SCoT和ISSR分析金针菇单核体遗传差异的研究

Analysis of Genetic Variation of Flammulina velutipes Strains by SCOT and ISSR Marker

[目的]研究金针菇单核体的遗传差异.[方法]采用常规稀释分离法分离获得金针菇F1代20个单核体,对15条scot和9条IS-SR引物进行了筛选,根据扩增结果采用软件NTsys 210e计算菌株间的遗传相似系数,并对供试菌株进行聚类分析.[结果]有6条引物能扩增出条带清晰且具丰富多态性的带谱,共计扩增出327条清晰易辨的DNA片段条带,其中多态性条带287条,占总条带数的87.77％.菌株的SCoT分析一致度GI值为0.187 5 ～0.937 5,其中W2和W3,W11和W12的一致度最大,为0.937 5;W15和W18的一致度最小,为0.1875.菌株的ISSR分析一致度GI值为0.250 0 ～ 1.000 0,W3、W4和W9,W15和W17,W16和W19的一致度最大,为1.000 0;W14和W18的一致度最小,为0.250 0.这种低的遗传一致度充分表明了金针菇菌株系列单核体间具有的较大遗传差异.聚类分析结果表明,当相似性达到0.55的水平时,SCoT标记的20个菌株可以明显地分为3组;当相似性达到0.66的水平时,ISSR标记的20个菌株也可以分为3组,其中W11、W18、W203个菌株是1组里共有的菌株.当聚到亚组时,W15和W17是共有的1组.[结论]该方法采用2种标记对单核体菌株进行遗传多样性分析,对金针菇育种中杂交亲本的选择具有重要的科研以及实践意义.

[Objective] To study genetic variation of Flammulina velutipes strains.[Method] 20 sporulated monokaryon have been developed from F1 generation of Flammulina velutipes.15 SCoT primers and 9 ISSR primers were screened.According to the results of amplification using NTsys software 2.10 e to calculate the genetic similarity coefficient between strains.[Result] There are 6 primers could amplify the stripe clear and rich polymorphism band spectrum.A total of 327 fragments were amplified,among them 287 （accounting for 87.77% of the total） were polymorphic.Strains SCoT analysis consistent GI value is 0.187 5-0.937 5,W2 and W3,W11 and W12 consistent GI value is maximum,0.937 5 ; W15 and W18 consistent GI value is minimum,0.187 5.Strains ISSR analysis consistent GI value is 0.250 0-1.000 0,W3,W4 and W9,W15 and W17,W16 and W19 consistent GI value is maximum,1.000; W14 and W18 consistent GI value is minimum,0.2500.This low genetic consistency fully shows that the Flammulina velutipes strains series Ssporulated monokaryon with large genetic differences.Strains for clustering analysis,the results showed that when the similarity at 0.55 level,20 strains of SCoT markers can be clearly divided into three groups,when similarity at 0.66 level,20 strains of ISSR markers can also be divided into three groups,including W1 I,W18 and W20 three strains is a set of shared strains.When together into subgroups,W15 and W17 is shared by a group.[Conclusion] The genetic diversity analysis was conducted with 2 markers in the study,which has important scientific and practice meaning in selection of hybrid parents in Flammulina velutipes breeding.