Detection Method for Unapproved Genetically Modified Rose Plants in Korea Using Duplex Polymerase Chain Reaction

A duplex PCR method was developed to detect a transformation vector pSPB130 used in the development of a genetically modified (GM) rose plant. To detect a GM rose plant, the anthocyanin synthase (ANS) was used as an endogenous reference gene of rose in PCR detection. The primer pair RHANS-KF/KR. producing 107 bp amplicon was used to amplify the ANS gene and no amplified product was observed in any of the 9 different plants used as a template. The primer pair GMRH-KF/KR was designed to amplify the junction sequence between 35S promoter and flavonoid 3',5'-hydroxylase (F3 '5 'H) gene in pSPB130. The detection limit of the duplex PCR method is approximately 0.5%. This result indicates that this duplex PCR method could be useful for monitoring unauthorized GM rose in Korea.