苹果RAPD分析体系的建立

Establishment of RAPD Analysis System in Apples

对富士苹果（Ｍａｌｕｓ ｐｕｍｉｌａ Ｍｉｌｌ．ｃｖ．Ｆｕｊｉ）ＲＡＰＤ（Ｒａｎｄｏｍ Ａｍｐｌｉｆｉｅｄ Ｐｏｌｙｍｏｒｐｈｉｃ ＤＮＡ）分析体系的优化研究表明，ＲＡＰＤ反应体系中，ＤＮＡ、Ｔａｑ酶、引物和Ｍｇ２＋４种主要成分的最适用量分别为：２０ｎｇ、１．０ Ｕ、０．２ｕｍｏｌ·Ｌ－１和３．０ｕｍｏｌ·Ｌ－１。采用该优化体系，以 ＯＰＪ０３为引物，构建了我国及世界范围内苹果生产中重要的２８个品种的ＲＡＰＤ指纹图谱，分析了其遗传多样性，区分了供试的２８个苹果品种中的１５个，区分率达５３．６％。讨论了ＲＡＰＤ鉴定苹果品种的应用及其主要影响因素。

The optimal reaction of RAPD in Fuji apple variety(Malus pumila Mill. )was studied.The results showed that the optimum concentrations of four important components i. e. DNA, Taq DNase, primer, and Mg2+ were 20ng, 1.0 U,0.2 umol.1-l, and 3.0umol.l-1 respectively. The RAPD fingerprints of 28 apple varieties grown world-widely were established by using the optimum system by primer OPJ03. The genetic diversity was analyzed,and 15 out of 28 apple varieties were identified based on the fingerprints. The identification rate was 53 .6%. The RAPD application in apple variety identification and the important influence factors on RAPD reac- tion were discussed.