【作者】 满玉萍； 张蕾； 王彦昌； 李作洲； 姜正旺； 孙晓荣； 刘长江；

【Author】 Yuping Man~(1,2),Lei Zhang~1,Yanchang Wang~1,Zuozhou Li~2, Zhengwang Jiang~1,Xiaorong Sun~3,Changjiang Liu~3 (1.Wuhan Botanical Garden,Chinese Academy of Sciences,Wuhan,430074; 2.School of Life Sciences,South-central University for Nationalities,Wuhan,430074,China; 3.Shenyang Agricultural University,Shenyang,110866)

【机构】 中国科学院武汉植物园； 中南民族大学生命科学学院； 沈阳农业大学；

【摘要】 应用SSR Hunter软件对中华猕猴桃EST数据库进行搜索,查找微卫星(Microsatellite)序列。利用检索出的SSR-EST序列设计并合成了150对EST-SSR引物。利用这些引物及已发表的20对中华猕猴桃EST-SSR引物分别在中华猕猴桃、软枣猕猴桃各两个野生居群的基因组DNA中进行PCR扩增,其中有98、103、94和80对引物分别在平江中华猕猴桃、毛坝美味猕猴桃、东北居群的软枣猕猴桃和四川居群的软枣猕猴桃中有扩增产物,可扩增率分别为57.6%、60.6%、55.3%和47%。80对引物在平江中华猕猴桃中扩增出多态,占所用引物数的47%;71对引物在毛坝美味猕猴桃中扩增出多态,占所用引物数的41.8%;60对引物在东北居群的软枣猕猴桃中扩增出多态,占所用引物数的35.3%;47对引物在四川居群的软枣猕猴桃中扩增出多态,占所用引物数的27.7%。这些结果表明,中华猕猴桃EST-SSR在软枣猕猴桃中只具有一定的通用性,而不同地理位置的同物种居群多态性差异较大。本研究开发了数量较多、质量较好的软枣猕猴桃SSR,将为软枣猕猴桃居群遗传学、分子系统地理学以及分子育种提供有效工具。更多还原

【Abstract】 SSRs(Microsatellite) could be found out from EST(Expressed Sequence Tag) database of Actinidia Chinensis by using the SSRHunter software.150 EST-SSR markers were designed to sequence flanking SSRs from the selected SSR-ESTs.All these markers and 20 others from Actinidia Chinensis that had been reported were used to amplify the genomic DNA extracted from two natural populations of Actinidia Chinensis and two natural populations of Actinidia arguta,respectively.98,103,94 and 80 of 170 markers brought detectable amplification in Actinidia Chinensis from Pingjiang,A.deliciosa from Maoba,A.arguta from northeast and A.arguta from Sichuan,respectively,the transferability was 57.6%,60.6%,55.3%and 47%,respectively.Among the total SSR markers,47%have detected polymorphism in A.Chinensis from pingjiang,41.8%in A.deliciosa from maoba,35.3%in A.arguta from northeast and 27.7%in A.arguta from Sichuan,respectively.Our results indicate that EST-SSR markers from Actinidia Chinensis have a certain transferability to A.arguta,and the polymorphism in the same species from different geographical positions are of great differences.This study develop a large number of SSR makers with highly quality for A.arguta,and they will become effective tools in studying A.arguta genetics,molecular phylogeography and Molecular Breeding.更多还原