Emma K. Wear

Dissolved organic carbon (DOC) forms the largest reservoir of reduced carbon in the ocean, yet its bioavailability and the microbial processes governing its transformation remain poorly constrained. Traditional seawater batch cultures have been indispensable for measuring bioavailable DOC (BDOC), but their long incubation times and limited throughput constrain comparisons across environmental gradients and treatments. Here, we adapted marine plug flow bioreactors, glass columns packed with inert glass beads colonized by natural microbial assemblages, to provide a high‐throughput operational assay of potential DOC bioavailability. After approximately 6 months of conditioning, single‐column bioreactor assays consistently removed DOC from coastal surface waters, deep seawater, and surface seawater amended with diatom‐derived organic matter within 4–7 hr. Linking bioreactors in series increased cumulative DOC removal although the number of columns contributing additional removal varied among source waters. In paired experiments, terminal bioreactor DOC removal equaled or exceeded that measured at the final sampling point of seawater batch cultures incubated for weeks to months. Bioreactor‐associated microbial communities were enriched in Planctomycetota and Thermoproteota relative to the free‐living communities in batch cultures, reflecting the distinct microbial assemblages supported by the two approaches. Because bioreactors operate under high‐biomass, attached community conditions that differ from the open ocean, their measurements represent internally comparable estimates of potential DOC bioavailability rather than realized in situ utilization. Used alongside seawater batch cultures, marine bioreactors provide a complementary framework for distinguishing potential DOC bioavailability from realized DOC utilization.

Temperate lakes experience variation in mixing and stratification that affects the distributions, activities, abundances, and diversity of plankton communities. We examined temporal and vertical changes in the composition of planktonic microorganisms (including Bacteria and Archaea) in oligotrophic Flathead Lake, Montana. Using a combination of approaches that included 16S rRNA gene sequencing and flow cytometric determination of cell abundances, we found that the microbial community was responsive to variations in stratification and mixing at time scales ranging from episodic (scale of days) to seasonal. However, the impact of such physical dynamics varied among taxa, likely reflecting taxa‐specific responses to environmental changes that coincide with stratification and mixing (e.g., light availability and nutrient supply). During the early spring, periods of relatively short‐term (< 7 d) intermittency in stratification and mixing influenced the vertical distributions of specific microbial taxa, notably including the cyanobacteria. These events highlight time scales of biological responses to high‐frequency variations associated with lake stratification and mixing, particularly during the transition to the growing season in the early spring.

Quantifying phytoplankton composition is critical to predicting marine ecosystem structure and function. DNA meta‐barcoding and high‐performance liquid chromatography (HPLC) pigment analysis are two widely used methods for assessing phytoplankton composition; however, comparing their performance has been done only rarely. Here, we integrate DNA meta‐barcoding and HPLC pigment observations to determine eukaryotic phytoplankton composition in the Santa Barbara Channel, California. We find that both methods identify the same four dominant eukaryotic phytoplankton taxa (diatoms, dinoflagellates, chlorophytes, and prymnesiophytes), but inter‐ and intra‐lineage variability in biomarker pigmentation (associated with both a lack of taxonomic specificity of biomarker pigments and intrinsic differences in accessory pigmentation) drives substantial disagreement between the methods. Covariation network analysis circumvents this disagreement and reveals that diverse assemblages of phytoplankton and other protists covary with distinct suites of biomarker pigments. Our results highlight the strengths and weaknesses of each method in characterizing phytoplankton composition and reveal novel insights into phytoplankton physiology that could only be gained by integrating the two methods. Finally, we suggest a path to monitor eukaryotic plankton communities on unprecedented spatiotemporal scales based on the covariation of unique phytoplankton and protistan assemblages with remotely sensible phytoplankton pigment concentrations.

