中药材桔梗及其易混品的DNA条形码分子鉴定

Molecular identification of Platycodon grandiflorum and its common miscible using DNA barcording

为了特异性地鉴别桔梗药材Platycodon grandiflorum及其易混品,通过优化DNA提取方法,并基于ITS(internal transcribed spacer)序列上的特异性位点,设计特异性引物,进行特异性PCR扩增,以扩增成功率为判定指标快速鉴别桔梗药材真伪.研究结果表明,改良CTAB(cetyltrimethylammonium bromide)法提取DNA较纯且100%扩增成功;基于桔梗293位和538位的特异性位点,设计特异性引物U1/D1,且特异性PCR鉴别中,仅桔梗能扩增得到约264 bp的特异性条带,其他药材均为阴性扩增.ITS序列以及特异性引物可准确鉴别桔梗及其常见易混品,为桔梗药材的基原鉴定提供科学依据.

To specifically identify Platycodon grandiflorum and its miscible products,a method was established by optimizing DNA extraction method,designing specific primers based on the ITS(internal transcribed spacer)sequence,and carrying out specific PCR amplification.The authenticity of P.grandiflorum was quickly determined by the amplification success rate.The results showed that the modified CTAB(cetyltrimethylammonium bromide)method gave relatively pure DNA and amplification was 100%successful,specific primers U1/D1 and specific PCR were designed based on the specific sites of P.grandiflorum 293 and 538.In the identification,only P.grandiflorum can be amplified to obtain a 264 bp specific band,and other medicinal materials were negative.The ITS sequence and the designed specific primers can be applied to identification of the P.grandiflorum and its common miscible products,which should provide scientific evidence for origin identification of P.grandiflorum.