利用等位基因特异PCR鉴别虎掌南星及其部分混淆品

Authentication of Pinellia pedatisecta and their adulterants by AS-PCR

目的:建立中药材虎掌南星的分子鉴定方法,为天南星药材质量控制提供依据。方法:GenBank中下载半夏属、天南星属、犁头尖属物种的叶绿体rpl20基因序列,利用DNAMAN进行比对分析,发现半夏属和虎掌南星的SNP位点,并设计鉴别引物Ptrpl94F,Ptrpl708R和Pprpl148F,Pprpl484R,采用等位基因特异PCR方法对虎掌南星、半夏、水半夏3个物种的10个样品进行扩增。结果:引物Pprpl148F和Pprpl484R仅对虎掌南星扩增333 bp条带,半夏、水半夏均无扩增,能准确鉴别虎掌南星。引物Ptrpl94F和Ptrpl708R对半夏、虎掌南星均扩增611 bp条带,水半夏无扩增。结论:本实验建立的等位基因特异PCR方法能准确区别出虎掌南星和半夏、水半夏,为虎掌南星及半夏的正确用药提供了保障。

Objective： To establish a molecular method for the authentication of PineUia pedatisecta and its adulterants. Method： DNA sequences of some species from P. tenore, Typhonium and Ar/saema were down/oaded from GenBank, the sequences were aligned using DNAMAN. Allele-specific primers for P. pedatisecta and P. tenore were designed according to their SNPs in rpl 20 sequence. The designed primers were used to amplify 10 samples of P. pedatisecta, P. ternata and T. flageUiforme. Result： A 351 bp band was amplified from P. pedatisecta but not form P. ternata and T. flageUiforme by primer Pprpll49F and Pprpl484R. A 630 bp band was amplified from P. ternate and P. pedatisecta but not from T. flageUiforme by primer Ptrpl94F and Ptrp1699R. Conclution： AS-PCR has the advantages of highly specific and good reproducibility, by which P. pedatisecta can be identified from part of its adulterants quickly. It is a potential method to be used in the molecular identification of other materia medica.