猕猴桃AFLP分析体系的建立

Construction of the AFLP Analysis System for Actinidia Genome

从64对AFLP分析引物中随机选取了19对引物组合.经过跑小板的聚丙烯酰胺凝胶,银染显带AFLP条带,从中筛选出可做荧光的8对引物.再从中选取了扩增位点丰富,带型质量好,分辨率较高,条带信号强度一致性好,条带分布均匀,且条带较完整的4对引物:E-AAC+M-CAC、E-AAG+M-CTG、E-AAC+M-CAG、E-AAC+M-CTA进行荧光跑带、读带.共在156个位点上扩增出条带,4对引物共扩增出多态性条带132条,多态性比例平均为84.62%,4对引物对10份猕猴桃材料的区分率达100%.说明该4对引物用于猕猴桃属植物的AFLP分析是可行的.

19 pairs were chosen from 64 pairs of AFLP analysis primers. And 8 pairs of the 19 pairs suitable for fluorescent testing were screened through polyacrylamide gel electrophoresis and siliver-staining AFLP analysis. Of the 8pairs, 4 pairs that had abundant amplification sites, high band quality, high resolution, consistent band signal and evenly distributed and complete bands, were chosen as E-AAC＋M-CAC, E-AAG＋M-CTG, E-AAC＋M-CAG, and E-AAC＋M-CTA. They were tested for their fluorescent AFLP bands. 156 sites were found capable of producing amplifying bands; the 4 pairs of AFLP analysis primers totally produced 132 polymorphism bands with their average polymorphism rate at 84.62% and divided ten Actinidia materials with the accuracy rate of 100%.