Molecular genetic diversity of bacteria in the bottom section of arctic sea ice from the Canada Basin

PCR-DGGE approach was used to analyze bacterial diversity in the bottom section of seven arctic sea ice samples colleted from the Canada Basin. Thirty-two 16S rDNA sequences were obtained from prominent DGGE bands. The closest relatives of these sequences are found to be those of cultivated or uncultured bacteria from antarctic or arctic sea ice. Phylogenetic analysis clustered these sequences or phylotypes within α- proteobacteria, γ-proteobacteria and CFB (cytophaga-flexibacter-bacteroides) group. Sequences belonging to γ-proteobacteria were dominant and members of the CFB group were highly abundant. It was suggested that the CFB group was the representative of the bottom section of sea ice samples.     

Bacterioplankton responses to iron enrichment during the SAGE experiment

We studied the microbial food web in the upper 100 m of the water column in iron-limited sub-Antarctic HNLC waters south-east of New Zealand in the SAGE experiment in 2004, with focus on bacterioplankton. Samples were collected daily from inside and outside the iron enriched patch. Short term enrichment experiments were conducted on board in 4 L polycarbonate bottles with water outside the iron enriched patch to study single and combined effects of micronutrient additions on microbial food web. Low bacterial growth was recorded in the study area with community turnover times of 50 h or more during the study period. Measurements of bacterial standing stocks and production rates in the study show minor responses to the large scale iron enrichment, with increase in rates and stocks after the first enrichment and at the end of the study period after the third iron enrichment when solar radiation increased and wind mixing decreased. The average daily bacterial production rates were 31.5 and 33.7 mgCm⁻² d⁻¹ for the OUT and IN stations, respectively; thus overall there was not a significant difference between the control and the iron-enriched patch. In the bottle experiments bacterial thymidine incorporation showed responses to single iron and silicic acid enrichments and a major growth response to the combined iron and sucrose enrichments. Phytoplankton chlorophyll-a showed clear stimulation by single additions of iron and silicic acid and silicic acid enhanced the iron impact. Cobalt additions had no effect on bacteria growth and a negative effect on phytoplankton growth. Low bacterial in situ growth rates and the enrichment experiments suggest that bacteria are co-limited by iron and carbon, and that bacterial iron uptake is dependent on carbon supply by the food web. With the high iron quota (??mol Fe mol C⁻¹) bacteria may scavenge considerable amounts of the excess iron, and thus influence the relative importance of the microbial food web as a carbon sink.     

Community structure and photosynthetic physiology of phytoplankton in the northwest subarctic Pacific during an in situ iron fertilization experiment (SEEDS-II)

Temporal changes in the abundance, community composition, and photosynthetic physiology of phytoplankton in surface waters were investigated during the second in situ iron (Fe) fertilization experiment in the NW subarctic Pacific (SEEDS-II). Surface chlorophyll a concentration was 0.75 mg m⁻³ on the day before the first Fe enrichment (i.e. Day 0), increased ca. 3-fold until Day 13 after two Fe additions, and thereafter declined with time. The photochemical quantum efficiency (F_v/F_m) and functional absorption cross-section (σPSII) of photosystem II for total phytoplankton in surface waters increased and decreased inside the Fe-enriched patch through Day 13, respectively. These results indicate that the photosynthetic physiological condition of the phytoplankton improved after the Fe infusions. However, the maximum F_v/F_m value of 0.43 and the maximum quantum yield of carbon fixation (φ_max) of 0.041 mol C (mol photon)⁻¹ during the development phase of the bloom were rather low, compared to their theoretical maximum of ca. 0.65 and 0.10 mol C (mol photon)⁻¹, respectively. Diatoms, which were mainly composed of oceanic species, did not bloom, and autotrophic nanoflagellates such as cryptophytes and prasinophytes became predominant in the phytoplankton community inside the Fe-enriched patch. In ferredoxin/flavodoxin assays for micro-sized (20–200 μm in cell length) diatoms, ferredoxin was not detected but flavodoxin expressions consistently occurred with similar levels both inside and outside the Fe-enriched patch, indicating that the large-sized diatoms were stressed by Fe bioavailability inside the Fe-enriched patch even after the Fe enrichments. Our data suggest that the absence of a Fe-induced large-sized diatom bloom could be partly due to their Fe stress throughout SEEDS-II.     

