Production and Characterization of the Intra‐ and Extracellular Carbohydrates and Polymeric Substances (EPS) of Three Sea‐Ice Diatom Species,and Evidence for a Cryoprotective Role for EPS

Diatoms and their associated extracellular polymeric substances (EPS) are major constituents of the microalgal assemblages present within sea ice. Yields and chemical composition of soluble and cell‐associated polysaccharides produced by three sea‐ice diatoms, Synedropsis sp., Fragilariopsis curta, and F. cylindrus, were compared. Colloidal carbohydrates (CC) contained heteropolysaccharides rich in mannose, xylose, galactose, and glucose. Synedropsis sp. CC consisted mainly of carbohydrates <8 kDa size, with relatively soluble EPS, compared to high proportions of less‐soluble EPS produced by both Fragilariopsis spp. F. curta colloidal EPS contained high concentrations of amino sugars (AS). Both Fragilariopsis species had high yields of hot bicarbonate (HB) soluble EPS, rich in xylose, mannose, galactose, and fucose (and AS in F. cylindrus). All species had frustule‐associated EPS rich in glucose–mannose. Nutrient limitation resulted in declines in EPS yields and in glucose content of all EPS fractions. Significant similarities between EPS fractions from cultures and different components of natural EPS from Antarctic sea ice were found. Increased salinity (52) reduced growth, but increased yields of EPS in Fragilariopsis cylindrus. Ice formation was inhibited byF. cylindrus, EPS, and by enhanced EPS content (additional xanthan gum) down to ?12°C, with growth rate reduced in the presence of xanthan. Differences in the production and composition of EPS between Synedropsis sp. and Fragilariopsis spp., and the association between EPS, freezing and cell survival, supports the hypothesis that EPS production is a strategy to assist polar ice diatoms to survive the cold and saline conditions present in sea ice.     

Protists in the marine ice of the Amery Ice Shelf, East Antarctica

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.     

Antifreeze proteins in polar sea ice diatoms: diversity and gene expression in the genus Fragilariopsis

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.     

Agglutinated loricae of some Baltic and Antarctic Tintinnina species (Ciliophora)

The abundance and species composition of diatoms adhering tothe loricae of four agglutinated Tintinnina species, Laackmanniellanaviculaefera, Codonellopsis gaussi, Cd.balechi and Tintinnopsislobiancoi, were determined. Diatoms from the Fragdariopsis group,F.cylindrus and F.pseudonana, dominated on tintinnid loricaefrom the Antarctic waters, whilst Thalassiosira spp. were predominanton loricae from the Baltic Sea. Although tintinnids utilizeddiatoms in the environment, it is not a rule that they use onlythese which are dominant. Our results suggest that certain diatomsare actively selected and agglutinated by particular tintinnidspecies.     

Summer phytoplankton distributions in the Weddell Sea

Summary The quantitative composition of phytoplankton was studied along a transect of 14 hydrographic stations, between the southern coast of the Weddell Sea and the Antarctic Peninsula, during the austral summer of 1984–1985. The most apparent feature of the phytoplankton distribution was the presence of a bloom of Phaeocystis at a frontal zone over the shelf break, and the marked contrast between a southern region, with high phytoplankton biomass, and a poorer region north of the shelf break. The most widely distributed diatom genus was Nitzschia (Fragilariopsis section). The phytoplankton assemblage of the southern region included the silicoflagellate Distephanus speculum, the diatom Rhizosolenia alata and several heterotrophic dinoflagellates such as Protoperidinium antarcticum and P. applanatum. The northern assemblage could be characterized by the abundance of flagellates and small dinoflagellates, and by diatoms such as Chaetoceros criophilum, Corethron criophilum Nitzschia kerguelensis and other Nitzschia species of the Fragilariopsis section.     

