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1.
The structure of bacterial communities in first‐year spring and summer sea ice differs from that in source seawaters, suggesting selection during ice formation in autumn or taxon‐specific mortality in the ice during winter. We tested these hypotheses by weekly sampling (January–March 2004) of first‐year winter sea ice (Franklin Bay, Western Arctic) that experienced temperatures from ?9°C to ?26°C, generating community fingerprints and clone libraries for Bacteria and Archaea. Despite severe conditions and significant decreases in microbial abundance, no significant changes in richness or community structure were detected in the ice. Communities of Bacteria and Archaea in the ice, as in under‐ice seawater, were dominated by SAR11 clade Alphaproteobacteria and Marine Group I Crenarchaeota, neither of which is known from later season sea ice. The bacterial ice library contained clones of Gammaproteobacteria from oligotrophic seawater clades (e.g. OM60, OM182) but no clones from gammaproteobacterial genera commonly detected in later season sea ice by similar methods (e.g. Colwellia, Psychrobacter). The only common sea ice bacterial genus detected in winter ice was Polaribacter. Overall, selection during ice formation and mortality during winter appear to play minor roles in the process of microbial succession that leads to distinctive spring and summer sea ice communities.  相似文献   

2.
Summary Factors influencing the fate of ice algae released from melting sea ice were studied during a R V Polarstern cruise (EPOS Leg 2) to the northwestern Weddell Sea. The large-scale phytoplankton distribution patterns across the receding ice edge and small-scale profiling of the water column adjacent to melting ice floes indicated marked patchiness on both scales. The contribution of typical ice algae to the phytoplankton was not significant. In experiments simulating the conditions during sea ice melting, ice algae revealed a strong propensity to form aggregates. Differences in the aggregation potential were found for algal assemblages collected from the ice interior and the infiltration layer. Although all algal species collected from the ice were also found in aggregates, the species composition of dispersed and aggregated algae differed significantly. Aggregates were of a characteristic structure consisting of monospecific microaggregates which are likely to have formed in the minute brine pockets and channels within the ice. Sinking rates of aggregates were three orders of magnitude higher than those of dispersed ice algae. These observations, combined with the negligible seeding effect of ice algae found during this study, suggest that ice algae released from the melting sea ice are subject to rapid sedimentation. High grazing pressure at the ice edge of the investigation area is another factor eliminating ice algae released during melting.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation  相似文献   

3.
The effect of nutrient limitation on Baltic Sea ice algae, and substrate and nutrient limitation on ice bacteria, was studied in a series of in situ -experiments conducted during the winter of 2002 in northern Baltic Sea. Community level changes in algal biomass (chlorophyll a) and productivity, and bacterial thymidine and leucine incorporation were followed for one week after the addition of nutrient and/or organic carbon rich filtered seawater to the experimental units. The results showed the major contribution of snow cover to the algal responses during the beginning of the ice-covered season. Algal communities were able to grow even in January if no snow was present. Nutrient addition did occasionally have an effect on algal biomass and productivity in the ice. Surprisingly, seeding effect from the ice to the underlying water was negatively affected by the nutrient availability in March. Bacterial limitation varied between nutrient (phosphorus) and substrate limitations. The results showed, that limitation in both algal and bacterial communities changed periodically in the northern Baltic Sea ice.  相似文献   

4.
D. Delille 《Polar Biology》1992,12(2):205-210
Summary In the eastern Weddell Sea on several transects from ice-covered, through ice melt, to open-ocean stations, total and heterotrophic bacteria were estimated to document an enhanced bacteriological biomass expected near the ice edge. The highest numbers of bacteria were found in melted ice cores, with 4.2·103 CFUml–1 and 1.1·107 Cells ml–1. Although brine from pore water samples average more than one order of magnitude less cells per ml, the highest bacterial production, 2.2·107 cells l–1 day–1, was recorded in brine samples. All quantitatively studied bacterial parameters were lower under the ice than in the ice samples but there were no clear vertical gradients in the water column. In the studied spring situation, sea ice occurrence seems to play only a minor role in the general distribution of the seawater bacterioplankton. The bacterial community structure was investigated by carrying out 29 morphological and biochemical tests on 118 isolated strains. The bacterial communities inhabiting Antarctic pack ice differ from those found in underlying seawater. Although non fermentative Gram-negative rods were always dominant in seawater, Vibrio sp. represented more than 25% of the strains isolated from some ice samples. The results clearly indicated that a large majority of the bacteria isolated from seawater must be considered psychrotrophic but that truly psychrophilic strains occurred in melted ice and brine samples.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation  相似文献   

