应用RAPD技术探讨穿龙薯蓣遗传多样性及亲缘关系

The Application of RAPD Technology in Genetic Diversity Detection of Dioscorea nipponica Makino

目的分析不同居群的穿龙薯蓣及其近缘品种山药、盾叶薯蓣的亲缘关系，鉴别中药材穿龙薯蓣及其近缘品种。方法采用随机引物扩增多态性DNA（RAPD）技术扩增植物基因组DNA样品，利用NTSYS软件，建立了UPGMA聚类图结果从50条随机引物中筛选出清晰且多态性高的引物7条。共扩增出62条DNA片段。其中54条具有遗传多样性，多态率为92．71％。遗传相似系数在0．6595～0．875；应用引物S75，S313可有效鉴别穿龙薯蓣、盾叶薯蓣、山药。结论不同居群的穿龙薯蓣具有丰富的遗传多样性，DNA带型差异可准确区分穿龙薯蓣、盾叶薯蓣及山药，RAPD方法可以准确、快速简便地鉴定中药。尤其在正确引种方面具有很高的价值。

Objective To analyze different samples from different habitats and to identify Chinese traditional medicine Dioscorea nipponica Makino and its counterfeits. Methods Total genomic DNA sample of different plant pieces were amplified by randomly amplified polymorphic DNA （RAPD）. The data of amplified bands were analyzed by the software NTSYS . The cluster map including all samples were obtained by UPGMA. Results Seven random primers of high stable quality were screened from 50 primers. The cluster analysis results of 62 amplified DNA fragments were indicated. 54 were polymorphic and thepolymorphism rate was 92.17% , genetic similarity coefficient was from 0. 659 5 to 0. 875. Application of 575,5313 could effectively identify Dioscorea nipponica Makino, Dioscorea opopsita Thunb. and Dioscorea zingiberensis C. H. Wright. Conclusion The genetic diversity exists in population of Dioscorea nipponica Makino. RAPD is able to identify Dioscorea nipponica Makino, Dioscorea opopsita Thunb. and Dioscorea zingiberensis C. H. Wright quickly and truly, which is also quite valuable for correctly introducing plant.