Use of PCR-RFLP for the discrimination of Japanese Porphyra and Pyropia species (Bangiales, Rhodophyta)

In order to distinguish 18 Porphyra and Pyropia species, the present study employed polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis using mitochondrial DNA related to the ATP synthase F0 subunit 6 (ATP6) gene and partial mitochondrial DNA including trnC, rps11, sdh3, trnG, trnN, trnP, and rns. The two primer sets on the mitochondrial DNA used in this study were able to amplify single fragments with PCR in 16 Japanese and 2 non-Japanese Porphyra and Pyropia species. Lengths of partial mitochondrial DNA related to ATP6 gene and trnC-rns ranged from 664 bp (Py. dentata and Py. haitanensis) to 677 bp (Py. lacerata and Py. kurogii) and from 1,292 bp (Py. seriata) to 1,343 bp (Py. kurogii and Py. moriensis), respectively. All 18 species were successfully distinguished using a combination of five restriction enzymes (TaqI, SspI, AciI, Cfr13I, and AluI). It was therefore concluded that PCR-RFLP analysis is a valuable tool for discrimination of wild strains of Porphyra and Pyropia species for potential use in mariculture.