莪术基原植物 DNA 条形码序列的筛选与鉴定.

In order to find a rapid and efficient identification method for three original species of Curcumae Rhizoma, identification efficiency of different DNA barcoding sequences were evaluated in present study. Totally nine samples of three different species of Curcuma were collected. After DNA extraction, PCR amplification and sequencing, seven kinds of DNA barcoding sequences, including ITS, ITS2, matK, psbA-trnH, trnL-trnF, rpoB and atpB-rbcL, were firstly compared in terms of success rate of PCR amplification and characteristics of sequences. Then, by means of variation site analysis and genetic distance calculation, these seven kinds of DNA barcoding sequences were further evaluated. Finally, the unidentified samples were identified by the phylogenetic tree based on the chosen DNA barcoding sequences. The results were as follows:(1)ITS, ITS2 and matK barcoding sequences were inapplicable due to the low success rate of PCR amplification and sequencing; The variation information of psbA-trnH, trnL-trnF and rpoB was insufficient to distinguish three different original species of Curcumae Rhizoma; Only atpB-rbcL barcoding sequence was 642-645 bp in length with 29.0%-29.9% GC content and 11 variation sites, for which, the three species of Curcumae Rhizoma could be distinguished only by atpB-rbcL barcoding sequence.(2)According to the phylogenetic tree based on atpB-rbcL sequence, the unidentified samples were identified as Curcuma wenyujin. In conclusion, atpB-rbcL sequence could be used as a standard sequence for the rapid and efficient identification of original species of Curcumae Rhizoma. [ABSTRACT FROM AUTHOR]