nrDNA ITS和cpDNA rpl16基因测序法对六种鼠尾草的检测

为从鼠尾草属植物中鉴别丹参品种,采用基因测序方法,用核糖体核酸内转录间隔区基因(nrDNA ITS),编码核蛋白体大亚基多肽L16的基因(rpl16)及叶绿体DNA上包含trnL以及trnL和trnF间隔区的区域基因(trnL-trnF)的序列,检测六种鼠尾草属新鲜植物.由于nrDNA ITS和rpl16突变率较高,可以做为6种鼠尾草的基源鉴定标记,依此设计了两对特异引物,从6种鼠尾草中鉴定出丹参(Salvia miltiorrhiza)和云南鼠尾草(S.yunnanensis).但trnL-trnF突变率太低,未能用于鉴别.商品干燥中药材因加工和储藏的方式致使DNA降解严重,基因测序法难于应用.

Identification of Salvia Species by nrDNA ITS and cpDNA rpl16 Sequence Analyses

Three DNA regions were sequenced for testing six fresh plant samples of Salvia species. These three DNA regions were nrDNA ITS (nuclear ribosomal DNA internal transcribed spacer), chloroplast rpl16 (the gene encoding ribosomal protein L16), and trnL-trnF (the cpDNA region comprising the trnL and the intergenic spacer between trnL and trnF). The results showed that the nrDNA ITS and rpl16 genes could provide novel information for origin identification of Salvia species. Due to their higher mutation rates of these 2 gene markers, Salvia species-specific primers were designed and S.miltiorrhiza and S.yunnanensis were identified. The trnL-trnF gene expressed low mutation rate, it could not identify the species. Since the damage of DNA by the pretreatments of the dry roots of Chinese herbs, it is hard to apply the molecular markers to commercial samples for identification.