摘要
目的研究海风藤原植物风藤和山蒟的分子鉴定方法。方法提取31份来自全国各地药材植物及不同医药企业生产饮片的基因组,对psbA-trnH、matK、petA-psbJ等叶绿体基因片段以及核糖体间隔序列ITS进行PCR扩增测序及进化树构建。从叶绿体基因组的简单重复序列中筛选4对特异性引物,对其扩增产物进行毛细管电泳分析。结果所有样品的psbA-trnH序列高度一致,其它DNA条形码的序列均有一定差异。进化树的聚类分析结果matK和petA-psbJ片段相近,且与SSR分子标记结果一致,但与ITS聚类结果有差异。结论matK和petA-psbJ片段对风藤和山蒟的鉴别能力优于ITS和psbA-trnH,DNA条形码和SSR分子标记法均可为中药材海风藤的种质资源鉴定提供分子依据。
Objective To identify Piper kadsura and Piper hancei through molecular methods.Methods The total DNA were extracted from 31 samples from different geographical populations and Chinese medicine manufacturers.PsbA-trnH,matK,petA-psbJ and ITS were amplified,sequenced,and phylogenetic trees were constructed.Moreover,4 pairs of SSR primers were designed to amplify the chloroplast genomes and the results were analyzed by capillary electrophoresis.Results Among the 4 DNA barcodes,PsbA-trnH sequences had high similarity while others had some discrimination.The phylogenetic trees of matK and petA-psbJ barcodes were quiet similar,which shared the same clustering results with SSR,but different from ITS.Conclusion MatK and petA-psbJ are better than ITS and psbA-trnH to discriminate Piper kadsura and Piper hancei.The DNA barcode and SSR methods can be used to identify the germ plasm resource of Chinese medicine HaiFengTeng.
作者
严卓彦
夏瑛瑛
崔洁
李彬
YAN Zhuoyan;XIA Yingying;CUI Jie;LI Bin(Zhoushan Institute for Food and Drug Control(National Ocean Food Quality Inspection Center),Zhoushan 316012,China)
出处
《现代中药研究与实践》
CAS
2023年第2期23-27,共5页
Research and Practice on Chinese Medicines
基金
浙江省药监局科技计划项目(2022014)。
