Variety identification and comparative analysis of genetic diversity in yardlong bean (Vigna unguiculata spp. sesquipedalis) using morphological characters,SSR and ISSR analysis

Genetic diversity and relatedness of 23 yardlong bean (Vigna unguiculata spp. sesquipedalis) accessions and 7 accessions of a hybrid between cowpea (V. unguiculata spp. unguiculata) and yardlong bean (dwarf yardlong bean) in Thailand were estimated using morphological characters, simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers. In addition, two mungbean (Vignaradiata (L.) Wilczek) and two blackgram (Vigna mungo (L.) Hepper) accessions were also used as outgroup species for molecular analysis. Five morphological characters were diverse among most accessions. However, five groups of 2–3 accessions could not be distinguished from one another based on these morphological characters alone. Unweighted pair-group arithmetic average (UPGMA) analysis of these characters separated these 30 accessions into 2 major groups; the yardlong bean group and the dwarf yardlong bean group. Eleven of the sixteen SSR primers yielded clear SSRs, ten of which were polymorphic (90.91% polymorphism), detecting a total of 54 alleles with an average of 4.91 alleles per locus. These 10 polymorphic SSR markers successfully distinguished 28 yardlong bean and dwarf yardlong bean accessions. The polymorphic information content (PIC) among genotypes varied from 0.251 to 0.752 with an average of 0.597. Among the 16 ISSR primers used, a total of 312 ISSR fragments were amplified for these three Vigna species, revealing the polymorphism percentage of 91.03%. The average ISSR PIC value (0.197) with the range of 0.137–0.276 was lower than that of SSR. Nevertheless, the average marker index of this multilocus marker was 3.495, which was higher than that of SSR (0.669), owing to the differences in the effective multiplex ratio. In addition, Mantel test cophenetic correlation coefficient was higher for ISSR (0.566) than that of SSR (0.198). These results indicated higher efficiency of ISSR for estimating the levels of genetic diversity and relationships among yardlong beans and dwarf yardlong beans in this study. Pair-wise coefficients of SSR- and ISSR-based genetic similarity among all yardlong bean and dwarf yardlong bean accessions averaged 0.87 and 0.91, respectively, suggesting a narrow genetic base that emphasizes the need to broaden genetic diversity to ensure continued breeding success. Clustering of genotypes within groups was not similar when SSR and ISSR derived dendrograms from UPGMA analysis were compared. It appeared that ISSR was the most effective marker system in determining the genetic variability and relationships among yardlong bean and dwarf yardlong bean accessions and differentiating three Vigna species. In addition, ISSR was also most useful for variety identification since all 30 yardlong beans and dwarf yardlong bean accessions can be effectively distinguished by only four ISSR primers with the highest PIC values.     

基于SSR标记的清凉峰地区三叶海棠遗传多样性研究

【目的】评价清凉峰地区三叶海棠的遗传多样性水平,为其保护提供理论依据。【方法】利用10对SSR引物,对清凉峰地区的36份三叶海棠样品进行了遗传多样性分析,并从中随机抽取6,9,12,15,18,27,34株大小不同的样本,组成7个群体进行遗传多样性特征比较。【结果】(1)10对引物在36份样品中的多态性等位基因数(Na)平均值为7.1(5.0~10.0),有效等位基因数(Ne)平均值为3.954(1.527～5.786),平均观察杂合度(Ho)为0.781(0.194～1.000),平均期望杂合度(He)为0.699(0.345～0.827),香农多样性指数(I)平均值为1.458(0.662～1.918);(2)采用UPGMA法构建的系统树中,很明显地将36份供试样品划分为两组,与用贝叶斯聚类方法得到的结果一致,对三叶海棠所有样品进行的主成分分析进一步证实了以上结果;(3)群体样本量≥15株时,样本量就不会对群体内遗传多样性水平、遗传一致度及群体内等位基因数目产生明显的影响。【结论】清凉峰地区三叶海棠的遗传多样性水平较高,其群体明显分化为2个基因源,三叶海棠群体遗传多样性研究和种质收集保存中的适宜样本量为≥15株,研究供试样品可以反映清凉峰地区三叶海棠遗传多样性的总体水平。