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生防菌株ZA1的GFP基因标记及其功能稳定性测定
  • ISSN号:0577-7518
  • 期刊名称:《植物保护学报》
  • 时间:0
  • 分类:S436.341.1[农业科学—农业昆虫与害虫防治;农业科学—植物保护]
  • 作者机构:[1]甘肃农业大学植物保护学院,甘肃省农作物病虫害生物防治工程实验室,兰州730070, [2]甘肃农业大学,甘肃省作物遗传改良与种质创新重点实验室,兰州730070
  • 相关基金:国家现代农业(马铃薯)产业技术体系(CARS-10-P18),国家自然科学基金(31660148)
中文摘要:

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p GFP78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p GFP78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p GFP78,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。

英文摘要:

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 tech- nique were used to test the functional stability of strain ZA1. The results showed that pGFP78 has trans- formed into strain ZA1 successfully, there was no difference in morphology between positive strain ZAl-gfp and strain ZA1, the ZAl-gfp appeared bright green under the fluorescence microscope, and ob- tained promoter 78 of pGFP78 by PCR after plasmid extraction from ZAl-gfp. The stability turned out to 100% and there was no obvious difference was found in the growth curves of strains ZA1 and ZAI- 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 ZAl-gfp against F. avenaceum, C. coccodes and P. foveata were 67.46%, 76.56% and 66.75%, respectively, which indicat- ed that genetic transformation had no impact on the ability of biocontrol B. mojavensis.

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期刊信息
  • 《植物保护学报》
  • 中国科技核心期刊
  • 主管单位:中国科学技术学会
  • 主办单位:中国植物保护协会
  • 主编:彩万志
  • 地址:北京圆明园西路中国农业大学植保学院
  • 邮编:100094
  • 邮箱:zbxb@cau.edu.cn
  • 电话:010-62732528
  • 国际标准刊号:ISSN:0577-7518
  • 国内统一刊号:ISSN:11-1983/S
  • 邮发代号:82-620
  • 获奖情况:
  • 曾获中国科协优秀期刊奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:14691