RAPD技术筛选麦冬分子鉴定标记的研究

The screening of Ohpiopogon japonicus molecular marker by RAPD analysis

应用RAPD技术筛选三种药用麦冬品种湖北麦冬,川麦冬和浙麦冬的分子鉴定标记,为构建麦冬特异性PCR鉴定方法奠定基础.对RAPD反应的关键影响因子进行优化,确立最佳反应条件.以麦冬嫩叶中提取的基因组DNA作为RAPD反应模板,逐一使用40条随机引物对三种麦冬基因组DNA进行RAPD分析,从引物S86,S65,S64,S22和S20结合下的RAPD图谱中找到了8条麦冬分子鉴定标记,其中湖北麦冬分子鉴定标记4条,川麦冬和浙麦冬各2条.这些分子鉴定标记在琼脂糖凝胶中的分子量介于500～2 000 bp之间,条带明亮清晰,可供回收进行序列分析,以便设计出特异性PCR引物,实现三种麦冬特异性PCR鉴定的目标。

Molecular markers of three Ophiopogon japonicas from Hubei,Sichuan,Zhejiang were screened by RAPD analysis to further applying for specific PCR identification.RAPD conditions were established by optimizing several key factors.The genomic DNA isolated from the tender leaves was as the template of RAPD.RAPD analysis was carried out using forty random primers one by one.From the RAPD results of primer S86,S65,S64,S22 and S20,eight Ophiopogon japonicas molecular identification markers were found,including four markers from Liriope spicatavar and two Oghiopogon japonicns marker from each province of Sichuan and Zhejiang.The bands of molecular markers in the agarose gel were bright and clear,with the appropriate molecular weight from 500 to 2 000 base pairs.These molecular markers could be recovered and sequenced,which will enable us to design specific PCR primers for the specific PCR identification of Ophiopogon japonicas.