5.
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.  相似文献   

6.
To investigate the responses of Baltic Sea wintertime bacterial communities to changing salinity (5 to 26 practical salinity units), an experimental study was conducted. Bacterial communities of Baltic seawater and sea ice from a coastal site in southwest Finland were used in two batch culture experiments run for 17 or 18 days at 0°C. Bacterial abundance, cell volume, and leucine and thymidine incorporation were measured during the experiments. The bacterial community structure was assessed using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rRNA genes with sequencing of DGGE bands from initial communities and communities of day 10 or 13 of the experiment. The sea ice-derived bacterial community was metabolically more active than the open-water community at the start of the experiment. Ice-derived bacterial communities were able to adapt to salinity change with smaller effects on physiology and community structure, whereas in the open-water bacterial communities, the bacterial cell volume evolution, bacterial abundance, and community structure responses indicated the presence of salinity stress. The closest relatives for all eight partial 16S rRNA gene sequences obtained were either organisms found in polar sea ice and other cold habitats or those found in summertime Baltic seawater. All sequences except one were associated with the α- and γ-proteobacteria or the Cytophaga-Flavobacterium-Bacteroides group. The overall physiological and community structure responses were parallel in ice-derived and open-water bacterial assemblages, which points to a linkage between community structure and physiology. These results support previous assumptions of the role of salinity fluctuation as a major selective factor shaping the sea ice bacterial community structure.  相似文献   

7.
Samples of marine ice were collected from the Amery Ice Shelf, a large embayed ice shelf in East Antarctica, during the Austral summer of 2001–2002. The samples came from a site ∼90 km from the iceberg calving front of the shelf, where the ice is 479 m thick and the lower 203 m is composed of accreted marine ice. Protists identified within the marine ice layer of the Amery Ice Shelf include diatoms, chrysophytes, silicoflagellates and dinoflagellates. The numerical dominance of sea ice indicator diatoms such as Fragilariopsis curta, Fragilariopsis cylindrus, Fragilariopsis rhombica and Chaetoceros resting spores, and the presence of cold open water diatoms such as Fragilariopsis kerguelensis and species of Thalassiosira suggest the protist composition of the Amery marine ice is attributable to seeding from melting pack and/or fast ice protist communities in the highly productive waters of Prydz Bay to the north.  相似文献   

8.
To investigate the responses of Baltic Sea wintertime bacterial communities to changing salinity (5 to 26 practical salinity units), an experimental study was conducted. Bacterial communities of Baltic seawater and sea ice from a coastal site in southwest Finland were used in two batch culture experiments run for 17 or 18 days at 0 degrees C. Bacterial abundance, cell volume, and leucine and thymidine incorporation were measured during the experiments. The bacterial community structure was assessed using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rRNA genes with sequencing of DGGE bands from initial communities and communities of day 10 or 13 of the experiment. The sea ice-derived bacterial community was metabolically more active than the open-water community at the start of the experiment. Ice-derived bacterial communities were able to adapt to salinity change with smaller effects on physiology and community structure, whereas in the open-water bacterial communities, the bacterial cell volume evolution, bacterial abundance, and community structure responses indicated the presence of salinity stress. The closest relatives for all eight partial 16S rRNA gene sequences obtained were either organisms found in polar sea ice and other cold habitats or those found in summertime Baltic seawater. All sequences except one were associated with the alpha- and gamma-proteobacteria or the Cytophaga-Flavobacterium-Bacteroides group. The overall physiological and community structure responses were parallel in ice-derived and open-water bacterial assemblages, which points to a linkage between community structure and physiology. These results support previous assumptions of the role of salinity fluctuation as a major selective factor shaping the sea ice bacterial community structure.  相似文献   

9.
Summary The amphipod Gammarus wilkitzkii does not survive being frozen totally into solid sea ice. When the animals are cooled in air or freezing seawater, they will freeze and die at a temperature of about-4° C. However, during sea ice growth, the amphipods may tolerate to stay in the vicinity of the ice by conforming to the ambient brine in a salinity range of 34 ppt to about 60 ppt. A passive relationship between the concentrations of the haemolymph and seawater Na+ and Cl-, lowers the melting point of the body fluids of the animals, thus preventing internal ice formation at low temperatures.  相似文献   

