Metabarcoding and metabolome analyses of copepod grazing reveal feeding preference and linkage to metabolite classes in dynamic microbial plankton communities.

In order to characterize copepod feeding in relation to microbial plankton community dynamics, we combined metabarcoding and metabolome analyses during a 22-day seawater mesocosm experiment. Nutrient amendment of mesocosms promoted the development of haptophyte (Phaeocystis pouchetii)- and diatom (Skeletonema marinoi)-dominated plankton communities in mesocosms, in which Calanus sp. copepods were incubated for 24 h in flow-through chambers to allow access to prey particles (<500 µm). Copepods and mesocosm water sampled six times spanning the experiment were analysed using metabarcoding, while intracellular metabolite profiles of mesocosm plankton communities were generated for all experimental days. Taxon-specific metabarcoding ratios (ratio of consumed prey to available prey in the surrounding seawater) revealed diverse and dynamic copepod feeding selection, with positive selection on large diatoms, heterotrophic nanoflagellates and fungi, while smaller phytoplankton, including P. pouchetii, were passively consumed or even negatively selected according to our indicator. Our analysis of the relationship between Calanus grazing ratios and intracellular metabolite profiles indicates the importance of carbohydrates and lipids in plankton succession and copepod-prey interactions. This molecular characterization of Calanus sp. grazing therefore provides new evidence for selective feeding in mixed plankton assemblages and corroborates previous findings that copepod grazing may be coupled to the developmental and metabolic stage of the entire prey community rather than to individual prey abundances.

Virioplankton and virus-induced mortality of prokaryotes in the Kara Sea (Arctic) in summer.

Among the Arctic seas, the largest volume of river runoff (~45% of the total river-water inflow into the Arctic Ocean) enters the Siberian Kara Sea. The viral communities of the Kara Sea are important for the functioning of the marine ecosystem. Studies of virus-prokaryote interactions on the Kara Sea shelf have been conducted only in spring and autumn. Here, we investigated the abundance of free viruses, viruses attached to prokaryotes, and pico-sized detrital particles; the morphology (shape and size) of the viruses, viral infection and virus-mediated mortality of prokaryotes in early summer, i.e., during a seasonal ice melting period and maximum inflow of river-water volumes with high concentrations of dissolved and suspended organic carbon. Seawater samples for microbial analyses were collected across the Kara Sea shelf zone on board the Norilskiy Nickel as a research platform from June 29 to July 15, 2018. Abundances of prokaryotes (range (0.6-25.3) × 105 cells mL-1) and free viruses (range (10-117) × 105 viruses mL-1) were correlated (r = 0.63, p = 0.005) with an average virus: prokaryote ratio of 23.9 ± 5.3. The abundance of free viruses and viral-mediated mortality of prokaryotes were significantly higher in early summer than in early spring and autumn. Free viruses with a capsid diameter of 16-304 nm were recorded in the examined water samples. Waters in the Kara Sea shelf contained high concentrations of suspended organic particles 0.25-4.0 µm in size (range (0.6-25.3) × 105 particles mL-1). The proportions of free viruses, viruses attached to prokaryotes, and viruses attached to pico-sized detrital particles were 89.8 ± 6.0%, 2.2 ± 0.6% and 8.0 ± 1.3%, respectively, of the total virioplankton abundance (on average (61.5 ± 6.2) × 105 viruses mL-1). Viruses smaller than 60 nm clearly dominated at all studied sites. The majority of free viruses were not tailed. We estimated that an average of 1.4% (range 0.4-3.5%) of the prokaryote community was visibly infected by viruses, suggesting that a significant proportion of prokaryotic secondary production, 11.4% on average (range 4.0-34.0%), was lost due to viral lysis. There was a negative correlation between the abundance of pico-sized detrital particles and the frequency of visibly infected prokaryotic cells: r = -0.67, p = 0.0008.

Intense zonal freshwater transport in the Eurasian Arctic during ice-covered season revealed by in situ measurements.

The Kara Sea receives ~ 1/3 of total freshwater discharge to the Arctic Ocean, mainly from the large Ob and Yenisei rivers. The Ob-Yenisei plume covers wide area in the central part of the Kara Sea during ice-free season (June-October) and accumulates ~ 1000 km3 of freshwater volume. In late autumn, the Kara Sea becomes covered by ice, which hinders in situ measurements at this area. As a result, the fate of the Ob-Yenisei plume below sea ice during winter and spring remains unclear. In this study, we report multiple in situ measurements performed in the Kara Sea shortly before and during ice-covered season. We demonstrate that late autumn convection in the plume shortly before ice formation significantly reduces friction between the plume and the subjacent sea. The subsequent formation of solid sea ice coverage isolates the plume from wind forcing. These two factors precondition the Ob-Yenisei plume to form an intense buoyancy-driven coastal current below sea ice. As a result, the plume advects eastward to the Laptev Sea through the Vilkitsky Strait during several months in November-February. Eventually, by late winter this huge freshwater volume disappears from the Kara Sea and contributes to freshwater content of the Laptev Sea. The obtained result improves our understanding of freshwater balance of the Kara and Laptev seas, as well as provides an important insight into the large-scale freshwater transport in the Eurasian Arctic, which remain largely unknown during ice-covered season.

Metabolomics-derived marker metabolites to characterize Phaeocystis pouchetii physiology in natural plankton communities.

