Trichodesmium IMS101 Research Articles

Correlation of methane production with physiological traits in Trichodesmium IMS 101 grown with methylphosphonate at different temperatures.

The diazotrophic cyanobacterium Trichodesmium has been recognized as a potentially significant contributor to aerobic methane generation via several mechanisms including the utilization of methylphophonate (MPn) as a source of phosphorus. Currently, there is no information about how environmental factors regulate methane production by Trichodesmium. Here, we grew Trichodesmium IMS101 at five temperatures ranging from 16 to 31°C, and found that its methane production rates increased with rising temperatures to peak (1.028 ± 0.040 nmol CH4 μmol POC-1 day-1) at 27°C, and then declined. Its specific growth rate changed from 0.03 ± 0.01 d-1 to 0.34 ± 0.02 d-1, with the optimal growth temperature identified between 27 and 31°C. Within the tested temperature range the Q10 for the methane production rate was 4.6 ± 0.7, indicating a high sensitivity to thermal changes. In parallel, the methane production rates showed robust positive correlations with the assimilation rates of carbon, nitrogen, and phosphorus, resulting in the methane production quotients (molar ratio of carbon, nitrogen, or phosphorus assimilated to methane produced) of 227-494 for carbon, 40-128 for nitrogen, and 1.8-3.4 for phosphorus within the tested temperature range. Based on the experimental data, we estimated that the methane released from Trichodesmium can offset about 1% of its CO2 mitigation effects.

Environmental and nutrient controls of marine nitrogen fixation

Biological dinitrogen (N2) fixation by diazotrophic cyanobacteria has great biogeochemical implications in nitrogen (N) cycling in the ocean as this process represents the major source of new N input to the oceans, thereby controlling the marine primary productivity. Numerous factors can affect the extent of N2 fixation. A better understanding of the major environmental and nutrient factors governing this process is highly required. Iron (Fe) and/or phosphorus (P) are thought to be limiting factors in most oceanic regions. Special attention has been given in the present study to evaluate the effects of mineral dust deposition which is believed to stimulate N2 fixation as it increases the availability of both Fe and P. Through three laboratory bioassays (+Fe, +P, +Dust) via incubation experiments performed on Trichodesmium IMS101, we found that each addition of Fe, P or desert dust could stimulate the growth and N2 fixation of Trichodesmium IMS101. Several adaptive nutrient utilization strategies were observed, such as a Fe luxury uptake mechanism, a P-sparing effect and colony formation. In addition, during a field study using natural phytoplankton assemblages from the temperate Northeast Atlantic Ocean the critical role of dissolved Fe (DFe) was again highlighted by the remarkably enhanced N2 fixation rate observed after the addition of DFe under low temperature and P-depleted conditions. At the time of our study no Trichodesmium filaments were found in the studied region, the diazotrophic community was dominated by unicellular cyanobacteria symbiont (prymnesiophyte-UCYN-A1) and heterotrophic diazotrophs, therefore demonstrating that DFe could as well be the ultimate factor limiting N2 fixation of these smaller diazotrophs. Recently, the effects of ongoing climate change (ocean warming and acidification) on N2 fixation drew much attention, but various studies led to controversial conclusions. Semi-continuous dilution growth experiments were conducted on Trichodesmium IMS101 under present-day and future high pCO2 (400 and 800 µatm, respectively) and warming seawater (24 and 28 °C) conditions. The results indicate that higher pCO2 and therefore ocean acidification may be beneficial for Trichodesmium growth and N2 fixation. However, our observations suggest that Fe or P limitation in oligotrophic seawaters may offset the stimulation induced on Trichodesmium IMS101 resulting from ocean acidification. In contrast, ocean warming may not play an important role in Trichodesmium growth and N2 fixation with a 4 °C increase from 24 °C to 28 °C. Nevertheless, ocean warming is predicted to cause a shift in the geographical distribution of Trichodesmium species toward higher latitudes, extending its niche to subtropical ocean regions and potentially reducing its coverage in tropical ocean basins.

