Abstract
The Brassicaceae family comprises a diverse group of species important for agriculture, medicine, and ecology. This study assessed the effectiveness of DNA barcoding using rbcL and matK gene regions to identify 12 Brassicaceae species collected across Kazakhstan. Both loci were successfully amplified and sequenced; however, matK exhibited significantly higher nucleotide diversity, greater polymorphism, and superior phylogenetic resolution compared to rbcL. Phylogenetic trees based on maximum likelihood analysis demonstrated finer species discrimination with matK, while rbcL provided broader clade separation. Combining both loci enhanced species-level identification reliability. These results underscore the value of multi-locus barcoding for robust taxonomic resolution and highlight the genetic richness of Kazakhstan’s Brassicaceae flora. Findings from this study contribute to ongoing efforts in biodiversity conservation and molecular systematics of economically and ecologically significant plant groups.