Eveline Pinseel

Molecular and morphological characterization of the Achnanthidium minutissimum complex (Bacillariophyta) in Petuniabukta (Spitsbergen, High Arctic) including the description of A. digitatum sp. nov.

ABSTRACT Recent morphology-based investigations of freshwater Arctic diatoms suggest that many species remain to be discovered, reflecting a unique polar flora. During a survey of the freshwater diatom flora of northern Billefjorden, including the Petuniabukta fjord region (Spitsbergen), several morphodemes belonging to the Achnanthidium minutissimum species complex were recorded. Molecular phylogenies based on rbcL, 28S and 18S sequences, including single cells from Canada and strains from Marion Island (sub-Antarctica) and GenBank revealed the presence of 12 distinct A. minutissimum complex lineages, of which three contained strains from Spitsbergen. One Arctic lineage is described as a new species. Achnanthidium digitatum sp. nov. is morphologically characterized by narrow, linear to only slightly lanceolate valves and usually two areolae per stria. The two remaining Arctic lineages are in need of a more complete morphological and molecular comparison with other representatives of the A. minutissimum complex to clarify their taxonomic identity. It is argued that implementation of molecular data in the taxonomy of Achnanthidium will be essential to solve the taxonomic problems associated with this group, eventually resulting in a better understanding of the biogeography and niche differentiation of different species belonging to the A. minutissimum complex. Ideally, this should be based on more variable genes than the currently widely used 18S, which does not have a species level resolution in the A. minutissimum complex.

Pinnularia catenaborealis sp. nov. (Bacillariophyceae), a unique chain-forming diatom species from James Ross Island and Vega Island (Maritime Antarctica)

Abstract: A recent detailed survey of the Maritime Antarctic diatom flora using a fine-grained taxonomy resulted in the description of many new species of Pinnularia in general and the section Distantes, including the P. borealis species complex, in particular. Moreover, DNA-based studies of P. borealis revealed that many more species need to be described within this complex. During a survey of the freshwater littoral diatom flora of James Ross Island (Ulu Peninsula) and Vega Island in Maritime Antarctica, a previously unknown chain-forming species in the P. borealis species complex of section Distantes was cultured from three different localities. Molecular phylogenies based on the nuclear-encoded D1–D3 large-subunit ribosomal DNA and plastid rbcL genes revealed that all cultures belong to a distinct highly supported lineage within the P. borealis species complex. Pinnularia catenaborealis sp. nov. is characterised by the presence of small spines located on a raised, thin silica ridge that almost entirely surrounds the valve face near the valve face/mantle junction, and the presence of small silica plates near the apices. In culture, P. catenaborealis formed chains of several tens of cells and in oxidised natural material, chains up to seven frustules were observed. Pinnularia catenaborealis is described from the littoral zone of freshwater Black Lake (Ulu Peninsula, James Ross Island) and has also been observed on nearby Vega Island. Although P. borealis is generally regarded as a (semi-)terrestrial diatom complex mainly occurring in (moist) soils and mosses, P. catenaborealis was found in freshwater habitats with an alkaline pH and low conductivity.

Diversity, ecology and community structure of the freshwater littoral diatom flora from Petuniabukta (Spitsbergen)

Despite the frequent application of diatoms in palaeoecological research in the High Arctic, our knowledge concerning the diversity, ecology, community associations, community dynamics and survival strategies of High Arctic freshwater littoral diatom floras is still rather limited. In the present study, the diversity, ecological characteristics and community structures of the freshwater littoral diatom flora from ponds and lakes of Petuniabukta (Spitsbergen, Svalbard Archipelago) were studied. In total, 87 samples from the littoral zone of 53 freshwater ponds and lakes were examined using light and scanning electron microscopy. A highly diverse diatom flora dominated by Achnanthidium and Nitzschia was observed. In total, 310 diatom taxa belonging to 59 genera were found, of which about one-third could not be identified beyond the genus level. Based on species composition, four main diatom communities were distinguished. Differences in environmental characteristics, such as habitat type, habitat diversity, habitat stability, streams, water pH and influence of the fjord, biota and human activities, appeared to be important factors in determining the observed diatom communities. The high number of unidentified taxa in this study indicates that a profound revision of the Arctic diatom flora is highly desired, not only to improve our fundamental knowledge concerning the diversity, ecology, community associations and biogeography of Arctic diatoms but also to aid applied (palaeo)environmental sciences.

