枯草芽孢杆菌ge25对两种人参病原菌的抑制作用及脂肽类抑菌代谢产物的鉴定  

Antagonism of Bacillus subtilis ge25 against Two Kinds of Ginseng Pathogens and Identification of Antifungal Lipopeptide Metabolites

作　　者：胡陈云 李勇[1] 刘敏[1] 秦民坚[2] 丁万隆[1] 

机构地区：[1]中国医学科学院药用植物研究所,北京100193 [2]中国药科大学,南京210009

出　　处：《中国生物防治学报》2015年第3期386-393,共8页Chinese Journal of Biological Control

基　　金：中央公益性科研院所基金项目(2012D13);协和青年科研基金项目(3332013113)

摘　　要：为明确枯草芽孢杆菌ge25抑菌代谢产物的组成,采用层析柱分离结合抑菌活性追踪方法分离、纯化菌株发酵液中的抑菌脂肽,使用UPLC-ESI-Q-TOF对抑菌活性组分进行鉴定。层析柱分离共获得28个组分,其中G23~G27有抑菌活性,且G25和G26的抑菌活性最强;G25和G26经硅胶柱层析及TLC薄层层析分离,共获得3个组分L1、L2和L3,均有显著抑菌活性。UPLC-ESI-Q-TOF分析发现,L2和L3中包括1个fengycin A和6个fengycin B同系物或其衍生物;L1中发现2个化合物,但其分子量及碎片裂解规律与已知脂肽类成分存在明显差异,且未能获得明确的氨基酸裂解顺序,推测其可能不同于已报道的抑菌脂肽。扫描电镜观察发现,菌株ge25分泌的抑菌脂肽严重影响人参黑斑菌和人参锈腐菌菌丝体的正常生长,导致菌丝体形态异常、发育畸形。In order to identify the antagonistic materials produced by Bacillus subtilis ge25, we used column chromatography and activity tracking methods to isolate and purify the lipopeptides from fermentation broth, and UPLC-ESI-Q-TOF to understand their chemical structure. Results indicated that, in total, 28 components were isolated by column chromatography. Components G23 to G27 were of antagonistic activity, and G25 and G26 expressed the highest. Three components, L1, L2 and L3 were identified from G25 and G26 by Silica gel column chromatography and thin-layer chromatography(TLC). Finally, analytic results showed that, one fengycin A and six fengycin B homologues or their derivatives from L2 and L3, two kinds of unknown chemicals from L1 were identified by UPLC-ESI-Q-TOF. As that the molecular weight and debris pyrolysis were different from known lipopeptides, and no clear amino acid sequence could be retrieval, we deduced that two lipopeptides in L1 were probably different from antagonistic lipopeptides reported. Scanning electron microscopy results indicated that antagonistic lipopeptides secreted by B. subtilis ge25 significantly led to abnormal mycelia growth of ginseng pathogens Alternaria panax and Cylindrocarpon destructans.

关 键 词：人参 枯草芽孢杆菌 抑菌脂肽 FENGYCIN UPLC-ESI-Q-TOF 

分 类 号：S476[农业科学—农业昆虫与害虫防治] S435.675[农业科学—植物保护]