一株产巴卡亭Ⅲ红豆杉内生真菌的分离与鉴定  

Identification and Isolation of A Baccatin III-producing Endophytic Fungus

作　　者：郑文龙[1] 周选围[2,3] 朱慧芳[2,3] 王玉亮[2,3] 刘战民[1] 尹京苑[4] 

机构地区：[1]上海大学生命科学学院,上海200444 [2]上海交通大学农业与生物学院 [3]复旦-交大-诺丁汉植物生物技术研发中心,上海200030 [4]上海大学计算机工程与科学学院,上海200444

出　　处：《安徽农业科学》2010年第25期13612-13616,共5页Journal of Anhui Agricultural Sciences

基　　金：上海市科委中药现代化专项基金项目(07DZ19724)

摘　　要：[目的]分离鉴定产巴卡亭Ⅲ红豆杉内生真菌。[方法]采用常规分离方法,从东北红豆杉(TaxuscuspidataSieb.etZucc.)树皮中分离并筛选内生真菌35株;通过高效液相色谱(HPLC)技术对菌株发酵物进行检测,并结合选择性离子质谱(MS)进行确认,从中获得1株能产紫杉醇前体物质巴卡亭Ⅲ的菌株Z-1;利用ITSrDNA通用引物对产紫杉烷类物质的内生真菌进行分子生物学鉴定,并结合菌落特征、孢子与分生孢子梗的形态特征进行分类鉴定。[结果]在未经诱导的情况下,Z-1产巴卡亭Ⅲ的量约为39μg/L;形态和分子鉴定分析判定Z-1属于木霉属(Trichoderma sp.)真菌。[结论]该研究可为植物内生真菌产紫杉烷类化合物的种类多样性和产紫杉烷类内生真菌的生物多样性提供依据。[Objective]The research aimed to identify and isolate baccatin Ⅲ-producing endophytic fungus.[Method]35 strains of endophytic fungi had been isolated from the bark of Taxus cuspidata Sieb.et Zucc.using conventional isolation method.One strain Z-1 was obtained which was a baccatin Ⅲ-producing endophytic fungus,detected by HPLC and affirmed by MS.Phenotypic and biologic（ITS rDNA PCR） analysis methods were used to identify Z-1.[Result]The results showed that the volume of baccatin Ⅲ production of strain Z-1 was about 39 μg/L under non induced circumstance.Z-1 was high similarity with genus Trichoderma sp..[Conclusion]It is a dependable evidence to biological diversity of taxane-producing endophytic fungus.And it is a new source for production of baccatin Ⅲ by fermenting.

关 键 词：巴卡亭Ⅲ 木霉属 ITS rDNA分析 高效液相色谱 质谱 

分 类 号：S718.43[农业科学—林学]