Multiple rDNA ITS-types within the diatom Pseudo-nitzschia delicatissima (Bacillariophyceae) and their relative abundances across a spring bloom in the Gulf of Naples

The genus Pseudo-nitzschia includes a number of species responsible for blooms in coastal and open waters worldwide. P. delicatissima, a species reported as a potential source of amnesic shellfish poisoning (ASP), reaches high concentrations in the Gulf of Naples (Mediterranean Sea), where it regularly blooms in spring and, at times, in autumn. We assessed both intra- and inter-individual genetic diversity of this species before and during a bloom (February to April 2001) by sequencing the internal transcribed spacer regions (ITS1 and ITS2) and the 5.8S gene of the nuclear ribosomal DNA. PCR products obtained from 70 strains were cloned and several ITS copies were sequenced for each strain to assess intra-individual polymorphism. Phylogenies showed the presence of 5 distinct, well-supported lineages within what was considered to be a single morphospecies. Genetic diversity was higher in pre-bloom conditions, while all strains collected at the height of the bloom clustered within a single major clade. Ultrastructural investigations carried out on selected strains revealed morphological features slightly different from the ones typical for F delicatissima only in 1 strain, outside the major clade. Our results, supported by the analysis of the hypervariable domains of LSU (large subunit) rDNA carried out on selected strains, suggest the presence of cryptic diversity within P. delicatissima. Such diversity could in fact explain the existence of toxic and nontoxic strains within the same species and the occasional mismatches between 'species-specific' molecular probes and target species.