广西产鸡血藤遗传多样性的RAPD与ISSR标记方法的比较研究

摘要

目的:比较随机扩增多态性DNA分子标记技术(RAPD)和简单重复序列间区扩增技术(ISSR)两种标记方法,研究广西不同居群鸡血藤遗传多样性的优劣.方法:分别采用RAPD和ISSR标记方法,利用POPGENE 32软件、Ntsys软件、SPSS 17.0软件,对广西不同采集地的9份鸡血藤的遗传多样性进行研究.结果:筛选出的3条RAPD引物及4条ISSR引物扩增后,共得到198和315个位点,多态性位点37和80个,多态性位点比率为18.7%和25.4%,有效等位基因数为1.416 8、1.584 0,基因多样性指数为0.269 4、0.351 3,Shannon多样性指数为0.431 6、0.529 9.ISSR标记均高于RAPD标记,RAPD和ISSR的平均遗传相似系数为0.757 64、0.683 80,表明ISSR检测遗传多样性更灵敏.二者的聚类结果相近,相关系数r为0.847,说明其在0.001水平呈极显著正相关.结论:ISSR标记方法比RAPD标记反映出更多的遗传多样性信息,更适合于广西产不同居群鸡血藤的遗传多样性研究.%OBJECTIVE:To compare genetic diversity of Spatolobi caulis from different areas of Guangxi by random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR). METHODS:Through using POPGENE 32 software,Ntsys software and SPSS 17.0 software,RAPD and ISSR methods were used to study genetic diversity of 9 samples of S. caulis from dif-ferent areas of Guangxi. RESULTS:After amplification of screened 3 RAPD primers and 4 ISSR primers,and there were 198 and 315 locus,and 37 and 80 polymorphism locus. Rates of polymorphism locus were 18.7% and 25.4%;the number of effective al-leles were 1.416 8 and 1.584 0;genetic diversity index were 0.269 4 and 0.351 3;Shannon diversity index were 0.431 6 and 0.529 9. All the values of ISSR marker were higher than RAPD marker. The average genetic similarity coefficient of ISSR and RAPD were 0.757 64 and 0.683 80,indicating ISSR was more sensitive for the detection of genetic diversity. The clustering result of them was close to each other. The correlation coefficient of them were 0.847,indicating very significant positive correlation at the level of 0.001. CONCLUSIONS:ISSR could reflect more information of genetic diversity than RAPD,and is more suitable for research of genetic diversity of S. caulis from different areas of Guangxi.