Characterization of the molecular mechanisms of silicon uptake in coccolithophores.

Coccolithophores are an important group of calcifying marine phytoplankton. Although coccolithophores are not silicified, some species exhibit a requirement for Si in the calcification process. These species also possess a novel protein (SITL) that resembles the SIT family of Si transporters found in diatoms. However, the nature of Si transport in coccolithophores is not yet known, making it difficult to determine the wider role of Si in coccolithophore biology. Here, we show that coccolithophore SITLs act as Na+ -coupled Si transporters when expressed in heterologous systems and exhibit similar characteristics to diatom SITs. We find that CbSITL from Coccolithus braarudii is transcriptionally regulated by Si availability and is expressed in environmental coccolithophore populations. However, the Si requirement of C. braarudii and other coccolithophores is very low, with transport rates of exogenous Si below the level of detection in sensitive assays of Si transport. As coccoliths contain only low levels of Si, we propose that Si acts to support the calcification process, rather than forming a structural component of the coccolith itself. Si is therefore acting as a micronutrient in coccolithophores and natural populations are only likely to experience Si limitation in circumstances where dissolved silicon (DSi) is depleted to extreme levels.

Diminished carbon and nitrate assimilation drive changes in diatom elemental stoichiometry independent of silicification in an iron-limited assemblage.

In the California Current Ecosystem, upwelled water low in dissolved iron (Fe) can limit phytoplankton growth, altering the elemental stoichiometry of the particulate matter and dissolved macronutrients. Iron-limited diatoms can increase biogenic silica (bSi) content >2-fold relative to that of particulate organic carbon (C) and nitrogen (N), which has implications for carbon export efficiency given the ballasted nature of the silica-based diatom cell wall. Understanding the molecular and physiological drivers of this altered cellular stoichiometry would foster a predictive understanding of how low Fe affects diatom carbon export. In an artificial upwelling experiment, water from 96 m depth was incubated shipboard and left untreated or amended with dissolved Fe or the Fe-binding siderophore desferrioxamine-B (+DFB) to induce Fe-limitation. After 120 h, diatoms dominated the communities in all treatments and displayed hallmark signatures of Fe-limitation in the +DFB treatment, including elevated particulate Si:C and Si:N ratios. Single-cell, taxon-resolved measurements revealed no increase in bSi content during Fe-limitation despite higher transcript abundance of silicon transporters and silicanin-1. Based on these findings we posit that the observed increase in bSi relative to C and N was primarily due to reductions in C fixation and N assimilation, driven by lower transcript expression of key Fe-dependent genes.

Diatom populations in an upwelling environment decrease silica content to avoid growth limitation.

A mix of adaptive strategies enable diatoms to sustain rapid growth in dynamic ocean regions, making diatoms one of the most productive primary producers in the world. We illustrate one such strategy off coastal California that facilitates continued, high, cell division rates despite silicic acid stress. Using a fluorescent dye to measure single-cell diatom silica production rates, silicification (silica per unit area) and growth rates we show diatoms decrease silicification and maintain growth rate when silicon concentration limits silica production rates. While this physiological response to silicon stress was similar across taxa, in situ silicic acid concentration limited silica production rates by varying degrees for taxa within the same community. Despite this variability among taxa, silicon stress did not alter the contribution of specific taxa to total community silica production or to community composition. Maintenance of division rate at the expense of frustule thickness decreases cell density which could affect regional biogeochemical cycles. The reduction in frustule silicification also creates an ecological tradeoff: thinner frustules increase susceptibility to predation but reducing Si quotas maximizes cell abundance for a given pulse of silicic acid, thereby favouring a larger eventual population size which facilitates diatom persistence in habitats with pulsed resource supplies.

Taxon-specific contributions to silica production in natural diatom assemblages.

