基于ITS序列对青海不同地区中国沙棘遗传变异的分析

Genetic Variation Analysis of Hippophae rhamnoides in Different Regions of Qinghai Province Based on ITS Sequences

为研究青海省不同地区中国沙棘的遗传变异,本研究采集青海省不同地区野生的中国沙棘、西藏沙棘和肋果沙棘叶片作为实验材料,提取沙棘叶片的DNA,对其内转录间隔区(ITS)进行PCR扩增和扩增产物测序检测。对中国沙棘的ITS序列进行同源性和差异性分析,以西藏沙棘和肋果沙棘为外类群构建系统发育树。结果表明,不同地区中国沙棘ITS序列的同源性很高,均在99.5%以上。聚类分析结果显示中国沙棘、肋果沙棘和西藏沙棘各自分为一组,与形态学分类中归属于三个不同的种的分类结果一致。11个中国沙棘样品的ITS1区序列长度均为307 bp,5.8S区均为167 bp,ITS2区均为264 bp,3个区域分别存在3个、1个和5个变异位点。说明青海省不同地区中国沙棘的ITS区基因发生了不同程度的变异,本研究为进一步研究沙棘ITS基因提供了理论依据。

In order to study the genetic variation of Hippophae rhamnoides in different areas of Qinghai Province.In this study,the leaves of wild seabuckthron(Hippophae rhamnoides subsp.sinensis,H.tibetana and H.neurocarpa)collected from different areas of Qinghai Province were used as experimental materials,the DNA was ex tracted from the leaves,the internal transcribed spacer(ITS)were amplified by PCR and sequencing,and the sequence homology and diversity were analyzed.In the case of H.tibetana and H.neurocarpa as the outgroups,the phylogenetic tree was constructed.The results showed the Hippophae rhamnoides had high homology,the homolo gy is above 99.5%.The cluster analysis divided Hippophae rhamnoides subsp.sinensis,H.tibetana and H.neurocarpa into different groups,the results of clustering analysis are consistent with those of morphological classification.The lengths of ITS1,5.8 S and ITS2 from 11 samples were 307 bp,167 bp and 264 bp,ITS1,5.8 S,and ITS2 had 3,1,and 5 mutation points.The results showed that the ITS of Hippophae rhamnoides in different areas of Qinghai Province had different degrees of variation.These findings provide a theoretical basis for further research on Hippophae rhamnoides ITS genes.