Identification of Cuscutae Semen and adulterants using DNA barcode
Objective To investigate the purity and species of adulterants in commercially available Tusizi(Cuscutae Semen)in order to provide a basis for the improvement of the quality of Cuscuta and its safe application..Methods Genomic DNA was extracted from a total of 25 Cuscutae Semen samples collected from markets.ITS2 sequences were amplified,sequenced and then annotated by HMMer.MEGA6.0 was used to calculate genetic distance and construct a NJ phylogenetic tree.The functional domain,haplotype and secondary structures of ITS2 sequences were analyzed using DNAsp5.0.The obtained sequences were transformed into chromatic barcode images and two-dimensional DNA barcodes using the R Program.Results The adulteration of commercially available Cuscutae Semen was serious.All Cuscuta samples contained at least one adulterant.Seeds of Chenopodium album,Amaranthus hybridus,Corispermum hyssopifolium,Acalypha australis,Echinochloa crus-galli,Persicaria lapathifolia and cracked soybean(Glycine max(Linn.)Merr.)were successfully identified in Cuscutae Semen as adulterants.Conclusion This study confirmed the serious adulteration of Cuscutae Semen and clarified the species of these adulterants.ITS2 sequence barcode is able to identify Cuscuta and other common adulterants,providing a method for identifying tiny seeds.In addition,a combination of ITS2 sequences and two-dimensional code technology enables standardization and informationization in the management of Cuscuta market.
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