Fuminori Hashihama

Macro-scale exhaustion of surface phosphate by dinitrogen fixation in the western North Pacific

[1]xa0In the subtropical oceans, nutrient concentrations are frequently below the detection limits of standard analytical methods. We applied a highly sensitive method to the surface water of the western and central Pacific between 42°N and 40°S and between 141°E and 158°W except in the equatorial zone, and detected overall depletion of nitrate + nitrite and an excess of SRP. However, a remarkable exception was found: an almost complete exhaustion of SRP ( 2000 km in the western subtropical North Pacific in both summer and winter. The SRP exhaustion was a consequence of an elevated dinitrogen fixation, which occurred in areas with high dust deposition from the Asian continent that likely enhanced SRP consumption. A coupling among nutrient dynamics, dinitrogen fixation and dust deposition produces the extremely low P availability spanning a large area, which appears to be unique to the western North Pacific.

Occurrence of rain‐origin nitrate patches at the nutrient‐depleted surface in the East China Sea and the Philippine Sea during summer

[1]xa0Nutrient concentrations at the nanomolar level were monitored in the East China Sea and the Philippine Sea during summer using a continuous underway system with a highly sensitive colorimetric method. Concentrations of both nitrate and soluble reactive phosphorus (SRP) varied generally at 1000 nM at a horizontal scale of <10 km to tens of kilometers compared with neighboring waters. These nitrate patches were grouped into three types according to the associated environmental conditions with (1) lowered salinity and no changes in SRP and in vivo chlorophyll fluorescence, (2) lowered temperature and elevations of SRP and in vivo chlorophyll fluorescence, and (3) no consistent environmental variations. The first type was ascribed to rainfall, which is rich in nitrate but phosphate free. The second type was formed near islands/landmasses and was ascribed to upwelling or intense vertical mixing. The third type was not characterized and was considered to include several sources. Nine out of 20 nitrate patches belonged to the first type. Within each of these nine patches, nitrate concentration was negatively correlated with salinity. By extrapolating this correlation, nitrate concentrations in the rainwater were estimated to be within 0.14–6.2 μM, which is well within the reported concentrations of nitrate in rainwater. On the basis of the estimated nitrate concentrations and amount of precipitation, nitrogen supply from rainfall in the study area was evaluated to be in the same order as that of diffusive upward flux and N2 fixation in summer.

Dissolved Phosphorus Pools and Alkaline Phosphatase Activity in the Euphotic Zone of the Western North Pacific Ocean

We measured pools of dissolved phosphorus (P), including dissolved inorganic P (DIP), dissolved organic P (DOP) and alkaline phosphatase (AP)-hydrolyzable labile DOP (L-DOP), and kinetic parameters of AP activity (APA) in the euphotic zone in the western North Pacific Ocean. Samples were collected from one coastal station in Sagami Bay, Japan, and three offshore stations between the North Pacific subtropical gyre (NPSG) and the Kuroshio region. Although DIP concentrations in the euphotic zone at all stations were equally low, around the nominal method detection limit of 20u2009nmolu2009L-1, chlorophyll a (Chl a) concentrations were one order of magnitude greater at the coastal station. DOP was the dominant P pool, comprising 62–92% of total dissolved P at and above the Chl a maximum layer (CML). L-DOP represented 22–39% of the total DOP at the offshore stations, whereas it accounted for a much higher proportion (about 85%) in the coastal surface layers. Significant correlations between maximum potential AP hydrolysis rates and DIP concentrations or bacterial cell abundance in the offshore euphotic zone suggest that major APA in the oligotrophic surface ocean is from bacterial activity and regulated largely by DIP availability. Although the range of maximum potential APA was comparable among the environmental conditions, the in situ hydrolysis rate of L-DOP in the coastal station was 10 times those in the offshore stations. L-DOP turnover time at the CML ranged from 4.5u2009days at the coastal station to 84.4u2009days in the NPSG. The ratio of the APA half-saturation constant to the ambient L-DOP concentration decreased markedly from the NPSG to the coastal station. There were substantial differences in the rate and efficiency of DOP remineralization and its contribution as the potential P source between the low-phosphate/high-biomass coastal ecosystem and the low-phosphate/low biomass oligotrophic ocean.

