ALBERT

Description: Protists (microbial eukaryotes) are diverse, major components of marine ecosystems, and are fundamental to ecosystem services. In the last 10 years, molecular studies have highlighted substantial novel diversity in marine systems including sequences with no taxonomic context. At the same time, many known protists remain without a DNA identity. Since the majority of pelagic protists are too small to identify by light microscopy, most are neither comprehensively or regularly taken into account, particularly in Long-term Ecological Research Sites. This potentially undermines the quality of research and the accuracy of predictions about biological species shifts in a changing environment. The ICES Working Group for Phytoplankton and Microbial Ecology conducted a questionnaire survey in 2013–2014 on methods and identification of protists using molecular methods plus a literature review of protist molecular diversity studies. The results revealed an increased use of high-throughput sequencing methods and a recognition that sequence data enhance the overall datasets on protist species composition. However, we found only a few long-term molecular studies and noticed a lack of integration between microscopic and molecular methods. Here, we discuss and put forward recommendations to improve and make molecular methods more accessible to Long-term Ecological Research Site investigators.

Description: Phytoplankton community analysis is important with respect to natural or humaninduced changes in the marine environment. Because of the efforts involved and the limitations of traditional methods, molecular sensing approaches are becoming more popular. Among others, microarray techniques targeting ribosomal 18S sequences have been successfully applied for phytoplankton investigation. In this contribution, we compared the results of two microarray methods targeting 18S rDNA and 18S rRNA with the results obtained from microscopy, HPLC and flow cytometry. On a qualitative basis, the microarrays showed similar or potentially better performance than the non-molecular methods. Quantitatively, our data suggest that microarray signals obtained from 18S rDNA provide relatively rough estimates of phytoplankton abundance. In contrast, when targeting 18S rRNA instead, a robust linear relationship (r2 ¼ 0.68) between molecular sensing signal and microscopic cell counts could be demonstrated using a probe specific to the genus Pseudo-nitzschia as an example. Thus, for both qualitative and quantitative purposes, microarray techniques can be valuable additions to traditional methods for phytoplankton analysis. Routine monitoring approaches in particular could benefit from advantages like reduced effort, higher taxonomic resolution and a potential for automation.

Description: Knowledge about the protist diversity of the Pacific sector of the Southern Ocean is scarce. We tested the hypothesis that distinct protist community assemblages characterize large-scale water masses. Therefore, we determined the composition and biogeography of late summer protist assemblages along a transect from the coast of New Zealand to the eastern Ross Sea. We used state of the art molecular approaches, such as automated ribosomal intergenic spacer analysis and 454-pyrosequencing, combined with high-performance liquid chromatography pigment analysis to study the protist assemblage. We found distinct biogeographic patterns defined by the environmental conditions in the particular region. Different water masses harbored different microbial communities. In contrast to the Arctic Ocean, picoeukaryotes had minor importance throughout the investigated transect and showed very low contribution south of the Polar Front. Dinoflagellates, Syndiniales, and small stramenopiles were dominating the sequence assemblage in the Subantarctic Zone, whereas the relative abundance of diatoms increased southwards, in the Polar Frontal Zone and Antarctic Zone. South of the Polar Front, most sequences belonged to haptophytes. This study delivers a comprehensive and taxon detailed overview of the protist composition in the investigated area during the austral summer 2010.

Description: In the Arctic Ocean, sea-ice decline will significantly change the structure of biological communities. At the same time, changing nutrient dynamics can have similarly strong and potentially interacting effects. To investigate the response of the taxonomic and trophic structure of planktonic and ice-associated communities to varying sea-ice properties and nutrient concentrations, we analysed four different communities sampled in the Eurasian Basin in summer 2012: (1) protists and (2) metazoans from the under-ice habitat, and (3) protists and (4) metazoans from the epipelagic habitat. The taxonomic composition of protist communities was characterised with 18S meta-barcoding. The taxonomic composition of metazoan communities was determined based on morphology. The analysis of environmental parameters identified (i) a ‘shelf-influenced’ regime with melting sea ice, high-silicate concentrations and low NOx (nitrate + nitrite) concentrations; (ii) a ‘Polar’ regime with low silicate concentrations and low NOx concentrations; and (iii) an ‘Atlantic’ regime with low silicate concentrations and high NOx concentrations. Multivariate analyses of combined bio-environmental datasets showed that taxonomic community structure primarily responded to the variability of sea-ice properties and hydrography across all four communities. Trophic community structure, however, responded significantly to NOx concentrations. In three of the four communities, the most heterotrophic trophic group significantly dominated in the NOx-poor shelf-influenced and Polar regimes compared to the NOx-rich Atlantic regime. The more heterotrophic, NOx-poor regimes were associated with lower productivity and carbon export than the NOx-rich Atlantic regime. For modelling future Arctic ecosystems, it is important to consider that taxonomic diversity can respond to different drivers than trophic diversity.