【摘要】 SSR作为一种微卫星分子标记方法,被广泛用于遗传多样性相关的研究,而SSR引物是否高效是影响整个实验结果的重要因素。本研究利用生物信息学手段,对马铃薯甲虫基因组数据进行分析,搜索基因组SSR位点,并设计引物和验证SSR引物的效率。最终在马铃薯甲虫基因组中共检测出SSR位点81 937个,且这些位点中以单核苷酸、二核苷酸、三核苷酸重复为主;从SSR位点的平均分布距离来看,单、三、四核苷酸重复在马铃薯甲虫基因组中平均分布距离较小,分别为2.21 kb、8.39 kb、36.21 kb。马铃薯甲虫基因组SSR位点中,优势基序总数为68 388个,比例高达83.46%,其中,单核苷酸以A/T重复基序占绝对优势,二核苷酸重复的SSR位点中,以AG/CT重复基序为主,三核苷酸重复的位点中,以AAT/ATT重复基序为主;在ClassⅠ长度的SSR位点中,以二核苷酸、三核苷酸重复的数量最多,比例高达82.32%;最后设计19对SSR引物测试效率,其中14对成功扩增条带,11对为多态性引物,其平均多态性条带6.27条。多态性引物的PIC值在0.375-0.794,平均值为0.600,大于平均值的引物为6对,占有效扩增引物的54.5%。研究表明,利用生物信息发掘SSR位点的方法简便高效,这为后续进一步利用SSR引物研究马铃薯甲虫的遗传多样性,及在其它昆虫中开发SSR引物开发奠定研究基础。更多还原

【Abstract】 As a microsatellite molecular marker method, SSR is widely used in genetic diversity research,and primers’ efficiency is fundamental to the experiment’s results. By the use of bioinformatics methods,SSR loci in colorado potato beetle’s genomes were analyzed,SSR primers were designed and the efficiency were verified. As results,81 937 SSR loci were detected in the genomes,most of which were single nucleotide,dinucleotide and trinucleotide repeats,and the average distribution distance of SSRloci were 2. 21 kb,8. 39 kb and 36. 21 kb,respectively. Of the CPB genome SSR loci,advantage motifs account for a total of 68 388,up to 83. 46%,among which the single nucleotide type as A/T repeats motif dominates,the dinucleotide type AG/CT repeats motif ranks secondly,and trinucleotide types AAT/ATT repeats ranks thirdly. Among the Class Ⅰ length type of SSR loci,the largest number were dinucleotide and trinucleotide repeats and the proportion is 82. 32%. 19 primers were designed to test efficiency,among which 14 primers were amplified successfully, 11 primers were comfirmed as polymorphism primers with averaged 6. 27 bands and the PIC value range between 0. 375 to 0. 794,the average was 0. 600,of which larger than 0. 600 was 6 pairs,accounting for 54. 5%. The results showed that,it was more efficient for developing SSR primers by bioinformatics method. This result will help to study Colorado potato beetle’s genetic diversity and lay foundation of developing SSR primers in insects.更多还原