---

作 者 ：陈随清;潘成学;卢小蕾;王利丽

期 刊 ：武汉植物学研究 2010年 28卷 1期 页码：72-75

关键词：山茱萸；栽培品种；rDNA ITS序列；

Keywords:Cornus officinalis, Cultivated varieties, rDNA ITS sequence,

摘 要 ：

为测定山茱萸(Cornus officinalis Sieb.et.Zucc.)核糖体DNA的ITS序列,对山茱萸不同栽培品种进行了ITS序列分析。通过实验筛选出一对引物,进行PCR扩增,对扩增产物提取纯化,双脱氧链终止法DNA测序。然后,利用DNAssist Version 2.0软件加手工校正确定ITS1-5.8S-ITS2序列,并进行ITS序列分析。获得了山茱萸的ITS1-5.8S-ITS2完全序列,ITS1为253bp,5.8S为156bp,ITS2为273bp,总共682bp。7种果型的山茱萸其5.8S基因序列显示高度的一致性,圆柱形果型、长梨形果型、椭圆形果型和纺锤形果型的ITS区序列完全一致,短圆柱形果型在ITS1区3′端及ITS2区5′端各有1个变异位点;短梨形果型在ITS1区5′端有3个变异位点;长圆柱形果型在ITS1区有5个变异位点。结果表明,ITS序列在山茱萸种内比较保守,有的栽培品种之间有较小的差异,此研究为中药山茱萸分子鉴定提供了科学依据。

Abstract:To study rDNA ITS sequence of Comus officinalis Sieb. Et. Zucc., and analyzed the rDNA ITS sequence of its different cultivated varieties. A pair of primers was selected by experi-ment. The ITS gene fragment was amplified using this pair of primers, and PCR products were purified with DNA gel extraction kit (AxyPrep~(TM)) and sequenced by the method of Sanger dideoxy. Then the ITS1-5. 8S-ITS2 sequences were analyzed by the software of DNAssist Ver-sion 2. 0. The ITS1-5. 8S-ITS2 sequences was found whole ITS sequence consisting of 682 bp, ITS1 253 bp,5.8S 156 bp and ITS2 273 bp. The 5.8S gene sequences of 7 fruit shapes show highly coincidence. The ITS1 and ITS2 gene sequences of cylinderic shape, long pear-shape, el-liptic shape and spindly shape are in coincidence. The short cylinderic shape have 1 variable site in 3’end of ITS1 and 5’end of ITS2. The short pear-shape have 3 variable site in 5’end of ITS1 and the long clinderic shape have 5 variable site in ITS1. The ITS1 and ITS2 gene fragments are conservative at specific level in C. Officinalis. While they are less difference among some culti-vars. So they can being molecular marker for authentication of C. Officinalis Sieb. Et. Zucc.

---