Responses of diatoms to iron-enrichment (SEEDS) in the western subarctic Pacific, temporal and spatial comparisons

Phytoplankton species composition was analyzed inside and outside of the iron-enriched patch during the SEEDS experiment. Before the iron-enrichment, the phytoplankton community consisted of similar proportions of pico-, nano- and micro-sized phytoplankton, and the micro-phytoplankton was dominated by the pennate diatom Pseudo-nitzschia turgidula. Although all the diatoms, except the nano-sized Fragilariopsis sp., increased during the two weeks of the observation period, the flora in the patch dramatically changed with the increase of phytoplankton biomass to a centric diatom-dominated community. Neritic diatoms, especially Chaetoceros debilis, showed higher growth rates than other diatoms, without any delay in the initiation of growth after the enrichment, and accounted for 90% of the micro-phytoplankton after day 9. In contrast, the oceanic diatoms showed distinct delays in the initiation of growth. We conclude that the responses of the diatoms to the manipulation of iron concentration were different by species, and the fast and intensive response of the phytoplankton to iron-enrichment resulted from the presence of a small amount of neritic diatoms at the study site. The important factors that determine the dominant species in the bloom are the potential growth rates under an iron-replete condition and the growth lag. Abundant species in the patch are widely distributed in the North Pacific and their relative contributions in the Oyashio area and at Stn KNOT are high from spring to summer. However, a characteristic difference of species composition between the SEEDS bloom and natural blooms was the lack of Thalassiosira and Coscinodiscus species in the patch, which usually account for a major part of the phytoplankton community under blooming conditions in the western North Pacific.     

Rare earth elements in seawater during an iron-induced phytoplankton bloom of the western subarctic Pacific (SEEDS-II)

During an iron-enrichment experiment in the western subarctic Pacific (SEEDS-II), concentrations of dissolved and acid-soluble rare earth elements (REEs) were determined in shallow waters. Detailed vertical profiles of dissolved REEs were compared with those of nutrients in a preliminary survey. The results showed good correlation, except for Ce and Y. Along with phytoplankton growth within the iron-enriched patch, dissolved REEs were decreased in surface waters. The changes in REE concentrations were small but systematic: light REEs were adsorbed preferentially over heavy REEs. This systematic trend is consistent with results of earlier studies of REE adsorption onto biogenic particle surfaces. However, we were unable to detect a decrease of acid-soluble REEs in surface waters. During the phytoplankton bloom decline phase, dissolved REEs were elevated to concentration levels at the preliminary survey, although nutrients were decreased. The liberation of REEs from the suspended particles might be related to disaggregation from large particles to colloid particles, desorption from the particulate phase, and remineralization from organic matter. Outside the Fe-patch, time variations of REEs were also observed during SEEDS-II, indicating that both artificially induced phytoplankton blooms and natural variation of biogenic particles can affect the biogeochemical cycles of REEs in shallow waters within a short time.     

Iron and mixing affect biological carbon uptake in SOIREE and EisenEx,two Southern Ocean iron fertilisation experiments

