基于丁香假单胞菌基因中断技术的烟草 野火病菌基因敲除  

Gene knockout in Pseudomonas amygdalipv.tabaci based on Pseudomonas syringae gene disruption technique

作　　者：赫英宇 吴琳 李尊强[3] 乔婵[3] 万秀清[3] 潘洪杏 郭兆奎[3] 李若[3] HE Yingyu;WU Lin;LI Zunqiang;QIAO Chan;WAN Xiuqing;PAN Hongxing;GUO Zhaokui;LI Ruo(Mudanjiang Tobacco Leaf Company,Heilongjiang Provincial Tobacco Company,Mudanjiang 157000,Heilongjiang,China;Shuangcheng Branch of Harbin Tobacco Company,Harbin 150100,China;Institute of Tobacco Science,Heilongjiang Provincial Tobacco Company of CNTC,Harbin 150076,China)

机构地区：[1]黑龙江省烟草公司牡丹江烟叶公司,黑龙江省牡丹江市157000 [2]哈尔滨市烟草公司双城分公司,哈尔滨市150100 [3]中国烟草总公司黑龙江省公司烟草科学研究所,哈尔滨市150076

出　　处：《烟草科技》2025年第4期26-33,共8页Tobacco Science & Technology

基　　金：黑龙江省烟草专卖局科技开发项目“抗角斑病烟草材料的创制”(2022230000200045);“烟草野火病抗病基因定位及龙江911抗性改良”(2023230000200148)。

摘　　要：为建立烟草野火病菌基因敲除技术体系,以烟草野火病菌0号生理小种菌株D151为材料,选择质粒pEX18Gm为出发载体,构建pEX18Gm-HopF1-KO自杀质粒,经过接合转移与多轮抗性筛选以敲除烟草野火病菌0号生理小种中的HopF1效应蛋白基因。结果表明,通过菌液PCR鉴定,获得5个阳性突变菌株,阳性转化率为50%,测序结果进一步验证了HopF1基因敲除菌株构建成功。本研究中建立了烟草野火病菌基因敲除技术体系,为培育野火病菌生理小种专化型抗病烟草和克隆烟草中生理小种专化型抗病基因奠定了基础。To establish the gene knockout technical system for Pseudomonas amygdalipv.tabaci(Pta),Pta strain D151(race 0)was used as material and plasmid pEX18Gm was selected as the starting vector to construct the pEX18Gm-HopF1-KO suicide plasmid.After conjugation transfer and several rounds of resistance screening,the effector HopF1 was knocked out in the Pta strain(race 0).The results showed that five positive mutant strains were obtained by PCR identification of the bacterial solution,and the positive transformation rate was 50%.Sequencing results further confirmed that the HopF1 gene knockout strains were successfully constructed.This study showed that the technical system of Pta gene knockout was successfully established,which lay the foundation for the cultivation of Pta-race-specialized resistant tobacco varieties and the cloning of Pta-race-specialized resistant genes in tobacco.

关 键 词：烟草 野火病菌 效应蛋白HopF1 基因敲除 

分 类 号：TS413[农业科学—烟草工业]