Abstract: In this study, plant DNA barcode (matK, psbA-trnH, psbK-psbI, and rbcL) candidate sequences were used for the PCR amplification and sequencing of 104 samples from 19 Aspidistra species. The amplification success rate, sequencing success rate, and intra- and inter-species genetic variation of each barcode sequence were investigated, with BLAST searching adopted to evaluate the identification efficiency of each sequence. Results showed that the identification success rate of psbK-psbI was 88.7%, which was the highest for a single sequence. The identification efficiency of multiple combinational sequences was also compared and found to be significantly higher than that of the single sequences, with the success rate of matK+(psbK-psbI) reaching 100%. In addition, the neighbor-joining tree constructed based on the matK+(psbK-psbI) combinational sequence showed that samples of the same species had better aggregation and were mostly monophyletic. Therefore, matK+(psbK-psbI) appears to be the best barcode sequence for interspecific identification of Aspidistra.