枇杷核基因组DNA提取方法的改良及其SSR分析  

The Nuclear DNA Isolation from the Leaves of Loquat and Analysis of Its SSR

作　　者：仲艳[1] 杜青珍[1] 李海芬[1] 孟祥勋[1] 袁卫民[2,3] 王化坤[2,3] 郗红丽[2,3] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123 [2]江苏省常绿果树研究中心,江苏苏州215123 [3]苏州农业技术学院,江苏苏州215123

出　　处：《安徽农业科学》2010年第18期9419-9422,9434,共5页Journal of Anhui Agricultural Sciences

基　　金：江苏省高新技术项目(BG2007312)

摘　　要：[目的]获得高质量基因组DNA,探讨并改进快速提取枇杷基因组DNA的方法。[方法]比较了3种传统的基因组DNA提取方法并对CTAB法进行改良,同时以27对适用苹果属扩增的SSR特异引物,对15种枇杷品种和2个组合的杂交后代进行PCR扩增,分析品种扩增片断多态性构建聚类关系树,统计了杂种后代等位性位点。[结果]改良CTAB法所提取的DNA具有良好的质量;27对引物15个品种中扩增出33个多态性片段,用NTYST软件进行分析聚为6类;一个杂种杂交后代扩增出32条片段,确定22个等位位点。[结论]改良CTAB适合于快速提取枇杷基因组DNA,SSR在枇杷基因组中具有多态性。[Objective] The aim was to improve the method of DNA extraction for high quality genomic DNA from loquat leaves.[Method] Three kinds of methods of extracting DNA were compared and the CTAB method of DNA extraction for high quality genomic DNA from loquat leaves was improved.Twenty-seven primers flank microsatellite sequences previously identified in Malus × domestica（Borkh.） were assayed in fifteen loquat cultivars and progenies of two hybridization crosses.The amplification fragment polymorphism in the different cultivars was analyzed to construct the phylogenic tree and the numbers of allele sites were estimated in the hybridization progenies.[Result] By the improved CTAB,high quality DNA needed for PCR was obtained.Thirty-three polymorphic bands were amplified with the twenty-seven primers.Unweighted Pair-group Method（UPGMA） analysis based on Nei＇s genetic distance showed that the 15 genotypes were clustered into six groups.Thirty-two polymorphic amplifications were obtained from progenies of the crosses in loquat and twenty-two of them were allelic.[Conclusion] The improved CTAB method could extract high quality DNA from loquat leaves quickly and SSR showed polymorphic in loquat genomes.

关 键 词：枇杷 DNA提取 引物 SSR 微卫星序列 

分 类 号：S667.3[农业科学—果树学]