【摘要】 为明确生防莫海威芽胞杆菌Bacillus mojavensis菌株ZA1在作物体内定殖与作物和病原物的作用机理,运用电击转化法将绿色荧光蛋白表达载体pGFP78导入菌株ZA1体内,并采用细菌培养法、继代培养法和平板抑制法检测其功能稳定性。结果表明,质粒pGFP78成功导入菌株ZA1,标记后菌株ZA1-gfp经形态学和分子生物学验证均证明菌株ZA1已成功标记,标记菌株ZA1-gfp与菌株ZA1形态一致,在荧光显微镜下呈现明亮的绿色,并且能够提取导入质粒pGFP78,扩展出质粒所携带的启动子78序列。功能稳定性鉴定结果表明,标记菌株ZA1-gfp遗传稳定性达100%;与菌株ZA1的生长速率差异不显著,且不影响菌株的运动性;菌株ZA1对马铃薯枯萎病菌、马铃薯炭疽病菌和马铃薯坏疽病菌的抑制率分别为64.50%、68.35%和66.55%,标记菌株ZA1-gfp对3种病原真菌的抑制率分别为67.46%、76.56%和66.75%,表明外源质粒的导入,不影响菌株的抑菌能力。更多还原

【Abstract】 In order to confirm the interactions between Bacillus mojavensis strain ZA1 with plant-host and pathogens after strain ZA1 was transformed into plant, and the vector of pGFP78 was transformed into biocontrol strain ZA1 by electroporation, and the germiculture, subculture and dual-culture technique were used to test the functional stability of strain ZA1. The results showed that p GFP78 has transformed into strain ZA1 successfully, there was no difference in morphology between positive strain ZA1-gfp and strain ZA1, the ZA1-gfp appeared bright green under the fluorescence microscope, and obtained promoter 78 of pGFP78 by PCR after plasmid extraction from ZA1-gfp. The stability turned out to 100% and there was no obvious difference was found in the growth curves of strains ZA1 and ZA1-gfp. In addition, the results revealed genetic transformation had no effect on the motility of strain ZA1.The inhibition rates of strain ZA1 against Fusarium avenaceum, Colletotrichum coccodes and Phoma foveata were 64.50%, 68.35% and 66.55%, respectively, and the inhibition rates of ZA1-gfp against F.avenaceum, C. coccodes and P. foveata were 67.46%, 76.56% and 66.75%, respectively, which indicated that genetic transformation had no impact on the ability of biocontrol B. mojavensis.更多还原