This study explores the changes in the surface water fugacity of carbon dioxide (fCO₂) and biological carbon uptake in two Southern Ocean iron fertilisation experiments with different hydrographic regimes. The Southern Ocean Iron Release Experiment (SOIREE) experiment was carried out south of the Antarctic Polar Front (APF) at 61°S, 141°E in February 1999 in a stable hydrographic setting. The EisenEx experiment was conducted in a cyclonic eddy north of the APF at 48°S, 21°E in November 2000 and was characterised by a rapid succession of low to storm-force wind speeds and dynamic hydrographic conditions. The iron additions promoted algal blooms in both studies. They alleviated algal iron limitation during the 13-day SOIREE experiment and probably during the first 12 days of EisenEx. The fCO₂ in surface water decreased at a constant rate of 3.8 μatm day⁻¹ from 4 to 5 days onwards in SOIREE. The fCO₂ reduction was 35 μatm after 13 days. The evolution of surface water fCO₂ in the iron-enriched waters (or ‘patch’) displayed a saw tooth pattern in EisenEx, in response to algal carbon uptake in calm conditions and deep mixing and horizontal dispersion during storms. The maximum fCO₂ reduction was 18–20 μatm after 12 and 21 days with lower values in between. The iron-enriched waters in EisenEx absorbed four times more atmospheric CO₂ than in SOIREE between 5 and 12 days, as a result of stronger winds. The total biological uptake of inorganic carbon across the patch was 1389 ton C (±10%) in SOIREE and 1433 ton C (±27%) in EisenEx after 12 days (1 ton=10⁶ g). This similarity probably reflects the comparable size of the iron additions, as well as algal growth at a similar near-maximum growth rate in these regions. The findings imply that the different mixing regimes had less effect on the overall biological carbon uptake across the iron-enriched waters than suggested by the evolution of fCO₂ in surface water.     

Enhanced carbohydrate production by Southern Ocean phytoplankton in response to in situ iron fertilization

Storage carbohydrates (e.g., water-extractable β-1,3-d-glucan in diatoms) are of key importance for phytoplankton growth in a variable light climate, because they facilitate continued growth of the cells in darkness by providing energy and carbon skeletons for protein synthesis. Here, we tested the hypothesis that synthesis of storage carbohydrates by phytoplankton in the Southern Ocean is reduced by low iron and light availability. During the EisenEx/CARbon dioxide Uptake by the Southern Ocean (CARUSO) in situ iron enrichment experiment in the Atlantic sector of the Southern Ocean in November 2000, we studied the dynamics of water-extractable carbohydrates in the particulate fraction over the period of 3 weeks following the iron release. The areal amount (integral between 0- and 100-m depth) of carbohydrates increased from 1400 to 2300 mg m⁻² inside the iron-enriched patch, while remaining roughly constant in the surrounding waters. Most of the increase inside the patch was associated with the fraction of large (>10 μm) phytoplankton cells, consistent with the shift in the community structure towards larger diatoms. Deck incubations at 60% of the ambient irradiance revealed that the diurnal chlorophyll a (Chl a)-specific production rates of water-extractable polysaccharides were significantly higher for “in-patch” than for “out-patch” samples (0.5 vs. 0.3 μg C [μg Chl a]⁻¹ h⁻¹, respectively). Together with the higher photochemical efficiency of photosystem II (F_v/F_m), this indicates enhanced photosynthetic performance in response to iron fertilization. In addition, the nocturnal polysaccharide consumption rates were also enhanced by iron release, causing a striking increase in the diel dynamics of polysaccharide concentration. An iron-stimulated increase in diel dynamics was also observed in the fluorescence and size of pico- and nanophytoplankton cells (measured by flow cytometry) and is indicative of enhanced phytoplankton growth. Diurnal polysaccharide production by phytoplankton inside the patch was light-limited when they were incubated at intensities below ca. 200 μmol m⁻² s⁻¹ (daytime average). These irradiance levels correspond to those at 20- to 30-m depth in situ, whereas the upper mixed layer was frequently several-fold deeper due to storms. Therefore, these first measurements of phytoplankton carbohydrates during an in situ iron release experiment have revealed that both light and iron availability are the key factors controlling the synthesis of storage carbohydrates in phytoplankton and, hence, the development of diatom blooms in the Southern Ocean.     

