RAPD标记在山葡萄种质鉴定中的应用

采用修改后的CTAB 法获得了高质量的基因组DNA 。利用RAPD 标记技术对山葡萄7 份种质进行鉴定, 用4 个引物(从30 个引物中筛选)对试材进行PCR 扩增, 共扩增出30 条谱带, 平均每条引物产生7.5 条谱带, 其中21 条谱带为多态性谱带, 占总谱带数的70%。不同引物扩增的谱带数不同, 范围在6~9 条之间。利用4 个引物扩增出的多态性谱带可以将7 份山葡萄种质区分。     

Genetic Diversity in Wild Grapes Native to China Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis

The use of the RAPD technique was investigated on a set of 73 genotypes of 18 wild grape species native to China, and one interspecific hybrid, seven Vitis vinifera L. cultivars, one rootstock cultivar and one strain of V. riparia L. Genetic diversity among these grapes was investigated based on RAPD analysis. The screening of 280 decamer oligonucleotides allowed the selection of 20 primers used for the analysis. A total of 191 RAPD markers were produced from the 20 selected primers. Relationships among the 83 clones or accessions based on their genetic distances were clustered using unweighted pair-group method arithmetic average (UPGMA) analysis in a dendrogram. Twenty-two clusters which fortunately adapted to 22 grape species level were clearly resolved on the dendrogram. The 18 wild grape species native to China were grouped into ten subclusters. The largest distance was found between V. riparia L., V. vinifera L., interspecific hybrid ( V. vinifera L.× V. larbrusca L.) and the wild grapes native to China. Among the wild grapes native to China, the largest distance was found between V. hancockii Hance and the other wild species. V. qinlingensis P.C.He was the second. Large genetic variation occurred among the different flower-type clones in one species.