Assessment of the dinitrogen released as ammonium and dissolved organic nitrogen by unicellular and filamentous marine diazotrophic cyanobacteria grown in culture

The dinitrogen (N2) released as dissolved nitrogen (DN) has been compared in batch cultures of four marine diazotrophic cyanobacteria: the colony forming Trichodesmium IMS101 and the unicellular strains Cyanothece ATCC51142, Crocosphaera watsonii WH8501 and WH0003. Two approaches were conducted for this purpose. The first approach consisted in the comparison of the total accumulation of fixed N2 in the culture (both in dissolved and particulate pools) with the net N2 fixation rates (i. e. the fixed N2 incorporated only in the particulate fraction after 15N2 incubation). The difference between the two measures accounted for the fixed N2 released as DN. The second approach consisted in the direct measure of the 15N-enrichment of ammonium (NH4+) and dissolved organic N (DON) following the 15N2 incubations. The N2 released as DN accounted for ~0 - 20 % and ~1 % of the N2 fixed after 24 h in the first and second approach, respectively. We show that the recent methodological improvements in the net N2 fixation determination applied in this study tend to reconcile the two approaches that formerly led to contrasted values. However, the large analytical uncertainties of the first approach limit its reliability. Thus, the direct determination of the 15N-enrichment of the dissolved pool remains the best tool to assess the fixed N2 released in the DN pool, in particular as it allows shorter incubation times. There were no clear patterns detected between the filamentous Trichodesmium and unicellular strains, neither in terms of the amount of N2 released as DN nor in terms of the proportion of NH4+ relative to DON. This suggests that N2 release processes are shared among the filamentous and free living diazotrophs.

Levels of daily light doses under changed day-night cycles regulate temporal segregation of photosynthesis and N2 Fixation in the cyanobacterium Trichodesmium erythraeum IMS101.

While the diazotrophic cyanobacterium Trichodesmium is known to display inverse diurnal performances of photosynthesis and N2 fixation, such a phenomenon has not been well documented under different day-night (L-D) cycles and different levels of light dose exposed to the cells. Here, we show differences in growth, N2 fixation and photosynthetic carbon fixation as well as photochemical performances of Trichodesmium IMS101 grown under 12L:12D, 8L:16D and 16L:8D L-D cycles at 70 μmol photons m-2 s-1 PAR (LL) and 350 μmol photons m-2 s-1 PAR (HL). The specific growth rate was the highest under LL and the lowest under HL under 16L:8D, and it increased under LL and decreased under HL with increased levels of daytime light doses exposed under the different light regimes, respectively. N2 fixation and photosynthetic carbon fixation were affected differentially by changes in the day-night regimes, with the former increasing directly under LL with increased daytime light doses and decreased under HL over growth-saturating light levels. Temporal segregation of N2 fixation from photosynthetic carbon fixation was evidenced under all day-night regimes, showing a time lag between the peak in N2 fixation and dip in carbon fixation. Elongation of light period led to higher N2 fixation rate under LL than under HL, while shortening the light exposure to 8 h delayed the N2 fixation peaking time (at the end of light period) and extended it to night period. Photosynthetic carbon fixation rates and transfer of light photons were always higher under HL than LL, regardless of the day-night cycles. Conclusively, diel performance of N2 fixation possesses functional plasticity, which was regulated by levels of light energy supplies either via changing light levels or length of light exposure.

Electron transport kinetics in the diazotrophic cyanobacterium Trichodesmium spp. grown across a range of light levels.

The diazotrophic cyanobacterium Trichodesmium is a major contributor to marine nitrogen fixation. We analyzed how light acclimation influences the photophysiological performance of Trichodesmium IMS101 during exponential growth in semi-continuous nitrogen fixing cultures under light levels of 70, 150, 250, and 400 μmol photons m(-2) s(-1), across diel cycles. There were close correlations between growth rate, trichome length, particulate organic carbon and nitrogen assimilation, and cellular absorbance, which all peaked at 150 μmol photons m(-2) s(-1). Growth rate was light saturated by about 100 μmol photons m(-2) s(-1) and was photoinhibited above 150 μmol photons m(-2) s(-1). In contrast, the light level (I k) to saturate PSII electron transport (e (-) PSII(-1) s(-1)) was much higher, in the range of 450-550 μmol photons m(-2) s(-1), and increased with growth light. Growth rate correlates with the absorption cross section as well as with absorbed photons per cell, but not to electron transport per PSII; this disparity suggests that numbers of PSII in a cell, along with the energy allocation between two photosystems and the state transition mechanism underlie the changes in growth rates. The rate of state transitions after a transfer to darkness increased with growth light, indicating faster respiratory input into the intersystem electron transport chain.

