南苍术与苍术素相关的ISSR-SCAR分子标记筛选

Screening of ISSR-SCAR Molecular Markers associated with atractylodin in Atractylodes lancea

为了探索鉴别茅山地区野生南苍术中高苍术素含量植株的分子学方法,为快速筛选高苍术素含量的植株提供理论依据,在茅山地区野外采集30株南苍术,提取根茎中挥发油,根据液相色谱法测定的挥发油中苍术素含量,筛选出5个高含量和5个低含量的单株,分别构建高含量和低含量植株DNA混合池,用于南苍术与苍术素相关的ISSR-SCAR标记研究。结果显示,在所选的29条ISSR引物中,引物ISSR-846(5’-CACACACACACACACART-3’)可以在苍术素含量高的南苍术中扩增出片段长度为948 bp的特异性条带。针对该特异性条带的序列,设计了正向引物(5’-AAGAGATCATTGGTATTAAAATTAAGTGTTATTAGTAG-3’)和反向引物(5’-CATAGATTCAATGGCAATCCTCACATCAACTTCAGATTCA-3’),验证发现,设计的ISSR-SCAR引物仅能在茅山南苍术高苍术素含量个体中扩增出条带,其他含量个体均未扩增出条带。认为本研究筛选的位点特异性引物,可用来快速鉴别高苍术素含量的南苍术植株。

To explore the molecular methods for the identification of Maoshan wild Atractylodes lancea with high atractylodin and to provide a theoretical basis for rapid screening of the plants with high atractylodin,thirty Atractylodes lancea individu-als were collected in Maoshan area and their essential oil was extracted to measure the content of atractylodin. Five individu-als with higher atractylodin as well as another five with lower atractylodin,were got to use for exploring the ISSR-SCAR markers associated with atractylodin content. Among 29 primers used, IS SR -8 4 6 （5 ’ -CACACACACACACACART-3’） could be used to amplify a specific band in higher atractylodin individuals. The SCAR primers（F-primer： 5 ’ -AAGAGAT- CATTGGTATTAAAATTAAGTGTTATTAGTAG-3’ ） and（R-primer： 5 ’ -CATAGATTCAATGGCAATCCTCACATCAACT- TCAGATTCA-3’）,designed according to the sequence of the specific bands,could be only used to amplify bands in high-er atractylodin individuals while could not be used in others. Therefore, the above-mentioned site-specific primers could be used to identify Atractylodes lancea individuals with higher atractylodin.