甜瓜SRAP多态性分析及在品种鉴定中的应用

以国家蔬菜工程技术研究中心甜瓜单倍体育种组提供的12份薄皮甜瓜,30份厚皮甜瓜,2份野生甜瓜为试材,该研究优化了甜瓜SRAP反应体系,筛选出10对引物组合,分析了甜瓜多态性并对品种进行鉴定.结果表明:10μL反应体系为1×Buffer lμL,引物组合各20 ng,模板DNA 20 ng,dNTPs 2.0 mmol/L,Taq DNA聚合酶0.2U.10对引物组合在2份甜瓜上共扩增出166条带,其中多态性条带104条,每对引物组合平均能扩增出10.4个多态性位点.甜瓜间遗传距离在0.08～0.79(平均为0.357),54份甜瓜被分为二大类:Ⅰ类包括1份印度野生甜瓜和3份哈密瓜；Ⅱ类可再分为2个亚类:Ⅱ-1,是12份薄皮甜瓜；Ⅱ-2,包括1份中国野生甜瓜和37份厚皮甜瓜.引物组合m21e01、m21e10和m21e11区分率分别高达79.6％、81.5％和79.6％,有助于品种的高效鉴定.

Analysis of SRAP Polymorphism and Application of Variety Identification in Cucumis melo L.

The 12 striffen muskmelon,30u thick-skin muskmelon,2 wild melon as test materials from melon haploid breeding group of National Engineering Research Center of vegetables SRAP reaction system in C.melon were optimized,10 pairs of filtered primer combinations analyzed polymorphism and identified varieties in C.melon.The results showed that the optimum concentration of five important components,i.e.1×Buffer,primer combinations,template DNA,dNTPs,and Taq DNA polymerase in 10 μL reaction system were 1 μL,20 ng,20 ng,2.0 mmol/L and 0.2 U respectively.10 pairs of primer combinations amplified 166 bands altogether in two C.melon,104 bands were polymorphic,each pair of primer combinations could amplified 10.4 polymorphic locus.The genetics distances(GD)of C.melon were between 0.08~0.79(average GD was 0.375),the 54 C.melon were divided two kinds:one concluded 1 India wild melon and 3 Hami melons;the other could divided two sub-kinds,the first was 12 C.melo sp.conomom,the second was 1 China wild melon and 37 C.melo sp.melo.The discriminated ratio of primer combinations m21e01,m21e10 and m21e11 were 79.6%,81.5% and 79.6% in proper order,which would be help for melons to identify high effectively.