特殊环境巨大芽孢杆菌LB01抗菌活性成分的分离鉴定及抗病机理  

Isolation and Identification of Antifungal Substance Synthesized by Bacillus megaterium LB01 from Special Environment and Its Action Mechanism

作　　者：丁从文 冯群 李春焕 DING Congwen;FENG Qun;LI Chunhuan(Key Laboratory of Regional Ecological Environment Analysis and Pollution Control of West Guangxi,Baise 533000,China;College of Chemistry and Environment Engineering,Baise University,Baise 533000,China)

机构地区：[1]桂西区域生态环境分析和污染控制重点实验室,广西百色533000 [2]百色学院化学与环境工程学院,广西百色533000

出　　处：《食品科学》2020年第17期75-82,共8页Food Science

基　　金：百色学院高层次人才科研启动基金项目(080107022);广西自然科学基金项目(2018GXNSFAA281178)。

摘　　要：以对采后芒果炭疽病菌(Colletotrichum gloeosporioides)的抗菌能力为活性追踪指标,以纸片扩散法(K-B法)测量的抑菌圈直径为活性评价指标,通过色谱分离、活性追踪自广西岩溶区特殊环境巨大芽孢杆菌(Bacillus megaterium LB01)发酵液中分离的抗菌活性成分,对抗病效果显著的活性成分进行结构鉴定,并对其抗病机理进行初步研究。结果表明:从发酵液中共分离出4种单体化合物,筛选到1种拮抗炭疽病菌效果良好的单体,编号为001,经电喷雾离子源质谱、核磁共振氢谱和碳谱解析,鉴定为12-羟基茉莉酸(12-hydroxyjasmonic acid,TA),此化合物的抗病机理是通过诱导活性氧产生、损害线粒体、破坏孢子细胞膜完整性、抑制菌丝生长及孢子萌发而达到抑菌效果。本研究也证实了TA通过直接抑制炭疽菌菌丝生长和孢子萌发而抑制采后芒果炭疽病,该结果为芒果采后炭疽病害控制提供参考。In order to isolate and identify highly bioactive compounds against anthracnose disease caused by Colletotrichum gloeosporioides in postharvest mangos and to investigate their mechanism of action,bioassay-guided fractionation was used to isolate antifungal substances from the fermented broth of Bacillus megaterium LB01 and the antifungal activity was evaluated by measuring the inhibition zone diameter using the disk diffusion test method.The results showed that four monomeric compounds were separated from the fermented broth,and only one monomer with strong antifungal activity,numbered 001,was obtained and identified as 12-hydroxyjasmonic acid(TA)by electrospray ionization mass spectrometry(ESI-MS),1 H nuclear magnetic resonance(NMR)and 13 C NMR analysis.Its mechanism of action was by inducing reactive oxygen production to cause mitochondrial damage,destroy the integrity of spore membrane,and finally significantly inhibit spore germination and mycelium growth.This study confirmed that TA can inhibit the anthracnose disease of postharvest mangos by directly inhibiting the mycelium growth and spore germination of Colletotrichum gloeosporioides,which will provide reference for controlling anthracnose disease in postharvest mangos.

关 键 词：特殊环境 采后芒果炭疽菌 巨大芽孢杆菌LB01 12-羟基茉莉酸 结构鉴定 抗病机理 

分 类 号：Q939.92[生物学—微生物学] TS255.36[轻工技术与工程—农产品加工及贮藏工程]