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基于ISSR技术对藏药“蒂达”3种基源植物的亲缘关系分析
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| 篇名： | 基于ISSR技术对藏药“蒂达”3种基源植物的亲缘关系分析 |
| TITLE： | |
| 摘要： | 目的：对藏药“蒂达”的3种基源植物紫红獐牙菜、青叶胆和椭圆叶花锚进行亲缘关系研究。方法：利用简单重复序列区间（ISSR）分子标记技术对紫红獐牙菜9个样品（样品ZT-1～ZT-5采自大理苍山感通寺，样品ZC-1～ZC-4采自大理宾川县）、青叶胆2个样品（QYD-1～QYD-2）和椭圆叶花锚2个样品（HM-1～HM-2）进行聚合酶链式反应（PCR）扩增。采用青叶胆的DNA基因组为模板DNA进行引物筛选，对筛选得到的8条引物进行PCR反应；对PCR扩增产物进行人工读带，建立原始数据矩阵，并计算多态性条带比率；同时，采用NTSYS 2.1软件计算遗传相似系数，并选择UPGMA法绘制聚类图。结果：通过8条ISSR引物共反应获得113条清晰可辨的PCR扩增产物条带，多态性位点百分率为100％。紫红獐牙菜、青叶胆和椭圆叶花锚3个品种共13个样品的遗传相似系数在0.301～0.500之间；紫红獐牙菜9个样品的种内遗传相似系数在0.752～0.929之间。聚类分析结果显示，当距离线为0.410时，可将13个样本分为3个类群，即青叶胆、椭圆叶花锚、紫红獐牙菜；当距离线为0.780时，可将紫红獐牙菜的9个样品分为2个亚类，一个亚类只有苍山感通寺采集的样品ZT-1，另一个亚类含有其余8个样品。结论：利用ISSR分子标记技术能从分子水平对紫红獐牙菜、青叶胆和椭圆叶花锚进行鉴别。紫红獐牙菜与青叶胆、椭圆叶花锚存在一定的亲缘关系，但亲缘关系相对较远、遗传差异较大；大理地区两个不同地点产紫红獐牙菜的遗传差异较小、亲缘关系较近，但表现出丰富的遗传多样性。 |
| ABSTRACT： | OBJECTIVE： To study the phylogenetic relationships of 3 basic plants of Tibetan medicine “Dida”， such as Swertia puricea， Wertia mileensis， Halenia elliptica. METHODS： ISSR technology was used for PCR amplification of 9 samples of S. puricea （ZT-1 to ZT-5 from Gantongsi in Dali Cangshan， ZC-1 to ZC-4 from Binchuan county of Dali）， 2 samples of W. mileensis （QYD-1 to QYD-2） and 2 samples of H. elliptica （HM-1 to HM-2）. Using DNA genome of S. puricea as template， 8 primers were screened and used for PCR reaction. The PCR amplification products were read by hand， the original data matrix was established， and the polymorphic band ratio was calculated. At the same time， genetic similarity coefficient was calculated by using NTSYS 2.1 software， and UPGMA method was used to draw cluster diagram. RESULTS： A total of 113 clear and identifiable amplification product bands were obtained by 8 ISSR primers. The rate of polymorphic site was 100％. The genetic similarity coefficients for totally 13 samples of S. puricea， W. mileensis and H. elliptica ranged 0.301-0.500. Intraspecific genetic similarity coefficients for 9 samples of S. puricea ranged from 0.752 to 0.929. The cluster analysis showed， when the range line was 0.410， 13 samples could be divided into three groups， i.e. S. puricea， W. mileensis， H. elliptica； when the range line was 0.780， 9 samples of S. purpurea could be divided into 2 subgroups， one of which was only sample ZT-1 collected from Gantongsi in Cangshan， and the other contained the remaining 8 samples. CONCLUSIONS： ISSR technology can be used to identify S. punicea， S. glabra and H. elliptica at the molecular level. S. punicea has some genetic relationship with S. glabra and H. elliptica， but the genetic relationship is relatively distant and the genetic difference is large. S. punicea from two different locations in Dali area has little genetic difference and close relationship， but it shows abundant genetic diversity. |
| 期刊： | 2019年第30卷第12期 |
| 作者： | 李水仙，夏从龙，陈丽元 |
| AUTHORS： | LI Shuixian，XIA Conglong，CHEN Liyuan |
| 关键字： | ISSR分子标记技术；紫红獐牙菜；青叶胆；椭圆叶花锚；鉴定；亲缘关系；遗传多样性 |
| KEYWORDS： | ISSR； Swertia puricea； Swertia mileensis； Halenia elliptica； Identification； Phylogenetic relationship； Genetic diversity |
| 阅读数： | 274 次 |
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