The response of microbial communities to diverse organic matter sources in the Arctic Ocean

The response of Arctic microbial communities to a variety of natural organic matter substrates, including peat, ice algae and ice-rafted debris was examined using bacterial regrowth experiments and compared to unamended controls. Bacterial growth and production were followed together with the phylogenetic composition using length-heterogeneity polymerase chain reaction (LH-PCR), and 16S rRNA gene cloning and sequencing. Intact phospholipids (IPLs) and fatty acids evaluated the relationship between lipids and bacterial community structure and the impact of varied organic substrates on microbial lipid synthesis. Differential responses to organic matter sources were observed, with ice algae supporting both higher bacterial growth and production than terrestrial-derived peat. In spite of disparate growth kinetics, the community composition remained similar in all amended incubations as was confirmed by automated ribosomal intergenic spacer analysis (ARISA). Gammaproteobacteria dominated the initial incubations, whereas in extended incubations with terrestrial peat Alphaproteobacteria dominated; in particular Sulfitobacter phylotypes closely related (>99%) to an Arctic sea-ice-associated member of the Roseobacter clade (ARK10278). Arctic bacterioplankton preferentially synthesized two phospholipids, phosphatidylethanolamine (PE) and phosphatidylglygerol (PG), with 18:0n, 18:1Δ11, 16:0n and 16:1Δ9 as the primary fatty acids. Overall, results show that organic matter source plays an important role in structuring bacterioplankton community composition, with similar IPL and fatty acid lipid distributions observed among phylogenetically distinct bacteria.     

Responses of phytoplankton and heterotrophic bacteria in the northwest subarctic Pacific to in situ iron fertilization as estimated by HPLC pigment analysis and flow cytometry

To verify the hypothesis that the growth of phytoplankton in the Western Subarctic Gyre (WSG), which is located in the northwest subarctic Pacific, is suppressed by low iron (Fe) availability, an in situ Fe fertilization experiment was carried out in the summer of 2001. Changes over time in the abundance and community structure of phytoplankton were examined inside and outside an Fe patch using phytoplankton pigment markers analyzed by high-performance liquid chromatography (HPLC) and flow cytometry (FCM). In addition, the abundance of heterotrophic bacteria was also investigated by FCM. The chlorophyll a concentration was initially ca. 0.9 μg l⁻¹ in the surface mixed layer where diatoms and chlorophyll b-containing green algae (prasinophytes and chlorophytes) were predominant in the chlorophyll biomass. After the iron enrichment, the chlorophyll a concentration increased up to 9.1 μg l⁻¹ in the upper 10 m inside the Fe patch on Day 13. At the same time, the concentration of fucoxanthin (a diatom marker) increased 45-fold in the Fe patch, and diatoms accounted for a maximum 69% of the chlorophyll biomass. This result was consistent with a microscopic observation showing that the diatom Chaetoceros debilis had bloomed inside the Fe patch. However, chlorophyllide a concentrations also increased in the Fe patch with time, and reached a maximum of 2.2 μg l⁻¹ at 5 m depth on Day 13, suggesting that a marked abundance of senescent algal cells existed at the end of the experiment. The concentration of peridinin (a dinoflagellate marker) also reached a maximum 24-fold, and dinoflagellates had contributed significantly (>15%) to the chlorophyll biomass inside the Fe patch by the end of the experiment. Concentrations of 19′-hexanoyloxyfucoxanthin (a prymnesiophyte marker), 19′-butanoyloxyfucoxanthin (a pelagophyte marker), and alloxanthin (a cryptophyte marker) were only incremented a few-fold increment inside the Fe patch. On the contrary, chlorophyll b concentration reduced to almost half of the initial level in the upper 10 m water column inside the Fe patch at the end of the experiment. A decrease with time in the abundance of eukaryotic ultraphytoplankton (<ca. 5 μm in size), in which chlorophyll b-containing green algae were possibly included was also observed by FCM. Overall, our results indicate that Fe supply can dramatically alter the abundance and community structure of phytoplankton in the WSG. On the other hand, cell density of heterotrophic bacteria inside the Fe patch was maximum at only ca. 1.5-fold higher than that outside the Fe patch. This indicates that heterotrophic bacteria abundance was little respondent to the Fe enrichment.     

