RAPD方法鉴定油菜“杂油59”种子纯度的研究

IDENTIFICATION OF GENETIC PURITY OF SEED OF HYBRID RAPE “ZAYOU No.59” WITH RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD)

本文采用RAPD分子标记技术对我国育成的油菜新品种“杂油 5 9”的亲本和F1代种子纯度进行了技术鉴定 .用 40个 10mer的随机引物对“杂油 5 9”的 2个亲本“垦C8”和“陕 3A”进行了RAPD分析 ,共出现 2 90条带 ,分布于 35 30～ 2 2 0bp之间 ,大部分条带在双亲之间是相同的 .引物Opa 0 6 ,Opc 12 ,Opc 2 0 ,Opk 0 3,opk 13,Opj 12出现差异扩增带 ,经实验确认Opk 0 3的PCR结果重复性较好 .用它对两个亲本进行RAPD分析 ,检测出 5条强带 ,其中 5 10bp是母本特征带 ,2 6 0bp是父本的特征带 ,在F1代中这两条特征带互补 ,且结果稳定 .本文还对RAPD方法鉴定种子纯度的样品选择问题进行了探讨 .图 3参

The identification of the genetic purity of F1 hybrid seeds in new variety, rape “Zayou No59” was made via poly morphic RAPD (Random Amplified polymorphic DNA) markers. Using 40 ten mer arbitrary oligonucheotide primers the parents of “Zayou No. 59” were analyzed will RAPD. Approximately 290 products, ranging from 220～3530 base pairs(bp), were amplified. The majority of the products were identical in both “Ken C8” and “Shan 3A”. Primer Opa 06, Opc 12, Opc 20, Opk 03, Opk 13, Opj 12 generated polymorphic products. It was found that the reproducibility of primer Opk 03 was steady after repeating the tests. It could produce 5 bright bands, the fragment at 510 bp unique to the female line, and the fragment at 260 bp unique to the male line. The two fragments were complementary and stable in the F1 generation.The sampling selection of DNA sample used in identification of the purity in F1 was discussed in this report. Fig 3, Ref 11