Nitrification and its influence on biogeochemical cycles from the equatorial Pacific to the Arctic Ocean

We examined nitrification in the euphotic zone, its impact on the nitrogen cycles, and the controlling factors along a 7500u2009km transect from the equatorial Pacific Ocean to the Arctic Ocean. Ammonia oxidation occurred in the euphotic zone at most of the stations. The gene and transcript abundances for ammonia oxidation indicated that the shallow clade archaea were the major ammonia oxidizers throughout the study regions. Ammonia oxidation accounted for up to 87.4% (average 55.6%) of the rate of nitrate assimilation in the subtropical oligotrophic region. However, in the shallow Bering and Chukchi sea shelves (bottom ⩽67u2009m), the percentage was small (0–4.74%) because ammonia oxidation and the abundance of ammonia oxidizers were low, the light environment being one possible explanation for the low activity. With the exception of the shallow bottom stations, depth-integrated ammonia oxidation was positively correlated with depth-integrated primary production. Ammonia oxidation was low in the high-nutrient low-chlorophyll subarctic region and high in the Bering Sea Green Belt, and primary production in both was influenced by micronutrient supply. An ammonium kinetics experiment demonstrated that ammonia oxidation did not increase significantly with the addition of 31–1560u2009nm ammonium at most stations except in the Bering Sea Green Belt. Thus, the relationship between ammonia oxidation and primary production does not simply indicate that ammonia oxidation increased with ammonium supply through decomposition of organic matter produced by primary production but that ammonia oxidation might also be controlled by micronutrient availability as with primary production.

Liquid waveguide spectrophotometric measurement of nanomolar ammonium in seawater based on the indophenol reaction with o-phenylphenol (OPP)

We describe a highly sensitive colorimetric method for the determination of nanomolar concentrations of ammonium in seawater based on the indophenol reaction with o-phenylphenol [(1,1-biphenyl)-2-ol, abbreviated as OPP]. OPP is available as non-toxic, stable flaky crystals with no caustic odor and has some advantages over phenol in practical use. The method was established by using a gas-segmented continuous flow analyzer equipped with two types of long path liquid waveguide capillary cell, LWCCs (100 cm and 200 cm) and an UltraPath (200 cm), which have inner diameters of 0.55 mm and 2 mm, respectively. The reagent concentrations, flow rates of the pumping tubes, and reaction path and temperature were determined on the basis of a manual indophenol blue method with OPP (Kanda, Water Res. 29 (1995) 2746-2750). The sample mixed with reagents that form indophenol blue dye was measured at 670 nm. Aged subtropical surface water was used as a blank, a matrix of standards, and the carrier. The detection limits of the analytical systems with a 100 cm LWCC, a 200 cm LWCC, and a 200 cm UltraPath were 6, 4, and 4 nM, respectively. These systems had high precision (<4% at 100 nM) and a linear dynamic range up to 200 nM. Non-linear baseline drift did not occur when using the UltraPath system. This is due to the elimination of cell clogging because of the larger inner diameter of the UltraPath compared to the LWCCs. The UltraPath system is thus more suitable for long-term measurements compared with the LWCC systems. The results of the proposed sensitive colorimetry and a conventional colorimetry for the determination of seawater samples showed no significant difference. The proposed analytical systems were applied to underway surface monitoring and vertical observation in the oligotrophic South Pacific.

Sensitive determination of enzymatically labile dissolved organic phosphorus and its vertical profiles in the oligotrophic western North Pacific and East China Sea

Trace concentrations of labile dissolved organic phosphorus (LDOP) in oligotrophic seawater were measured by use of an enzymatic procedure and a nanomolar phosphate analytical system consisting of a gas-segmented continuous flow analyzer with a liquid waveguide capillary cell. LDOP, defined as DOP hydrolyzed by alkaline phosphatase (AP) from Escherichia coli, was quantified as the difference between the phosphate concentrations of the seawater sample with and without AP treatment. For sensitive measurement of LDOP, we found that phosphate contamination derived from commercially available AP must be corrected, and azide treatment before AP treatment proved effective in removing biological effect that occurs during DOP hydrolysis. Field observations at six stations of the western North Pacific and the East China Sea during the boreal summer revealed that, in the upper 200xa0m of the water column, LDOP concentrations ranged from the detection limit (3xa0nM) to 243xa0nM, and phosphate concentrations ranged from 5 to 374xa0nM. The spatial distribution patterns of LDOP were similar to those of phosphate. Most of the depth profiles for LDOP and phosphate showed concentrations were extremely low, <25xa0nM, between the surface and the deep chlorophyll maximum layer (DCML) and increased below the DCML. Strongly depleted LDOP and phosphate above the DCML suggest that LDOP is actively hydrolyzed under phosphate-depleted conditions and utilized by microbes.

Basin scale variability of active diazotrophs and nitrogen fixation in the North Pacific, from the tropics to the subarctic Bering Sea

Nitrogen-fixing micro-organisms (diazotrophs) provide biologically available nitrogen to plankton communities and thereby greatly influence the productivity in many marine regions. Various cyanobacterial groups have traditionally been considered the major oceanic diazotrophs, but later non-cyanobacterial and presumably heterotrophic diazotrophs were also found to be widespread and potentially important in nitrogen fixation. However, the distribution and activity of different diazotroph groups is still poorly constrained for most oceanic ecosystems. Here, we examined diazotroph community structure and activity along a 7,500-km south-north transect between the central equatorial Pacific and the Bering Sea. Nitrogen fixation contributed up to 84% of new production in the upper waters of the subtropical gyre, where the diazotroph community included the gammaproteobacterium γ-24774A11 and highly active cyanobacterial phylotypes (>50% of total nifH transcript abundance). Nitrogen fixation was sometimes detectable down to 150u2009m depth and extended horizontally to the edge of the gyre at around 35°N. Nitrogen fixation was even detected far north on the Bering Sea shelf. In the Alaskan Coastal Waters on the Bering Sea shelf, low nitrate together with high dissolved iron concentrations seemed to foster diazotroph growth, including a prominent role of UCYN-A2, which was abundant near the surface (1.2u2009×u2009105 nifH gene copies L-1). Our study provides evidence for nitrogen fixation in the Bering Sea and suggests a clear contrast in the composition of diazotrophs between the tropical/subtropical gyre and the separate waters in the cold northern regions of the North Pacific.

