青枯病生防菌蜡状芽孢杆菌(ANTI-8098A)的绿色荧光蛋白基因(gfp)转导及其生物学特性的变化  

Tagging of Bacterial-wilt-disease Biocontrol Bacteria ANTI-8098A(Bacillus cereus) with the Green Fluorescence Protein Gene and its Biological Characteristics

作　　者：车建美[1,2] 刘波[1] 张彦[1,3] 蓝江林[1] 

机构地区：[1]福建省农业科学院农业生物资源研究所,福州350003 [2]福建农林大学生物农药与化学生物学教育部重点实验室,福州350002 [3]福建农林大学植物保护学院,福州350002

出　　处：《农业生物技术学报》2010年第2期337-345,共9页Journal of Agricultural Biotechnology

基　　金：国家自然基金(30871667);国家高技术研究与发展计划项目(2006AA10A211);福建省自然基金项目(2008J0054);福建省财政专项-福建省农业科学院科技创新团队建设基金(STIF-Y03);福建省农科院青年人才创新基金项目(B2007QJ07)共同资助

摘　　要：采用质粒pCM20进行青枯病生防菌蜡状芽孢杆菌(Bacillus cereus,ANTI-8098A)的电击转化,对电击转化条件的优化结果表明,当电击条件为2.5kV,200Ω,25μF时,转化的最佳条件为质粒DNA含量118ng,感受态细胞浓度为1.14×108CFU/mL。转化子ANTI-8098A:pCM20在无红霉素选择压力的LB培养基转接16次后,绿色荧光的表达仍为100%。对其生长曲线的测定结果表明,青枯病生防菌ANTI-8098A的生长速度和ANTI-8098A:pCM20基本一致。不同培养温度和培养转速对ANTI-8098A:pCM20生长的影响较大,当培养温度为35℃时,ANTI-8098A:pCM20的含量为60.0×108CFU/mL,当培养转速为250r/min时,ANTI-8098A:pCM20的含菌量为50.33×108CFU/mL。青枯病生防菌ANTI8098A:pCM20和ANTI8098A对不同作物分离的青枯雷尔氏菌(Ralstonia solanacearum)具有明显的抑制作用,抑菌效果无显著差异。The transformation syst em by electroporation of the biocontrol bacteria ANTI-8098A（Bacillus cereus） was optimized in this paper.The results showed that the system was best for the electroporation when the concentration of compentent cell was 1.14×108 CFU/mL and the content of plasmid was 118 ng.The percent of the green cells was 100% when the strains of ANT8098A：pCM20 was reinculated in the non-selective medium for 16 times.And the growth speed of gfp-tagged strains was almost the same to the wild type strain ANTI-8098A.The best culture conditions of wild type strain and gfp-tagged one including the culture temperature of 35℃ and rotation speed of 250 r/min were almost the same.Both strains ANTI8098A and ANTI8098A：pCM20 showed the same in vitro inhibition of Ralstonia solanacearum from different crops.

关 键 词：青枯病 生防菌 蜡状芽孢杆菌 绿色荧光蛋白 

分 类 号：S432.42[农业科学—植物病理学]