Modern distribution of diatoms in sediments from the George V Coast, Antarctica

Currently, little information exists concerning high frequency climate change in the Antarctic, data that are essential to gaining a more complete understanding of global climate change. Sediments from the George V Coast continental margin potentially contain a high resolution record of the late Holocene. The relatively high rate of sedimentation ( 3 mm/year) and the presence of laminated intervals which contain undisturbed sequences make these sediments valuable for future downcore work. In order to utilize diatom assemblages as proxies of paleoclimatic and paleoceanographic parameters, a preliminary study of the surface sediment distribution of diatoms from the George V Coast continental margin was completed. Nitzschia curta (Van Heurck) Hasle and Nitzschia kerguelensis (O'Meara) Hasle present great potential for tracking the Holocene retreat of ice across the continental shelf of this region. Concentrations of N. curta, an “ice-related” form, decreases in an offshore direction, whereas N. kerguelensis, an “oceanic” species associated with circumpolar flow, displays an increase in sediments offshore. The genus Thalassiosira Cleve displays a clear association with areas of open water primary production, as has been noted in others parts of the Antarctic. Downcore variability in relative abundance of this genus will be useful in understanding the history of sea ice versus open water production on the George V Coast shelf. In contrast, the distribution of Chaetoceros Ehrenberg resting spores appears to be more complex, but may be related to the position of the stationary ice edge associated with the annual minimal extent of sea ice. Finally, an understanding of the downcore variability of Thalassiothrix antarctica Schimper ex Karsten, a species which is very abundant in the laminated sections of the sediment cores, will have to rely on studies of its distribution in other areas of the Antarctic, as it is sparsely distributed along the George V Coast today.     

Fragilariopsis cylindrus (Grunow) Krieger: The most abundant diatom in water column assemblages of Antarctic marginal ice-edge zones

Summary Planktonic diatoms were sampled in the ice-edge zone of the Bellingshausen Sea during the early austral spring of 1990 and of the Weddell Sea during the late spring of 1983, the autumn of 1986, and the winter of 1988. The four cruises in the Antarctic marginal ice edge zones, combined with the summer cruise in Prydz Bay during a brief ice-free period (1988) provided us with opportunities for spatial and seasonal studies of diatom abundance and distribution in the water column. Cells from discrete water samples from 73 stations near the marginal ice-edge zones during all seasons were counted to gain quantitative information on the composition, abundance, and distribution of diatoms. Diatom abundance was dominated by the pennate diatom, usually nanoplanktonic, Fragilariopsis cylindrus (Grunow) Krieger, during all five cruises. The highest integrated numbers of F. cylindrus were found during the summer cruise with 7.9 × 10¹⁰ cells m^–2 and the lowest numbers were found during the winter cruise with 1.1 × 10⁸ cells m^–2. The average integrated abundance of F. cylindrus from the five cruises was about 35% of the total diatom abundance. The overall spatial pattern of F. cylindrus near the marginal ice-edge zones during the five seasonal cruises were similar with the highest number of cells in open waters compared to ice-covered waters. When all 73 stations during the five cruises were included in the correlation analysis, the abundance of total diatoms was positively correlated with the abundance of F. cylindrus, suggesting that the ice-edge pulses of diatom assemblages in the water column largely reflected its abundance. Cluster analysis revealed that the stations in marginal ice-edge zones were not only separated by seasons and locations, but they also separated based on location of stations in relation to the ice edge (open water stations vs. ice-covered stations).     

Unmasking Antarctic mollusc lineages: novel evidence from philinoid snails (Gastropoda: Cephalaspidea)

Since its introduction, the genus Philine has epitomized numerous mollusc snails with strong morphological convergence. Recently, a molecular analysis including a wide taxon sampling split this group into four non-sister families. Although they are especially diverse in cold and deep waters, no comprehensive studies are available for the Antarctic counterparts. Here, our morpho-anatomical and molecular data suggest major changes in the systematics of the group. From the eight known species, two are synonymized, Antarctophiline amoena with A. alata, and A. gouldi with A. gibba, and two are transferred to the genus Antarctophiline, namely A. apertissima comb.n. and A. falklandica comb.n. Two new species are described, A. easmithi sp.n. and A. amundseni sp.n. from different depths in the eastern Weddell Sea. The elusive P. antarctica from the Ross Sea was found in the Weddell Sea and Waegelea gen.n. is erected to place this species. Both phylogenetic and morphological data support the erection of Antarctophilinidae fam.n. to embrace most of the Philinoidea species described in the Southern Ocean. Only two species of Philinidae are found in Antarctic waters, Spiraphiline bathyalis gen. et sp.n. from bathyal depths in the Weddell Sea and S. kerguelensis comb.n. from the Kerguelen Islands. In light of the new data provided for all described species and the phylogenetic framework proposed herein, we briefly discuss the diversification and biogeographical patterns of Antarctic philinoid snails. Overall, antarctophilinid species seem to have restricted and grossly nonoverlapping distributions suggesting allopatric speciation connected possibly to geographical or bathymetric isolation.     

