中国中药杂志

2014, v.39(14) 2624-2628

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枯草芽孢杆菌ge25菌株发酵及其抑菌活性成分的初步研究
Fermentation of Bacillus subtilis ge25 strain and preliminary study on its antagonistic substances

胡陈云;李勇;刘敏;丁万隆;秦民坚;
HU Chen-yun;LI Yong;LIU Min;DING Wan-long;QIN Min-jian;China Pharmaceutical University;Institute of Medical Plant Development,Chinese Academy of Medicinal Sciences and Peking Union College;

摘要(Abstract):

人参是我国传统名贵中药材,病害问题严重影响人参的产量和品质。在前期分离到对多种人参致病菌有理想抑菌活性的内生枯草芽孢杆菌ge25菌株基础上,研究采用盐析和酸沉淀法从LB和Landy发酵液中制备蛋白粗提物和脂肽粗提物,用人参黑斑病菌检测粗提物的抑菌活性及其对高温和蛋白酶消解的稳定性;研究发现,LB发酵物中的抑菌成分主要是蛋白类物质,不耐高温和蛋白酶K;Landy发酵物中的抑菌成分主要是脂肽类物质,其对高温和蛋白酶K稳定。原位酸水解-茚三酮显色和TLC-生物自显影分析发现,脂肽粗提物中至少含有1种抗菌环肽。鉴于源自ge25菌株的脂肽类物质的高稳定性及理想的抑菌活性,对其开展深入研究将有助于推动人参病害的生物防治。
Panax ginseng is one of the most important traditional Chinese herbal medicine,soil borne diseases influenced the yield and quality severely. In our previous work,endophytic Bacillus subtilis ge25 strain was isolated from ginseng root,and which showed significant antagonistic activity against several most destructive ginseng phytopathogens. In the present work,crude protein and lipopeptid extracts were prepared from LB and Landy supernate by salting out,acid precipitation methods respectively. The antagonistic activity of crude extracts and stability to temperature and protease digestion were examined by ginseng phytopathogen Alternaria panax.Results showed that,the antagonistic activity of crude protein extracts from LB culture was complete and partially lost when treated by high temperature and proteinase K. However,crude lipopeptid from Landy culture showed significant stabile antagonistic activity to them. Acid-hydrolyzation and TLC-bioautography analysis showed,that the crude lipopeptide contained at least one cyclic lipopeptide.In consideration of the stability and perfect antagonistic activity of ge25,further researches will promote the biocontrol of ginseng diseases in the field.

关键词(KeyWords): 人参;内生菌;枯草芽孢杆菌;抑菌物质;脂肽
Panax ginseng;endophyte;B.subtilis;antagonistic substance;lipopeptid

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基金项目(Foundation): 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)

作者(Author): 胡陈云;李勇;刘敏;丁万隆;秦民坚;
HU Chen-yun;LI Yong;LIU Min;DING Wan-long;QIN Min-jian;China Pharmaceutical University;Institute of Medical Plant Development,Chinese Academy of Medicinal Sciences and Peking Union College;

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