Identification of medicinal plants of Elsholtzia based on ITS2 sequence
The internal transcribed spacer 2(ITS2)sequences in DNA barcoding were employed in DNA barcoding to identify Elsholtzia ciliata and its closely related species,as well as to develop a reliable meth-od for distinguishing E.ciliata.ITS2 sequences were amplified and sequenced from 14 Elsholtzia plants of diverse origins,and 21 ITS2 sequences from the same genus were retrieved from GenBank.In total,35 ITS2 sequences were obtained.The Kimura-2-Parameter(K2P)model was employed to analyze intraspecific and interspecific genetic distance,the Neighbor-joining(NJ)method was used to construct a phylogenetic tree,and ITS2 database was utilized to predict the secondary structure of ITS2.The results indicated that ITS2 sequence length in Elsholtzia ranged from 219 to 233 bp.The interspecific genetic distance of Elsholtzia was larger than the intraspecific genetic distance,with a clear barcoding gap observed between the species.The NJ phylogenetic tree demonstrated clear differentiation among species.Significant differences in arm length,inner ring size and the number of inner ring were observed between E.ciliata and other species within four spiral regions of the ITS2 secondary structure.This study demonstrates that DNA barcoding technology based on ITS2 sequence can effectively identify E.ciliata and its related species.
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