Remineralization of phytoplankton-derived organic matter by natural populations of heterotrophic bacteria

The relative lability, elemental stoichiometry, and remineralization rates of various particulate organic matter (POM) substrates by natural heterotrophic marine microorganisms were investigated. POM was harvested from laboratory cultures of a marine diazotroph (Trichodesmium IMS101), a cosmopolitan diatom (Thalassiosira weissflogii), a common marine cyanobacteria (Prochlorococcus MED4), and from surface waters off the Oregon coast. These POM resources were used as inoculants in a field experiment conducted at the Hawaii Ocean Time-series Station ALOHA in the North Pacific Subtropical Gyre. POM from these various sources was added to seawater collected from below the surface mixed layer, incubated in the dark, and remineralization rates were quantified via high-frequency measurement of soluble phosphorus (P) and nitrogen (N) concentrations over a 6-d period. Rapid solubilization and near complete remineralization of particulate P (PP) occurred in all treatments where cultured POM was used, with lesser relative mobilization of P from a ‘natural’ POM sample isolated from surface seawater off the Oregon coast. Soluble P pools, likely consisting of surface-adsorbed inorganic P and inorganic P liberated from cells during harvesting of biomass accounted for 28% of natural PP pools and 80±32% of cultured PP. 31P nuclear magnetic resonance (NMR) confirmed that PP was predominately present as orthophosphate in all POM types. By the end of the incubation period, all added P from cultured material had been converted to dissolved inorganic P. This finding may be a caveat of our utilization of laboratory cultures and natural POM which has been exposed to high inorganic P concentrations (0.8–5.0μmolL−1), albeit it is consistent with previous reports of significant contributions of surface-adsorbed P to total particulate P. In contrast, over the course of these experiments, only 37–40% of added N had been remineralized to ammonium (NH4+). In general, N remineralization rates of cultured material increased with the amount of N added (per gram of dry material). The net yield of bacterial cells was also positively correlated to the initial amount of C and N added. Most notably, when corrected for non-biological turnover (i.e. removal of soluble pools), the N:P remineralization ratio of cultured material (8.5±1.3) was independent of the N:P of added organic material (5–23).

Decoupling of ammonium regulation and ntcA transcription in the diazotrophic marine cyanobacterium Trichodesmium sp. IMS101.

Nitrogen (N) physiology in the marine cyanobacterium Trichodesmium IMS101 was studied along with transcript accumulation of the N-regulatory gene ntcA and of two of its target genes: napA (nitrate assimilation) and nifH (N(2) fixation). N(2) fixation was impaired in the presence of nitrite, nitrate and urea. Strain IMS101 was capable of growth on these combined N sources at <2 μM but growth rates declined at elevated concentrations. Assimilation of nitrate and urea was impaired in the presence of ammonium. Whereas ecologically relevant N concentrations (2-20 μM) suppressed growth and assimilation, much higher concentrations were required to affect transcript levels. Transcripts of nifH accumulated under nitrogen-fixing conditions; these transcript levels were maintained in the presence of nitrate (100 μM) and ammonium (20 μM). However, nifH transcript levels were below detection at ammonium concentrations >20 μM. napA mRNA was found at low levels in both N(2)-fixing and ammonium-utilizing filaments, and it accumulated in filaments grown with nitrate. The positive effect of nitrate on napA transcription was abolished by ammonium additions of >200 μM. This effect was restored upon addition of the glutamine synthetase inhibitor L-methionin-DL-sulfoximine. Surprisingly, ntcA transcript levels remained high in the presence of ammonium, even at elevated concentrations. These findings indicate that ammonium repression is decoupled from transcriptional activation of ntcA in Trichodesmium IMS101.

The influence of pCO2 and temperature on gene expression of carbon and nitrogen pathways in Trichodesmium IMS101.

