DNA barcoding and phylogenetic relationships in marine toxic dinoflagellate genus Ostreopsis based on mithocondrial genes

The marine dinoflagellate genus Ostreopsis includes species producing potent toxic compounds, such as palytoxin and palytoxin analogs, which cause toxic blooms in Mediterranean and tropical or other temperate areas. Phylogenetic and phylogeographical analyses based on ribosomal data set revealed the existence of distinct species and, within them, clades related to geographical distribution. Due to a high morphological variability, species belonging to this genus are very difficult to identify and cryptic species could be present. Thus, developing a standardized DNA barcode approach for this marine dinoflagellate can allow confirming known species and uncover hidden variability with consequent description of new species. These information, besides leading to a better understanding of species diversity in natural environment, could assist in identification and detection of different species directly in field sample. This study represents the first attempt to assess the suitability of mitochondrial COI (cytochrome c oxidase 1) and cob (cytochrome b) as an identification tool for Ostreopsis species. For this purpose, we designed specific primers to amplify and sequence mitochondrial COI and cob genes from several Ostreopsis spp. isolates from worldwide areas. Phylogenetic analyses of single and concatenated mtDNA genes within genus Ostreopsis, as well as their intra and inter-specific divergences, were compared to those based on nuclear ribosomal genes LSU and 5.8S-ITS regions. Phylogenetic analyses of the genus Ostreopsis using mtDNA genes resulted uninformative, being not able to distinguish different species. Moreover, the so called 'barcode gap' that is interspecific variation exceeding intraspecific variation, was detected only in ribosomal genes. Thus, our original goal to apply the mtDNA barcode to recognize species within this toxic marine dinoflagellate was not accomplished