Simple differentiation of two closely related species Porphyra tenera and Porphyra yezoensis (Bangiophyceae, Rhodophyta) based on length polymorphism of actin-related protein 4 gene (ARP4)

Genetic polymorphisms were investigated to develop a simple and rapid method to differentiate between the two closely related species, Porphyra tenera and Porphyra yezoensis. Polymerase chain reaction (PCR) using the specific primer pair of the ARP4 gene gave length polymorphic single fragments of genomic DNAs from five strains of P. tenera (Japan T-8, JTW; Korea KTY1, KTY2, KTY3) and seven strains of P. yezoensis (Japan TU-1, TU-2, TUH-25, JHU, JA-1; Korea KGJ, KPH). All strains of P. yezoensis had introns 60 bp longer than that of P. tenera. Multiple cleaved amplified polymorphic sequence (CAPS) markers were also developed to differentiate P. tenera and P. yezoensis. This is the first report of length polymorphisms that can be used to differentiate between the two species using only PCR amplification with agarose gel electrophoresis. It is expected that the length polymorphism and plentiful CAPS profiles obtained in this study will be useful in the assessment of genetic diversity within P. tenera and P. yezoensis as well as in breeding science that requires the collection of various strains of the two species.