Growth, protein amount, and activity levels of metabolic pathways in Trichodesmium are influenced by environmental changes such as elevated pCO2 and temperature. This study examines changes in the expression of essential metabolic genes in Trichodesmium grown under a matrix of pCO2 (400 and 900 µatm) and temperature (25 and 31°C). Using RT-qPCR, we studied 21 genes related to four metabolic functional groups: CO2 concentrating mechanism (bicA1, bicA2, ccmM, ccmK2, ccmK3, ndhF4, ndhD4, ndhL, chpX), energy metabolism (atpB, sod, prx, glcD), nitrogen metabolism (glnA, hetR, nifH), and inorganic carbon fixation and photosynthesis (rbcL, rca, psaB, psaC, psbA). nifH and most photosynthetic genes exhibited relatively high abundance and their expression was influenced by both environmental parameters. A two to three orders of magnitude increase was observed for glnA and hetR only when both pCO2 and temperature were elevated. CO2 concentrating mechanism genes were not affected by pCO2 and temperature and their expression levels were markedly lower than that of the nitrogen metabolism and photosynthetic genes. Many of the CO2 concentrating mechanism genes were co-expressed throughout the day. Our results demonstrate that in Trichodesmium, CO2 concentrating mechanism genes are constitutively expressed. Co-expression of genes from different functional groups were frequently observed during the first half of the photoperiod when oxygenic photosynthesis and N2 fixation take place, pointing at the tight and complex regulation of gene expression in Trichodesmium. Here we provide new data linking environmental changes of pCO2 and temperature to gene expression in Trichodesmium. Although gene expression indicates an active metabolic pathway, there is often an uncoupling between transcription and enzyme activity, such that transcript level cannot usually be directly extrapolated to metabolic activity.

Combined effects of CO2 and light on the N2-fixing cyanobacterium Trichodesmium IMS101: physiological responses.

Recent studies on the diazotrophic cyanobacterium Trichodesmium erythraeum (IMS101) showed that increasing CO(2) partial pressure (pCO(2)) enhances N(2) fixation and growth. Significant uncertainties remain as to the degree of the sensitivity to pCO(2), its modification by other environmental factors, and underlying processes causing these responses. To address these questions, we examined the responses of Trichodesmium IMS101 grown under a matrix of low and high levels of pCO(2) (150 and 900 microatm) and irradiance (50 and 200 micromol photons m(-2) s(-1)). Growth rates as well as cellular carbon and nitrogen contents increased with increasing pCO(2) and light levels in the cultures. The pCO(2)-dependent stimulation in organic carbon and nitrogen production was highest under low light. High pCO(2) stimulated rates of N(2) fixation and prolonged the duration, while high light affected maximum rates only. Gross photosynthesis increased with light but did not change with pCO(2). HCO(3)(-) was identified as the predominant carbon source taken up in all treatments. Inorganic carbon uptake increased with light, but only gross CO(2) uptake was enhanced under high pCO(2). A comparison between carbon fluxes in vivo and those derived from (13)C fractionation indicates high internal carbon cycling, especially in the low-pCO(2) treatment under high light. Light-dependent oxygen uptake was only detected under low pCO(2) combined with high light or when low-light-acclimated cells were exposed to high light, indicating that the Mehler reaction functions also as a photoprotective mechanism in Trichodesmium. Our data confirm the pronounced pCO(2) effect on N(2) fixation and growth in Trichodesmium and further show a strong modulation of these effects by light intensity. We attribute these responses to changes in the allocation of photosynthetic energy between carbon acquisition and the assimilation of carbon and nitrogen under elevated pCO(2). These findings are supported by a complementary study looking at photosynthetic fluorescence parameters of photosystem II, photosynthetic unit stoichiometry (photosystem I:photosystem II), and pool sizes of key proteins in carbon and nitrogen acquisition.

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: A Mechanistic View

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO(2) partial pressure (pCO(2)) with higher N(2) fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO(2) (150 and 900 microatm) and light (50 and 200 micromol photons m(-2) s(-1)) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO(2) enhanced N(2) fixation and growth, oxygen evolution and carbon fixation increased mainly as a response to high light. Here, we investigated changes in the photosynthetic fluorescence parameters of photosystem II, in ratios of the photosynthetic units (photosystem I:photosystem II), and in the pool sizes of key proteins involved in the fixation of carbon and nitrogen as well as their subsequent assimilation. We show that the combined elevation in pCO(2) and light controlled the operation of the CO(2)-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO(2) and high light decreased the amounts of several key proteins (NifH, PsbA, and PsaC), while amounts of AtpB and RbcL did not significantly change. Reduced investment in protein biosynthesis, without notably changing photosynthetic fluxes, could free up energy that can be reallocated to increase N(2) fixation and growth at elevated pCO(2) and light. We suggest that changes in the redox state of the photosynthetic electron transport chain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enable Trichodesmium to flourish in future surface oceans characterized by elevated pCO(2), higher temperatures, and high light.

