贡嘎蝠蛾幼虫肠道真菌多样性分析  

Analysis of fungal diversity in intestines of Hepialus gonggaensis larvae

作　　者：俞和韦[1] 王中康[1] 刘莉[1] 夏玉先[1] 殷幼平[1] 袁青[1] 曹月青[1] 彭国雄[1] 

机构地区：[1]重庆大学生物工程学院,重庆市功能基因及调控技术重点实验室,重庆市杀虫真菌农药工程技术中心,重庆400030

出　　处：《微生物学报》2008年第4期439-445,共7页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30572325)~~

摘　　要：【目的】分析贡嘎蝠蛾肠道(Hepialus gonggaensis)幼虫肠道真菌多样性。【方法】采用常规分离与分子鉴定的方法和基于ITS(internal transcribed spacer)基因的RFLP方法,建立贡嘎蝠蛾幼虫肠道真菌的ITS克隆文库,分别用MspI、HaeⅢ和TaqⅠ对205个阳性克隆的质粒酶切指纹图谱分析,结果显示有23个不同的RFLP操作分类单元(OTU),对这23个操作分类单元的阳性克隆子进行测序并绘制系统进化树。【结果】结果显示贡嘎蝠蛾幼虫肠道内存在8个属的真菌类群。其中被孢霉属(Mortierella)和丝孢酵母属(Trichosporon)的丰度最高,分别占克隆文库的46.34%和40.00%,鉴定为肠道内的优势真菌类群。用常规分离与分子鉴定方法只获得隐球酵母(Cryptococcus magnus)、Geomyces sp和丝孢酵母(Trichosporon porosum)3个类群的真菌。结合常规分离法与RFLP法能够更有效的分析肠道微生物的多样性,获得更多更全面的微生物多样性信息。[Objective] Studied fungi diversity in the guts of larval Hepialus gonggaensis using culture-independent and traditional culturing methods. [Methods]For the culture-independent method, the total DNA of fungus was extracted from the intestinal contents and internal transcribed spacer （ITS） regions were amplified with fungal universal primers. A near-full length ITS gene library was constructed. Subsequently, the fingerprints of the microorganisms were analyzed by isolated plasmid and digestion with Msp Ⅰ, Hae Ⅲ and Taq Ⅰ enzymes, respectively. Restriction fragment length polymorphism （RFLP） analysis based on the fungal ITS sequences indicated that the library established includes 23 operational taxonomic units （OTUS） and a phylogenetic tree depicted the linkage of the isolated fungi. [Results]Abundant fungi were in the intestines of H. gonggaensis larvae, but their abundance was very different. The dominant fungi belonged to Mortierellaone and Trichosporon and accounted for 46.34 % and 39.02 % of the total ITS clones, respectively. Only three genera of fungi were identified from eight isolated fungal populations by traditional culturing methods. [Conclusion] We could get more information by combined traditional culturing and molecular biology methods.

关 键 词：贡嘎蝠蛾 ITS序列 真菌多样性 ITS克隆文库 RFLP分析 

分 类 号：Q93[生物学—微生物学]