Objective To screen species and identify DNA molecular markers based on the chloroplast whole genome data of four species of the genus Bupleurum for medicinal use. Methods Based on the published chloroplast genome sequences of Bupleurum plants, the chloroplast genome was analyzed using bioinformatics methods. A specific DNA barcode for accurately identifying the four original species of Bupleurum medicinal materials was developed based on the hypervariable regions of the chloroplast genome, and then verified. Results The chloroplast genomes of four species ranged in size from 155 478—178 668 bp. All the four genomes were represented as typical quadripartite structures. Two pairs of primers based on the hypervariable regions were designed to efficiently amplify 55 samples. The analysis of sequencing results confirmed that two hypervariable regions, rps16-psbK and petN-psbM, can be used as specific DNA barcodes for the identification of Bupleurum chinense, B. scorzonerifolium, B. smithii and B. marginatum var. stenophyllum. Conclusion The specific DNA barcodes rps16-psbK and petN-psbM, screened by comparative chloroplast genomics, could achieve precise classification and identification of the four original species of Bupleurum medicinal materials, providing important scientific support for ensuring the safety of clinical use and regulating the market order of Bupleurum medicinal materials.

Bupleuri Radix  /  Bupleurum L.  /  chloroplast genome  /  original species identification  /  specific DNA barcode