Phosphonate metabolism by Trichodesmium IMS101 and the production of greenhouse gases

A series of laboratory experiments were conducted to investigate (1) the capacity of Trichodesmium IMS101 to hydrolyze phosphonates as a source of phosphorus (P) for growth, (2) the stoichiometric relationship between phosphonate use and biogenic gas production, and (3) the potential inhibition of phosphonate hydrolysis by additions of dissolved inorganic phosphorus (DIP). Trichodesmium IMS101 is capable of cleaving the carbon‐P bond found in methylphosphonate (MPn) and ethylphosphonate (EPn), and the decomposition of these particular phosphonates results in the stoichiometric production of the greenhouse gases methane (CH4) and ethane (C2H6), respectively. Growth on 2‐aminoethylphosphonate (2‐AEP) led to modest ethylene (C2H4) production. Normalized to rates of Trichodesmium carbon (C) fixation, biogenic gas production as a result of either MPn or EPn hydrolysis (0.95 ± 0.04 mmol CH4 (mol C)−1 and 1.18 ± 0.11 mmol C2H6 (mol C)−1, respectively) approximates rates of DIP use (1.11 ± 0.05 P mmol P (mol C)−1) measured in parallel cultures. DIP, MPn, and EPn can be used by Trichodesmium IMS101 as a sole source of P with equal metabolic efficiency. Additionally, neither MPn hydrolysis nor the production of CH4 was significantly inhibited by additions of up to 30 mmol DIP L−1. These results imply that Trichodesmium can use multiple P resources simultaneously for growth and that the production of greenhouse gases occurs during decomposition pathways of select phosphonates.

Phosphonate metabolism by Trichodesmium IMS101 and the production of greenhouse gases

Herein we present the results of a series of laboratory experiments conducted to delineate the relationship between dissolved organic phosphorus (DOP) utilization and organic matter production by the model diazotroph Trichodesmium IMS101. The rate and ratio of Trichodesmium carbon (C) and nitrogen (N) production driven by phosphorus (P) derived from select phosphonates and P monoesters were not found to be significantly different than growth rates based on dissolved inorganic phosphorus (DIP). Conversely, despite the observed similarities in C and N fixation rates between parallel DIP and DOP incubations, cultures grown solely on methylphosphonate (MPn) had reduced maximal P content per unit chlorophyll a or C relative to cultures grown on P monoesters or DIP. So while Trichodesmium IMS101 appears to be able to maintain similar rates of growth on DIP and select DOP substrates, the P uptake efficiency and, hence, the P content per cell is greater for growth on DIP or tested P monoesters relative to growth on the phosphonate MPn. The primary conclusion arising from these results is that neither the observation of P monoester hydrolysis nor highly P‐deficient organic matter or expression of phosphonate hydrolyzing genes can be interpreted to indicate P limitation of C or N fixation of Trichodesmium spp. under light‐saturated, DIP‐deficient conditions.

Carbon acquisition by Trichodesmium: the effect of pCO2 and diurnal changes

We investigated carbon acquisition by the N2‐fixing cyanobacterium Trichodesmium IMS101 in response to CO2 levels of 15.1, 37.5, and 101.3 Pa (equivalent to 150, 370, and 1000 ppm). In these acclimations, growth rates as well as cellular C and N contents were measured. In vivo activities of carbonic anhydrase (CA), photosynthetic O2 evolution, and CO2 and HCO3‐ fluxes were measured using membrane inlet mass spectrometry and the 14C disequilibrium technique. While no differences in growth rates were observed, elevated CO2 levels caused higher C and N quotas and stimulated photosynthesis and N2 fixation. Minimal extracellular CA (eCA) activity was observed, indicating a minor role in carbon acquisition. Rates of CO2 uptake were small relative to total inorganic carbon (Ci) fixation, whereas HCO{3 contributed more than 90% and varied only slightly over the light period and between CO2 treatments. The low eCA activity and preference for HCO3‐ were verified by the 14C disequilibrium technique. Regarding apparent affinities, half‐saturation concentrations (K1/2) for photosynthetic O2 evolution and HCO3‐ uptake changed markedly over the day and with CO2 concentration. Leakage (CO2 efflux : Ci uptake) showed pronounced diurnal changes. Our findings do not support a direct CO2 effect on the carboxylation efficiency of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (RubisCO) but point to a shift in resource allocation among photosynthesis, carbon acquisition, and N2 fixation under elevated CO2 levels. The observed increase in photosynthesis and N2 fixation could have potential biogeochemical implications, as it may stimulate productivity in N‐limited oligotrophic regions and thus provide a negative feedback in rising atmospheric CO2 levels.

