Characterization of Ice-Binding Proteins from Sea Ice Algae

ICE BINDING PROTEINS FROM SEA ICE ALGAE Sea ice is mainly a two-phase system, and its porous structure is largely determinant for biological activity within ice. During ice formation, solutes in the seawater are excluded from the ice matrix and segregate into brine droplets or brine channels, generally defined as brine inclusions inside sea ice. Outflow of high salinity brine and inflow of seawater of lower salinity, as well as further cooling, cause brine inclusions to narrow and eventually separate into individual pockets divided by ice bridges. Despite the harsh conditions that govern within sea ice, where temperatures range from about -1.8°C on the bottom to -20°C or less on the top, and brine salinities can be as high as 200 on the Practical Salinity Scale, brine inclusions offer a habitat for a variety of microalgae. These algae play a crucial role for the ecology of the Polar Oceans, since they represent a concentrated food source in the low-productivity ice-covered sea, and in the months of melting they initiate blooms by seeding the water column. Algae have been found distributed within brine inclusions throughout the entire thickness of the ice column. The strategies adopted by ice microorganisms to cope with conditions in sea ice remain to be unraveled. Recent studies showed that several organisms that populate sea ice, spreading from bacteria to diatoms and a crustacean species, have ice binding proteins (IBPs). These proteins are common in polar species, but lack in temperate organisms, suggesting that IBPs play a key role in adaptation to subzero conditions. The nomenclature of these proteins varies, depending on authors, from ice binding to antifreeze or ice structuring. In the generally accepted adsorption–inhibition model describing the mechanism of action of IBPs, proteins bind to the ice lattice and locally inhibit ice growth by the Gibbs-Thomson effect. Recent publications showed that some IBPs organize water molecules into an ice-like structure that matches defined planes of the ice crystal and is then gradually frozen to the ice lattice. One of the most prominent and best described effects of IBPs is thermal hysteresis, which describes the lowering of the freezing point of a solution below the melting point. Another effect which defines IBPs is inhibition of recrystallization, which is the grain boundary migration resulting in a growth of larger crystals at the expenses of small grains. The biological role of IBPs from sea ice microalgae remains an open question. The importance of some IBP families, as observed in fishes or insects, lies in lowering the freezing point below environmental temperature, in order to avoid ice formation in cells or organs. Other IBPs have the function to inhibit recrystallization, as it has been suggested for plant IBPs. In the context of sea ice, it seems unlikely that the biological role of IBPs may be thermal hysteresis (measured in the order of 1°C) or recrystallization inhibition. Most of the IBPs from sea ice algae are active extracellularly. It has been suggested that they are trapped and accumulate within a layer of extracellular polysaccharide substances (EPS) secreted by several sea ice organisms. Microalgal IBPs produced recombinantly or collected from spent growth medium affect the structure of ice surface, causing pitting and characteristic microstructural features. This suggests that the proteins shape their frozen environment in order to increase their habitable space within sea ice. However, the characterization of IBPs is of relevance not only to understand their functional role in sea ice, but also in the frame of possible applications of IBPs in the medical field, in the food industry and in other fields related to a control of ice crystals. In the following we present some standard techniques to determine the protein activity in terms of thermal hysteresis (TH) and recrystallization inhibition (RI), which define the proteins as ice binding. Also, we present further methods (ice pitting assay, determination of the nucleating temperature) to characterize the activity of IBPs

Biogenic silica recycling in sea ice inferred from Si-isotopes: constraints from Arctic winter first-year sea ice

We report silicon isotopic composition (d30Si vs. NBS28) in Arctic sea ice, based on sampling of silicic acid from both brine and seawater in a small Greenlandic bay in March 2010. Our measurements show that just before the productive period, d30Si of sea-ice brine similar to d30Si of the underlying seawater. Hence, there is no Si isotopic fractionation during sea-ice growth by physical processes such as brine convection. This finding brings credit and support to the conclusions of previous work on the impact of biogenic processes on sea ice d30Si: any d30Si change results from a combination of biogenic silica production and dissolution. We use this insight to interpret data from an earlier study of sea-ice d30Si in Antarctic pack ice that show a large accumulation of biogenic silica. Based on these data, we estimate a significant contribution of biogenic silica dissolution (D) to production (P), with a D:P ratio between 0.4 and 0.9. This finding has significant implications for the understanding and parameterization of the sea ice Sibiogeochemical cycle, i.e. previous studies assumed little or no biogenic silica dissolution in sea ice.BELCANT

Effect of melting Antarctic sea ice on the fate of microbial communities studied in microcosms

Although algal growth in the iron-deficient Southern Ocean surface waters is generally low, there is considerable evidence that winter sea ice contains high amounts of iron and organic matter leading to ice-edge blooms during austral spring. We used field observations and ship-based microcosm experiments to study the effect of the seeding by sea ice microorganisms, and the fertilization by organic matter and iron on the planktonic community at the onset of spring/summer in the Weddell Sea. Pack ice was a major source of autotrophs resulting in a ninefold to 27-fold increase in the sea ice-fertilized seawater microcosm compared to the ice-free seawater microcosm. However, heterotrophs were released in lower numbers (only a 2- to 6-fold increase). Pack ice was also an important source of dissolved organic matter for the planktonic community. Small algae (<10 μm) and bacteria released from melting sea ice were able to thrive in seawater. Field observations show that the supply of iron from melting sea ice had occurred well before our arrival onsite, and the supply of iron to the microcosms was therefore low. We finally ran a “sequential melting” experiment to monitor the release of ice constituents in seawater. Brine drainage occurred first and was associated with the release of dissolved elements (salts, dissolved organic carbon and dissolved iron). Particulate organic carbon and particulate iron were released with low-salinity waters at a later stage