DEVELOPMENT OF IMMUNOASSAYS FOR THE IRON‐REGULATED PROTEINS FERREDOXIN AND FLAVODOXIN IN POLAR MICROALGAE1

While the growth of Southern Ocean phytoplankton is often limited by iron availability, there are no comparable experiments on sea‐ice algae. Here we assess the use of ferredoxin and flavodoxin to investigate the iron nutritional status of sea‐ice algae and describe the development of a quantitative immunoassay for both proteins in marine diatoms. High‐affinity monoclonal antibodies toward both proteins were produced from Cylindrotheca closterium (Ehrenb.) J. M. Lewin et Reimann, and these were used to develop Western blots. Western blots run on whole protein extracts detected both proteins with little cross‐reactivity toward other proteins. The two proteins could be successfully quantitated when applied to gels at between 5 and 50 ng in a volume of 25 μL (0.2–2 μg · mL^?1). Flavodoxin and ferrodoxin expression was examined in the Antarctic diatoms Entomoneis kjellmannii (Cleve) Poulin et Cardinal, Navicula directa (W. Sm.) Ralfs, Fragilariopsis curta (Van Heurck) Hust., Pseudo‐nitzschia sp., Porosira glacialis (Grunow) E. G. Jørg., Fragilariopsis cylindrus (Grunow) Willi Krieg., Fragilariopsis sublinearis (Van Heurck) Heiden et Kolbe, C. closterium, Nitzschia lecointei Van Heurck, and the dinoflagellate Polarella glacialis Montresor, Procaccini et Stoecker. Two Arctic isolates were also examined, Nitzschia frigida (Grunow) and Fragilariopsis oceanica (Cleve) Hasle. Significant heterogeneity of protein expression was observed despite all cultures being grown in iron‐replete f/2 medium. Only one species, F. cylindrus, displayed the expected expression of ferredoxin only in iron‐replete medium. Four were observed to produce both proteins under iron‐replete conditions. Ferredoxin was not detected at all in F. curta and Pseudo‐nitzschia sp., but distinct flavodoxin bands were observed in both of these organisms. All species examined were observed to express either flavodoxin or ferredoxin or both of the proteins as determined by Western immunoblotting.     

Nitzschia cylindroformis sp. nov., a common and abundant nanoplankton diatom of the eastern subarctic Pacific

A new species of diatoms, intermediate between Nitzschia cylindrus(Grun.) Hasle and Nitzschia pseudonana Hasle is described. Differencesin morphology of the three species are outlined. The new specieshas been recorded only between 37°N and 53°N in thePacific Ocean while N. cylindrus is bipolar and N. pseudonanais cosmopolitan in distribution.     

Two Southern Ocean diatoms are more sensitive to ocean acidification and changes in irradiance than the prymnesiophyte Phaeocystis antarctica

To better understand the impact of ocean acidification (OA) and changes in light availability on Southern Ocean phytoplankton physiology, we investigated the effects of pCO₂ (380 and 800 µatm) in combination with low and high irradiance (20 or 50 and 200 µmol photons m^?2 s^?1) on growth, particulate organic carbon (POC) fixation and photophysiology in the three ecologically relevant species Chaetoceros debilis, Fragilariopsis kerguelensis and Phaeocystis antarctica. Irrespective of the light scenario, neither growth nor POC per cell was stimulated by OA in any of the tested species and the two diatoms even displayed negative responses in growth (e.g. C. debilis) or POC content (e.g. F. kerguelensis) under OA in conjunction with high light. For both diatoms, also maximum quantum yields of photosystem II (F_v/F_m) were decreased under these conditions, indicating lowered photochemical efficiencies. To counteract the negative effects by OA and high light, the two diatoms showed diverging photoacclimation strategies. While cellular chlorophyll a (Chl a) and fucoxanthin contents were enhanced in C. debilis to potentially maximize light absorption, F. kerguelensis exhibited reduced Chl a per cell, increased disconnection of antennae from photosystem II reaction centers and strongly lowered absolute electron transport rates (ETR). The decline in ETRs in F. kerguelensis might be explained in terms of different species‐specific strategies for tuning the available flux of adenosine triphosphate and nicotinamide adenine dinucleotide phosphate. Overall, our results revealed that P. antarctica was more tolerant to OA and changes in irradiance than the two diatoms, which may have important implications for biogeochemical cycling.     