The metabolic activity and growth of phytoplankton taxa drives their ecological function and contribution to biogeochemical processes. We present the first quantitative, taxon-resolved silica production rates, growth rates, and silica content estimates for co-occurring diatoms along two cross-shelf transects off the California coast using the fluorescent tracer PDMPO (2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole), and confocal microscopy. Taxon contribution to total diatom community silica production was predominantly a function of the surface area of new frustule that each taxon created as opposed to cell abundance or frustule thickness. The influential role of surface area made large diatoms disproportionately important to community silica production over short time scales (<1 d). In some cases, large taxa that comprised only ~15% of numerical cell abundance accounted for over 50% of total community silica production. Over longer time scales relevant to bloom dynamics, the importance of surface area declines and growth rate becomes the dominant influence on contribution to production. The relative importance of surface area and growth rate in relation to silica production was modeled as the time needed for a smaller, faster-growing taxon to create more surface area than a larger, slower-growing taxon. Differences in growth rate between the taxa effected the model outcome more than differences in surface area. Shifts in relative silica production among taxa are time restricted by finite resources that limit the duration of a bloom. These patterns offer clues as to how taxa respond to their environment and the consequences for both species succession and the potential diatom contribution to elemental cycling.

Evaluating Carbonate System Algorithms in a Nearshore System: Does Total Alkalinity Matter?

Ocean acidification is a threat to many marine organisms, especially those that use calcium carbonate to form their shells and skeletons. The ability to accurately measure the carbonate system is the first step in characterizing the drivers behind this threat. Due to logistical realities, regular carbonate system sampling is not possible in many nearshore ocean habitats, particularly in remote, difficult-to-access locations. The ability to autonomously measure the carbonate system in situ relieves many of the logistical challenges; however, it is not always possible to measure the two required carbonate parameters autonomously. Observed relationships between sea surface salinity and total alkalinity can frequently provide a second carbonate parameter thus allowing for the calculation of the entire carbonate system. Here, we assessed the rigor of estimating total alkalinity from salinity at a depth <15 m by routinely sampling water from a pier in southern California for several carbonate system parameters. Carbonate system parameters based on measured values were compared with those based on estimated TA values. Total alkalinity was not predictable from salinity or from a combination of salinity and temperature at this site. However, dissolved inorganic carbon and the calcium carbonate saturation state of these nearshore surface waters could both be estimated within on average 5% of measured values using measured pH and salinity-derived or regionally averaged total alkalinity. Thus we find that the autonomous measurement of pH and salinity can be used to monitor trends in coastal changes in DIC and saturation state and be a useful method for high-frequency, long-term monitoring of ocean acidification.

Quantifying diatom silicification with the fluorescent dye, PDMPO.

Diatoms require silicic acid to construct ornately detailed cell walls called frustules. The growth and geographic distribution of diatoms is often controlled by the availability of silicic acid. Analytical methods exist to assess diatom community biogenic silica (bSiO2) production, but partitioning production among taxa has been largely qualitative. We present a method for the quantitative analysis of taxa-specific silica production through labeling diatoms with the fluorescent dye PDMPO [2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole]. To make PDMPO a quantitative tool: diatom frustules were solubilized to assess the total diatom community incorporation by quantitation of PDMPO fluorescence using a fluorometer, and laser confocal microscopy was used to quantify the fluorescence of PDMPO in single diatom cells. We created a fluorescence standard to intercalibrate the raw fluorescence signals of the fluorometer and microscope and to determine the fluorescence per mole of PDMPO. PDMPO incorporation was converted to silica production using diatom bSiO2:PDMPO incorporation ratios which varied systematically with silicic acid concentration. Above 3 µM Si(OH)4, bSiO2:PDMPO was constant and PDMPO incorporation was converted to silica production using a mole ratio of 2,916 as determined from cultures. Below 3 µM, the ratio was a linear function of [Si(OH)4] (bSiO2:PDMPO = 912.6 × [Si(OH)4]), as determined using data from two oceanographic cruises. Field evaluation of the method showed that total community PDMPO incorporation generally agreed to within 30% of radioisotope-determined silica production. This PDMPO method has the potential to be a powerful tool for understanding physiology, silicification and resource competition among diatom taxa.