Summary: Coral reefs are highly productive ecosystems with distinct biogeochemistry and biology nestled within unproductive oligotrophic gyres. Coral reef islands have often been associated with a nearshore enhancement in phytoplankton, a phenomenon known as the Island Mass Effect (IME). Despite being documented more than 60 years ago, much remains unknown about the extent and drivers of IMEs. Here we utilized 16S rRNA gene metabarcoding as a biological tracer to elucidate horizontal and vertical influence of an IME around the islands of Mo′orea and Tahiti, French Polynesia. We show that those nearshore oceanic stations with elevated chlorophyll a included bacterioplankton found in high abundance in the reef environment, suggesting advection of reef water is the source of altered nearshore biogeochemistry. We also observed communities in the nearshore deep chlorophyll maximum (DCM) with enhanced abundances of upper euphotic bacterioplankton that correlated with intrusions of low‐density, O ₂ rich water, suggesting island influence extends into the DCM.

In stratified oligotrophic waters, phytoplankton communities forming the deep chlorophyll maximum (DCM) are isolated from atmospheric iron sources above and remineralized iron sources below. Reduced supply leads to a minimum in dissolved iron (dFe) near 100 m, but it is unclear if iron limits growth at the DCM. Here, we propose that natural iron addition events occur regularly with the passage of mesoscale eddies, which alter the supply of dFe and other nutrients relative to the availability of light, and can be used to test for iron limitation at the DCM. This framework is applied to two eddies sampled in the North Pacific Subtropical Gyre. Observations in an anticyclonic eddy center indicated downwelling of iron‐rich surface waters, leading to increased dFe at the DCM but no increase in productivity. In contrast, uplift of isopycnals within a cyclonic eddy center increased supply of both nitrate and dFe to the DCM, and led to dominance of picoeukaryotic phytoplankton. Iron addition experiments did not increase productivity in either eddy, but significant enhancement of leucine incorporation in the light was observed in the cyclonic eddy, a potential indicator of iron stress among Prochlorococcus. Rapid cycling of siderophores and low dFe:nitrate uptake ratios also indicate that a portion of the microbial community was stressed by low iron. However, near‐complete nitrate drawdown in this eddy, which represents an extreme case in nutrient supply compared to nearby Hawaii Ocean Time‐series observations, suggests that recycling of dFe in oligotrophic ecosystems is sufficient to avoid iron limitation in the DCM under typical conditions.

In stratified oligotrophic waters, phytoplankton communities forming the deep chlorophyll maximum (DCM) are isolated from atmospheric iron sources above and remineralized iron below. Reduced supply leads to a minimum in dissolved iron (dFe) near 100 m, but it is unclear if iron limits growth at the DCM. Here, we propose that natural iron addition events occur regularly with the passage of mesoscale eddies, which alter the supply of dFe and other nutrients relative to the supply of light, and can be used to test for iron limitation at the DCM. This framework is applied to two eddies sampled in the North Pacific Subtropical Gyre. Observations in an anticyclonic eddy center indicated downwelling of iron-rich surface waters, leading to increased dFe at the DCM but no increase in productivity. In contrast, uplift of isopycnals within a cyclonic eddy center increased supply of both nitrate and dFe to the DCM, and led to dominance of picoeukaryotic phytoplankton. Iron addition experiments did not increase productivity in either eddy, but did enhance leucine incorporation at ambient light in the cyclonic eddy, a potential indicator of iron stress among Prochlorococcus . Rapid cycling of siderophores and low dFe:nitrate uptake ratios also indicate that a portion of the microbial community was stressed by low iron. However, near-complete nitrate drawdown in this eddy, which represents an extreme case in nutrient supply compared to nearby Hawaii Ocean Time-series observations, suggests that recycling of dFe in oligotrophic ecosystems is sufficient to avoid iron limitation in the DCM under typical conditions.

