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Fragilariopsis is a dominating psychrophilic diatom genus in polar sea ice. The two species Fragilariopsis cylindrus and Fragilariopsis curta are able to grow and divide below freezing temperature of sea water and above average sea water salinity. Here we show that antifreeze proteins (AFPs), involved in cold adaptation in several psychrophilic organisms, are widespread in the two polar species. The presence of AFP genes (afps) as a multigene family indicated the importance of this group of genes for the genus Fragilariopsis, possibly contributing to its success in sea ice. Protein phylogeny showed the potential mobility of afps, which appear to have crossed kingdom and domain borders, occurring in Bacteria, diatoms, crustaceans and fungi. Our results revealed a broad distribution of AFPs not only in polar organisms but also in taxa apparently not related to cold environments, suggesting that these proteins may be multifunctional. The relevance of AFPs to Fragilariopsis was also shown by gene expression analysis. Under stress conditions typical for sea ice, with subzero temperatures and high salinities, F. cylindrus and F. curta strongly expressed selected afps. An E/G point mutation in the Fragilariopsis AFPs may play a role in gene expression activity and protein function.

ABSTRACT:The low temperatures of polar regions and high altitude environments, especially icy habitats, present challenges for many microorganisms. Their ability to live under subfreezing conditions implies the production of compounds conferring cryotolerance. Colwellia psychrerythraea 34H, a -proteobacterium isolated from subzero Arctic marine sediments, provides a model for the study of life in cold environments. We report here the identification and detailed molecular primary and secondary structures of capsular polysaccharide from C. psychrerythraea 34H cells. The polymer was isolated in the water layer when cells were extracted by phenol/water and characterized by one-and two-dimensional NMR spectroscopy together with chemical analysis. Molecular mechanic and dynamic calculations were also performed. The polysaccharide consists of a tetrasaccharidic repeating unit containing two amino sugars and two uronic acids bearing threonine as substituent. The structural features of this unique polysaccharide resemble those present in antifreeze proteins and glycoproteins. These results suggest a possible correlation between the capsule structure and the ability of C. psychrerythraea to colonize subfreezing marine environments.

Abstract. Impurities control a variety of physical properties of polar ice. Their impact can be observed at all scalesfrom the microstructure (e.g., grain size and orientation) to the ice sheet flow behavior (e.g., borehole tilting and closure). Most impurities in ice form micrometer-sized inclusions. It has been suggested that these µ inclusions control the grain size of polycrystalline ice by pinning of grain boundaries (Zener pinning), which should be reflected in their distribution with respect to the grain boundary network. We used an optical microscope to generate high-resolution large-scale maps (3 µm pix −1 , 8×2 cm 2 ) of the distribution of micro-inclusions in four polar ice samples: two from Antarctica (EDML, MIS 5.5) and two from Greenland (NEEM, Holocene). The in situ positions of more than 5000 µ inclusions have been determined. A Raman microscope was used to confirm the extrinsic nature of a sample proportion of the mapped inclusions. A superposition of the 2-D grain boundary network and µ-inclusion distributions shows no significant correlations between grain boundaries and µ inclusions. In particular, no signs of grain boundaries harvesting µ inclusions could be found and no evidence of µ inclusions inhibiting grain boundary migration by slow-mode pinning could be detected. Consequences for our understanding of the impurity effect on ice microstructure and rheology are discussed.

Ice‐binding proteins (IBPs) control the growth and shape of ice crystals to cope with subzero temperatures in psychrophilic and freeze‐tolerant organisms. Recently, numerous proteins containing the domain of unknown function (DUF) 3494 were found to bind ice crystals and, hence, are classified as IBPs. DUF3494 IBPs constitute today the most widespread of the known IBP families. They can be found in different organisms including bacteria, yeasts and microalgae, supporting the hypothesis of horizontal transfer of its gene. Although the 3D structure is always a discontinuous β‐solenoid with a triangular cross‐section and an adjacent alpha‐helix, DUF3494 IBPs present very diverse activities in terms of the magnitude of their thermal hysteresis and inhibition of ice recrystallization. The proteins are secreted into the environments around the host cells or are anchored on their cell membranes. This review covers several aspects of this new class of IBPs, which promise to leave their mark on several research fields including structural biology, protein biochemistry and cryobiology.