10.
K. Kivi  H. Kuosa 《Polar Biology》1994,14(6):389-399
Microbial communities in the water column and sea ice were studied during the EPOS-cruise on R/V Polarstern in the western Weddell Sea in late winter (October–November 1988). Samples were taken from four transects from heavy pack-ice to open water. The results indicated the important role of protozoans especially in the ice-edge area. Heterotrophic nanofiagellates, dinoflagellates, ciliates and sarcodines showed significant positive correlations with chlorophyll a. Autotrophic picoplankton and autotrophic flagellates, which were probably motile zooids of Phaeocystis pouchetii (up to 3×106 cells 1–1), were most abundant in the areas of low or medium chlorophyll a concentration. Sea ice contained high numbers of heterotrophic organisms, and the distribution of the different groups showed distinct vertical zonation. At two sites, the microbial assembly beneath the ice was clearly influenced by communities from the melting ice.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation  相似文献   

11.
Ocean acidification substantially alters ocean carbon chemistry and hence pH but the effects on sea ice formation and the CO2 concentration in the enclosed brine channels are unknown. Microbial communities inhabiting sea ice ecosystems currently contribute 10–50% of the annual primary production of polar seas, supporting overwintering zooplankton species, especially Antarctic krill, and seeding spring phytoplankton blooms. Ocean acidification is occurring in all surface waters but the strongest effects will be experienced in polar ecosystems with significant effects on all trophic levels. Brine algae collected from McMurdo Sound (Antarctica) sea ice was incubated in situ under various carbonate chemistry conditions. The carbon chemistry was manipulated with acid, bicarbonate and bases to produce a pCO2 and pH range from 238 to 6066 µatm and 7.19 to 8.66, respectively. Elevated pCO2 positively affected the growth rate of the brine algal community, dominated by the unique ice dinoflagellate, Polarella glacialis. Growth rates were significantly reduced when pH dropped below 7.6. However, when the pH was held constant and the pCO2 increased, growth rates of the brine algae increased by more than 20% and showed no decline at pCO2 values more than five times current ambient levels. We suggest that projected increases in seawater pCO2, associated with OA, will not adversely impact brine algal communities.  相似文献   

12.
The diversity of diatom communities found epiphytically on red macroalgae collected under the sea ice at Cape Evans, Antarctica, was studied microscopically. Comparison of communities along a depth profile showed that species diversity decreased as depth below sea ice increased. Dominant taxa also changed with depth, with Cocconeis fasciolata dominant at 10 and 15 m, Porosira glacialis at 20 m and Eunotogramma marginopunctatum at 25 m. Epiphytic communities were also compared to sympagic and sediment-associated communities collected from a single depth. Species richness was greatest for communities associated with surface sediments while the poorest was found in sympagic, or sea-ice, communities. E. marginopunctatum, previously only described from fossil material, was found associated epiphytically on macroalgae from 20 to 25 m below sea ice.  相似文献   

13.
The structure, productivity and heterotrophic potential of an extensive microalgal community growing on the underside of sea ice near the Australian Antarctic Station of Casey, are described. Underwater observations made near the Australian Antarctic stations of Davis and Mawson are also reported. This community develops during September, is largely suspended from the bottom surface of annual sea ice and often extends into the underlying water column as conspicuous strands up to 15 cm long. The algal community structure in the strands is dominated by an unidentified tube diatom belonging to the Amphipleura/Berkeleya group and chains of a species of Entomoneis cf. Amphiprora paludosa var. hyperborea (Grunow) Cleve. Unlike previously described bottom ice environments, a brash ice layer under the hard sea ice is absent. Living cells, predominantly Nitzschia frigida Grunow, also occur in microbrine channels in the bottom 3 cm of the ice. Maximal primary production rates of 81 μg C · L-1· h-1 occurred during November, then began declining near the end of December. Minimal rates (2.8 μg C · L-1· h-1) were reached in mid-January and coincided with changes in the physical structure of the sea ice and in the stability of the water column. An abundant epibacterial community associated with the microalgal strands assimilated 3H-labelled amino acids suggesting significant heterotrophic recycling of dissolved organic matter. Turnover times of assimilated amino acids in the bottom ice community averaged 55 h during November while negligible turnover of these substrates occurred in the water column 1.5 m below the ice. These bottom ice communities have higher primary productivity than typical brash ice communities; they are also accessible to marine herbivores and so may be more important to the Antarctic marine food chain than previously supposed.  相似文献   

