四川汉源铅锌矿区植物内生真菌的遗传多样性初步研究  

Genetic Diversity of the Plant Endophytic Fungi in a Pb-Zn Mine Area in Hanyuan,Sichuan Province,China

作　　者：郑有坤[1] 陈诚[1] 任丹[1] 辜运富[1] 

机构地区：[1]四川农业大学资源环境学院,四川温江611130

出　　处：《四川农业大学学报》2013年第3期308-313,共6页Journal of Sichuan Agricultural University

基　　金：四川农业大学双支计划(01423700);四川省教育厅青年基金(SC200902158)

摘　　要：【目的】了解铅锌矿区植物内生真菌的分布特点和遗传多样性。【方法】采用传统组织块培养法对四川汉源铅锌矿区6种常见植物进行内生真菌分离，根据茵落形态特征挑选了27株代表菌株，采用随机扩增多态性DNA（RAPD）指纹图谱分析技术进行了遗传多样性分析，并在RAPD分析的基础上测定了8株代表菌株的rDNA-ITS序列，利用MEGA4．0构建了供试菌株的系统发育树。【结果】从6种植物的300个组织块中分离获得内生真菌186株，内生真菌的分离频率在0．43～O．85之间，平均为0．62，所有植物茎内生真菌的分离频率最高，叶部次之，根部最低。RAPD分析表明，在76％的相似性水平处，27株供试菌株被分为8个遗传群。系统发育表明，这8个代表菌株分别分布于镰刀菌属（Fusarium），青霉属（Penicillium），座囊菌纲（Dothideomycetes），小不整球壳属（Plecto—spha8rella），漆斑菌属（Myrothecium），格孢腔菌属（Pleosporales），链格孢属（Alternaria）和拟盘多毛孢属（Pestalotiopsis）8个系统发育分支。【结论】铅锌矿区植物内生真菌具有一定的多样性，值得进一步研究。[Objective] To analyze the distribution characteristics and genetic diversity of plant en- dophytie fungi in Pb-Zn mine area. [Method] Using the tissue cultivation method, endophytic fungi were isolated from six plant species collected from one Pb-Zn mine area in Hanyuan, Si- chuan province. Eased on their morphological characteristics, 27 endophytic fungal strains were selected and carried out the genetic diversity analysis by using RAPD fingerprinting technique, and then one representative strain from each genetic cluster according to the RAPD result was used for rDNA-ITS sequencing, by which phylogenetic tree was constructed using neighbor-joining method with MEGA 4.0 software. [Results] A total of 186 endophytic fungal strains were i- solated from 300 tissue segments of the six different plant species. The isolation rate ranged from 0.43 to 0.85 with the mean rate of 0.62. Stem was found having the highest isolation rate in all six plant species, and leaf came second and root was the lowest. The RAPD analysis showed that 27 strains were genetically clustered into eight different groups at the level of 76%similarity. Phylogenetic analysis based on the ITS sequences of representative strains of eight groups showed that these eight endophytic fungi were closely related to Fusariurn, Penicillium, Dothideomyce tes, Plectosphaerella, Myrothecium, Pleosporales, Alternaria and Pestalotiopsis, respectively. [Conclusion] The study suggested that plants in this Pb-Zn mine area had diverse endophytic fun gi to some extent. However, the ecological function of them in such extreme environment is still not clear, which is worthy to be studied further.

关 键 词：铅锌矿区 植物内生真菌 分离频率 遗传多样性 

分 类 号：Q938.1[生物学—微生物学]