Sea ice seasonality during the Holocene, Adélie Land, East Antarctica

Thin sections of laminated cores from different Antarctic coastal areas have demonstrated the potential of diatom species to document climate change at the seasonal scale. Here we present the relative abundances of four diatom species and species groups (Fragilariopsis curta group as a proxy for yearly sea ice cover, F. kerguelensis as a proxy for summer sea-surface temperature, Chaetoceros Hyalochaete resting spores as a proxy for spring sea ice melting and the Thalassiosira antarctica group as a proxy for autumn sea ice formation) in core MD03-2601 retrieved off Adélie Land on the Antarctic continental shelf. These abundances were compared to surface temperatures and sea ice cover modelled over the last 9000 years. Both the marine records and the simulated climate demonstrated a cooler Early Holocene (9000–7700 years BP), a warmer Mid-Holocene (7700–4000 years BP) and a colder Late Holocene (4000–1000 years BP). Yearly sea ice cover followed an inverse pattern to temperatures with less sea ice during the Mid-Holocene Hypsithermal than during the Late Holocene Neoglacial. However, diatom census counts and model output indicate that sea ice spring melting happened earlier in the season, as expected, but that autumn sea ice formation also occurred earlier in the season during the Hypsithermal than during the colder Neoglacial, thereby following seasonal changes in local insolation.     

Filling biodiversity gaps: benthic hydroids from the Bellingshausen Sea (Antarctica)

The Bellingshausen Sea constitutes the third largest sea in the Southern Ocean, though it is widely recognized as one of the less-studied Antarctic areas. To reduce this lack of knowledge, a survey to study the biodiversity of its marine benthic communities was carried out during the Bentart 2003 and Bentart 2006 Spanish Antarctic expeditions. The study of the hydroid collection has provided 27 species, belonging to ten families and 15 genera. Twenty-one out of the 27 species constitute new records for the Bellingshausen Sea, raising the total number of known species to 37, as also do nine out of the 15 genera. Candelabrum penola, Lafoea annulata, and Staurotheca juncea are recorded for the second time. Most species belong to Leptothecata. Sertulariidae with 13 species (48%) is by far the most speciose family, and Symplectoscyphus with seven species (26%), including S. bellingshauseni sp. nov. and S. hesperides sp. nov., the most diverse genus. Considering the whole benthic hydroid fauna of the Bellingshausen Sea, 18 species (69%) are endemic to Antarctic waters, either with a circum-Antarctic (12 species, 46%) or West Antarctic (6 species, 23%) distribution, 23 (88%) are restricted to Antarctic or Antarctic/sub-Antarctic waters, and only three species have a wider distribution. Bellingshausen Sea hydroid fauna is composed of a relatively high diversity of typical representatives of the Antarctic benthic hydroid fauna, though with a surprisingly low representation of some of the most diverse and widespread Antarctic genera (Oswaldella and Schizotricha), what could be related to the fact that its shelf-inhabiting hydroid fauna remains practically unknown.     

THE EFFECT OF IRON LIMITATION ON THE PHOTOPHYSIOLOGY OF PHAEOCYSTIS ANTARCTICA (PRYMNESIOPHYCEAE) AND FRAGILARIOPSIS CYLINDRUS (BACILLARIOPHYCEAE) UNDER DYNAMIC IRRADIANCE1

The effects of iron limitation on photoacclimation to dynamic irradiance were studied in Phaeocystis antarctica G. Karst. and Fragilariopsis cylindrus (Grunow) W. Krieg. in terms of growth rate, photosynthetic parameters, pigment composition, and fluorescence characteristics. Under dynamic light conditions mimicking vertical mixing below the euphotic zone, P. antarctica displayed higher growth rates than F. cylindrus both under iron (Fe)–replete and Fe‐limiting conditions. Both species showed xanthophyll de‐epoxidation that was accompanied by low levels of nonphotochemical quenching (NPQ) during the irradiance maximum of the light cycle. The potential for NPQ at light levels corresponding to full sunlight was substantial in both species and increased under Fe limitation in F. cylindrus. Although the decline in F_v/F_m under Fe limitation was similar in both species, the accompanying decrease in the maximum rate of photosynthesis and growth rate was much stronger in F. cylindrus. Analysis of the electron transport rates through PSII and on to carbon (C) fixation revealed a large potential for photoprotective cyclic electron transport (CET) in F. cylindrus, particularly under Fe limitation. Probably, CET aided the photoprotection in F. cylindrus, but it also reduced photosynthetic efficiency at higher light intensities. P. antarctica, on the other hand, was able to efficiently use electrons flowing through PSII for C fixation at all light levels, particularly under Fe limitation. Thus, Fe limitation enhanced the photophysiological differences between P. antarctica and diatoms, supporting field observations where P. antarctica is found to dominate deeply mixed water columns, whereas diatoms dominate shallower mixed layers.     

