番茄青枯病植物疫苗工程菌的构建及其防效评价.docVIP

番茄青枯病植物疫苗工程菌的构建及其防效评价 郑雪芳1,2，刘波1，林乃铨2，朱育菁1，车建美1，林抗美1 （1福建省农业科学院农业生物资源研究所 福州 350003; 2 350002） 摘 要：利用青枯雷尔氏菌无致病力菌株作为植物疫苗防治番茄青枯病具有很好的应用潜力。本研究通过自发突变、60Co辐射诱变和EZ-Tn5插入诱变来构建青枯雷尔氏菌无致病力突变株体库，结果分别从番茄青枯发病田块分离到3株自发致弱的无致病力青枯雷尔氏菌，从60Co辐射诱变获得12株无致病力青枯雷尔氏菌，从EZ-Tn5插入诱变获得40株无致病力青枯雷尔氏菌。比较不同方式致弱的无致病力青枯雷尔氏菌对番茄青枯病的温室控病效果，结果表明自发致弱的无致病力青枯雷尔氏菌FJAT-1458对番茄青枯病的控病效果最好，接种致病性青枯雷尔氏菌后第10d，植株发病率和病情指数均为0，对照组发病率和病情指数均达到100%。进一步研究菌株FJAT-1458 在番茄根系土壤和植株体内的定殖特性及对番茄青枯病的免疫抗病效果，结果表明，菌株FJAT1458能在番茄植株根系土壤及根部和茎部定殖，不能在叶片定殖，定殖数量均表现为“由增到减”的趋势。接种浓度和接种苗龄均会影响菌株FJAT1458在番茄植株根系土壤和体内的定殖数量，接种浓度越大、接种苗龄越小，定殖数量越大；控病试验表明，不同接种浓度的无致病力青枯雷尔氏菌FJAT1458对番茄青枯病均具有控病效果，接种浓度越大，番茄植株发病率越低，防治效果越好，接种浓度为108cfu/ml，其防治效果可达60%。从构建的番茄青枯病的控病模型可以看出，菌株FJAT1458在不同接种浓度条件下，植株发病率随时间变化符合的回归方程不同，相关系数R值也不同，接种浓度越大，R值越小。 关键词：植物疫苗，青枯雷尔氏菌，番茄青枯病，定殖，生物防治 Colonization and control efficiency of plant vaccine with avirulent Ralstonia solanacearum FJAT1458 on tomato bacterial wilt disease Zheng Xuefang1,2, Liu Bo1, Lin Neiquan2, Zhang Wenzhou1, Zhu Yujing1, Lin Kangmei1,Che Jianmei1 （1Biotechnology Institution, Fujian Academy of Agricultural Sciences, Fuzhou 350003 2Biological Research Institute, Fujian Agriculture and Forestry University, Fuzhou 350002） Abstract: Avirulent Ralstonia solanacearum strain FJAT1458 was isolated from tomato, which was an efficient biological control agent on tomato bacterial wilt. In this paper, the colonizing ability of avirulent Ralstonia solanacearum strain FJAT1458 in rhizosphere soil and plant of tomato were tested by the different inoculated concentration and the different seedling ages, in addition, the control effect of the plant vaccine on the bacterial wilt disease in tomato was evaluated. The result of colonization test showed that rhizosphere soil, root and stem, but not leaf, could be colonized by the plant vaccine (strain FJAT1458). The population of strain FJAT1458 showed the trend of “in-crease at beginning and decrease afterward” during 1-25d after inoculation. Both inoculated concentration and seedling ages had