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Quantification of grazing losses of marine heterotrophic bacteria is critical for understanding nutrient and carbon pathways in aquatic systems. The dilution method is a commonly used experimental approach for quantifying bacterivory. However, valid estimates of grazing rates obtained using this method depend on several methodological assumptions including that the method does not influence specific growth rates of bacteria. Here, we hypothesize that filtration during the set-up of a dilution experiment has the potential to release allelochemicals from phytoplankton cells and thereby stimulate or inhibit bacterial growth with the consequence of biased grazing estimates. We tested this hypothesis during a natural Phaeocystis pouchetii bloom at two different locations within an Arctic fjord. Results from the dilution experiments suggest higher gross growth rate and grazing impact for bacteria in the outer fjord compared with the inner fjord. However, specific growth rates estimated by bacterial production cell−1 were significantly elevated in dilutions of water from the outer fjord but not the inner fjord. The analysis of dissolved metabolites in the seawater from both experiments prior and after filtration revealed altered metabolic profiles after filtration at both stations. As unaffected specific growth of prey on dilution is one of three fundamental assumptions of the dilution method, we conclude that it is important that empirically estimated bacterial specific growth rates be routinely included when using the dilution method to quantify bacterivory. [ABSTRACT FROM AUTHOR]

Microalgae in the division Haptophyta play key roles in the marine ecosystem and in global biogeochemical processes. Despite their ecological importance, knowledge on seasonal dynamics, community composition and abundance at the species level is limited due to their small cell size and few morphological features visible under the light microscope. Here, we present unique data on haptophyte seasonal diversity and dynamics from two annual cycles, with the taxonomic resolution and sampling depth obtained with high-throughput sequencing. From outer Oslofjorden, S Norway, nano- and picoplanktonic samples were collected monthly for 2 years, and the haptophytes targeted by amplification of RNA/ cDNA with Haptophyta-specific 18S rDNA V4 primers. We obtained 156 operational taxonomic units (OTUs), from c. 400.000 454 pyrosequencing reads, after rigorous bioinformatic filtering and clustering at 99.5%. Most OTUs represented uncultured and/or not yet 18S rDNA-sequenced species. Haptophyte OTU richness and community composition exhibited high temporal variation and significant yearly periodicity. Richness was highest in September-October (autumn) and lowest in April-May (spring). Some taxa were detected all year, such as Chrysochromulina simplex, Emiliania huxleyi and Phaeocystis cordata, whereas most calcifying coccolithophores only appeared from summer to early winter. We also revealed the seasonal dynamics of OTUs representing putative novel classes (clades HAP-3-5) or orders (clades D, E, F). Season, light and temperature accounted for 29% of the variation in OTU composition. Residual variation may be related to biotic factors, such as competition and viral infection. This study provides new, in-depth knowledge on seasonal diversity and dynamics of haptophytes in North Atlantic coastal waters. [ABSTRACT FROM AUTHOR]

Cells maintain an osmotic pressure essential for growth and division, using organic compatible solutes and inorganic ions. Mg2+, which is the most abundant divalent cation in living cells, has not been considered an osmotically important solute. Here we show that under carbon limitation or dormancy native marine bacterial communities have a high cellular concentration of Mg2+ (370-940 mM) and a low cellular concentration of Na+ (50-170 mM). With input of organic carbon, the average cellular concentration of Mg2+ decreased 6-12-fold, whereas that of Na+ increased ca 3-4-fold. The concentration of chlorine, which was in the range of 330-1200 mM, and was the only inorganic counterion of quantitative significance, balanced and followed changes in the concentration of Mg2++Na+. In an osmotically stable environment, like seawater, any major shift in bacterial osmolyte composition should be related to shifts in growth conditions, and replacing organic compatible solutes with inorganic solutes is presumably a favorable strategy when growing in carbon-limited condition. A high concentration of Mg2+ in cells may also serve to protect and stabilize macromolecules during periods of non-growth and dormancy. Our results suggest that Mg2+ has a major role as osmolyte in marine bacteria, and that the [Mg2+]/[Na+] ratio is related to its physiological condition and nutritional status. Bacterial degradation is a main sink for dissolved organic carbon in the ocean, and understanding the mechanisms limiting bacterial activity is therefore essential for understanding the oceanic C-cycle. The [Mg2+]/[Na+]-ratio in cells may provide a physiological proxy for the transitions between C-limited and mineral nutrient-limited bacterial growth in the ocean's surface layer. [ABSTRACT FROM AUTHOR]

