一株萎缩芽胞杆菌YL3的鉴定及其脂肽类化合物分析

doi:10.16409/j.cnki.2095-039x.2017.01.020

中国生物防治学报 ›› 2017, Vol. 33 ›› Issue (1): 142-150.DOI: 10.16409/j.cnki.2095-039x.2017.01.020

• 研究简报 • 上一篇

一株萎缩芽胞杆菌YL3的鉴定及其脂肽类化合物分析

刘邮洲¹, 陈夕军², 梁雪杰¹, 钱亚明³, 乔俊卿¹, 刘永锋¹

1. 江苏省农业科学院植物保护研究所, 南京 210014;
2. 扬州大学园艺与植保学院, 扬州 220009;
3. 江苏省农业科学院园艺研究所, 南京 210014

收稿日期:2016-05-26 出版日期:2017-02-08 发布日期:2017-01-24
通讯作者: 刘永锋,博士,研究员,E-mail:liuyf@jaas.ac.cn
作者简介:刘邮洲,博士,研究员,E-mail:shitouren88888@163.com
基金资助:
江苏省农业科技自主创新基金（CX-15-1037）；江苏省科技支撑计划项目（BE2014386）

Identification of Bacillus atrophaeus YL3 and Detection of Its Lipopeptide Compounds

LIU Youzhou¹, CHEN Xijun², LIANG Xuejie¹, QIAN Yaming³, QIAO Junqing¹, LIU Yongfeng¹

1. Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. College of Horticulture and Plant Protection, Yangzhou University, Yangzhou 220009, China;
3. Institute of Horticulture, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2016-05-26 Online:2017-02-08 Published:2017-01-24

摘要/Abstract

摘要： 本研究通过生理生化特征、16S rDNA基因和gyrB基因序列分析对菌株YL3进行种属鉴定，采用室内平板对峙生长法测定其抑菌活性，对草莓枯萎病的田间防效。以选择性培养基检测其分泌的多种胞外酶，以结晶紫染色法检测生物膜的形成，采用酸沉淀法提取菌株YL3脂肽类化合物并进行高效液相色谱串联质谱分析（LC-MS）。研究表明，菌株YL3被鉴定为萎缩芽胞杆菌Bacillus atrophaeus，其对多种植物病原真菌菌丝生长和细菌都具有明显的抑制作用，稀释10和100倍的发酵液对“宁玉”草莓枯萎病的防效分别为82.86%和67.62%。菌株YL3能产生蛋白酶、几丁质酶和嗜铁素，培养3 d后形成大量生物膜。菌株YL3发酵液中脂肽类粗提物对草莓枯萎病菌菌丝生长有抑制作用，随着剂量加大，抑制效果显著。LC-MS检测到菌株YL3脂肽类粗提物中含有表面活性素和泛革素。因此，菌株YL3是一株具有较好生防应用潜力的拮抗菌株。

关键词: 萎缩芽胞杆菌, 草莓枯萎病, 脂肽类化合物

Abstract: In order to investigate the biological characteristics and evaluate the antagonistic activity of the strain YL3, the biochemical characteristics, 16S rDNA and gyrB sequences were used to identify the species. The antagonistic activity of strain YL3 in vitro and control effect on strawberry fusarium wilt in field were investigated in this study. The selective medium and crystal violet staining method were used to detect the extracellular enzymes and biofilm formation of the strain YL3. The lipopeptide compounds produced by strain YL3 was extracted by acid precipitation method and analyzed by LC-MS. The results showed that strain YL3 was identified as Bacillus atrophaeus. Strain YL3 had strong antagonistic activities against the growth of pathogenic fungi such as Fusarium oxysporum, Colletotrichum fragariae, Alternaria brassicae, Sclerotinia sclerotiorumErwinia amylovora, Xanthomonas oryzae pv. oryzicola and Bacillus megaterium. The 10 times dilution and 100 times dilution of strain YL3 fermented liquid could effectively control fusarium wilt on strawberry, "Ningyu" with the control efficiency of 82.86% and 67.62%, respectively. Strain YL3 had the ability to produce proteases, chitinases and siderophores, and form and retain a thick biofilm on the well wall after culturing for 3 days. The lipopeptide compounds produced by strain YL3 could inhibit more effectively the mycelia growth of Fusarium oxysporum along with the dosage increased. Surfactins and fengycins were detected from the lipopeptide compounds of strain YL3 by LC-MS. Therefore, the strain YL3 was a prospective antagonistic bacterium.

Key words: Bacillus atrophaeus, strawberry fusarium wilt, lipopeptide compounds

中图分类号:

S476.2

刘邮洲, 陈夕军, 梁雪杰, 钱亚明, 乔俊卿, 刘永锋. 一株萎缩芽胞杆菌YL3的鉴定及其脂肽类化合物分析[J]. 中国生物防治学报, 2017, 33(1): 142-150.

LIU Youzhou, CHEN Xijun, LIANG Xuejie, QIAN Yaming, QIAO Junqing, LIU Yongfeng. Identification of Bacillus atrophaeus YL3 and Detection of Its Lipopeptide Compounds[J]. journal1, 2017, 33(1): 142-150.

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一株萎缩芽胞杆菌YL3的鉴定及其脂肽类化合物分析

Identification of Bacillus atrophaeus YL3 and Detection of Its Lipopeptide Compounds

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