Sensitive determination of total particulate phosphorus and particulate inorganic phosphorus in seawater using liquid waveguide spectrophotometry.

Determining the total particulate phosphorus (TPP) and particulate inorganic phosphorus (PIP) in oligotrophic oceanic water generally requires the filtration of a large amount of water sample. This paper describes methods that require small filtration volumes for determining the TPP and PIP concentrations. The methods were devised by validating or improving conventional sample processing and by applying highly sensitive liquid waveguide spectrophotometry to the measurements of oxidized or acid-extracted phosphate from TPP and PIP, respectively. The oxidation of TPP was performed by a chemical wet oxidation method using 3% potassium persulfate. The acid extraction of PIP was initially carried out based on the conventional extraction methodology, which requires 1M HCl, followed by the procedure for decreasing acidity. While the conventional procedure for acid removal requires a ten-fold dilution of the 1M HCl extract with purified water, the improved procedure proposed in this study uses 8M NaOH solution for neutralizing 1M HCl extract in order to reduce the dilution effect. An experiment for comparing the absorbances of the phosphate standard dissolved in 0.1M HCl and of that dissolved in a neutralized solution [1M HCl: 8M NaOH=8:1 (v:v)] exhibited a higher absorbance in the neutralized solution. This indicated that the improved procedure completely removed the acid effect, which reduces the sensitivity of the phosphate measurement. Application to an ultraoligotrophic water sample showed that the TPP concentration in a 1075mL-filtered sample was 8.4nM with a coefficient of variation (CV) of 4.3% and the PIP concentration in a 2300mL-filtered sample was 1.3nM with a CV of 6.1%. Based on the detection limit (3nM) of the sensitive phosphate measurement and the ambient TPP and PIP concentrations of the ultraoligotrophic water, the minimum filtration volumes required for the detection of TPP and PIP were estimated to be 15 and 52mL, respectively.

Comparison of community structures between particle-associated and free-living prokaryotes in tropical and subtropical Pacific Ocean surface waters

The subtropical and tropical regions of the Pacific Ocean are less productive than other oceanic regions. Although particle association should be an important strategy for heterotrophic prokaryotes to survive in such environments, we have little information on particle-associated (PA) prokaryotes in these regions. The specific aim of this study was to determine bacterial and archaeal community structures in the PA assemblage in comparison to the free-living (FL) assemblage in the North Pacific Subtropical Gyre, the South Pacific Subtropical Gyre, and an eastern equatorial region of the Pacific Ocean. Community profiles and phylogenetic identities were obtained by denaturing gradient gel electrophoresis, 454-pyrosequencing, and cloning followed by Sanger sequencing of 16Sr RNA gene amplicons. The distribution patterns of some abundant groups in three regions and two lifestyles (PA and FL) are shown in this study. Also, the PA community structures of bacteria differed from the FL ones and exhibited higher diversity than the FL ones, while the archaeal community structures did not show significant differences between PA and FL assemblages. We found that specific phylotypes of Gammaproteobacteria and Flavobacteria were abundant in PA bacterial assemblages, suggesting that they prefer to attach and consume particulate organic matter. In summary, the surface seawater PA assemblages represent very different bacterial and archaeal community structures between three different oceanic regions, each of which had distinct PA and FL community structures. These results imply that environmental factors determine microbial community structures.

Basin-scale distribution of NH4+ and NO2− in the Pacific Ocean

We used more than 25,000 nutrient samples to elucidate for the first time basin-scale distributions and seasonal changes of surface ammonium (NH4+) and nitrite (NO2−) concentrations in the Pacific Ocean. The highest NH4+, NO2−, and nitrate (NO3−) concentrations were observed north of 40°N, in the coastal upwelling region off the coast of Mexico, and in the Tasman Sea. NH4+ concentrations were elevated during May–October in the western subarctic North Pacific, May–December in the eastern subarctic North Pacific, and June–September in the subtropical South Pacific. NO2− concentrations were highest in winter in both hemispheres. The seasonal cycle of NH4+ was synchronous with NO2−, NO3−, and satellite chlorophyll a concentrations in the western subtropical South Pacific, whereas it was synchronous with chlorophyll-a but out of phase with NO2− and NO3− in the subarctic regions.