中文摘要
从NCBI数据库中的猕猴桃EST中，随机抽取56,400条序列，应用SSRHunter软件查找SSR重复序列。结果表明，从猕猴桃EST序列中获得了7939条SSR，其中包括二核苷酸重复5131条（64.63%），三核苷酸重复1237条（15.58%），四核苷酸重复284条（3.58%）,五核苷酸重复397条（5.00%）， 六核苷酸重复890条（11.21%）。利用设计合成的6对引物在DNA测序仪上对猕猴桃种质的多态性进行了PCR检测，结果表明，该6对引物在两个亲本与矮型猕猴桃和普通型猕猴桃F1个体间均能清晰产生多态性分离。用EST-SSR分子标记技术结合BSA分析，并对亲本和F1分离群体进行验证，共筛选了85对EST-SSR引物，发现了其中一个与中华猕猴桃矮型基因Ad连锁的EST-SSR标记（EST-Ad042），其片段大小为285bp。通过EST-SSR分子标记，采用MAPMAKER/EXE 3.0软件对其分子标记的分离数据进行连锁分析，利用Kosamli函数将重组值转化为遗传图距（cM),构建出目标基因的局部分子连锁图，EST-SSR标记与Ad基因的遗传距离为8.8cM。
英文摘要
56400 EST were selected randomly from NCBI database presented recently and search the SSR with SSRHunter software, the results showed that 7939 bands of SSR were picked out in EST, which contained 5131 bands of dinucleotide repeat (64.64%),1237 trinucleotide repeat (15.58%) , 284 tetranucleotide repeat (3.58%), five and six nucleotide repeat 397(5.00%), 890 (11.21%) respectively. Using the 6 pairs of EST-SSR fluorescence-labeled primers and PCR detection of species polymorphism with DNA sequencer, the results indicated that the 6 pairs of primers were able to produce a clear separation polymorphism in the two parental-type and dwarf types, and between F1 population of normal type Actinidia. Application of EST-SSR molecular marker technology coordinated with BSA analysis, and parents and F1 separation groups was confirmed, we screened a total of 85 pairs of EST-SSR primers, and found EST-SSR marker ( EST-Ad042), linked to Ad dwarf gene, which might be associated with the dwarfing gene fragments with 285bp of size. Linkage analysis of separation of molecular marker data through the EST-SSR and MAPMAKER / EXE 3.0 software, reorganization value was transformmed into genetic distance with the Kosamli Function, as well as constructing local molecular linkage map of a target gene. Finally, the genetic distance between
结题摘要
从NCBI数据库中的猕猴桃EST中，随机抽取56,400条序列，应用SSRHunter软件查找SSR重复序列。结果表明，从猕猴桃EST序列中获得了7939条SSR，其中包括二核苷酸重复5131条（64.63%），三核苷酸重复1237条（15.58%），四核苷酸重复284条（3.58%）,五核苷酸重复397条（5.00%）， 六核苷酸重复890条（11.21%）。利用设计合成的6对引物在DNA测序仪上对猕猴桃种质的多态性进行了PCR检测，结果表明，该6对引物在两个亲本与矮型猕猴桃和普通型猕猴桃F1个体间均能清晰产生多态性分离。用EST-SSR分子标记技术结合BSA分析，并对亲本和F1分离群体进行验证，共筛选了85对EST-SSR引物，发现了其中一个与中华猕猴桃矮型基因Ad连锁的EST-SSR标记（EST-Ad042），其片段大小为285bp。通过EST-SSR分子标记，采用MAPMAKER/EXE 3.0软件对其分子标记的分离数据进行连锁分析，利用Kosamli函数将重组值转化为遗传图距（cM),构建出目标基因的局部分子连锁图，EST-SSR标记与Ad基因的遗传距离为8.8cM。