Phytoplankton species composition and abundance in the Indian sector of the Antarctic Ocean

Summary Water samples collected in the southwestern Indian Ocean between Africa and Antarctica in March 1980 were analyzed quantitatively for phytoplankton. Diatoms dominate the phytoplankton in this region and their numbers generally increase southward with peaks of abundance in both the northern Antarctic Zone and south of the Antarctic Divergence. Average cell numbers (i.e., 6.1×10⁵ diatoms l^-1 in the Antarctic Zone) are comparable to maximum numbers previously reported for the Southern Ocean. Dinoflagellates, flagellates and monads occur in highest concentrations north of the Polar Front. Their numbers are somewhat reduced south of the Antarctic Divergence, and are lowest in the Antarctic Zone. Various diatom assemblages are characteristic of different latitudinal zones. Waters north of and in the vicinity of the Polar Front are rich in the Nitzschia, Pseudonitzschia group of species. In the Antarctic Zone, Nitzschia nana and Dactyliosolen tenuijunctus dominate. Nitzschia species of the Fragilariopsis group are most numerous at stations south of the Antarctic Divergence. Striking differences are noted between the species compositions of quantitative and net-haul samples. A few nanoplanktonic diatoms (e.g. Nitzschia nana and single cells of Chaetoceros spp.) and the weakly silicified Dactyliosolen tenuijunctus, which are dominant in the quantitative samples, are either entirely absent or present only as solitary cells in the net collections.     

EFFECT OF HYPEROXIA ON THE GROWTH AND PHOTOSYNTHESIS OF POLAR SEA ICE MICROALGAE1

Sea ice algal communities are naturally exposed to very high concentrations of dissolved oxygen, which are likely to lead to increasing stress levels and declines in productivity. To test this hypothesis, cultures of Fragilariopsis cylindrus (Grun?) Hasle, Pseudo‐nitzschia sp., Fragilariopsis curta (Van Heurch), Porosira glacialis (Grunow), and Entomoneis kjellmannii (Cleve) from Antarctic sea ice and Nitzschia frigida from Arctic sea ice were exposed to elevated dissolved oxygen levels, and their growth, maximum quantum yield, relative maximum electron transport rate, and photosynthetic efficiency were measured. At oxygen concentrations equivalent to approximately four times air saturation (89% oxygen), the growth rate and maximum quantum yield were significantly reduced in all taxa. When the oxygen concentration was regularly allowed to drop, the effect on growth and quantum yield was reduced. At lower dissolved oxygen concentrations (52%), the declines in growth and quantum yield were reduced but were still mostly significantly different from the controls (21% oxygen). It is likely that the generation of excess active oxygen radicals in the presence of free oxygen is responsible for most of the decline in growth, maximum quantum yield, relative maximum electron transport rate, and photosynthetic efficiency in all species.     

Benthic hydroids (Cnidaria: Hydrozoa) from the Spanish Antarctic expedition Bentart 95

A total of 61 species of hydroids, belonging to 13 families and 23 genera, were found during the Spanish Antarctic expedition Bentart 95 with the RV Hespérides. Ten of the species were identified only to generic level. The dominance of the subclass Leptothecatae, with 57 species, was remarkable. The remaining four species belong to the subclass Anthoathecatae. By far the most diverse family was the Sertulariidae, with 25 species (41%), followed by Haleciidae with nine species (15%) and Kirchenpaueriidae with six (10%). The family Plumulariidae, represented by one species of Nemertesia, is recorded for the first time from Antarctic waters. Eudendrium scotti, Perarella clavata and Symplectoscyphus hero are each recorded for the second time. Symplectoscyphus with 11 species was the dominant genus. Almost 60% of the species diversity is concentrated in just a little more than the 20% of genera. Nearly 70% of the species are endemic to Antarctic waters and 90% of them are restricted to Antarctic or Antarctic/sub-Antarctic waters.