The CBOL Plant Working Group has evaluated seven leading candidate plastid DNA regions for use in DNA barcoding of land plants. The regions assessed include the atpF–atpH spacer, matK gene, rbcL gene, rpoB gene, rpoC1 gene, psbK–psbL spacer, and trnH–psbA spacer. Based on criteria such as universality, sequence quality, and species discrimination, the group recommends the combination of the rbcL and matK genes as the standard plant barcode. This 2-locus combination is expected to provide a universal framework for the routine use of DNA sequence data to identify specimens and contribute to the discovery of overlooked species of land plants. The rbcL gene offers high universality and good discrimination power, while the matK gene provides higher resolution but requires further development. The recommended barcode is a pragmatic solution that balances universality, sequence quality, discrimination, and cost.The CBOL Plant Working Group has evaluated seven leading candidate plastid DNA regions for use in DNA barcoding of land plants. The regions assessed include the atpF–atpH spacer, matK gene, rbcL gene, rpoB gene, rpoC1 gene, psbK–psbL spacer, and trnH–psbA spacer. Based on criteria such as universality, sequence quality, and species discrimination, the group recommends the combination of the rbcL and matK genes as the standard plant barcode. This 2-locus combination is expected to provide a universal framework for the routine use of DNA sequence data to identify specimens and contribute to the discovery of overlooked species of land plants. The rbcL gene offers high universality and good discrimination power, while the matK gene provides higher resolution but requires further development. The recommended barcode is a pragmatic solution that balances universality, sequence quality, discrimination, and cost.