Clustering of ice nucleation protein correlates with ice nucleation activity

Antibodies raised against a synthetic peptide specifically detect ice nucleation proteins from Pseudomonas species in Western blots. In immunofluorescent staining of whole bacteria, the antibodies reveal the protein in clusters, as indicated by patches of intense fluorescence in Escherichia coli cells heterologously expressing Pseudomonas ice nucleation genes. The abundance, size, and brightness of the clusters vary considerably from cell to cell. Their varying sizes may explain the variability in activity of bacterial ice nuclei. Growth at lower temperatures produces more ice nuclei, and gives brighter and more frequent patches, than growth at 37 degrees C. The observed clustering may thus reflect formation of functional ice nucleation sites in vivo. The presence of ice nucleation protein in clusters is also correlated with alterations in cell morphology.     

Intercellular ice propagation: experimental evidence for ice growth through membrane pores

Propagation of intracellular ice between cells significantly increases the prevalence of intracellular ice in confluent monolayers and tissues. It has been proposed that gap junctions facilitate ice propagation between cells. This study develops an equation for capillary freezing-point depression to determine the effect of temperature on the equilibrium radius of an ice crystal sufficiently small to grow through gap junctions. Convection cryomicroscopy and video image analysis were used to examine the incidence and pattern of intracellular ice formation (IIF) in the confluent monolayers of cell lines that do (MDCK) and do not (V-79W) form gap junctions. The effect of gap junctions on intracellular ice propagation was strongly temperature-dependent. For cells with gap junctions, IIF occurred in a directed wave-like pattern in 100% of the cells below -3 degrees C. At temperatures above -3 degrees C, there was a marked drop in the incidence of IIF, with isolated individual cells initially freezing randomly throughout the sample. This random pattern of IIF was also observed in the V-79W monolayers and in MDCK monolayers treated to prevent gap junction formation. The significant change in the low temperature behavior of confluent MDCK monolayers at -3 degrees C is likely the result of the inhibition of gap junction-facilitated ice propagation, and supports the theory that gap junctions facilitate ice nucleation between cells.     

Ecology under lake ice

Winter conditions are rapidly changing in temperate ecosystems, particularly for those that experience periods of snow and ice cover. Relatively little is known of winter ecology in these systems, due to a historical research focus on summer ‘growing seasons’. We executed the first global quantitative synthesis on under‐ice lake ecology, including 36 abiotic and biotic variables from 42 research groups and 101 lakes, examining seasonal differences and connections as well as how seasonal differences vary with geophysical factors. Plankton were more abundant under ice than expected; mean winter values were 43.2% of summer values for chlorophyll a, 15.8% of summer phytoplankton biovolume and 25.3% of summer zooplankton density. Dissolved nitrogen concentrations were typically higher during winter, and these differences were exaggerated in smaller lakes. Lake size also influenced winter‐summer patterns for dissolved organic carbon (DOC), with higher winter DOC in smaller lakes. At coarse levels of taxonomic aggregation, phytoplankton and zooplankton community composition showed few systematic differences between seasons, although literature suggests that seasonal differences are frequently lake‐specific, species‐specific, or occur at the level of functional group. Within the subset of lakes that had longer time series, winter influenced the subsequent summer for some nutrient variables and zooplankton biomass.     

Protein stability in ice

This study presents an experimental approach, based on the change of Trp fluorescence between native and denatured states of proteins, which permits to monitor unfolding equilibria and the thermodynamic stability (DeltaG degrees ) of these macromolecules in frozen aqueous solutions. The results obtained by guanidinium chloride denaturation of the azurin mutant C112S from Pseudomonas aeruginosa, in the temperature range from -8 to -16 degrees C, demonstrate that the stability of the native fold may be significantly perturbed in ice depending mainly on the size of the liquid water pool (V(L)) in equilibrium with the solid phase. The data establish a threshold, around V(L)=1.5%, below which in ice DeltaG degrees decreases progressively relative to liquid state, up to 3 kcal/mole for V(L)=0.285%. The sharp dependence of DeltaG degrees on V(L) is consistent with a mechanism based on adsorption of the protein to the ice surface. The reduction in DeltaG degrees is accompanied by a corresponding decrease in m-value indicating that protein-ice interactions increase the solvent accessible surface area of the native fold or reduce that of the denatured state, or both. The method opens the possibility for examining in a more quantitative fashion the influence of various experimental conditions on the ice perturbation and in particular to test the effectiveness of numerous additives used in formulations to preserve labile pharmaco proteins.     

Organism losses during ice melting: A serious bias in sea ice community studies

Summary When ice samples are melted, microorganisms living within the brine inclusions are subjected to rapid and extreme changes in salinities. This procedure results in substantial losses of flagellates and ciliates. Most of these losses can be prevented if ice samples are melted in larger volumes of sterile sea water to buffer salinity and osmotic changes. Since most studies on the ice biota have ignored, or have been unable to avoid this bias, current views of the composition and activity of sea ice communities are based on assemblages over-representing organisms with rigid cell material.     