The response of phytoplankton to iron enrichment in Sub-Antarctic HNLCLSi waters: Results from the SAGE experiment

Areas of high nutrients and low chlorophyll a comprise nearly a third of the world’s oceans, including the equatorial Pacific, the Southern Ocean and the Sub-Arctic Pacific. The SOLAS Sea-Air Gas Exchange (SAGE) experiment was conducted in late summer, 2004, off the east coast of the South Island of New Zealand. The objective was to assess the response of phytoplankton in waters with low iron and silicic acid concentrations to iron enrichment. We monitored the quantum yield of photochemistry (F_v/F_m) with pulse amplitude modulated fluorometry, chlorophyll a, primary productivity, and taxonomic composition. Measurements of F_v/F_m indicated that the phytoplankton within the amended patch were relieved from iron stress (F_v/F_m approached 0.65). Although there was no significant difference between IN and OUT stations at points during the experiment, the eventual enhancement in chlorophyll a and primary productivity was twofold by the end of the 15-day patch occupation. However, no change in particulate carbon or nitrogen pools was detected. Enhancement in primary productivity and chlorophyll a were approximately equal for all phytoplankton size classes, resulting in a stable phytoplankton size distribution. Initial seed stocks of diatoms were extremely low, <1% of the assemblage based on HPLC pigment analysis, and did not respond to iron enrichment. The most dominant groups before and after iron enrichment were type 8 haptophytes and prasinophytes that were associated with ∼75% of chlorophyll a. Twofold enhancement of biomass estimated by flow cytometry was detected only in eukaryotic picoplankton, likely prasinophytes, type 8 haptophytes and/or pelagophytes. These results suggest that factors other than iron, such as silicic acid, light or physical disturbance limited the phytoplankton assemblage during the SAGE experiment. Furthermore, these results suggest that additional iron supply to the Sub-Antarctic under similar seasonal conditions and seed stock will most likely favor phytoplankton <2 ??m. This implies that any iron-mediated gain of fixed carbon will most likely be remineralized in shallow water rather than sink and be sequestered in the deep ocean.     

An in situ iron-enrichment experiment in the western subarctic Pacific (SEEDS): Introduction and summary

To test the iron hypothesis in the subarctic Pacific Ocean, an in situ iron-enrichment experiment (SEEDS) was performed in the western subarctic gyre in July–August 2001. About 350 kg of iron (as acidic iron sulfate) and 0.48 mol of the inert chemical tracer sulfur hexafluoride were introduced into a 10-m deep surface mixed layer over an 80 km² area. This single iron infusion raised dissolved iron levels to 2.9 nM initially. Dissolved iron concentrations rapidly decreased after the infusion, but levels remained close to 0.15 nM even at the end of the 14-day experimental period. During SEEDS there were iron-mediated increases in chlorophyll a concentrations (up to 20 μg l⁻¹), primary production rates, biomass and photosynthetic energy conversion efficiency relative to waters outside the iron-enriched patch. The rapid and very high accumulation of phytoplankton biomass in response to the iron addition appeared to be partly attributable to shallow mixed-layer depth and moderate water temperature in the western subarctic Pacific. However, the main reason was a floristic shift to fast-growing centric diatom Chaetoceros debilis, unlike the previous iron-enrichment experiments in the equatorial Pacific and the Southern Ocean, in both of which iron stimulated the growth of pennate diatoms. The iron-mediated blooming of diatoms resulted in a marked consumption of macronutrients and drawdown of pCO₂. Biological and physiological measurements indicate that phytoplankton growth in the patch became both light- and iron-limited, making phytoplankton biomass relatively constant after day 9. The increase in microzooplankton grazing rate after day 9 also influenced the net growth rate of phytoplankton. There was no significant increase in the export flux of carbon to depth during the 14-day occupation of the experimental site. The export flux between day 4 and day 13 was estimated to be only 13% of the integrated primary production in the iron-enriched patch. The major part of the carbon fixed by the diatom bloom remained in the surface mixed layer as biogenic particulate matter. Our findings support the hypothesis that iron limits phytoplankton growth and biomass in a ‘bottom up’ manner in this area, but the fate of algal carbon remains unknown.     

