桂花品种的ISSR-PCR分析

利用ISSR—PCR方法对桂花的54个品种进行了基因组多态性分析,从70个ISSR引物中筛选出13个多态性引物用于正式扩增,共扩增出90条DNA片段,其中多态性DNA带79条,占总扩增片段的87.8％,平均每个引物扩增的DNA带的数目为6.92条。依据扩增结果,应用RAPD Distans软件进行Nei相似性系数和遗传距离计算,利用UPGMA法构建聚类树状图。结果表明:ISSR分析中产生了一些品种特有的指纹图谱,因此ISSR技术在桂花品种和品种群(品系)的鉴定和阐明遗传关系中有很大的应用潜力。利用DNA扩增结果进行聚类分析,把供试桂花的54个品种分为7个大类,并对品种进行了处理及种下品种群的遗传关系进行了探讨。

A ISSR-PCR Analyze of the Osmanthus fragrans Cultivars

In this study, inter-simple sequence repeat (ISSR) was evaluated for its potential use in the identification of 54 Osmanthus fragrans cultivars. 13 out of 70 (18. 6%) ISSR primers could generate reproducible polymorphic fragments. The ISSR-PCR assay revealed a total number of 90 DNA bands, of which 79 bands were polymorphic (the percentage of polymorphic bands, PPB=89. 9%). Optimized ISSR primers amplified 4 to 10 bands ranging in size from 300 bp to 2000 bp, with an overall average of 6. 92 amplified bands per primer. Additionally, ISSR produced 1 cultivar-specific molecular marker. AMOVA(Analysis of molecular variance) software was used to calculate the Nei's genetic distance and a dendrogram was constructed based on UPGMA cluster analysis. These 54 sweet osmanthus cultivars surveyed were classified into 7 major groups, and each cultivar in this study could be distinguished from others, suggesting that PCR-based ISSR analysis was an efficient method for cultivar identification. The efficiency of ISSR analysis for cultivar identification and classification of sweet olive were also discussed.