柿EST-SSR引物的开发及筛选  

Development and screening of EST-SSR markers in Diospyros kaki Thunb.

作　　者：吴硕[1] 傅建敏[1] 乌云塔娜[2] 梁玉琴[1] 李芳东[1] 

机构地区：[1]中国林业科学研究院经济林研究开发中心,河南郑州450003 [2]中南林业科技大学经济林育种与栽培国家林业局重点实验室,湖南长沙410004

出　　处：《经济林研究》2012年第3期27-31,共5页Non-wood Forest Research

基　　金：林业公益性行业科研专项项目"柿属植物种质资源保护与选育技术研究"(200904041)

摘　　要：为丰富柿树SSR引物来源,利用从NCBI下载的9 474条柿属EST序列开发柿EST-SSR引物,并用于分析7个柿品种的遗传多样性。EST序列经去除低质量及冗余片段后拼接成1 390条重叠序列和4 097条无重复序列。共搜索到601条EST序列中含有540个SSR。SSR出现频率为10.96%,其中以2核苷酸为主(61.06%)。利用检出的SSR-ESTs序列设计SSR引物100对,对7个柿样品进行了PCR扩增,31对引物有扩增带,占设计引物的31%,其中14对引物有多态性,占可扩增引物的45.2%。同时发现引物68是临潼板柿的特异引物。引物68的P68-115、P68-160和P68-210处为临潼板柿的3个特异性标记位点,而引物28的P28-225是麻城秋艳的特异性标记位点。In order to enrich SSR primer sources of Diospyros kaki Thunb.,SSR makers were developed from 9 474 ESTs of Diospyros L.in NCBI database and they were used to analyze diversities among 7 cultivar of D.kaki.1 390 repeated sequences and 4 097 non-repeated sequences were assembled by removing low-quality and redundant fragments in ESTs sequences.In total,540 SSRs in 601 EST sequences were searched out,accounting for 10.96% of ESTs,among which 61.06% of SSRs was dinucleotides.One hundred primer pairs were designed for PCR amplification of 7 cultivars.Thirty one primer pairs show the successful amplified bands,accounting for 31% of the designed primers,and 14 primer pairs show polymorphisms,accounting for 45.2% of available primers.Meanwhile,the primer 68 is the specific primer of Lintong-banshi,and the bands of P68-115,P68-160 and P68-210 in the primer 68 are the specific marker sites in Lintong-banshi.P28-225 in the primer 28 is the specific marker sites of Macheng-qiuyan.

关 键 词：柿 EST SSR标记 

分 类 号：S665.2[农业科学—果树学]