Ding G, Zhang D, Yu Y, Zhang B, Zhao L
Chemical and Biological Engineering College, Yancheng Institute of Technology, Jiangsu Yancheng 224003, China.
Genetika. 2013 May;49(5):626-31. doi: 10.7868/s0016675813050044.
The single nucleotide polymorphism (SNP) and amplification refractory mutation system (ARMS) have been applied to authenticate Limonium species and their corresponding herb samples. One species specific primer was designed and the amplification product is 200 bp (Limonium sinense) by using this primer. No band was observed with other Limonium species. The phylogenetic relationship of Limonium species were studied using ribosomal DNA ITS and the adulterants (L. bicolor, L. aureum and L. wrightii) were clustered with L. sinense in NJ tree. Inter simple sequence repeat (ISSR) was used to assess genetic diversity and population structure of L. sinense and a high level of genetic diversity was detected (H(E) = 0.2573, PPB = 85.71%) with POPGENE. Based on AMOVA analysis, there was moderate variation between pairs of populations with phi(ST) from 0.1744 to 0.5131 and on average 28.81% of the genetic variation occurred among populations. Five main clusters were shown in UPGMA dendrogram using TFPGA. The results showed that SNP and ARMS could be used to authenticate not only Limonium species but related herbs on rDNA internal transcribed spacer region. Possible strategies should be implemented for conservation of this endemic herb.
单核苷酸多态性（SNP）和扩增阻滞突变系统（ARMS）已被应用于对补血草属植物及其相应的草药样本进行鉴定。设计了一种物种特异性引物，使用该引物扩增产物为200 bp（中华补血草）。其他补血草属物种未观察到条带。利用核糖体DNA ITS研究了补血草属植物的系统发育关系，在邻接树上，掺杂物（二色补血草、金色补血草和黄花补血草）与中华补血草聚类在一起。利用简单重复序列区间（ISSR）评估了中华补血草的遗传多样性和种群结构，通过POPGENE检测到较高水平的遗传多样性（H(E)=0.2573，PPB=85.71%）。基于分子方差分析（AMOVA），种群间存在中等程度的变异，φ(ST)为0.1744至0.5131，平均28.81%的遗传变异发生在种群间。使用TFPGA在非加权组平均法（UPGMA）聚类图中显示了五个主要聚类。结果表明，SNP和ARMS不仅可用于鉴定补血草属物种，还可用于鉴定rDNA内转录间隔区的相关草药。应实施可能的策略来保护这种特有草药。
