【作者】 李旻；

【导师】 樊美珍； 黄勇平；

【作者基本信息】 安徽农业大学 ， 微生物学， 2006， 硕士

【摘要】 本文应用SSR(微卫星)标记和ISSR(简单重复序列间隔区)标记对在宣城、牯牛降、大别山、琅琊山4个地区采集的122株白僵菌菌株进行遗传多样性分析,探讨了白僵菌不同地区之间和每个地区内部的遗传多样性,获得了如下结果:1.引物筛选:分别对15对SSR引物和20个ISSR引物进行了筛选,获得了13对微卫星引物和15个ISSR引物,这些引物进行PCR扩增均可获得清晰的DNA条带,且多态性较高。2.分别确立了SSR和ISSR的最佳反应体系,在各自的反应体系下,均可以获得清晰的图谱。3.在ISSR实验中,15个ISSR引物共扩增出244条带,多态位点百分率(PPL)为97.95%。总的Nei’s基因多样性指数(H)为0.1659,Shannon’s信息多样性指数(I)为0.2732。居群内基因多样度(Hs)为0.1697,总基因多样度(Ht)为0.1836。各个居群之间的基因分化系数(Gst)是0.0757。在群体间的遗传相似度的聚类分析中,宣城和牯牛降的遗传一致度最高为0.9962,宣城和琅琊山的遗传一致度最低为0.9600。从Shannon指数来看,群体内变异程度最大的是大别山地区。4.在SSR实验中,13对SSR引物共扩增出293条带,多态位点百分率(PPL)为98.98%。总的Nei’s基因多样性指数(H)为0.0408,Shannon’s信息多样性指数(I)为0.0891。居群内基因多样度(Hs)为0.0391,总基因多样度(Ht)为0.0407。各个居群之间的基因分化系数(Gst)是0.0384。在群体间的遗传相似度的聚类分析中,宣城和牯牛降的遗传一致度最高为0.9990,宣城和琅琊山的遗传一致度最低为0.9965。从Shannon指数来看,群体内变异程度最大的是大别山地区。与之前ISSR分析得出的结果相一致。5.由SSR和ISSR计算得到的两组相似系数之间的相关系数r=0.9889>r0.01,达到极显著水平,表明SSR标记和ISSR标记这两种技术对白僵菌遗传多样性的分析具有较高的一致性和可信度。以上结果表明:安徽地区白僵菌具有较高的遗传多样性,不同居群的遗传多样性存在一定的差异。基因分化系数表明居群间遗传变异较小,居群内表现出较高水平的遗传分化。更多还原

【Abstract】 In this paper, the genetic diversity of 122 Beauveria bassiana collected from 4 different mountainous areas in Anhui province was tested using inter-simple sequence repeat (ISSR) markers and simple sequence repeat (SSR) markers. The major results are listed as follows:1. 13 SSR locl and 15 ISSR locl were selected from 15 SSR locl and 20 ISSR locl respectively considering their reproducllbility and polymorphic rate via PCR amplifications.2. The optimal reactive conditions were determined for better using these markers.3. With 13 pairs of SSR primers, a total of 293 fragments were amplified, in which the polymorphic rate was 98.98%. Nei’s genetic diversity index (H) was 0.0408, Shannon’s information diversity index (I) 0.0891. The inner-population genetic diversity (Hs) was 0.0391, and the genetic diversity (Ht) 0.0407. And the inter-population genetic differentiation (Gst) was 0.0384. The cluster analysis of the inter-population genetic differentiation showed that the genetic similarity in Xuancheng and Guniujiang was 0.9990 at the highest point whereas the genetic similarity in Xuancheng and Langya Mountain was 0.9965 at the lowest point. Measured by Shannon’s genetic diversity index (I) it showed that the biggest inner-population variations were in the Dabieshan mountainous area.4. With 15 ISSR primers, a total of 244 fragments were amplified, in which the polymorphic rate was 97.95%. Nei’s genetic diversity index (H) was 0.1659 and Shannon’s information diversity index (I) 0.2732. The inner-population genetic diversity (Hs) was 0.1697 and the total genetic diversity (Ht) 0.1836. And the inter-population genetic differentiation (Gst) was 0.0757. The cluster analysis of the inter-population genetic differentiation showed that the genetic similarity in Xuancheng and Guniujiang was 0.9962 at the highest point whereas the genetic similarity in Xuancheng and Langya Mountain was 0.9600 at the lowest point. Shannon’s genetic diversity index (I) showed that the biggest inner-populations variations were in the Dabieshan mountainous area. This test also showed that the ISSR analytical outcomes are consistent with the SSR outcomes listed above.5. There was a high correlation between the two sets of similarity parameters (r=0.9889>r0.01) calculated with SSR and ISSR markers respectively, indicating that the high consistency and confidence for detecting the genetic diversity in B.bassiana. All of the above result showed that the B.bassiana in Anhui Province has a high degree of genetic diversity where there exists a certain degree of differences among populations though and that a low degree of inter-population genetic variations and a high degree of inner-population genetic differentiation occurred in the B.bassiana.更多还原