基于rDNA ITS序列SNP位点的特异性聚合酶链反应引物鉴别青椒和花椒

Identification of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. based on SNP in rDNA ITS sequence by specific PCR primers

目的设计专用于花椒与青椒鉴别的特异性聚合酶链反应(PCR)引物,建立花椒与青椒的分子鉴别方法。方法本研究提取花椒与青椒样品的DNA,并利用rDNA ITS(核糖体DNA内部转录间隔区)序列通用型引物分别对样品进行PCR扩增,PCR产物经双向测序、比对,寻找单核苷酸多态性(SNP)特异性位点,并分别针对花椒与青椒的SNP位点设计出花椒与青椒的特异性PCR引物,用于花椒与青椒的特异性PCR引物鉴别。结果所设计的特异性引物为花椒和青椒的鉴别提供了分子鉴定的依据,可以快速、准确地检测出花椒和青椒。结论本文所设计的特异性PCR引物能有效地鉴别出花椒与青椒,该方法具有准确、高效、灵敏、简便等特点,具有较好的市场应用前景。

Objective To design specific PCR primers for identification of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim., and to establish a molecular identification method for Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. Methods The DNA of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. samples were extracted in the study. The samples were amplified by PCR using ribosome DNA internal transcriptional spacer（rDNA ITS） sequence universal primers. The PCR products were determined by bi-directional sequencing and compared to find out the single nucleotid polymorphism（SNP） loci. The specific PCR primers of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. were designed for the SNPs loci of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim., respectively. It was used to identify the specific PCR primers of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. Results The specific primers designed for the identification of Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. provided the basis for molecular identification, which could quickly and accurately detect Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim. Conclusion The specific PCR primers designed in this paper can effectively identify Zanthoxylum schinifolium Sieb. et Zucc. and Zanthoxylum bungeanum Maxim., which is accurate, efficient, sensitive and simple with a good prospect of market application.