Phaeocystis pouchetii (Hariot) Lagerheim, 1893 regularly dominates phytoplankton blooms in higher latitudes spanning from the English Channel to the Arctic. Through zooplankton grazing and microbial activity, it is considered to be a key resource for the entire marine food web, but the actual relevance of biomass transfer to higher trophic levels is still under discussion. Cell physiology and algal nutritional state are suggested to be major factors controlling the observed variability in zooplankton grazing. However, no data have so far yielded insights into the metabolic state of Phaeocystis populations that would allow testing this hypothesis. Therefore, endometabolic markers of different growth phases were determined in laboratory batch cultures using comparative metabolomics and quantified in different phytoplankton blooms in the field. Metabolites, produced during exponential, early and late stationary growth of P. pouchetii, were profiled using gas chromatography-mass spectrometry. Then, metabolites were characterized that correlate with the growth phases using multivariate statistical analysis. Free amino acids characterized the exponential growth, whereas the early stationary phase was correlated with sugar alcohols, mono- and disaccharides. In the late stationary phase, free fatty acids, sterols and terpenes increased. These marker metabolites were then traced in Phaeocystis blooms during a cruise in the Barents Sea and North Norwegian fjords. About 50 endometabolites of P. pouchetii were detected in natural phytoplankton communities. Mannitol, scyllo-inositol, 24-methylcholesta-5,22-dien-3ß-ol, and several free fatty acids were characteristic for Phaeocystis-dominated blooms but showed variability between them. Distinct metabolic profiles were detected in the nutrient-depleted community in the inner Porsangerfjord (< 0.5 µM NO3-, < 0.1 µM PO 4 3- ), with high relative amounts of free mono- and disaccharides indicative for a limited culture. This study thereby shows how the variable physiology of phytoplankton can alter the metabolic landscape of entire plankton communities.

Dataset of phytoplankton productive parameters and environmental forces in autumn in the Kara Sea.

This data article contains data on the photosynthetic activity, shares of the main algae groups in the total phytoplankton biomass and the main environmental forces in the Kara Sea. The data were collected from 17 to 29 of September, 2011 in the following different areas: the Kara Sea Shelf with and without river runoff influence, the shelf edge and continental slope. The photosynthetic activity parameters include the relative electron transport rate (rETR) values, the maximum quantum efficiency of PSII (Fv/Fm) and the PB/rETR ratio as photosynthetic performance. The main environmental forces include the average day incident light, salinity and water temperature in the upper mixed layer and dissolved nutrients. The data presented in this article are associated with the research article entitled "Assessment of phytoplankton photosynthetic efficiency based on measurement of fluorescence parameters and radiocarbon uptake in the Kara Sea" (Mosharov et al., 2019). The related research article examines the relationships between biological parameters and with environmental characteristics such as temperature, salinity, incident photosynthetically active radiation (PAR) and nutrient concentration.

A light-induced shortcut in the planktonic microbial loop.

Mixotrophs combine photosynthesis with phagotrophy to cover their demands in energy and essential nutrients. This gives them a competitive advantage under oligotropihc conditions, where nutrients and bacteria concentrations are low. As the advantage for the mixotroph depends on light, the competition between mixo- and heterotrophic bacterivores should be regulated by light. To test this hypothesis, we incubated natural plankton from the ultra-oligotrophic Eastern Mediterranean in a set of mesocosms maintained at 4 light levels spanning a 10-fold light gradient. Picoplankton (heterotrophic bacteria (HB), pico-sized cyanobacteria, and small-sized flagellates) showed the fastest and most marked response to light, with pronounced predator-prey cycles, in the high-light treatments. Albeit cell specific activity of heterotrophic bacteria was constant across the light gradient, bacterial abundances exhibited an inverse relationship with light. This pattern was explained by light-induced top-down control of HB by bacterivorous phototrophic eukaryotes (PE), which was evidenced by a significant inverse relationship between HB net growth rate and PE abundances. Our results show that light mediates the impact of mixotrophic bacterivores. As mixo- and heterotrophs differ in the way they remineralize nutrients, these results have far-reaching implications for how nutrient cycling is affected by light.

Future climate scenarios for a coastal productive planktonic food web resulting in microplankton phenology changes and decreased trophic transfer efficiency.

We studied the effects of future climate change scenarios on plankton communities of a Norwegian fjord using a mesocosm approach. After the spring bloom, natural plankton were enclosed and treated in duplicates with inorganic nutrients elevated to pre-bloom conditions (N, P, Si; eutrophication), lowering of 0.4 pH units (acidification), and rising 3°C temperature (warming). All nutrient-amended treatments resulted in phytoplankton blooms dominated by chain-forming diatoms, and reached 13-16 µg chlorophyll (chl) a l-1. In the control mesocosms, chl a remained below 1 µg l-1. Acidification and warming had contrasting effects on the phenology and bloom-dynamics of autotrophic and heterotrophic microplankton. Bacillariophyceae, prymnesiophyceae, cryptophyta, and Protoperidinium spp. peaked earlier at higher temperature and lower pH. Chlorophyta showed lower peak abundances with acidification, but higher peak abundances with increased temperature. The peak magnitude of autotrophic dinophyceae and ciliates was, on the other hand, lowered with combined warming and acidification. Over time, the plankton communities shifted from autotrophic phytoplankton blooms to a more heterotrophic system in all mesocosms, especially in the control unaltered mesocosms. The development of mass balance and proportion of heterotrophic/autotrophic biomass predict a shift towards a more autotrophic community and less-efficient food web transfer when temperature, nutrients and acidification are combined in a future climate-change scenario. We suggest that this result may be related to a lower food quality for microzooplankton under acidification and warming scenarios and to an increase of catabolic processes compared to anabolic ones at higher temperatures.