Prokaryotic community composition revealed by denaturing gradient gel electrophoresis in the East Sea

To understand the temporal and spatial variation of the prokaryotic community in the East Sea, their composition was determined by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE)-sequencing techniques. The investigations were conducted twice annually in 2007 and 2009 in coastal and offshore stations. Prokaryotic abundance (PA), leucine incorporation rate, and other environmental parameters were also measured. By using the DGGE approach, we obtained 283 bacterial sequences and 160 archaeal sequences. The most frequently detected bacterial phylotypes during the investigations belonged to Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes. However, their relative compositions differed in time and space. Although Alphaproteobacteria and Bacteroidetes were the dominant groups in the surface water in May 2007 and in May and October 2007, Gammaproteobacteria was dominant in mesopelagic samples. However, Gammaproteobacteria was overwhelmingly dominant in most samples in August 2009. Although Deltaproteobacteria was rarely found as a dominant bacterial group, it occupied the highest fraction in a mesopelagic sample in October 2007. Epsilonproteobacteria also showed a similar trend, although its maximal dominance was found in a mesopelagic sample in August 2009. The archaeal community was dominated overwhelmingly by members of the Euryarchaeota in most of the investigations. However, Nitrosopumilales was dominant in aphotic samples in August 2009. Further, their spatiotemporal composition at the family level changed more dynamically in the East Sea. These temporal and spatial distributions of the prokaryotic community were influenced mainly by seawater temperature and depth in the East Sea.     

Response of microzooplankton (protists and small copepods) to an iron-induced phytoplankton bloom in the Southern Ocean (EisenEx)

The dynamics, composition and grazing impact of microzooplankton were studied during the in situ iron fertilisation experiment EisenEx in the Antarctic Polar Frontal Zone in austral spring (November 2000). During the 21 day experiment, protozooplankton and small metazooplankton were sampled from the mixed layer inside and outside the patch using Niskin bottles. Aplastidic dinoflagellates increased threefold in abundance and biomass in the first 10 days of the experiment, but decreased thereafter to values twofold higher than pre-fertilisation values. The decline after day 10 is attributed to increasing grazing pressure by copepods. They also constrained ciliate abundances and biomass which were higher inside the fertilised patch than outside but highly variable. Copepod nauplii abundance remained stable whereas biomass doubled. Numbers of copepodites and adults of small copepod species (<1.5 mm) increased threefold inside the patch, but doubled in surrounding waters. Grazing rates estimated using the dilution method suggest that microzooplankton grazing constrained pico- and nanoplankton populations, but species capable of feeding on large diatoms (dinoflagellates and small copepods including possibly nauplii) were selectively predated by the metazoan community. Thus, iron fertilisation of a developing spring phytoplankton assemblage resulted in a trophic cascade which favoured dominance of the bloom by large diatoms.     

Microbial community structure and diversity in deep-sea hydrothermal vent sediments along the Eastern Lau Spreading Centre

The aim of this study is to investigate microbial structures and diversities in five active hydrothermal fields’ sediments along the Eastern Lau Spreading Centre (ELSC) in the Lau Basin (southwest Pacific). Microbial communities were surveyed by denatured gradient gel electrophoresis (DGGE) and clone library analysis of 16S rRNA genes. The differences in microbial community structures among sediment samples from the five deep-sea hydrothermal sites were revealed by DGGE profiles. Cluster analysis of DGGE profiles separated the five hydrothermal samples into two groups. Four different 16S rRNA gene clone libraries, representing two selected hydrothermal samples (19-4TVG8 and 19-4TVG11), were constructed. Twenty-three and 32 phylotypes were identified from 166 and 160 bacterial clones respectively, including Proteobacteria, Bacteroidetes, Firmicutes, Nitrospirae and Planctomycetes. The phylum Proteobacteria is dominant in both bacterial libraries with a predominance of Gamma-Proteobacteria. A total of 31 and 25 phylotypes were obtained from 160 and 130 archaeal clones respectively, including Miscellaneous Crenarchaeotic Group, Marine Group Ⅰ and Ⅲ, Marine Benthic Group E, Terrestrial Hot Spring Crenarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota. These results show a variety of clones related to those involved in sulfur cycling, suggesting that the cycling and utilization of sulfur compounds may extensively occur in the Lau Basin deep-sea hydrothermal ecosystem.     

