蔬菜保护地木霉菌rDNA-ITS序列和UP-PCR遗传多样性分析  

ITS sequence of Trichoderma species in soil planted vegetables in the greenhouse and UP-PCR analysis on genetic diversity

作　　者：贺字典[1,2] 高增贵[1] 高玉峰[2] 赵柏霞[1] 张小飞[1] 

机构地区：[1]沈阳农业大学植物免疫研究所,沈阳110161 [2]河北科技师范学院生命科技学院,秦皇岛066004

出　　处：《植物保护学报》2010年第5期459-465,共7页Journal of Plant Protection

基　　金：辽宁省科技攻关计划(2006215004);辽宁省自然科学基金(20062018);河北省科技支撑计划(07220304)

摘　　要：采用传统形态学分类和ITS序列比对的方法,研究蔬菜保护地土壤中木霉菌种群分布和遗传多样性。木霉菌分离培养结果显示,共获得397株木霉菌,鉴定出11个种,分别为:长枝木霉Trichoderma longibrachiatum、深绿木霉T.atroviride、哈茨木霉T.harzianum、粘绿木霉T.viren、微孢木霉T.minutisporum、拟康木霉T.pseudokoningii、黄绿木霉T.aureoviride、非钩木霉T.inhamatum、棘孢木霉T.asperellum、长孢木霉T.longipile和螺旋木霉T.helicum。经ITS序列建立系统发育树后,将木霉菌分为5个组。用5条通用引物经UP-PCR扩增后,扩增出46条谱带,其中多态性条带43条,占总条带数的93.5%。遗传多样性分析表明,当相似系数为0.80时,可将24个菌株划分为9个组。UP-PCR与ITS序列相比,更能体现木霉菌种间和种内的亲缘关系及遗传差异性,可以作为木霉菌分类的辅助方法。A total of 397 Trichoderma isolates were obtained from the different soil samples collected from some greenhouse vegetable fields of Hebei Province.Using traditional morphological classification and ITS sequences analysis methods,eleven Trichoderma species,Trichoderma longibrachiatum,T.atroviride,T.harzianum,T.viren,T.minutisporum,T.pseudokoningii,T.aureoviride,T.inhamatum,T.asperellum,T.longipile and T.helicum,were identified.Twenty strains were clustered into 5 groups based on rDNA-ITS sequences on phylogenetic tree.Additionally,genetic diversity of Trichoderma isolates was analyzed using universally primed PCR（UP-PCR）.The result indicated that a total of 46 bands appeared after amplification by using 5 primers,43 bands（93.5%） of which were polymorphic.Twenty-four Trichoderma isolates were classified into 9 groups at the similarity coefficient of 0.80 by UP-PCR cluster analysis.Our results indicated that UP-PCR could reflect the genetic relationships and differences of Trichoderma species,and could be used as an assistant method for Trichoderma classification.

关 键 词：木霉菌 ITS 通用引物PCR 遗传多样性 

分 类 号：S476[农业科学—农业昆虫与害虫防治]