一株多花黄精内生真菌的鉴别及其抗菌代谢产物  

Isolation and identification of an endophytic fungus of Polygonatum cyrtonema and its antifungal metabolites

作　　者：汪滢[1] 王国平 王丽薇[3] 徐旭辉[4] 夏静 黄雪芳 吴友贵 章初龙[1,2] 

机构地区：[1]浙江大学生物技术研究所,杭州310029 [2]浙江大化浙大生物药物研发中心,杭州311616 [3]杭州师范大学医药卫生管理学院,杭州310036 [4]浙江大学农药与环境毒理研究所,杭州310029 [5]浙江庆元百山祖国家级自然保护区,庆元323800

出　　处：《微生物学报》2010年第8期1036-1043,共8页Acta Microbiologica Sinica

基　　金：浙江省重大科技攻关项目(2006C12008)~~

摘　　要：【目的】药用植物内生真菌是一类重要的微生物资源,能代谢产生多种生物活性物质。本研究从浙江庆元百山祖自然保护区多花黄精(Polygonatum cyrtonema)分离获得1株具有抗菌活性的菌株zjqy610。【方法】通过形态和ITS rDNA序列分析,鉴定为变灰青霉(Penicillium canescens)。采用正相硅胶柱层析和凝胶(Sephadex LH-20)柱层析,以紫外光或碘蒸汽显迹,配合活性追踪等,从zjqy610发酵液中分离获得3个具抗真菌活性的化合物。【结果】通过质谱和核磁共振波谱技术分别将其结构鉴定为:乙基氧苯氨基亚胺乙酸(o-acetylbenzeneamidinocarboxylic acid)、灰黄霉素(griseofulvin)和[1,2-b]呋喃2-甲基3-羧甲基4-羟基-5-甲氧基萘(naphtho[1,2-b]furan-3-carboxylic acid,4-hydroxy-5-methoxy-2-methyl-)。抑菌活性测试表明,3个化合物对多种植物病原真菌具有抑制活性,其中化合物zjqy610D-4对番茄灰葡萄孢、圆形炭疽菌、泻根亚隔孢壳和核盘菌4种病原真菌的活性最强,半抑制浓度EC50分别为0.68、0.38、0.91和0.61mg/L。【结论】该化合物具有开发成农用抗生素的价值。[Objective] Endophytic fungi in medicinal plants produce a variety of bioactivity compounds.In this study,an endophytic fungus zjqy610 with antifungal activity was isolated from Polygonatum cyrtonema in Zhejiang Qingyuan Baishanzu Mountain nature reserve.[Methods]Strain zjqy610 was identified as Penicillium canescens based on the morphology and its rDNA sequence analysis.Three antifungal compounds were isolated from the fermentation broth of zjqy610 through normal-phase silica gel column chromatography and gel （ Sephadex LH-20 ） column chromatography,traced by ultraviolet light or iodine vapor with bioassay-guided fractionation.[Results]These three compounds were elucidated as o-acetylbenzeneamidinocarboxylic acid （ zjqy610B-g-3）,griseofulvin （ zjqy610D-4） and naphtho [1,2-b] furan-3 -carboxylic acid,4-hydroxy-5-methoxy-2-methyl-（zjqy610F-2） based on mass spectrometry and nuclear magnetic resonance spectroscopy.The antifungal activity assays showed that the three compounds had inhibitory to variety of plant pathogenic fungi.Compound zjqy610D-4 had strong antifungal activity against Botrytis cinerea,Colletotrichum orbiculare,Didymella bryoniae and Sclerotinia sclerotiorum.EC50 was 0.68,0.38,0.91 and 0.61 mg /L,respectively.[Conclusion]Zjqy610D-4 is deserved to develop an agricultural antibiotics.

关 键 词：多花黄精 植物内生菌 变灰青霉 抗真菌活性 结构鉴定 

分 类 号：S482.292[农业科学—农药学] O658.1[农业科学—植物保护]