Identification of north-western Atlantic Porphyra (Bangiaceae, Bangiales) based on sequence variation in nuclear SSU and plastid rbcL genes

Six species of Porphyra have commonly been recognized in the north-western Atlantic from Long Island Sound to the Canadian Maritimes: P. amplissima, P. leucosticta, P. linearis, P. miniata, P. purpurea, and P. umbilicalis. Distinguishing them with certainty has been problematic. A DNA-based system of molecular identification was developed using partial sequences of the nuclear small subunit ribosomal RNA gene (SSU) or the plastid ribulose-1,5-bisphosphate carboxylase-oxygenase large subunit gene (rbcL). Multiple samples of each taxon were surveyed for intraspecific variation. Intraspecific SSU divergences for Porphyra 'leucosticta', P. 'miniata', P. 'umbilicalis', and P. 'purpurea' ranged from 0% to 1%. There was more variation for P. 'amplissima' (0-2.1%) and P. 'linearis' (0-3.5%); however, each taxon was monophyletic. No intraspecific differences were observed for these taxa in rbcL (one to eight samples per taxon). These sequences were compared with P. yezoensis U51, introduced to Maine, and with P. 'dioica', a north-east Atlantic Porphyra easily confused with P. 'purpurea'. To discriminate between P. 'purpurea, P. 'umbilicalis', and A 'leucosticta', SSU variation was used to design primers for the Allele-Specific Polymerase Chain Reaction(TM). With molecular tools, we could classify over 80% of the monostromatic specimens surveyed, but the residue of unidentifiable specimens may indicate the existence of further monostromatic species in the north-west Atlantic. Porphyra 'purpurea' was found to occur further south than previously recorded. A morphologically cryptic Porphyra was discovered at Herring Cove, Nova Scotia, Canada. dagger Phylogenetic analyses using SSU or rbcL sequences showed 'soft incongruence' between gene trees, i.e. the topologies of the phylograms were similar but not identical, with only weak to moderate bootstrap support for the nodes that differed. Both trees strongly supported a clade including P. 'purpurea', P. 'umbilicalis', P. 'linearis', and P. 'dioica'. Porphyra sp. Herring Cove was allied with the remaining Porphyra taxa in the SSU tree. The rbcL phylogeny was less well resolved, consisting of a polytomy of a P. 'purpurea'-P. 'umbilicalis'-P. 'linearis'-P. 'dioica' clade, Porphyra sp. Herring Cove, a clade comprising P. 'amplissima' and P. 'miniata', and a P. 'suborbiculata'-P. 'leucosticta'-P. yezoensis clade.