14.
UniFrac: a new phylogenetic method for comparing microbial communities   总被引:9,自引:0,他引:9  
We introduce here a new method for computing differences between microbial communities based on phylogenetic information. This method, UniFrac, measures the phylogenetic distance between sets of taxa in a phylogenetic tree as the fraction of the branch length of the tree that leads to descendants from either one environment or the other, but not both. UniFrac can be used to determine whether communities are significantly different, to compare many communities simultaneously using clustering and ordination techniques, and to measure the relative contributions of different factors, such as chemistry and geography, to similarities between samples. We demonstrate the utility of UniFrac by applying it to published 16S rRNA gene libraries from cultured isolates and environmental clones of bacteria in marine sediment, water, and ice. Our results reveal that (i) cultured isolates from ice, water, and sediment resemble each other and environmental clone sequences from sea ice, but not environmental clone sequences from sediment and water; (ii) the geographical location does not correlate strongly with bacterial community differences in ice and sediment from the Arctic and Antarctic; and (iii) bacterial communities differ between terrestrially impacted seawater (whether polar or temperate) and warm oligotrophic seawater, whereas those in individual seawater samples are not more similar to each other than to those in sediment or ice samples. These results illustrate that UniFrac provides a new way of characterizing microbial communities, using the wealth of environmental rRNA sequences, and allows quantitative insight into the factors that underlie the distribution of lineages among environments.  相似文献   

15.
UniFrac: a New Phylogenetic Method for Comparing Microbial Communities   总被引:18,自引:8,他引:10       下载免费PDF全文
We introduce here a new method for computing differences between microbial communities based on phylogenetic information. This method, UniFrac, measures the phylogenetic distance between sets of taxa in a phylogenetic tree as the fraction of the branch length of the tree that leads to descendants from either one environment or the other, but not both. UniFrac can be used to determine whether communities are significantly different, to compare many communities simultaneously using clustering and ordination techniques, and to measure the relative contributions of different factors, such as chemistry and geography, to similarities between samples. We demonstrate the utility of UniFrac by applying it to published 16S rRNA gene libraries from cultured isolates and environmental clones of bacteria in marine sediment, water, and ice. Our results reveal that (i) cultured isolates from ice, water, and sediment resemble each other and environmental clone sequences from sea ice, but not environmental clone sequences from sediment and water; (ii) the geographical location does not correlate strongly with bacterial community differences in ice and sediment from the Arctic and Antarctic; and (iii) bacterial communities differ between terrestrially impacted seawater (whether polar or temperate) and warm oligotrophic seawater, whereas those in individual seawater samples are not more similar to each other than to those in sediment or ice samples. These results illustrate that UniFrac provides a new way of characterizing microbial communities, using the wealth of environmental rRNA sequences, and allows quantitative insight into the factors that underlie the distribution of lineages among environments.  相似文献   

16.
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, rETRmax 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.  相似文献   

17.
Extracellular polymeric substances (EPS) are known to help microorganisms to survive under extreme conditions in sea ice. High concentrations of EPS are reported in sea ice from both poles; however, production and dynamics of EPS during sea ice formation have been little studied to date. This investigation followed the production and partitioning of existing and newly formed dissolved organic matter (DOM) including dissolved carbohydrates (dCHO), dissolved uronic acids (dUA) and dissolved EPS (dEPS), along with bacterial abundances during early stages of ice formation. Sea ice was formed from North Sea water with (A) ambient DOM (NSW) and (B) with additional algal-derived DOM (ADOM) in a 6d experiment in replicated mesocosms. In ADOM seawater, total bacterial numbers (TBN) increased throughout the experiment, whereas bacterial growth occurred for 5d only in the NSW seawater. TBN progressively decreased within developing sea ice but with a 2-fold greater decline in NSW compared to ADOM ice. There were significant increases in the concentrations of dCHO in ice. Percentage contribution of dEPS was highest (63%) in the colder, uppermost parts in ADOM ice suggesting the development of a cold-adapted community, producing dEPS possibly for cryo-protection and/or protection from high salinity brines. We conclude that in the early stages of ice formation, allochthonous organic matter was incorporated from parent seawater into sea ice and that once ice formation had established, there were significant changes in the concentrations and composition of dissolved organic carbon pool, resulting mainly from the production of autochthonous DOM by the bacteria.  相似文献   

