Development of the specific molecular markers of the thin-blade strain (HR-5) in Pyropia haitanensis

Abstract

In order to establish a method for germplasm identification of the thin-blade strain (HR-5) of Pyropia haitanensis, the specific molecular markers of the conchocelis and blades of HR-5 were developed by using the techniques of insertion/deletion (InDel) and sequence-characterized amplified regions (SCAR). Results showed that a specific band was amplified by InDel-3 primer-pair (F: GGG GAT GGC AGT TTTAC; R: GAA TCT TCA TTT CGT GGT G), and a specific band was amplified by SCAR-170 primer-pair (F: GAC GCC GAG TCG TAG TGC; R: CCG TTG TTA GTG TGA TGG ) in conchocelis DNA of the HR-5 strain, which was significantly different from those in other four control strains of P. haitanensis and also present in the gametophytic blades of the HR-5 strain cultured in the laboratory. When those primer-pairs were applied to discriminate HR-5 blades from the other cultivars in the Pyropia sea area, the InDel-3 and SCAR-170 markers showed the same results. In addition, genetic analysis of the above specific markers showed that the InDel-3 marker was inherited from its female parent, while the SCAR-170 marker of HR-5 was inherited from its male parent. These results indicated that the specific bands amplified by InDel-3 and SCAR-170 primer-pairs could be considered as the specific markers of HR-5 strain and could be used for its germplasm identification.