Iron limitation in the marine cyanobacterium Trichodesmium reveals new insights into regulation of photosynthesis and nitrogen fixation

* As iron (Fe) deficiency is a main limiting factor of ocean productivity, its effects were investigated on interactions between photosynthesis and nitrogen fixation in the marine nonheterocystous diazotrophic cyanobacterium Trichodesmium IMS101. * Biophysical methods such as fluorescence kinetic microscopy, fast repetition rate (FRR) fluorimetry, and in vivo and in vitro spectroscopy of pigment composition were used, and nitrogenase activity and the abundance of key proteins were measured. * Fe limitation caused a fast down-regulation of nitrogenase activity and protein levels. By contrast, the abundance of Fe-requiring photosystem I (PSI) components remained constant. Total levels of phycobiliproteins remained unchanged according to single-cell in vivo spectra. However, the regular 16-kDa phycoerythrin band decreased and finally disappeared 16-20 d after initiation of Fe limitation, concomitant with the accumulation of a 20-kDa protein cross-reacting with the phycoerythrin antibody. Concurrently, nitrogenase expression and activity increased. Fe limitation dampened the daily cycle of photosystem II (PSII) activity characteristic of diazotrophic Trichodesmium cells. Further, it increased the number and prolonged the time period of occurrence of cells with elevated basic fluorescence (F(0)). Additionally, it increased the effective cross-section of PSII, probably as a result of enhanced coupling of phycobilisomes to PSII, and led to up-regulation of the Fe stress protein IsiA. * Trichodesmium survives short-term Fe limitation by selectively down-regulating nitrogen fixation while maintaining but re-arranging the photosynthetic apparatus.

Nitrogen fixation and growth rates of Trichodesmium IMS-101 as a function of light intensity

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 359:25-36 (2008) - DOI: https://doi.org/10.3354/meps07241 Nitrogen fixation and growth rates of Trichodesmium IMS-101 as a function of light intensity E. Breitbarth1,2,*, J. Wohlers1, J. Kläs1, J. LaRoche1, I. Peeken1 1Leibniz-Institute of Marine Sciences, IFM-GEOMAR, Düsternbrooker Weg 20, 24105 Kiel, Germany 2Present address: Department of Chemistry, Analytical and Marine Chemistry, University of Gothenburg, Kemivägen 10, 412 96 Göteborg, Sweden *Email: eike@chem.gu.se ABSTRACT: The diazotrophic cyanobacterium Trichodesmium is a significant contributor to marine nitrogen and carbon cycles and has been incorporated in biogeochemical ocean circulation models. To date, parameterization of light as a controlling factor for nitrogen fixation has been based on field observations, where factors other than light also affect Trichodesmium physiology. Here we present data on light-dependent (15 to 1100 µmol quanta m–2 s–1) diazotrophic growth from controlled laboratory experiments and their implications for modeling approaches. We supply a simple empirical model to describe nitrogen fixation by Trichodesmium in batch cultures. Diazotrophic growth of axenic Trichodesmium IMS-101 was light saturated at 180 µmol quanta m–2 s–1 and did not vary significantly at higher photon irradiances up to 1100 µmol quanta m–2 s–1 (μcarbon based ≈ 0.26 d–1). Chlorophyll a (chl a) normalized N2 fixation rates were significantly affected by light intensity during mid-exponential growth (0.74 to 4.45 mol N fixed mol chl a–1 h–1) over the range of photon irradiances tested. In contrast, nitrogen fixation rates normalized to the cellular carbon content were relatively unaffected by light intensity (0.42 to 0.59, averaging 0.5 mmol N mol particulate organic carbon [POC]–1 h–1). Trichodesmium carbon biomass can be used to estimate the nitrogen input by this diazotroph into the ocean; the maximum input rate is 350 nmol N fixed l–1 h–1. KEY WORDS: Trichodesmium · Light · Nitrogen fixation · Marine nitrogen cycle · Marine carbon cycle · Marine cyanobacteria · Diazotrophic growth Full text in pdf format Supplementary appendix PreviousNextCite this article as: Breitbarth E, Wohlers J, Kläs J, LaRoche J, Peeken I (2008) Nitrogen fixation and growth rates of Trichodesmium IMS-101 as a function of light intensity. Mar Ecol Prog Ser 359:25-36. https://doi.org/10.3354/meps07241Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 359. Online publication date: May 05, 2008 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2008 Inter-Research.

