黄花白及的SCAR分子标记转化研究

目的 建立黄花白及的SCAR标记,为黄花白及的分子鉴定提供科学依据.方法 用随机引物进行随机扩增多态性DNA (random amplified polymorphic DNA,RAPD)标记筛选,获得特异的RAPD标记条带,经回收、克隆、测序后,根据RAPD标记条带的两端序列设计特异引物进行常规PCR反应,以获得序列特征化扩增区(sequence characterizedamplified region,SCAR)标记.结果 利用50对随机引物筛选得到黄花白及的一条896 bp的特异片段,可以区别于小白及、独叶白芨和白及.该序列设计特异引物后,经过多次重复试验,该RAPD标记的特异片段已成功转化为SCAR标记.结论 建立的SCAR标记条带清晰明亮,结果稳定,可作为黄花白及分子鉴定的依据.

Research on SCAR molecular marker transformation of Bletilla ochracea

Objective In order to provide a scientific basis for molecular identification of Bletilla ochracea,the sequence characterized amplified region(SCAR) marker of B.ochracea was established.Methods The random primer was used for random amplified polymorphic DNA(RAPD) screening to obtain specific marker bands.Through separating,extracting,cloning,and sequencing,the specific primers were designed for normal PCR reaction based on the sequences of both ends of RAPD marker bands to obtain SCAR marker.Results A specific segment with 896 bp of B.ochracea was obtained by random screening of 50 pairs of primers,which was different from B.formosana,Pleione yunnanensis,and B.striata.A pair of specific primers was based on the sequence of RAPD marker bands and then the RAPD marker was successfully transformed into SCAR marker after repeated tests.Conclusion The established SCAR marker could make the band clear and bright and be used as the basis for molecular identification of B.ochracea with the stable results.