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In this study, ITS, ITS2,  matK ,  rbcL  and  psbA-trnH  in  Rehmannia  were successfully amplified and sequenced, but some ITS sequences need to be proofread according to ITS2 sequences. Compared with  rbcL ,  matK  and  psbA-trnH , ITS and ITS2 had higher mutation rate and more information sites, and ITS2 had higher interspecific diversity and lower intraspecific variation in  Rehmannia , but the interspecific genetic variation of  rbcL  and  matK  was lower. Furthermore, the obvious barcoding gap was found in  psbA-trnH  or ITS2 +  psbA-trnH , and the overlap between interspecific and intraspecific variation of ITS, ITS2 or  matK  was less. In addition, the phylogenetic tree based on ITS or ITS2 indicated that  R. glutinosa ,  R. chingii  or  R. henryi  with obvious monophyly could be successfully identified, but  R. piasezkii and R. elata  were clustered into one branch,  R. solanifolia  could not be distinguished from  R. glutinosa , and  R. chingii  was closer to  R. henryi . In phylogenetic tree based on  psbA-trnH  or ITS2 +  psbA-trnH , cultivars and wild varieties of  R. glutinosa  could be distinguished, were clearly separated from other  Rehmannia  species, and cultivars or wild varieties of  R. glutinosa  could be also distinguished by  matK . Taken together, ITS2 has great potential in systematic study and species identification of  Rehmannia , the combination of ITS2 and  psb A- trn H might be the most suitable DNA barcode for  Rehmannia  species.

The screening and identification of DNA barcode sequences for Rehmannia