Physiological constraints on the global distribution of &amp;lt;i&amp;gt;Trichodesmium&amp;lt;/i&amp;gt; – effect of temperature on diazotrophy

Abstract. The cyanobacterium Trichodesmium is an important link in the global nitrogen cycle due to its significant input of atmospheric nitrogen to the ocean. Attempts to incorporate Trichodesmium in ocean biogeochemical circulation models have, so far, relied on the observed correlation between temperature and Trichodesmium abundance. This correlation may result in part from a direct effect of temperature on Trichodesmium growth rates through the control of cellular biochemical processes, or indirectly through temperature influence on mixed layer depth, light and nutrient regimes. Here we present results indicating that the observed correlation of Trichodesmium with temperature in the field reflects primarily the direct physiological effects of temperature on diazotrophic growth of Trichodesmium. Trichodesmium IMS-101 (an isolate of Trichodesmium) could acclimate and grow at temperatures ranging from 20 to 34°C. Maximum growth rates (μmax=0.25 day–1) and maximum nitrogen fixation rates (0.13 mmol N mol POC−1 h–1) were measured within 24 to 30°C. Combining this empirical relationship with global warming scenarios derived from state-of-the-art climate models sets a physiological constraint on the future distribution of Trichodesmium that could significantly affect the future nitrogen input into oligotrophic waters by this diazotroph.

Elevated CO2 enhances nitrogen fixation and growth in the marine cyanobacterium Trichodesmium

AbstractThe increases in atmospheric pCO2 over the last century are accompanied by higher concentrations of CO2(aq) in the surface oceans. This acidification of the surface ocean is expected to influence aquatic primary productivity and may also affect cyanobacterial nitrogen (N)‐fixers (diazotrophs). No data is currently available showing the response of diazotrophs to enhanced oceanic CO2(aq). We examined the influence of pCO2 [preindustrial∼250 ppmv (low), ambient∼400, future∼900 ppmv (high)] on the photosynthesis, N fixation, and growth of Trichodesmium IMS101. Trichodesmium spp. is a bloom‐forming cyanobacterium contributing substantial inputs of ‘new N’ to the oligotrophic subtropical and tropical oceans. High pCO2 enhanced N fixation, C : N ratios, filament length, and biomass of Trichodesmium in comparison with both ambient and low pCO2 cultures. Photosynthesis and respiration did not change significantly between the treatments. We suggest that enhanced N fixation and growth in the high pCO2 cultures occurs due to reallocation of energy and resources from carbon concentrating mechanisms (CCM) required under low and ambient pCO2. Thus, in oceanic regions, where light and nutrients such as P and Fe are not limiting, we expect the projected concentrations of CO2 to increase N fixation and growth of Trichodesmium. Other diazotrophs may be similarly affected, thereby enhancing inputs of new N and increasing primary productivity in the oceans.

Flexible elemental stoichiometry in Trichodesmium spp. and its ecological implications

We conducted laboratory experiments to assess the bioelemental plasticity of cultures of Trichodesmium IMS101 under phosphorus (P)‐replete, P‐restricted, and light‐limited conditions. The results reveal a high degree of stoichiometric flexibility. Specifically, Trichodesmium IMS101 is capable of growth with carbon (C) : nitrogen (N) : P ratios of C585±56 : N90±10 : P1, approximately six times higher than would be predicted by the Redfield reference ratio (C106 : N16 : P1), thus signifying low cellular P quotas relative to C and N. Luxury consumption of P occurs rapidly after periods of prolonged P restriction, under both light and dark conditions, resulting in substantial increases in P quotas and reductions of C : N : P ratios (C96±8 : N16±1 : P1). Comparisons of laboratory culture data to our field observations from the Northwest Atlantic and the North Pacific indicate that, while natural populations of Trichodesmium exhibit persistently low P content relative to C and N (C290±15 : N53±3 : P1), the highest and lowest C : P and N : P ratios recorded in the laboratory are rarely observed in nature. We have also performed laboratory experiments intended to simulate the energetic and nutritional extremes that would occur as naturally migrating populations of Trichodesmium sink out of the euphotic zone into P‐rich regions of the upper disphotic zone. The duration of dark survival for this isolate is on the order of 3‐6 d, after which time cells are unable to recover from light deprivation. This finding provides a constraint on the temporal scale of vertical migration.

