Development of plastome-based HRM markers for the authentication of 12 major medicinal herbs in the Apiaceae family

Medicinal plants of the Apiaceae family often exhibit high morphological similarity, making accurate species identification challenging and leading to frequent cases of adulteration or substitution. To establish a systematic barcoding framework, we simultaneously assembled the chloroplast genomes (plastomes) and 45 S nuclear ribosomal DNA (nrDNA) sequences of 12 major medicinal herb species and performed comparative genomic analyses to identify interspecific variations in single-nucleotide polymorphisms (SNPs), insertions/deletions, nucleotide diversity (π), and simple sequence repeats (SSRs). Based on these genomic insights, seven high-resolution melting (HRM) markers were developed, which generated species-specific melting curve profiles and enabled unambiguous identification and discrimination of all 12 taxa. Compared with conventional DNA barcoding, HRM demonstrated advantages in cost-effectiveness, speed, and suitability for high-throughput analysis. Collectively, our study provides genomic resources and molecular tools that establish a comprehensive barcoding system for 12 medicinal herbs in the Apiaceae family, facilitating reliable species identification, genetic diversity assessment, and phylogenetic research, while contributing to the conservation and safe utilization of these plants.