Single locus microsatellites in Gracilariales (Rhodophyta): High level of genetic variability within Gracilaria gracilis and conservation in related species

Four single locus microsatellites identified in the red alga Gracilaria gracilis (Stackhouse) Steentoft, Irvine, et Farnham (Rhodophyta) were examined for allelic diversity at different spatial and taxonomic levels. First, because simple morphological diagnostic characters are often missing within the Gracilariacae, we developed a simple and rapid method based on rDNA ITS size variation in order to verify the taxonomic status of the samples used in this study. All European (including Mediterranean samples), Argentinian, and Namibian samples used in our study were confirmed to be a homogenous G. gracilis group. By contrast, our results on rDNA ITS sizes showed that Gracilaria from Japan, initially identified as G. gracilis, was different from the rest of the G. gracilis group. Secondly, microsatellite polymorphism and conservation at the species level was tested on the worldwide collection of G. gracilis and within a single population. The loci Gv1AAG and Gv1AAC showed no allelic variation, whereas two others, Gv1CT and Gv2CT, were highly polymorphic. All microsatellite loci were conserved within G. Gracilis, except in the sample from Japan. The taxonomic status of G. gracilis from Japan is thus questionable. This study revealed a high level of within-population polymorphism (52 alleles for Gv1CT and 12 for Gv2CT). Moreover; the combination of these two loci was shown to be very powerful for identifying individuals within a population, that is, 93% of the individuals were characterized by a unique genotype. Finally, conservation of the four loci was tested in taxonomically related species of Gracilaria (G. chilensis, G. pacifica, and G. tikvahiae) and two Gracilariopsis species (Gs. sp. and Gs. longissima). The results suggest that the polymorphic locus Gv2CT may provide a valuable genetic marker within the different species of the Gracilariaceae.