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  

The Identification of Antagonistic Bacteria B-04 against Botrytis cinerea and Improving Biocontrol Effect through Genetic Modification with β-1,3-glucanase Gene

作　　者：李桂霞[1] 马汇泉[1] 刘婧[1] 董瑾[1] 

机构地区：[1]山东理工大学生命科学学院,淄博255049

出　　处：《中国生物工程杂志》2007年第4期44-49,共6页China Biotechnology

基　　金：山东省教育厅研究基金资助项目(J04C08)

摘　　要：从淄博温室土壤中分离到菌株B-04,对灰霉病菌表现较高的拮抗作用。采用形态特征观察、生理生化特性测定以及16SrDNA序列分析对其进行鉴定,为蜡样芽孢杆菌(Bacillus cereus)。从质粒puc1940得到含有β-1,3-葡聚糖酶基因的4.1kb片段,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B-04菌株,获得工程菌株B-04-glu。与野生菌株相比,平板拮抗试验表明工程菌株对番茄灰霉病(Botrytis cinerea)抑菌效果明显增强。Bacillus cereus B-04 antagonist to Botrytis cinerea were isolated from samples of tomato soil infected by Botrytis cinerea in Zibo, which are identified through a series of morphological and biochemical characteristics and the sequence of 16SrDNA. Aiming at enhancing the inhibitory effect of this strain, a 4.1 kb DNA fragment containing β-1,3-glucanases gene from pUC1940 was inserted into vector pBE2 and pHY300PLK to construct recombination plasmids, PBE2-glu and pHY300PLK-glu, which were transferred into Bacillus cereus B-04, resulting in a new strain named B-04-glu. Restriction enzyme digestion and β-1,3-glucanases plate culture confirmed that B-04-glu contained a functional β-1,3-glucanases gene. Compared to the wild strain B-04, B-04- glu had an increased inhibitory effect against Botrytis cinerea on tomato.

关 键 词：蜡样芽孢杆菌 Β-1 3-葡聚糖酶基因 灰霉病 

分 类 号：Q78[生物学—分子生物学]