解淀粉芽孢杆菌Lx-11产脂肽类物质鉴定及表面活性素对水稻细菌性条斑病的防治作用.pdfVIP

中国农业科学 2013,46(10):2014-2021 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.10.006 解淀粉芽孢杆菌Lx-11 产脂肽类物质鉴定及 表面活性素对水稻细菌性条斑病的防治作用 张荣胜，王晓宇，罗楚平，刘永锋，刘邮洲，陈志谊 （江苏省农业科学院植物保护研究所，南京 210014 ） 摘要：【目的】对解淀粉芽孢杆菌Lx-11 产生的脂肽类物质进行分离鉴定和水稻条斑病防效测定。【方法】根 据脂肽类抗菌物质合成相关基因序列对Lx-11 进行PCR 扩增检测，采用MALDI-TOF-MS 对脂肽类物质进行分析。【结 果】通过3 对特异性引物对Lx-11 菌株检测，扩增产物经克隆、测序和BLAST 分析，表明解淀粉芽孢杆菌Lx-11 基因组中含有sfp、fenB、ituA 或bamA 基因。质谱分析发现提取的粗提物中含有surfactin、bacillomycin D 和 fengycin 等3 种脂肽类抗生素。通过构建surfactin 突变体菌株，平板和盆栽试验显示突变体菌株抑制条斑病菌 能力显著降低。【结论】解淀粉芽孢杆菌Lx-11 产生的surfactin 类物质对条斑病菌具有较强的抑制作用，是抑制 条斑病菌主要物质之一。 关键词：解淀粉芽孢杆菌；细菌性条斑病；脂肽类物质；生物防治 Identification of the Lipopeptides from Bacillus amyloliquefaciens Lx-11 and Biocontrol Efficacy of Surfactin Against Bacterial Leaf Streak ZHANG Rong-sheng, WANG Xiao-yu, LUO Chu-ping, LIU Yong-feng, LIU You-zhou, CHEN Zhi-yi (Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014) Abstract ：【Objective 】The objective of this study is to isolate and identify the lipopeptides produced by Bacillus amyloliquefaciens Lx-11 and their biocontrol efficacy against bacterial leaf streak (BLS). 【Method 】Three primers designed based on the known lipopeptide genes were used to amplify the corresponding genes from the genome of Lx-11 strain. Lipopeptides were identified by MALDI-TOF-MS techniques. 【Result 】The PCR products with three primer pairs were cloned and sequenced. The BLAST analysis showed that the sfp , fenB , ituA or bamA genes exist in the genome of B. amyloliquefaciens Lx-11. Three kinds of lipopeptides surfactin, bacillomycin D and fengycin were identified in the antimicrobial extract by the mass spectrum analysi s. The antibacterial activity c