解淀粉芽孢杆菌YN-1抑制植物病原真菌活性物质鉴定  

Identification of the antifungal substances from Bacillus amyloliquefaciens strain YN-1

作　　者：邓建良[1,2] 刘红彦[1] 刘玉霞[1] 刘新涛[1] 倪云霞[1] 高素霞[1] 李国庆[2] 

机构地区：[1]河南省农业科学院植物保护研究所,郑州450002 [2]华中农业大学植物科技学院,武汉430070

出　　处：《植物病理学报》2010年第2期202-209,共8页Acta Phytopathologica Sinica

基　　金：河南省重大科技攻关项目(082101110400);河南省财政预算外资金项目(豫财办教2005年291号);河南省农科院科研专项资金项目

摘　　要：为了鉴定解淀粉芽孢杆菌YN-1发酵液中抑制植物病原真菌物质的主要种类及其相对含量,本研究根据已知的脂肽抗生素合成相关基因序列设计了4对特异引物对YN-1菌株进行了检测,对PCR产物克隆、测序,然后采用酸沉淀法从菌株发酵液制备出抑菌物质粗提液,平板拮抗试验确定其抑菌活性,最后对活性粗提物进行HPLC-ESI-MS和MALDI-TOF-MS分析。3对引物的扩增产物经克隆、测序和BLAST分析,表明解淀粉芽孢杆菌YN-1基因组中含有sfp、fen B、itu A或bam A基因。YN-1菌株发酵液的粗提物对棉花枯萎病菌具有抑菌活性,质谱分析发现活性粗提物中含有C14-Iturin A、C15-Iturin A、C16-Iturin A、C14-Fengycin A、C15-Fengycin A、C16-Fengycin A、C17-Fengycin A、C16-Fengycin B和C17-Fengycin B9种脂肽抗生素,其中C16-Iturin A、C14-Fengycin A、C16-Fengycin A、C17-Fengycin A和C17-Fengycin B5种脂肽抗生素为活性粗提物中的主要成分。该研究结果为利用YN-1菌株开发成微生物杀菌剂奠定了基础。The lipopeptide-like antifungal substances produced by Bacillus amyloliquefaciens strain YN-1 were identified and relative quantified. Four primers designed based on the known lipopeptide genes were used to amplify the corresponding genes from YN-1. The PCR products were cloned and sequenced. The extract from YN-1 fermentation liquid was prepared by the acid precipitation, detected by the plate experiment of antagonism, and analyzed with HPLC-ESI-MS and MALDI-TOF-MS methods. 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 YN-1. The extract from YN-1 fermentation liquid could inhibit the mycelial growth of the pathogen Fusarium oxysporum f. sp. vasinfectum. Nine kinds of lipopeptides C14-Iturin A, C15-Iturin A, C16-Iturin A, C14-Fengycin A, C15-Fengycin A, C16-Fengycin A, C17-Fengycin A, C16- Fengycin B and C17-Fengycin B were identified in the antimicrobial extract by the mass spectrum analysis, from which five substances C16-Iturin A, C14-Fengycin A, C16-Fengycin A, C17-Fengycin A and C17- Fengycin B were the major components. Study on B. amyloliquefaciens YN-1 provided a foundation for the development of the micro-fungicide.

关 键 词：解淀粉芽孢杆菌 脂肽抗生素 鉴定 

分 类 号：S432[农业科学—植物病理学]