中药山茱萸SCAR标记的研究

目的:建立山茱萸的SCAR标记,为山茱萸药材分子鉴定提供科学依据.方法:用随机引物进行RAPD筛选,获取特异的RAPD标记条带,分离提取RAPD标记条带,进行克隆和测序,根据测定RAPD标记条带的两端序列设计1对特异引物进行常规PCR反应,获得SCAR标记.结果:根据RAPD标记条带的序列设计筛选了4对特异引物,其中YST38,YST43分别对7个山茱萸品种样本的DNA进行SCAR扩增,7个山茱萸栽培品种均能产生单一的PCR条带,可以用作山茱萸的鉴别;而YST38对圆柱形果型、长梨形果型还有1条特异性条带,可以做为这2个品种的鉴别依据;YST39对7个山茱萸品种进行扩增,纺锤形果型样本的谱带大小(350～400 bp)比其余6个果型的谱带大小(650～700 bp)短了300 bp左右,这条谱带可以做为纺锤形果型的鉴别依据;YST92对圆柱形果型、长梨形果型均产生了1条600～700 bp的条带,椭圆形果型、短圆柱形果型产生了1条200～300 bp的条带,长圆柱形果型、纺锤形果型、短梨形果型3个果型无扩增,YST92可用来筛选山茱萸的几种果型.结论:建立的SCAR标记,条带清晰明亮,结果稳定,可作为山茱萸品种选育和药材分子鉴定的依据.

Study on sequence characterized amplified region(SCAR) markers of Cornus officinalis

Objective:To establish sequence characterized amplified region markers of Cornus officinalis and provide a scientific basis for molecular identification of C.officinalis.Method: The random primer was screened through RAPD to obtain specific RAPD marker bands.The RAPD marker bands were separated,extracted,cloned and sequenced.Both ends of the sequence of RAPD marker bands were determined.A pair of specific primers was designed for conventional PCR reaction,and SCAR marker was acquired.Result: Four pairs of p...