【作者】 晏立英； 周乐聪； 谈宇俊； 单志慧； 沈明珍； Leonid Chernin；

【Author】 YAN Li-ying, ZHOU Le-cong, TAN Yu-jun, SHAN Zhi-hui, LEONID Chernin,SHEN Ming-zhen(Oil Crops Research Institute, CAAS, Wuhan 430062, Hubeiprovince, China, Faculty of Agricultural of Hebrew University of Jerusalem Rehovot, 71600, Israel)

【机构】 中国农业科学院油料作物研究所,农业部油料作物遗传改良重点实验室 武汉 430062； 以色列耶路撒冷希伯莱大学农学院；

【摘要】 利用PCR方法扩增生防细菌成团肠杆菌(Enterobacter agglomerans,IC1270)基因组调节基因GrrS,GrrA,rpoD,rpoS,插入质粒pGEMT-easy中;切下目的片段与酶切的穿梭质粒pJFF224-XN连接,经PCR方法及酶切鉴定,调节基因片段分别以正向或反向插入到质粒的T4启动子下;分别将带有调节基因的穿梭质粒导入IC1270的感受态细胞,得到带有同源调节基因不同插入方向的IC1270衍生物。抑菌实验结果表明,正向插入的调节基因未提高对病原真菌的生防效果,而反向插入的调节基因丧失对病原真菌的颉抗作用,推测与抑制了调节基因的转录有关。更多还原

【Abstract】 Regulatory genes derived from Enterobacter Agglomerans (IC 1270) genomic DNA were amplified by PCR, the fragments were inserted into pGEMT-esay vector. The target fragments were cut down from pGEMT-easy and ligated with shuttle vector pJFF224-XN/E.coRI. Through identification, the homogenous regulatory gene inserted in cis and trans direction under T4 promoter, separately. IC 1270 competent cells were transformed with these constructs. The results of the antagonistic assay are: in cis direction, there are no improving biocontrol effect comparing with parent bacteria; in trans direction, derivatives completely lost the antagonistic ability to pathogen, supposed suppressing the transcription of regulatory genes.更多还原