38种国兰亲缘关系的ISSR分析

ISSR Analysis of Genetic Relationship Among 38 China Orchid Cultivars

本研究以5种国兰的38个品种为实验材料,进行了ISSR分子标记分析。从100条随机引物中筛选出10条能扩增出清晰条带并具多态性的引物,共扩增出89条DNA片段,相对分子质量集中在150～2000bp之间,进而建立了国兰属植物的ISSR-PCR反应体系;利用UPMGA类平均聚类法对遗传相似系数进行分析并构建系统树,聚类结果与传统分类基本一致,初步证明利用ISSR分子标记技术对国兰进行亲缘关系分析的可靠性。根据聚类结果,选择遗传相似系数为0.67的来自浙江的下山春兰和来自福建的下山墨兰进行了杂交试验并获得了成形的蒴果,证明了利用ISSR分子标记指导兰属植物进行辅助育种的可行性。

We studied the inter-species relationship of 38 cultivars of 5 species in China orchid by using ISSR molecular markers.10 primers which could produce clear and polymorphic bands were screened out from 100 random primers,and then 89 DNA bands with a relative molecular weight,a range from 150 bp to 2000 bp were amplified in this paper,thus the reaction condition of RAPD-PCR amplification system of Cymbidium were established.Meanwhile,we calculated the genetic similarity coefficient by NTSYS-pc and constructed the dendrogram by UPMGA.The clustering results are basically consistent with the traditional classification,the preliminary proof proved that it was reliable to analyze the genetic relationship of China orchid with ISSR molecular markers.According to the clustering results,Xiashan Chuanlan from Zhejiang and Xiashan Molan from Fujian,which got tighter at 0.67 similarity level;were selected to do experimental hybrid and the capsule growing finally.The experiment demonstrates the feasibility of using ISSR molecular marker technology to guide the heredity and breeding of China orchid.