Diatoms in cryoconite holes and adjacent proglacial freshwater sediments, Nordenskiöld glacier (Spitsbergen, High Arctic)

Cryoconite holes are small, extreme habitats, widespread in the ablation zones of gla-ciers worldwide. They can provide a suitable environment for microorganisms including bacteria, cyanobacteria, algae, fungi, and invertebrates. Diatoms have been previously recovered from cryoconite holes of Greenland and of Svalbard, and recent findings from Antarctica suggest that cryoconite holes may harbor a unique diatom flora distinct from other aquatic habitats nearby. In the present study, we characterize the diatom communi-ties of Nordenskiold glacier cryoconite holes in Billefjorden (Svalbard, Spitsbergen), and multivariate approaches were used to compare them with three freshwater localities in the immediate vicinity to investigate possible sources of the species pool. We found cryoconite holes to have similar or greater average genus-richness than adjacent lake/ ponds habitats, even though lower numbers of valves were recovered. Overall, cryoconite hole diatom communities differed significantly from those observed in lakes, suggesting that other sources actively contribute to these communities than nearby lakes alone. This further suggests that (i) diatoms present in cryoconite might not exclusively originate from aquatic habitats, but also from (semi-) terrestrial ones; and (ii) that a much wider area than the immediate surroundings should be considered as a possible source for cryoconite diatom flora .

Incomplete Reproductive Isolation Between Genetically Distinct Sympatric Clades of the Pennate Model Diatom Seminavis robusta

Incomplete reproductive isolation between genetically distinct taxa provides an interesting opportunity for speciation and adaptation studies. This phenomenon is well-described in macro-organisms, but less experimental evidence is available for unicellular eukaryotes. Here, we document the sympatric occurrence of genetically differentiated populations of the pennate model diatom Seminavis robusta in coastal subtidal biofilm communities and show widespread potential for gene flow between them. Based on sequence variation in the plastid-encoded rbcL gene, three distinct clades were identified. Morphological variation between the clades reflected their phylogenetic relationships, with subtle differences in valve morphology in the most distant clade compared to the other two clades, which were indistinguishable. Using a large number of experimental crosses we showed that, although reproductive output was significantly lower compared to the majority of within-clade crosses, approximately 34.5% of the inter-clade crosses resulted in viable and fertile progeny. While the nature of the incomplete reproductive isolation remains unknown, its occurrence in natural diatom populations represents an additional mechanism contributing to population genetic structuring and adaptation and can spur further research into the mechanisms of species divergence and the maintenance of species identity in the presence of gene flow.

Expanding the toolbox for cryopreservation of marine and freshwater diatoms

Diatoms constitute the most diverse group of microalgae and have long been recognised for their large biotechnological potential. In the wake of growing research interest in new model species and development of commercial applications, there is a pressing need for long-term preservation of diatom strains. While cryopreservation using dimethylsulfoxide (DMSO) as a cryoprotective agent is the preferred method for long-term strain preservation, many diatom species cannot be successfully cryopreserved using DMSO. Therefore, in this study, we studied cryopreservation success in six different diatom species, representing the major morphological and ecological diatom groups, using a range of DMSO concentrations and Plant Vitrification Solution 2 (PVS2) as an alternative cryoprotectant to DMSO. In addition, we tested whether suppressing bacterial growth by antibiotics accelerates the post-thaw recovery process. Our results show that the effects of cryoprotectant choice, its concentration and the addition of antibiotics are highly species specific. In addition, we showed that PVS2 and antibiotics are useful agents to optimize cryopreservation of algae that cannot survive the traditional cryopreservation protocol using DMSO. We conclude that a species-specific approach will remain necessary to develop protocols for diatom cryopreservation and to increase their representation in public culture collections.