Biogeochemical conditions and ice algal photosynthetic parameters in Weddell Sea ice during early spring

Physical, biogeochemical and photosynthetic parameters were measured in sea ice brine and ice core bottom samples in the north-western Weddell Sea during early spring 2006. Sea ice brines collected from sackholes were characterised by cold temperatures (range −7.4 to −3.8°C), high salinities (range 61.4–118.0), and partly elevated dissolved oxygen concentrations (range 159–413 μmol kg⁻¹) when compared to surface seawater. Nitrate (range 0.5–76.3 μmol kg⁻¹), dissolved inorganic phosphate (range 0.2–7.0 μmol kg⁻¹) and silicic acid (range 74–285 μmol kg⁻¹) concentrations in sea ice brines were depleted when compared to surface seawater. In contrast, NH₄ ⁺ (range 0.3–23.0 μmol kg⁻¹) and dissolved organic carbon (range 140–707 μmol kg⁻¹) were enriched in the sea ice brines. Ice core bottom samples exhibited moderate temperatures and brine salinities, but high algal biomass (4.9–435.5 μg Chl a l⁻¹ brine) and silicic acid depletion. Pulse amplitude modulated fluorometry was used for the determination of the photosynthetic parameters F _v/F _m, α, rETR_max and E _k. The maximum quantum yield of photosystem II, F _v/F _m, ranged from 0.101 to 0.500 (average 0.284 ± 0.132) and 0.235 to 0.595 (average 0.368 ± 0.127) in the sea ice internal and bottom communities, respectively. The fluorometric measurements indicated medium ice algal photosynthetic activity both in the internal and bottom communities of the sea ice. An observed lack of correlation between biogeochemical and photosynthetic parameters was most likely due to temporally and spatially decoupled physical and biological processes in the sea ice brine channel system, and was also influenced by the temporal and spatial resolution of applied sampling techniques.     

Physical limnological processes under ice

Penetration of solar radiation through ice and snow covering northern lakes produces a gravity current between regions with varying depths. This baroclinic current is a dominant physical process in winter because ice cover insulates lakes from the usual turbulence sources such as breaking surface waves and near-surface shear produced by the wind. The current forms a directed circulation from the littoral zone to the centre of the lake that is an important distribution mechanism for nutrients and other chemical and biological constituents. Heat transported by the current degrades the ice cover and makes surface travel hazardous. The thinning of the ice cover is most severe at the inlet to isolated bays with mean depths that differ significantly from the lake. At the mouth of a bay, the gravity current takes the form of a two-layer flow with inflow to the bay occurring near the surface. The lower layer has the largest temperature gradients and is dominated by a succession of progressive internal bores which decrease in amplitude overnight and with increasing cloud cover. The repetition of the bores occurs very close to the period of the uninodal barotropic seiche which suggests that the internal bores are forced by the surface seiche.     

Protein interaction with ice.

Many organisms have evolved novel mechanisms to minimize freezing injury due to extracellular ice formation. This article reviews our present knowledge on the structure and mode of action of two types of proteins capable of ice interaction. The antifreeze proteins inhibit ice crystal formation and alter ice growth habits. The ice nucleation proteins, on the other hand, provide a proper template to stimulate ice growth. The potential applications of these proteins in different industries are discussed.     

Sea ice microbial dynamics over an annual ice cycle in Prydz Bay, Antarctica

Microbial community dynamics within the fast sea ice of Prydz Bay (68°S?78°E) were investigated over an annual cycle at two sites (1 and 3?km offshore) between April and November 2008. There are few long-term sea ice studies, and few that cover the phase of winter darkness when autotrophic processes are curtailed. Mean chlorophyll a concentrations in the ice column ranged between 0.76 and 44.8?μg?L^?1 at the 1-km site (Site 1) and 3.11–144.6?μg?L^?1 at the 3-km site (Site 2). Highest chlorophyll a usually occurred at the base of the ice. Bacterial concentrations ranged between 0.30 and 2.08?×?10⁸?cells?L^?1, heterotrophic nanoflagellates (HNAN) between 0.21?×?10⁵ and 2.98?×?10⁵?cells?L^?1 and phototrophic nanoflagellates (PNAN) 0–1.06?×?10⁵?cells?L^?1. While HNAN occurred throughout the year, PNAN were largely absent in winter. Dinoflagellates were a conspicuous and occasionally an abundant element of the community (maximum 17,460?cells?L^?1), while ciliates were sparse. The bacterial community showed considerable morphological diversity with a dominance of filamentous forms. Bacterial production continued throughout the year ranging between 0 and 22.92?μg?C?L^?1?day^?1 throughout the ice column. Lowest rates occurred between late June and early August. The sea ice sustained an active and diverse microbial community through its annual extent. The data suggest that during winter darkness the microbial community is dominated by heterotrophic processes, sustained by a pool of dissolved organic carbon.