Detection of ammonia-oxidizing Bacteria and Archaea within coral reef cyanobacterial mats

Coral-reef filamentous cyanobacterial mats are complex communities of microbes. We used DGGE and eubacterial and archaeal 16S rRNA and amoA ammonium monooxygenase subunit A primer sets to explore the presence of genes mediating nitrogen conversion within cyanobacterial mats from Hawaii. DGGE band analysis revealed the presence of halophiles such as Halococcus salifodinae and of Nitrosopumilus-like organisms. Twenty seven out of 31 clone sequences exhibited a 95 % or greater 16S rRNA gene identity with known archaeal ammonia oxidizers such as Nitrosopumilus maritimus. The presence of Archaea within those mats, as well as the co-occurrence of both ammonia-oxidizing Archaea and ammonia-oxidizing Bacteria suggests importance of the former in the cyanobacterial mat community, and suggests greater than anticipated diversity of nitrogen conversion processes and organisms in those systems.     

Diel microbial variations at a coastal Northern Adriatic station affected by Po River outflows

Diel sampling was performed during an early spring survey in the Northern Adriatic Sea at a coastal station off the Po River delta. Samples were taken every 6 h at spring tide maxima and minima in the sub-superficial layer, at the maximum fluorescence depth (∼3 m). Variations in microbial community structure and its processes were assessed by considering heterotrophic bacteria, picocyanobacteria, viruses, exoenzymatic activities, microphytoplankton, nanoplankton and bacterial/cyanobacterial Denaturing Gradient Gel Electrophoresis (DGGE) profiles. A considerable diatom bloom, mostly supported by Skeletonema marinoi was detected. All microbial parameters except viruses, showed a sinusoidal trend with a 12 h period; only picocyanobacteria expressed relative maxima during high tide, showing a phase in opposition to the other parameters. No substantial changes in DGGE band patterns were detected. Even though the results showed bacterial activities to be influenced by the phytoplankton bloom, all microbial parameters' diel trends (except viruses) preferentially followed the tidal fluctuation rather than the light:dark cycle.     

Processes governing the carbon chemistry during the SAGE experiment

Measurements of pCO₂, pH and alkalinity in the surface waters of an iron fertilised patch of sub-Antarctic water were made during SAGE (SOLAS SAGE: Surface-Ocean Lower Atmosphere Studies Air-Sea Gas Experiment). The iron addition induced a minor phytoplankton bloom, however the patch dynamics were dominated by physical processes which suppressed and masked the biological effects. The Lagrangian nature of the experiment allowed the carbonate chemistry in the patch to be followed for 15.5 days, and the relative importance of the biological and physical factors influencing the surface water pCO₂ was estimated. The pCO₂ of the surface waters of the patch increased from 327 ??atm prior to iron addition to 338 ??atm on Day 14, effects of vertical and horizontal mixing offset the 15 ??atm drawdown that would have occurred had the induced biological uptake been the sole factor to influence the pCO₂. The air-sea carbon flux calculated using the measured skin temperature and a piston velocity parameterisation determined during SAGE (Ho et al., 2006) was 98.5% of the flux determined using conventional bulk temperature measurement and the Wanninkhof (1992) piston velocity parameterisation. The skin temperature alone contributed to an 8% increase in the flux compared with that determined using bulk temperature.     