Ice-binding proteins (IBPs) affect ice crystal growth by attaching to crystal faces. We present the effects on the growth of an ice single crystal caused by an ice-binding protein from the sea ice microalga (IBP) that is characterized by the widespread domain of unknown function 3494 (DUF3494) and known to cause a moderate freezing point depression (below 1 °C). By the application of interferometry, bright-field microscopy, and fluorescence microscopy, we observed that the IBP attaches to the basal faces of ice crystals, thereby inhibiting their growth in the direction and resulting in an increase in the effective supercooling with increasing IBP concentration. In addition, we observed that theIBP attaches to prism faces and inhibits their growth. In the event that the effective supercooling is small and crystals are faceted, this process causes an emergence of prism faces and suppresses crystal growth in the direction. When the effective supercooling is large and ice crystals have developed into a dendritic shape, the suppression of prism face growth results in thinner dendrite branches, and growth in the direction is accelerated due to enhanced latent heat dissipation. Our observations clearly indicate that the IBP occupies a separate position in the classification of IBPs due to the fact that it suppresses the growth of basal faces, despite its moderate freezing point depression.

The morphology and growth kinetics of ice single crystals in aqueous solutions of type III antifreeze protein (AFP-III) have been studied in detail over a range of AFP-III concentrations and supercoolings. In pure water, the shape of ice crystals changes from the circular disklike to planar dendritic with increasing supercooling. In AFP-III solutions, ice crystals in the form of faceted plates, irregular dendrites with polygonized tips, and needles appear with increasing supercooling and AFP-III concentration. The growth rate of ice crystals in the crystallographic a direction is 2 orders of magnitude higher than that in the c direction. AFP-III molecules cause the stoppage of the growth of the prismatic and basal faces at low supercoolings. When supercooling exceeds the critical value, AFP-III favors the acceleration of the growth in both a and c directions. The observed behavior of AFP-III is explained in terms of the Cabrera-Vermilyea pinning model and the specificity of the dissipation of latent heat from the growing crystals with different shapes.

We examined the influence of small-scale turbulence and its associated shear on bacterioplankton abundance and cell size. We incubated natural microbial assemblages and bacteria-only fractions and subjected them to treatments with turbulence and additions of mineral nutrients and/or organic carbon. Bacterial abundance was not affected directly by turbulence in bacteria-only incubations. In natural microbial assemblage incubations, bacterial concentrations were higher under turbulence than in still-water controls when nutrients were added. In general, in the turbulence treatments bacteria increased significantly in size, mainly due to elongation of cells. The addition of inorganic nutrients had a negative effect on bacterial size, but a significantly positive effect on abundance independently of other factors such as turbulence and the presence of predators. Flagellate grazing did not trigger an increase in bacterial size as a grazing resistance response in unmixed containers. With the addition of organic carbon, bacteria elongated and partly settled to the bottom of the containers, in both the turbulent and still treatment, but bacterial abundance did not further increase. Furthermore, bacteria aggregated in the turbulence treatments after the second day of incubation even in the absence of other components of the microbial community. We found that turbulence and the associated shear increase bacterial size and change bacterial morphology, at least under certain nutrient conditions. This might be due to a physiological response (enhanced growth rate and/or unbalanced growth) or due to the selection of opportunistic strains when organic carbon is in excess compared to mineral nutrients. We suggest that shear associated with turbulent flow enhances the DOM flux to bacteria directly as well as indirectly through enhanced grazing activity and photosynthetic release. The formation of bacterial aggregates and filaments under turbulence might give selective advantage to bacteria in terms of nutrient uptake and grazing resistance.