青海产不同海拔唐古特大黄的遗传多样性分析

Genetic Diversity Analysis of Rheum tanguticum Maxim.ex Balf.Under Different Altitudes in Qinghai

目的探讨海拔高度对唐古特大黄遗传多样性的影响,为进一步研究其品质提供参考。方法采用(inter simple se-quence repeat,ISSR)分子标记技术,从97个ISSR引物中筛选出14个多态性引物用于不同海拔唐古特大黄样品的总DNA扩增,琼脂糖凝胶电泳检测扩增结果,GIS凝胶图像处理系统统计电泳结果,POPGENE 32分析遗传多样性,MVSP构建UPGMA聚类树。结果从14个ISSR引物中共扩增出213个位点,其中多态性位点208个,多态位点百分率(PPB)为97.7%,观测等位基因数(Na)1.976 5,有效等位基因数(Ne)1.533 5,Nei′s基因多样性(He)32.0%,Shannon′s指数(Ⅰ)0.487 5。大黄样品个体平均多态位点数随海拔的升高而上升。UPGMA聚类分析结果将所有大黄样品分为3类:海拔3 000 m以下的样品聚为一类;海拔3 000 m以上的样品也聚为一类;海拔3 000 m的样品单独聚为一类。结论唐古特大黄的遗传多样性随海拔升高而上升。在进行青海省不同海拔唐古特大黄的化学成分分析时,可将3 000 m海拔作为研究其化学成分差异的参照分界线,为进一步评价其品质提供参考依据。

OBJECTIVE Tn investigate the genetic diversity of Rheum tanguticum Maxim. ex Balf. distributed in different ahitude regions. METHODS 14 Primers screened from 97 ISSR primers were used for Inter Simple Sequence Repeat （ISSR） amplification of total DNA samples of R. tanguticum. The ISSR-PCR products were detected by agarose gel electrophoresis and Tanon GIS gel documentation ystem. POPGENE 32 was used to ealcalate the genetic diversity parameters and MVSP was used to cluster the samples. RESULTS 213 Discernible DNA fragments were produced, of which 208 documentations （ PPB = 97.7%） were polymorphic loci. The observed number of alleles （Na） was 1. 976 5. Effective number of alleles（Ne） was 1. 533 5. Nei＇s genediversity （He） was 32. 0%. Shannon＇s intormation index （ Ⅰ ） was 0. 487 5. The individual mean ISSR sites were increased with the raise of altitude. All the samples were divided into three types by cluster analysis （UPGMA）. Samples from altitude 3 000 m, below and above 3 000 m were grouped respcclively. CONCLUSION The genetic diversity of R. tanguticum was increased with the raise of altitude, and the critical altitude of clustering is 3 000 m, which may be also used as the critical line in its chemical analysis for further quality study.