基于叶绿体psbA-trnH基因间区序列鉴定山东丹参为新种  

Identification of Salvia shandongensis new species based on sequences of the plastid psbA-trnH intergenic region

作　　者：李晓娟[1] 韩建萍[2] 李建秀[1] 陈晓辰[2] 张龙霏[1] 李佳[1] 顾正位[1] 张永清[1] 

机构地区：[1]山东中医药大学,山东济南250355 [2]中国医学科学院、北京协和医学院药用植物研究所,北京100193

出　　处：《药学学报》2013年第8期1338-1344,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81001608);山东省科技发展计划项目(2008GG2NS02022);山东省农业良种工程课题项目(2009LZ01-03)

摘　　要：为从分子水平鉴定山东丹参(Salvia shandongensis)及其近缘物种,本研究采用psbA-trnH序列对山东丹参、丹参(S.miltiorrhiza)和白花丹参(S.miltiorrhiza f.alba)共计33份材料进行了PCR扩增并测序,获得了该间隔区的完整序列。采用CodonCodeAligner进行序列拼接,应用MEGA5.0计算山东丹参及其近缘种的种内、种间的K2P距离,构建UPGMA树。结果显示,山东丹参psbA-trnH序列长度约为391 bp,丹参和白花丹参psbA-trnH序列长度约为386 bp,psbA-trnH序列在山东丹参及其近缘种种间存在丰富的变异,山东丹参种内平均K2P遗传距离为0,丹参和白花丹参种内平均K2P遗传距离分别为0.002和0.001,山东丹参与丹参种间遗传距离为0.034～0.04,与白花丹参种间遗传距离为0.005～0.008,山东丹参种内平均K2P遗传距离均远远小于丹参和白花丹参种间平均K2P遗传距离;聚类结果显示山东丹参与丹参、白花丹参(变型)可明显区分,这与形态分类特征相符。本研究首次从分子水平确立了山东丹参在植物分类学上的地位,揭示了山东丹参与近缘种之间的亲缘关系,为山东丹参药用植物新资源的开发提供DNA遗传分子鉴定的科学依据。研究结果表明psbA-trnH基因间区可作为条形码来鉴定山东丹参及其近缘种。To identify Salvia shandongensis and its relatives at molecular level, the psbA-trnH intergenic region of three species including Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba were amplified and sequenced. Sequences were assembled with CodonCode Aligner. The K2P genetic distances between Salvia shandongensis and its relatives were calculated and UPGMA tree was performed by MEGA5.0. The results indicated that the lengths of psbA-trnH regions of Salvia shandongensis were about 391 bp, while the lengths of psbA-trnH regions of Salvia miltiorrhiza and S. miltiorrhiza f. alba were about 386 bp. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Salvia shandongensis and its relatives. The intra-specific genetic distances of Salvia shandongensis were 0, while the intra-specific genetic distances of Salvia miltiorrhiza and S. miltiorrhiza f. alba were 0.002 and 0.001 respectively. Additionally, the genetic distance of Salvia shandongensis and Salvia miltiorrhiza ranged from 0.034 to 0.04, and the genetic distance of Salvia shandongensis and S. miltiorrhiza f. alba ranged from 0.005 to 0.008, the intra-specific genetic distances of Salvia shandongensis were much smaller than that of Salvia miltiorrhiza and S. miltiorrhiza f. alba; clustering results showed that there were obvious differences between Salvia shandongensis, Salvia miltiorrhiza and S. miltiorrhiza f. alba, which was consistent with morphological characteristics. This study not only firstly provides the scientific basis for establishing the taxonomy position in molecular level and revealing their genetic relationships of S. shandongensis, S. miltiorrhiza and S. miltiorrhiza f. alba; but also provides DNA molecular identification scientific basis for the development of new medicinal plant resources of Salvia shandongensis. Our results suggest that the psbA-trnH intergenic spacer region can be used as a barcoding to identify Salvia shandongensis, Salvia m

关 键 词：psbA-trnH基因间区 山东丹参 丹参 白花丹参 DNA分子鉴定 

分 类 号：R931[医药卫生—生药学]