Abstract
BACKGROUND: Bread wheat (Triticum aestivum L.) is a major global food crop, and understanding its maternal lineage and genetic diversity is essential for breeding, authentication, and evolutionary studies. Chloroplast genomes provide valuable markers for phylogenetic inference and cultivar discrimination; however, conventional plant DNA barcodes often lack sufficient resolution within the genus Triticum. This study aimed to characterize the complete chloroplast genome of the Korean wheat cultivar 'Keumkang' and to develop effective chloroplast-based barcode markers for improved identification of Triticum species and cultivars. RESULTS: The complete chloroplast genome of T. aestivum cv. Keumkang was assembled using PacBio HiFi reads and determined to be 135 909 bp in length, exhibiting a typical quadripartite structure. Comparative analyses of 44 Triticum and related Aegilops chloroplast genomes revealed that Keumkang shared an identical chloroplast genome structure with several Korean cultivars, indicating a common maternal origin. Phylogenomic analysis placed T. aestivum in close association with T. turgidum subsp. durum, supporting its maternal derivation from the AABB genome lineage. Nucleotide diversity analysis identified six coding sequences and 11 intergenic regions with relatively high polymorphism. Based on these regions, 17 chloroplast-specific barcode markers were developed and experimentally validated. While conventional barcodes (matK, rbcL, trnL-F) achieved only approximately 18% cultivar discrimination, the combined use of the 17 newly developed markers improved identification accuracy to 50% among the examined accessions. CONCLUSION: The complete chloroplast genome of T. aestivum cv. Keumkang provides new insights into the maternal lineage and chloroplast diversity of wheat. The newly developed set of 17 chloroplast barcode markers substantially enhances cultivar-level discrimination within the genus Triticum and represents a useful molecular tool for wheat breeding, germplasm authentication, and evolutionary studies. © 2026 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.