18.
Exopolysaccharides (EPS) may have an important role in the Antarctic marine environment, possibly acting as ligands for trace metal nutrients such as iron or providing cryoprotection for growth at low temperature and high salinity. Ten bacterial strains, isolated from Southern Ocean particulate material or from sea ice, were characterized. Whole cell fatty acid profiles and 16S rRNA gene sequences showed that the isolates included representatives of the genera Pseudoalteromonas, Shewanella, Polaribacter, and Flavobacterium as well as one strain, which constituted a new bacterial genus in the family Flavobacteriaceae. The isolates are, therefore, members of the “Gammaproteobacteria” and Cytophaga-Flexibacter-Bacteroides, the taxonomic groups that have been shown to dominate polar sea ice and seawater microbial communities. Exopolysaccharides produced by Antarctic isolates were characterized. Chemical composition and molecular weight data revealed that these EPS were very diverse, even among six closely related Pseudoalteromonas isolates. Most of the EPS contained charged uronic acid residues; several also contained sulfate groups. Some strain produced unusually large polymers (molecular weight up to 5.7 MDa) including one strain in which EPS synthesis is stimulated by low temperature. This study represents a first step in the understanding of the role of bacterial EPS in the Antarctic marine environment.  相似文献   

19.
Summary The present paper describes the composition, abundance, biomass and diversity of the meso- and macrozooplankton in the epipelagic zone of the open water and under the ice of the northern Weddell Sea. Samples were collected in October/November 1988 with a multiple RMT1+8 net during the European Polarstern Study (EPOS). Multivariate analysis resulted in two distinct site clusters, a northern one mainly located in the open water/marginal ice zone and a southern one extending from the marginal ice zone into the consolidated pack-ice. Clusters were, however, faunistically coherent with a high degree in positive covariation of species. There was no basis for the separation into communities, but differences occurred on the population level in numerical abundances, biomass (wet weight) and in a shift in species dominance. Different ice zones and vertical layers were tested among each other with regard to their relative species abundance. Significant differences were found between the upper 60 m layer of the open sea, the upper 60 m layer of the closed pack-ice and the so called transitional zone. Species richness and diversity was lowest directly under the closed pack-ice. Abundance and biomass was highest in the surface layer of the open water, while both variablès decreased dramatically under the ice. Copepods dominated numerically in open water, while salps dominated in biomass. Euphausia superba and Thysanoessa macrura were the dominant species in the upper water column of the closed pack-ice zone. Krill was the only species with increasing abundance in the sub-ice area and a dominance in biomass of more than 91% demonstrated its unique importance for the sub-ice habitat.Data presented here were collected during the European Polarstern Study (EPOS) sponsored by the European Science Foundation  相似文献   

20.
The bacterial populations associated with sea ice sampled from Antarctic coastal areas were investigated by use of a phenotypic approach and a phylogenetic approach based on genes encoding 16S rRNA (16S rDNA). The diversity of bacteria associated with sea ice was also compared with the bacterial diversity of seawater underlying sea ice. Psychrophilic (optimal growth temperature, < or = 15 degrees C; no growth occurring at 20 degrees C) bacterial diversity was found to be significantly enriched in sea ice samples possessing platelet and bottom ice diatom assemblages, with 2 to 9 distinct (average, 5.6 +/- 1.8) psychrophilic taxa isolated per sample. Substantially fewer psychrophilic isolates were recovered from ice cores with a low or negligible population of ice diatoms or from under-ice seawater samples (less than one distinct taxon isolated per sample). In addition, psychrophilic taxa that were isolated from under-ice seawater samples were in general phylogenetically distinct from psychrophilic taxa isolated from sea ice cores. The taxonomic distributions of psychrotrophic bacterial isolates (optimal growth temperature, > 20 degrees C; growth can occur at approximately 4 degrees C) isolated from sea ice cores and under-ice seawater were quite similar. Overall, bacterial isolates from Antarctic sea ice were found to belong to four phylogenetic groups, the alpha and gamma subdivisions of the Proteobacteria, the gram-positive branch, and the Flexibacter-Bacteroides-Cytophaga phylum. Most of the sea ice strains examined appeared to be novel taxa based on phylogenetic comparisons, with 45% of the strains being psychrophilic. 16S rDNA sequence analysis revealed that psychrophilic strains belonged to the genera Colwellia, Shewanella, Marinobacter, Planococcus, and novel phylogenetic lineages adjacent to Colwellia and Alteromonas and within the Flexibacter-Bacteroides-Cytophaga phylum. Psychrotrophic strains were found to be members of the genera Pseudoalteromonas, Psychrobacter, Halomonas, Pseudomonas, Hyphomonas, Sphingomonas, Arthrobacter, Planococcus, and Halobacillus. From this survey, it is proposed that ice diatom assemblages provide niches conducive to the proliferation of a diverse array of psychrophilic bacterial species.  相似文献   

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