Identification of Pulsatillae Radix and Its Miscible Counterfeit Based on DNA Barcoding and ITS2 Secondary Structure
Objective:To identify Pulsatillae Radix and its miscible counterfeit(Anemone hupehensis var.japonica,Leontopodium leontopodioides,and Potentilla chinensis Ser.)based on DNA barcoding and ITS2 secondary structure,aiming to improve the accuracy and efficiency of Pulsatillae Radix identification.Methods:DNA extraction was performed on Pulsatillae Radix and its miscible counterfeit plants(Anemone hupehensis var.japonica,Leontopodium leontopodioides,and Potentilla chinensis Ser.),followed by PCR amplification and sequencing.Specific primers were designed,and molecular markers were utilized to extract DNA barcodes and ITS2(Internal Transcribed Spacer 2)region secondary structures.Analysis was conducted using MEGA 11.0 software to study and compare Pulsatillae Radix and its miscible counterfeit.Results:Neighbor-Joining(NJ)tree analysis revealed distinct clusters for Pulsatillae Radix,Anemone hupehensis var.japonica,Leontopodium leontopodioides,and Potentilla chinensis Ser.,with visibly different ITS2 secondary structures among them.Thus,identification of Pulsatillae Radix and its miscible counterfeit can be achieved based on their respective secondary structures.Conclusion:DNA barcoding and ITS2 secondary structure analysis provide an accurate method for identifying Pulsatillae Radix and its miscible counterfeit(Anemone hupehensis var.japonica,Leontopodium leontopodioides,and Potentilla chinensis Ser.).
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