杏EST-SSR标记的开发  

Development of EST-SSR Markers in Apricot

作　　者：上官凌飞[1] 李晓颖[1] 宁宁[1,2] 王玉柱[2] 章镇[1] 房经贵[1] 

机构地区：[1]南京农业大学园艺学院,南京210095 [2]北京市农林科学院林业果树研究所,北京100093

出　　处：《园艺学报》2011年第1期43-54,共12页Acta Horticulturae Sinica

基　　金：国家科技支撑计划项目(2008BAD92B02);林业公益性行业科研专项(200904032);北京市自然科学基金重点项目(6081001)

摘　　要：利用MISA软件对NCBI上杏的15387条EST序列进行SSR位点查找,发掘出含有SSR位点的序列1073条,共有1353个SSR位点,候选SSR位点出现的频率为8.79%。二核苷酸、三核苷酸和四核苷酸重复是最主要的重复类型,分别占23.50%、38.80%和23.43%。设计了50对EST-SSR引物并进行PCR扩增,发现40对引物能扩增出理想的PCR产物,其中有25对引物能够扩增出多态性带。随机对7对引物的部分杏扩增片段进行了测序分析,发现有57.1%的片段具有相应的SSR位点,对梅DNA指纹中部分谱带的测序结果也证明是杏扩增出的相应的SSR位点。根据推算,从杏EST中开发SSR标记的开发效率为3.98%。进一步选用20对扩增效果最好的引物对24个杏品种,16个花梅品种以及8个果梅品种进行DNA指纹构建与遗传多样性分析,结果表明,来源于杏的EST-SSR引物在梅中有很高的通用性,杏和梅是遗传差异明显的两种植物,果梅与花梅在遗传上是不能分为两类的同种植物。15 387 ESTs of apricot from NCBI were screened using MISA software.The results showed that 1 353 SSRs were mined from 1 073 ESTs with a frequency of 8.79%.Di-,tri-and tetra nucleotide repeats EST-SSRs were dominant,accounting for 23.50%,38.80% and 23.43%,respectively.Fifty primer pairs were designed for partial EST-SSRs and used for PCR amplification with 40 primer pairs showing amplifications while 25 were polymorphic.PCR products from 7 EST-SSR primer pairs were selected for sequence analysis where 57.1% of the fragments contained SSRs.EST-SSR motifs were conserved between mei and apricot.Accordingly,the proportion of developing efficient EST-SSR primers from apricot was 3.98%.The best twenty workable EST-SSR primer pairs were chosen to generate DNA fingerprints of 24 apricot,16 flower-mei and 8 fruit-mei cultivars.Using these EST-SSR primer pairs,mei and apricot were classified into 2 groups.The EST-SSR primers designed from apricot sequences showed high polymorphism in mei varieties and exhibited high transferability between mei and apricot cultivars;Fruit-mei and flower-mei were genetically the same plant.

关 键 词：杏 梅 EST-SSR 遗传差异分析 

分 类 号：S662.2[农业科学—果树学] S662.4[农业科学—园艺学]