原始菌株对番茄灰霉病（Botrytis cinerea）抑菌效果明显增强。

Improving Biocontrol Effect of Bacillus circus on Botrytis cinerea through Genetic Modification with β-1,3-glucanase

Bacillus cereus B-04 was isolated in zibo soil and proved as a biocontrol agent of Botrytis cinerea. The aim of this study was to enhance the biocontrol effect of this strain by introduction of a β-1,3-glucanase gene. A 4.1kb DNA fragment containing β-1,3-glucanase gene was inserted into vector pBE2 and pHY300PLK to construct a new plasmid, PBE2-glu and pHY300PLK-glu,which was introduced into Bacillus cereus B-04, resulting in a new strain named B-04-glu.Restriction digestion and β-1,3-glucanase plate culture and PCR experiment confirmed that B-04-glu contained a functional β-1,3-glucanase gene. Compared with wild type B-04 in pot experiment, B-04-glu had increased effect against Botrytis cinerea caused by tomato.