Population dynamics of phytoplankton, heterotrophic bacteria, and viruses during the spring bloom in the western subarctic Pacific

We characterized the community composition of phytoplankton in the western subarctic Pacific from the pre-bloom to the decline phase of the spring bloom with special reference to decreases in the silicic acid concentration in surface waters as an index for diatom bloom development. Furthermore, responses of heterotrophic bacteria and viruses to the spring bloom were also concomitantly investigated. Under pre-bloom conditions when nutrients were abundant but the surface mixed layer depth was relatively deep, chlorophyll (Chl) a concentrations were consistently low and green algae (chlorophytes and prasinophytes), cryptophytes, and diatoms were predominant in the phytoplankton assemblages as estimated by algal pigment signatures. Together with the shallowing of the mixed layer depth and the decrease in silicic acid concentration, diatoms bloomed remarkably in the Oyashio region, though the magnitude of the bloom in the Kuroshio-Oyashio transition (hereafter Transition) region was relatively small. A total of 77 diatom species were identified, with the bloom-forming diatoms mainly consisting of Thalassiosira, Chaetoceros, and Fragilariopsis species. It has become evident that the carotenoid fucoxanthin can serve as a strong indicator of the diatom carbon biomass during the spring diatom bloom. Differences in the species richness of diatoms among stations generally enabled us to separate the Oyashio bloom stations from the Transition and the Oyashio pre-bloom stations. Relatively high values of the Shannon-Wiener index for the diatom species were also maintained during the Oyashio bloom, indicating that a wide variety of species then shared dominance. In the decline phase of the Oyashio bloom when surface nutrient concentrations decreased, senescent diatom cells increased, as inferred from the levels of chlorophyllide a. Although the cell density of heterotrophic bacteria changed little with the development of the diatom bloom, viral abundance increased toward the end of the bloom, suggesting an increased likelihood of mortality among organisms including diatoms resulting from viral infection. This is the first report on the microbial trophodynamics, including viruses, during the spring diatom bloom in the western subarctic Pacific.     

Control of the phytoplankton response during the SAGE experiment: A synthesis

The SOLAS Air-Sea Gas Exchange (SAGE) experiment was conducted in Sub-Antarctic waters off the east coast of the South Island of New Zealand in the late summer of 2004. This mesoscale iron enrichment experiment was unique in that chlorophyll a (chl a) and primary productivity were only 2× OUT stations values toward the end of the experiment and this enhancement was due to increased activity of non-diatomaceous species. In addition, this enhancement in activity appeared to occur without a significant build up of particulate organic carbon. Picoeukaryotes (<2 ??m) were the only members of the phytoplankton assemblage that showed a statistically significant increase, a doubling in biomass. To better understand the controls of phytoplankton growth and biomass, we present results from a series of on-deck perturbation experiments conducted during SAGE. Results suggest that the pico-dominated phytoplankton assemblage was only weakly inhibited by iron. Diatoms with high growth rates comprised a small (<1%) fraction of the phytoplankton assemblage, were likely iron limited, and potentially further limited by silicic acid and therefore did not significantly contribute to bloom dynamics. On deck experiments and comparison of SAGE with other iron addition experiments suggested that neither light availability nor deep mixed layers limited phytoplankton growth. Although no substantial increase in grazing rate or specific phytoplankton growth rate was detected, microzooplankton biomass doubled over SAGE as a result of an increase in cell size. The importance of microzooplankton grazing was highlighted by the fact that they were capable of consuming 15-49% of the total phytoplankton production per day. Removal was highest on eukaryotic picophytoplankton production with a mean value of 72% (29-143%). Patch dilution played an important role during SAGE; the mean patch net algal growth:dilution rate, 1.13 (0.4-2.2) was the lowest reported for a mesoscale iron enrichment experiment. Phytoplankton biomass, estimated by chlorophyll a, only accumulated when phytoplankton growth exceeded grazing and when net algal growth exceeded dilution rate. The SAGE results highlight the function of the smallest phytoplankton size fraction described by the ecumenical Iron Hypothesis. Thus, adding iron to HNLC-low silicic acid regions during certain times of the year may simply transfer more carbon through the microbial food web. A primary implication of this study is that any iron-mediated gain in fixed carbon with this set of environmental conditions has a high probability of being recycled in surface waters.