Identification of diatom isolates from the Gulf of Gdansk: testing of species identifications using morphology, 18S rDNA sequencing and DNA barcodes of strains from the Culture Collection of Baltic Algae (CCBA)

Eighteen strains of diatoms isolated from different phytoplankton and microphytobenthic assemblages in the Gulf of Gdansk (Baltic Sea, Poland) and maintained at the Culture Collection of Baltic Algae (CCBA) were investigated for their species identifications. The latter were tested by phylogenetic analyses of nearly full 18S rDNA sequences as well as through sequence comparisons of 5.8S+ITS2 rDNA fragments for use as DNA barcode. The planktonic species were readily identified as a member of the Cyclotella meneghiniana species complex and Skeletonema marinoi, respectively, by both 18S phylogenics and DNA barcoding. In contrast, for the pennate diatom strains the suitability of DNA sequence comparisons for species identification appeared to be still rather limited. Only one strain could be identified to the species level (Navicula gregaria) using DNA barcodes, and no closest relative barcode sequences were available for the other strains. The 18S rDNA phylogenies supported species identification for only one strain (Bacillaria cf. paxillifera). In all other pennate strains identification was only possible to the genus level. Because for most species identified by morphology no closest neighbouring 18S rDNA sequences were available, the CCBA strains may serve as references to represent certain species which have been well characterized by morphology, 18S rDNA sequence analyses and DNA barcodes. Relatively high 18S rDNA differences between pairs of strains of Fistulifera saprophila and Navicula perminuta indicated that each species may represent a complex of several cryptic species. In contrast, the five isolates of Nitzschia microcephala had identical sequences as well as the two isolates of Nitzschia cf. fonticola. With the addition of new species to the 18S rDNA phylogeny of pennate raphid diatoms, the monophyly of the genera Fistulifera, Haslea and Navicula s. str. were confirmed, while Nitzschia appeared paraphyletic. The traditional family Naviculaceae was paraphyletic, in agreement with previous phylogenetic studies.