The isolation and characterization of a virus (designated EhV) that infects the marine coccolithophorid Emiliania huxleyi (Lohmann) Hay & Mohler are described. Three independent clones of EhV were isolated from Norwegian coastal waters in years 1999 and 2000. EhV is a double-stranded DNA-containing virus with a genome size of ∼415 kilo-base pairs. The viral particle is an icosahedron with a diameter of 160–180 nm. The virus particle contains at least nine proteins ranging from 10 to 140 kDa; the major capsid protein weighs ∼54 kDa. EhV has a latent period of 12–14 h and a burst size of 400–1000 (mean, 620) viral particles per cell. A phylogenetic tree based on DNA polymerase amino acid sequences indicates EhV should be assigned to the Phycodnaviridae virus family and that the virus is most closely related to viruses that infect Micromonas pusilla and certain Chlorella species. [ABSTRACT FROM AUTHOR]

In this study we investigated virus production in two marine phytoplankton species and how it relates to the host's cell cycle. Phaeocystis pouchetii (Hariot) Lagerheim and Pyramimonas orientalis McFadden, Hill & Wetherby, growing synchronously in batch cultures, were infected with their respective viruses (PpV and PoV) at four different stages in the cell cycle and the production of free virus was then measured for 30 h. The virus production in P. orientalis infected with PoV depended on the time of infection, whereas no such relation was found for P. pouchetii infected with PpV. The P. orientalis cultures infected at the end of the dark period and at the beginning of the light period produced three times more virus than those infected in the middle of the light period and eight times more virus than those infected at the beginning of the dark period. The latent periods for PpV and PoV were 12–14 h and 18–20 h, respectively, and in both cases were independent of the host cell cycle. The differences in virus production may be attributed to light or cell cycle dependent regulation of host infection, metabolism, or burst size. Regardless of the mechanism, these differences may be related to differences in the ecological strategies of the hosts and their ability to form blooms. [ABSTRACT FROM AUTHOR]

In this study we investigated virus production in two marine phytoplankton species and how it relates to the host's cell cycle. Phaeocystis pouchetii (Hariot) Lagerheim and Pyramimonas orientalis McFadden, Hill & Wetherby, growing synchronously in batch cultures, were infected with their respective viruses (PpV and PoV) at four different stages in the cell cycle and the production of free virus was then measured for 30 h. The virus production in P. orientalis infected with PoV depended on the time of infection, whereas no such relation was found for P. pouchetii infected with PpV. The P. orientalis cultures infected at the end of the dark period and at the beginning of the light period produced three times more virus than those infected in the middle of the light period and eight times more virus than those infected at the beginning of the dark period. The latent periods for PpV and PoV were 12–14 h and 18–20 h, respectively, and in both cases were independent of the host cell cycle. The differences in virus production may be attributed to light or cell cycle dependent regulation of host infection, metabolism, or burst size. Regardless of the mechanism, these differences may be related to differences in the ecological strategies of the hosts and their ability to form blooms. [ABSTRACT FROM AUTHOR]

The haptophytes Prymnesium parvum Carter and Prymnesium patelliferum Green, Hibberd, and Pienaar are two closely related species, which can only be distinguished by minor differences in the morphology of their organic body scales. The two Prymnesium species are reported to coexist at several locations, including the Sands fjord system in southwestern Norway. Comparisons of physiology and toxicity within the two species have failed to reveal differences that can add to the small morphological distributions used to separate them. To investigate the genetic relationship between the two species, we compared the sequence of the first internal transcribed spacer region (ITSI) and length variation in one intron separating calmodulin genes for four P. parvum strains and eight P. patelliferum strains. Both the ITSI sequence and the banding patterns obtained by PCR amplification of one intron in the calmodulin genes indicated that the Prymnesium isolates are related by their geographic origin instead of by their species affiliation. The results indicate that P. parvum and P. patelliferum are so closely related that they could be considered one species. Alternatively, we discuss the possibility that the two species might be joined in a heleionorphic haploid-diploid life cycle, as is now widely reported for other haptophycean algae. [ABSTRACT FROM AUTHOR]

The isolation and characterization of a virus (designated EhV) that infects the marine coccolithophorid Emiliania huxleyi (Lohmann) Hay & Mohler are described. Three independent clones of EhV were isolated from Norwegian coastal waters in years 1999 and 2000. EhV is a double-stranded DNA-containing virus with a genome size of ∼415 kilo-base pairs. The viral particle is an icosahedron with a diameter of 160–180 nm. The virus particle contains at least nine proteins ranging from 10 to 140 kDa; the major capsid protein weighs ∼54 kDa. EhV has a latent period of 12–14 h and a burst size of 400–1000 (mean, 620) viral particles per cell. A phylogenetic tree based on DNA polymerase amino acid sequences indicates EhV should be assigned to the Phycodnaviridae virus family and that the virus is most closely related to viruses that infect Micromonas pusilla and certain Chlorella species. [ABSTRACT FROM AUTHOR]

