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The roots and rhizomes of  Rhodiola crenulata  and  R. rosea  have been used worldwide as adaptogens for hundreds of years. However, rapid growth in demand has resulted in merchants using other species of  Rhodiola  as adulterants. Here, we surveyed 518 individuals representing 47 of the 55 species in the genus, including 253  R. crenulata  individuals from 16 populations and 98  R. rosea  individuals from 11 populations, to evaluate the utility of the internal transcribed spacer 2 (ITS2) barcode for identification of  Rhodiola  species. We detected six haplotypes in  R. crenulata  and only one haplotype in  R. rosea . An obvious overlap between intra- and inter-specific distance was detected, and the authentication efficacy of ITS2, which was assessed by BLAST1, a nearest distance method, and a tree test, was much lower than in other groups. However,  R. crenulata  and  R. rosea  could be exactly identified. Analysis showed that the secondary structure of ITS2 differs in  R. crenulata  and its closest relatives. Our results demonstrated that both a mini barcode from ITS2 and the structure of ITS2 are effective markers for the identification of  R. crenulata  and  R. rosea . This study represents the most comprehensive database of ITS2 barcodes in  Rhodiola  to date and will be useful in  Rhodiola  species identification.

Establishment of the most comprehensive ITS2 barcode database to date of the traditional medicinal plant Rhodiola (Crassulaceae)