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

doi:10.3864/j.issn.0578-1752.2013.10.006

中国农业科学 ›› 2013, Vol. 46 ›› Issue (10): 2014-2021.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

收稿日期:2013-01-08 出版日期:2013-05-15 发布日期:2013-03-28
通讯作者: 通信作者陈志谊，Tel：025-84390393；E-mail：chzy@jaas.ac.cn
作者简介:张荣胜，E-mail：r_szhang@163.com
基金资助:
国家公益性行业（农业）科研专项经费项目（201103002-3，201303015）、江苏省农业科技自主创新资金项目（CX（11）2045）

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

Received:2013-01-08 Online:2013-05-15 Published:2013-03-28

摘要/Abstract

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

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 analysis. The antibacterial activity could be associated with surfactin-lipopeptides, and practically abolished in surfactin-deficient mutants. Lx-11 strain significantly reduced disease incidence of BLS.【Conclusion】The antibacterial activity of the surfactin- deficient mutant strain was practically abolished, which suggested that surfactin might play an important role in control of BLS.

Key words: Bacillus amyloliquefaciens , bacterial leaf streak (BLS) , lipopeptides , biological control

张荣胜, 王晓宇, 罗楚平, 刘永锋, 刘邮洲, 陈志谊. 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[J]. 中国农业科学, 2013, 46(10): 2014-2021.

ZHANG Rong-Sheng, WANG Xiao-Yu, LUO Chu-Ping, LIU Yong-Feng, LIU You-Zhou, CHEN Zhi-Yi. Identification of the Lipopeptides from Bacillus amyloliquefaciens Lx-11 and Biocontrol Efficacy of Surfactin Against Bacterial Leaf Streak[J]. Scientia Agricultura Sinica, 2013, 46(10): 2014-2021.

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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

Identification of the Lipopeptides from Bacillus amyloliquefaciens Lx-11 and Biocontrol Efficacy of Surfactin Against Bacterial Leaf Streak

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