Luka Šupraha

Haptophyte Diversity and Vertical Distribution Explored by 18S and 28S Ribosomal RNA Gene Metabarcoding and Scanning Electron Microscopy.

Haptophyta encompasses more than 300 species of mostly marine pico‐ and nanoplanktonic flagellates. Our aims were to investigate the Oslofjorden haptophyte diversity and vertical distribution by metabarcoding, and to improve the approach to study haptophyte community composition, richness and proportional abundance by comparing two rRNA markers and scanning electron microscopy (SEM). Samples were collected in August 2013 at the Outer Oslofjorden, Norway. Total RNA/cDNA was amplified by haptophyte‐specific primers targeting the V4 region of the 18S, and the D1‐D2 region of the 28S rRNA. Taxonomy was assigned using curated haptophyte reference databases and phylogenetic analyses. Both marker genes showed Chrysochromulinaceae and Prymnesiaceae to be the families with highest number of Operational Taxonomic Units (OTUs), as well as proportional abundance. The 18S rRNA data set also contained OTUs assigned to eight supported and defined clades consisting of environmental sequences only, possibly representing novel lineages from family to class. We also recorded new species for the area. Comparing coccolithophores by SEM with metabarcoding shows a good correspondence with the 18S rRNA gene proportional abundances. Our results contribute to link morphological and molecular data and 28S to 18S rRNA gene sequences of haptophytes without cultured representatives, and to improve metabarcoding methodology.

Eco-physiological adaptation shapes the response of calcifying algae to nutrient limitation

The steady increase in global ocean temperature will most likely lead to nutrient limitation in the photic zone. This will impact the physiology of marine algae, including the globally important calcifying coccolithophores. Understanding their adaptive patterns is essential for modelling carbon production in a low-nutrient ocean. We investigated the physiology of Helicosphaera carteri, a representative of the abundant but under-investigated flagellated functional group of coccolithophores. Two strains isolated from contrasting nutrient regimes (South Atlantic and Mediterranean Sea) were grown in phosphorus-replete and phosphorus-limited batch cultures. While growing exponentially in a phosphorus-replete medium, the Mediterranean strain exhibited on average 24% lower growth rate, 36% larger coccosphere volume and 21% lower particulate inorganic carbon (PIC) production than the Atlantic strain. Under phosphorus limitation, the same strain was capable of reaching a 2.6 times higher cell density than the Atlantic strain due to lower phosphorus requirements. These results suggest that local physiological adaptation can define the performance of this species under nutrient limitation.

Morphology and phylogeny of four species from the genus Bacteriastrum (Bacillariophyta)

Abstract: Four colonial species of the multipolar centric diatom genus Bacteriastrum – B. furcatum, B. hyalinum, B. jadranum and B. mediterraneum – were investigated using field samples from the Adriatic Sea and cultured strains obtained from these samples. Morphology and frustule ultrastructure were examined using light and electron microscopy, and nuclear-encoded large-subunit (LSU) rDNA sequences were obtained from the strains. The description of B. jadranum has been emended based on a novel mechanism of colony formation in Bacteriastrum in which cells are joined through an organic cell jacket which holds together cells in chains. Descriptions for two of the three other species were supplemented; vegetative cells of B. hyalinum have short Y spinules on the fused part of the setae and T-shaped outgrowths, and the spores of this species possess a ring of puncta and a granular surface on the exterior side of the primary valve mantle. Specific for B. furcatum is the location of pores scattered around the setae bases. No unique ultrastructural character was found for B. mediterraneum. Phylogenetic results grouped the sequences of the four species in a clade in which B. furcatum (section Sagittata) and B. hyalinum (section Isomorpha) formed a clade as sister to a clade with B. jadranum (section Isomorpha) and B. mediterraneum (section Sagittata). Apparently, neither section was monophyletic, and the supposed defining features of isomorphic terminal setae in Isomorpha and dimorphic ones in Sagittata were not synapomorphies. Nonetheless, the genus Bacteriastrum itself remained monophyletic.

Bacterial diversity across a highly stratified ecosystem : A salt-wedge Mediterranean estuary

Highly stratified Mediterranean estuaries are unique environments where the tidal range is low and the tidal currents are almost negligible. The main characteristics of these environments are strong salinity gradients and other environmental parameters. In this study, 454 pyrosequencing of the 16S rRNA gene in combination with catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) was used to estimate the bacterial diversity across the Krka estuary in February and July 2013. The comparison of the data derived from these two techniques resulted in a significant but weak positive correlation (R=0.28) indicating a substantial difference in the bacterial community structure, depending on the applied method. The phytoplankton bloom observed in February was identified as one of the main factors shaping the bacterial community structure between the two environmentally contrasting sampling months. Roseobacter, Bacteroidetes and Gammaproteobacteria differed substantially between February and July. Typical freshwater bacterial classes (Actinobacteria and Betaproteobacteria) showed strong vertical distribution patterns depending on the salinity gradient. Cyanobacteria decreased in abundance in February due to competition with phytoplankton, while the SAR11 clade increased its abundance in July as a result of a better adaptation toward more oligotrophic conditions. The results provided the first detailed insight into the bacterial diversity in a highly stratified Mediterranean karstic estuary.