Objective To analyze the psbA-trnH sequences of Polygonaturn kingianum var.grandifolium plant and its related species in Chongqing based on DNA barcode molecular identification method of traditional Chinese medicine.Methods The DNA were extracted using the reagent kit method,and bidirectional sequencing was performed after amplification of their psbA-trnH sequences.And then the sequences were assembled by CodonCode Aligner,the phylogenetic trees were constructed after sequences alignment and computation in MEGA.Results A total of 77psbA-trnH sequences of 9 species were achieved(40 sequences of Polygonatum kingianum var.grandifolium plant and 24 sequences of its related species were obtained by self-testing,and the other 13 related sequences were downloaded from GenBank),with the length of 601 to 615 bp and the GC content of 35.16％to 35.49％.Among the above sequences,thepsbA-trnH sequence of master haplotype of P kingianum var.grandifolium was 602 bp in length and its GC content was 35.22％.The 12 haplotypes of the species were identified with the 19 variable sites.Based on Kimura 2-Parameter(K2-P)model,the genetic distance of the nine species was range from 0.001 7 to 0.024 9,with the overall average value of 0.012 1.The constructed UPGMA phylogenetic tree was an effective method in clustering and distinguishing between P kingianum var.grandifolium and the other species.Conclusion ThepsbA-trnH sequences of P kingianum var.grandifolium can use as an effective auxiliary tool for discriminating it from easily-confused species,providing a reference for ensuring the source of Polygonati Rhizoma in Chongqing.
维普
万方数据