Sinking particulate organic matter controls the flux of carbon (C) from the surface ocean to the deep sea. Microorganisms actively colonize particles, but the extent to which microbial metabolism influences particle export remains uncertain. We conducted experiments to quantify rates of bacterial production (derived based on ³H‐leucine incorporation) and dark C‐fixation (based on ¹⁴C‐bicarbonate assimilation) associated with sinking particles collected from the base of the euphotic zone (175 m) in the subtropical North Pacific Ocean. Seawater was amended with sinking particles and rates of filter size‐fractionated (0.2, 2, and 20 μm) bacterial production and dark C‐fixation were measured. Sequencing of 16S ribosomal RNA (rRNA) gene amplicons revealed that microorganisms in the particle‐amended treatments differed from those in the unamended seawater controls, with the particle treatments enriched in putative copiotrophic bacteria. The addition of sinking particles increased rates of bacterial production (by 6‐ to 9‐fold), and to a lesser extent dark C‐fixation (by 1.7‐ to 4.6‐fold), relative to unamended controls, with most of the production associated with filter pore sizes < 20 μm. Normalizing production to concentrations of particulate C yielded rates that were statistically indistinguishable between particle‐amended treatments and unamended controls. We then examined possible relationships between sinking particulate C flux attenuation and its supply to the mesopelagic waters, revealing that flux attenuation was positively related to increases in particulate C supply. Together with results from our experiments, we suggest processes that contribute to sinking particle disaggregation both increase flux attenuation and favor microbial mineralization of particle‐derived organic matter.

Mesoscale eddies, prominent physical processes in the stratified ocean gyres, affect community composition and metabolic rates of both phytoplankton and heterotrophic bacterioplankton (free‐living bacterial and archaeal) communities. We hypothesized that in situ differences in organic matter production would predispose bacterioplankton communities from cyclonic vs. anticyclonic eddies toward metabolic capabilities better suited to utilizing dissolved organic matter (DOM) from the phytoplankton groups commonly associated with each eddy polarity. To test this, we established dilution batch‐culture bioassay incubations along a cyclone to anticyclone spatial transect in the North Pacific Subtropical Gyre. Unamended incubations, to assess spatial variability in ambient DOM bioavailability, and incubations amended with lysates of phytoplankton cultures were established and community growth and metabolic responses were assessed. Over timescales of days, lysate type was more influential than incubation origin: Prochlorococcus lysate was rapidly remineralized, while Emiliania huxleyi lysate was efficiently incorporated into biomass and developed a unique community of copiotrophic bacteria. Over timescales of 1 week to 1 month, eddy effects were indirectly apparent in their potential to promote metabolic processes related to DOM production and consumption. Surface lysate incubations showed priming of ambient DOM, that is, the remineralization of DOM, which was otherwise not bioavailable, in the presence of labile substrates. Some incubations originating from the deep chlorophyll maximum demonstrated signatures of chemoautotrophy fueled by nitrification, coincident with eddy‐driven isopycnal uplift. We conclude that eddy polarity itself does not determine community‐level bacterioplankton metabolic capabilities; however, mesoscale processes may indirectly affect slower, semi‐labile organic matter processing in the oligotrophic ocean.

Summary: Primers targeting the 16S small subunit ribosomal RNA marker gene, used to characterize bacterial and archaeal communities, have recently been re‐evaluated for marine planktonic habitats. To investigate whether primer selection affects the ecological interpretation of bacterioplankton populations and community dynamics, amplicon sequencing with four primer sets targeting several hypervariable regions of the 16S rRNA gene was conducted on both mock communities constructed from cloned 16S rRNA genes and a time‐series of DNA samples from the temperate coastal Santa Barbara Channel. Ecological interpretations of community structure (delineation of depth and seasonality, correlations with environmental factors) were similar across primer sets, while population dynamics varied. We observed substantial differences in relative abundances of taxa known to be poorly resolved by some primer sets, such as Thaumarchaeota and SAR11, and unexpected taxa including Roseobacter clades. Though the magnitude of relative abundances of common OTUs differed between primer sets, the relative abundances of the OTUs were nonetheless strongly correlated. We do not endorse one primer set but rather enumerate strengths and weaknesses to facilitate selection appropriate to a system or experimental goal. While 16S rRNA gene primer bias suggests caution in assessing quantitative population dynamics, community dynamics appear robust across studies using different primers.