Traffic lights in trichodesmium. Regulation of photosynthesis for nitrogen fixation studied by chlorophyll fluorescence kinetic microscopy.

We investigated interactions between photosynthesis and nitrogen fixation in the non-heterocystous marine cyanobacterium Trichodesmium IMS101 at the single-cell level by two-dimensional (imaging) microscopic measurements of chlorophyll fluorescence kinetics. Nitrogen fixation was closely associated with the appearance of cells with high basic fluorescence yield (F(0)), termed bright cells. In cultures aerated with normal air, both nitrogen fixation and bright cells appeared in the middle of the light phase. In cultures aerated with 5% oxygen, both processes occurred at a low level throughout most of the day. Under 50% oxygen, nitrogen fixation commenced at the beginning of the light phase but declined soon afterwards. Rapid reversible switches between fluorescence levels were observed, which indicated that the elevated F(0) of the bright cells originates from reversible uncoupling of the photosystem II (PSII) antenna from the PSII reaction center. Two physiologically distinct types of bright cells were observed. Type I had about double F(0) compared to the normal F(0) in the dark phase and a PSII activity, measured as variable fluorescence (F(v) = F(m) - F(0)), similar to normal non-diazotrophic cells. Correlation of type I cells with nitrogen fixation, oxygen concentration, and light suggests that this physiological state is connected to an up-regulation of the Mehler reaction, resulting in oxygen consumption despite functional PSII. Type II cells had more than three times the normal F(0) and hardly any PSII activity measurable by variable fluorescence. They did not occur under low-oxygen concentrations, but appeared under high-oxygen levels outside the diazotrophic period, suggesting that this state represents a reaction to oxidative stress not necessarily connected to nitrogen fixation. In addition to the two high-fluorescence states, cells were observed to reversibly enter a low-fluorescence state. This occurred mainly after a cell went through its bright phase and may represent a fluorescence-quenching recovery phase.

Dinitrogen fixation and release of ammonium and dissolved organic nitrogen by Trichodesmium IMS101

Two methods used to measure dinitrogen (N 2 ) fixation (acetylene reduction and 15 N 2 uptake) often result in different N 2 fixation rates. Part of the discrepancy may arise from the observation that Trichodesmium can release a fraction of their recently fixed N 2 as dissolved organic nitrogen (DON) and/or ammonium (NH 4 + ). To resolve outstanding issues regarding N 2 fixation and the production of dissolved combined nitrogen (N) by Trichodesmium, we conducted a comprehensive analysis of N 2 fixation and the production of DON and NH 4 + in cultures of Trichodesmium IMS101. We performed 15 N 2 uptake experiments in parallel with acetylene (C 2 H 2 ) reduction assays, and measured production of 15 NH 4 + and DO 15 N from 15 N 2 , and 15 NH 4 + uptake and regeneration by isotope dilution. Four main results are highlighted. First, 15 N 2 uptake appears to provide a better approximation of net N-specific growth rates than N 2 fixation estimates made using C 2 H 2 reduction. Second, the C 2 H 2 reduction method provides a closer approximation of gross N 2 fixation. Third, simultaneous measurements of relevant N pools and pathways by several methods enabled us to rigorously evaluate deviations from theoretical conversion factors and to interpret the basis for those deviations. Our results suggest that a conversion ratio (mol C 2 H 2 reduced: mol N 2 reduced to PON, ammonium and DON) of 4:1 may be more appropriate for total N 2 fixation. Fourth, the difference between estimates of gross N 2 fixation, made using the C 2 H 2 reduction technique, and net 15 N 2 uptake into particulate N may be a good indicator of N release from N 2 fixation.