黄瓜叶绿体基因组全序列微卫星分布特征与标记开发  

Characterization of Microsatellites in Complete Chloroplast Genome of Cucumber (Cucumis sativus) and Marker Exploitation

作　　者：胡建斌[1] 李建吾[1] 梁芳芳[1] 刘颖[1] 

机构地区：[1]河南农业大学园艺学院,郑州450002

出　　处：《细胞生物学杂志》2009年第1期69-74,共6页Chinese Journal of Cell Biology

基　　金：国家高技术研究发展计划(863计划)(No.2007AA10Z100);河南省农业科技成果转化项目(No.072201110017);河南农业大学博士启动基金项目(No.30400247)资助~~

摘　　要：对黄瓜叶绿体基因组(chloroplast genomic DNA,cpDNA)全序列中微卫星(chloroplast simple sequence repeat,cpSSR)的分布特征进行了全面分析。结果发现,cpSSR主要分布在25～40kb和100～130kb两个区域,主要以完全重复和间隔重复为主。单碱基重复最多,占总数的79.3%,二碱基重复占10.1%,三碱基和三碱基以上重复仅占10.6%。单碱基重复主要以(A)n和(T)n为主,二者共占总数的78.8%,二碱基重复主要是(AT)n,占总数的9.3%。在重复次数分布上,完全重复的(A)n和(T)n均以最低重复次数为主,而间隔重复的(A)n和(T)n以14次或15次为多。设计了6对引物,对7份黄瓜自交系cpDNA进行扩增。结果发现,6对引物均有扩增产物,每对引物平均能检测到2.2个等位基因。几乎所有引物都能在甜瓜、西瓜、葫芦和栝楼中通用,5对引物在1～4种远缘种中有扩增产物。上述结果表明,从黄瓜cpDNA中发掘cpSSR标记是一条可行的途径。Characterization of microsatellites in complete sequence of chloroplast genome （cpDNA） of cucumber （Cucumis sativus） was analyzed in the round. The results showed that chloroplast simple sequence repeats （cpSSR） were mainly distributed in the two regions of cpDNA, 25-40 kb and 100-130 kb, most of the cpSSRs belonging to complete repeats and interrupted repeats. Of these cpSSRs, mononucleotide repeats were maximum types with the frequency of 79.3%, and dinucleotide repeats accounted for 10.1%, and all of the trinucle- otide repeats and those repeats over trinucleotide in length only accounted for 10.6%. （A）n and （T）n were major repeats in mononucleotides, totally accounting for 78.8%. （AT）n was dominant dinucleotide repeats with frequency of 9.3%. As for repeat number, most of the complete repeats of （A）n and （T）n had the lowest repeat mumber, while 14 or 15 repeats were major for interrupted repeats of （A）n and （T）n. Six primer pairs were designed and used for PCR amplification of the cpDNA of 7 cucumber inbred lines. The results showed that all primer pairs had amplified fragments in cucumber materials and each of the primers detected mean 2.2 alleles. All of the primer pairs were almost transferable to melon, watermelon, gourd and Trichosanthes kirilowii. Five primer pairs could successfully amplify in 1-4 distantly related species. These results indicated that exploitation of cpSSR markers from cucumber cpDNA is practicable.

关 键 词：黄瓜 叶绿体基因组 微卫星 标记 

分 类 号：Q943.2[生物学—植物学] S432.41[农业科学—植物病理学]