Identification of Rubus Chingii Hu and Its Related Confounders by PCR-RFLP

OBJECTIVE To establish a specific molecular identification method for Rubus chingii Hu. METHODS The ITS sequences of Rubus chingii Hu and its adulterants were sequenced and analyzed. A restriction enzyme was screened according to the variation sites of sequence analysis results. Subsequently, polymerase chain reaction restriction fragment length polymorphism PCR-RFLP method was established and optimized to distinguish Rubus chingii Hu and its adulterants, the stability and applicability of the method was investigated and verified. RESULTS The designed primers could amplify the sequences of Rubus chingii Hu and its adulterants to produce a 800 bp specific fragment. MboI restriction endonuclease could only digest the amplified fragment of Rubus chingii Hu into two smaller pieces and could not digest the amplified fragment of the adulterants. In this way, Rubus chingii Hu was identified specifically. CONCLUSION The PCR-RFLP method established in this study can be used to identify the authenticity of Rubus(PCR-RELP) chingii Hu.