Ocean acidification (OA), a consequence of anthropogenic carbon dioxide (CO 2 ) emissions, strongly impacts marine ecosystems. OA also influences iron (Fe) solubility, affecting biogeochemical and ecological processes. We investigated the interactive effects of CO 2 and Fe availability on the metabolome response of a natural phytoplankton community. Using mesocosms we exposed phytoplankton to ambient (390 μatm) or future CO 2 levels predicted for the year 2100 (900 μatm), combined with ambient (4.5 nM) or high (12 nM) dissolved iron (dFe). By integrating over the whole phytoplankton community, we assigned functional changes based on altered metabolite concentrations. Our study revealed the complexity of phytoplankton metabolism. Metabolic profiles showed three stages in response to treatments and phytoplankton dynamics. Metabolome changes were related to the plankton group contributing respective metabolites, explaining bloom decline and community succession. CO 2 and Fe affected metabolic profiles. Most saccharides, fatty acids, amino acids and many sterols significantly correlated with the high dFe treatment at ambient pCO 2 . High CO 2 lowered the abundance of many metabolites irrespective of Fe. However, sugar alcohols accumulated, indicating potential stress. We demonstrate that not only altered species composition but also changes in the metabolic landscape affecting the plankton community may change as a consequence of future high-CO 2 oceans., (© 2020 Society for Applied Microbiology and John Wiley & Sons Ltd.)

The Arctic marine environment experiences dramatic seasonal changes in light and nutrient availability. To investigate the influence of seasonality on Arctic marine virus communities, five research cruises to the west and north of Svalbard were conducted across one calendar year, collecting water from the surface to 1000 m in depth. We employed metabarcoding analysis of major capsid protein g23 and mcp genes in order to investigate T4-like myoviruses and large dsDNA viruses infecting prokaryotic and eukaryotic picophytoplankton, respectively. Microbial abundances were assessed using flow cytometry. Metabarcoding results demonstrated that seasonality was the key mediator shaping virus communities, whereas depth exerted a diversifying effect within seasonal virus assemblages. Viral diversity and virus-to-prokaryote ratios (VPRs) dropped sharply at the commencement of the spring bloom but increased across the season, ultimately achieving the highest levels during the winter season. These findings suggest that viral lysis may be an important process during the polar winter, when productivity is low. Furthermore, winter viral communities consisted of Operational Taxonomic Units (OTUs) distinct from those present during the spring-summer season. Our data provided a first insight into the diversity of viruses in a hitherto undescribed marine habitat characterized by extremes in light and productivity.

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Cells maintain an osmotic pressure essential for growth and division, using organic compatible solutes and inorganic ions. Mg(2+), which is the most abundant divalent cation in living cells, has not been considered an osmotically important solute. Here we show that under carbon limitation or dormancy native marine bacterial communities have a high cellular concentration of Mg(2+) (370-940 mM) and a low cellular concentration of Na(+) (50-170 mM). With input of organic carbon, the average cellular concentration of Mg(2+) decreased 6-12-fold, whereas that of Na(+) increased ca 3-4-fold. The concentration of chlorine, which was in the range of 330-1200 mM, and was the only inorganic counterion of quantitative significance, balanced and followed changes in the concentration of Mg(2+)+Na(+). In an osmotically stable environment, like seawater, any major shift in bacterial osmolyte composition should be related to shifts in growth conditions, and replacing organic compatible solutes with inorganic solutes is presumably a favorable strategy when growing in carbon-limited condition. A high concentration of Mg(2+) in cells may also serve to protect and stabilize macromolecules during periods of non-growth and dormancy. Our results suggest that Mg(2+) has a major role as osmolyte in marine bacteria, and that the [Mg(2+)]/[Na(+)] ratio is related to its physiological condition and nutritional status. Bacterial degradation is a main sink for dissolved organic carbon in the ocean, and understanding the mechanisms limiting bacterial activity is therefore essential for understanding the oceanic C-cycle. The [Mg(2+)]/[Na(+)]-ratio in cells may provide a physiological proxy for the transitions between C-limited and mineral nutrient-limited bacterial growth in the ocean's surface layer.