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贝莱斯芽胞杆菌ZF145抑菌物质提取方法比较及成分鉴定

引用本文：曾先锋,覃诗扬,谢学文,石延霞,柴阿丽,李宝聚,李磊.贝莱斯芽胞杆菌ZF145抑菌物质提取方法比较及成分鉴定.植物保护学报,2024,51(2):334-341

DOI：10.13802/j.cnki.zwbhxb.2024.2023050

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作者	单位	E-mail
曾先锋	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081
覃诗扬	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081 天津农学院园艺园林学院, 天津 300384
谢学文	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081
石延霞	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081
柴阿丽	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081
李宝聚	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081	lilei01@caas.cn
李磊	中国农业科学院蔬菜花卉研究所, 蔬菜生物育种全国重点实验室, 北京 100081	libaojuivf@163.com

中文摘要:为明确贝莱斯芽胞杆菌Bacillus velezensis菌株ZF145中可有效抑制黄瓜角斑病病原菌扁桃假单胞菌流泪致病变种Pseudomonas amygdali pv.lachrymans的主要物质，采用冻干法、酸沉法、絮凝法、萃取法和铵沉法提取菌株ZF145发酵液中的抑菌物质，通过室内盆栽试验验证粗提物对黄瓜角斑病的防治效果，综合比较不同方法的提取效果，并通过高效液相色谱-质谱（high performanceliquid chromatography-mass spectrometry，HPLC-MS）联用技术鉴定粗提物中的抑菌物质。结果表明，絮凝法最适用于贝莱斯芽胞杆菌菌株ZF145发酵上清液中抑菌物质的提取，通过絮凝法提取的粗提物对扁桃假单胞菌流泪致病变种的抑菌圈直径达到19.50 mm，对黄瓜角斑病的盆栽防治效果为42.85%，且粗提物中的主要抑菌物质为C₁₃~C₁₆的surfactin同系物，浓度和纯度均为最高，分别为2.77 g/L和69.28%。表明本研究优化的絮凝法可用于芽胞杆菌发酵液中抑菌物质的提取。

中文关键词:贝莱斯芽胞杆菌黄瓜角斑病絮凝法防治效果抑菌物质

Comparison of the extraction method and identification of antimicrobial substances from Bacillus velezensis strain ZF145

Author Name	Affiliation	E-mail
Zeng Xianfeng	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Qin Shiyang	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China College of Horticulture and Landscape Architecture, Tianjin Agricultural University, Tianjin 300384, China
Xie Xuewen	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Shi Yanxia	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Chai Ali	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
Li Baoju	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China	lilei01@caas.cn
Li Lei	State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China	libaojuivf@163.com

Abstract:In order to identify the substances with the main antibacterial effect in Bacillus velezensis strain ZF145, the effects of different extraction methods were comprehensively evaluated, and the bacteriostatic substances in the fermentation broth of B. velezensis strain ZF145 were extracted with lyophilization, acid precipitation, flocculation, extraction and ammonium precipitation, which verified the efficacy of crude extract on cucumber angular leaf spot, and identified the bacteriostatic substances in crude extract with high performance liquid chromatography-mass spectrometry (HPLC-MS). The results showed that the flocculation method was the most suitable for the extraction of bacteriostatic substances in the supernatant of B. velezensis strain ZF145 fermentation broth, the inhibition diameter of Pseudomonas amygdali pv. lachrymans by the crude extract prepared by the flocculation method reached 19.50 mm, the pot control effect on cucumber angular leaf spot was 42.85%, and the main bacteriostatic substance in the crude extract of flocculation was C₁₃-C₁₆ surfactin, and the concentration and-purity were the highest, reached 2.77 g/L and 69.28%. The flocculation method optimized in this study could be used for the extraction of bacteriostatic substances from bacillus fermentation broth.

keywords:Bacillus velezensis cucumber angular leaf spot flocculation method biocontrol effect bacteriostatic substance

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