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。
To study rDNA ITS sequence of Cornus officinalis Sieb.et.Zucc.,and analyzed the rDNA ITS sequence of its different cultivated varieties.A pair of primers was selected by experiment.The ITS gene fragment was amplified using this pair of primers,and PCR products were purified with DNA gel extraction kit（AxyPrepTM）and sequenced by the method of Sanger dideoxy.Then the ITS1-5.8S-ITS2 sequences were analyzed by the software of DNAssist Version 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,elliptic 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 cultivars.So they can being molecular marker for authentication of C.officinalis Sieb.et.Zucc.
出处
《武汉植物学研究》
CAS
CSCD
北大核心
2010年第1期72-75,共4页
Journal of Wuhan Botanical Research
基金
科技部"十一五"科技支撑计划项目(2006BAI09B05-8
2006BAI06A15-3)
关键词
山茱萸
栽培品种
RDNA
ITS序列
Cornus officinalis
Cultivated varieties
rDNA ITS sequence
