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

Trichodesmium's strategies to alleviate phosphorus limitation in the future acidified oceans

Global warming may exacerbate inorganic nutrient limitation, including phosphorus (P), in the surface-waters of tropical oceans that are home to extensive blooms of the marine diazotrophic cyanobacterium, Trichodesmium. We examined the combined effects of P limitation and pCO2, forecast under ocean acidification scenarios, on Trichodesmium erythraeum IMS101 cultures. We measured nitrogen acquisition, glutamine synthetase activity, C uptake rates, intracellular Adenosine Triphosphate (ATP) concentration and the pool sizes of related key proteins. Here, we present data supporting the idea that cellular energy re-allocation enables the higher growth and N2 fixation rates detected in Trichodesmium cultured under high pCO2. This is reflected in altered protein abundance and metabolic pools. Also modified are particulate organic carbon and nitrogen production rates, enzymatic activities, and cellular ATP concentrations. We suggest that adjusting these cellular pathways to changing environmental conditions enables Trichodesmium to compensate for low P availability and to thrive in acidified oceans. Moreover, elevated pCO2 could provide Trichodesmium with a competitive dominance that would extend its niche, particularly in P-limited regions of the tropical and subtropical oceans

CO2-dependend energy allocation in Trichodesmium

Recent studies on the diazotrophic cyanobacteria Trichodesmium erythraeum (IMS101) showed that increasing pCO2 availability enhances N2 fixation and growth. However, uncertainties remain regarding the overall processes causing these responses. We examined the responses of Trichodesmium IMS101 grown under a matrix of low and high levels of pCO2 (150 and 900 µatm) and irradiance (50 and 200 µmol photons m-2 s-1). Growth rates as well as cellular C and N content increased with increasing pCO2 and light levels in the cultures. The pCO2-dependent stimulation in organic C and N production was highest under low-light. High pCO2 stimulated rates of N2 fixation and prolonged the duration while high-light affected maximum rates only. Gross photosynthesis increased with light, but did not change with pCO2. HCO3- was identified as the predominant carbon source taken up in all treatments showing no trend with pCO2, while gross CO2 uptake was enhanced under elevated pCO2. A comparison between measured carbon fluxes and those derived from 13C fractionation revealed high internal carbon cycling within the high light and low pCO2 treatment. Our data confirm the pronounced pCO2 effect on N2 fixation and growth in Trichodesmium and indicate 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 C and N under elevated pCO2

CO2-dependent energy allocation in Trichodesmium

In view of the current increase in atmospheric pCO2 and concomitant changes in the marine environment, it is crucial to assess, understand and predict future responses of ecologically relevant phytoplankton species. The N2 fixing cyanobacteria Trichodesmium spp. contribute about half of all marine N2 fixation and thus can play a prominent role in the biogeochemical cycling of carbon and nitrogen. In this study, the effect of pCO2 and light on growth and production rates of Trichodesmium was investigated and underlying processes such as photosynthesis, C as well as N acquisition were studied

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: A Mechanistic View1

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO2 partial pressure (pCO2) with higher N2 fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO2 (150 and 900 μatm) and light (50 and 200 μmol photons m−2 s−1) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO2 enhanced N2 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 pCO2 and light controlled the operation of the CO2-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO2 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 N2 fixation and growth at elevated pCO2 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 pCO2, higher temperatures, and high light

CO2 and light effects on growth, photosynthesis, carbon acquisition and nitrogen fixation of the diazotrophic cyanobacteria Trichodemium

Recent studies on the diazotrophic cyanobacteria Trichodesmium showed a pronounced CO2 dependency in N2- and C-fixation. However, significant uncertainties remain as to the degree of sensitivity to CO2, modification of these responses by other environmental factors, and the underlying processes. To this end, we investigated carbon acquisition and nitrogen fixation in Trichodesmium IMS101 under different CO2 and light levels. In these acclimations, growth rates, cellular C and N content, and respective isotopic values were measured. In vivo activities of photosynthetic O2 evolution, O2 uptake, CO2 and HCO3- fluxes were obtained using membrane inlet mass spectrometry methods (MIMS). Nitrogen fixation rates were determined using the acetylene reduction assay, and chlorophyll a fluorescence was measured via fluorescence induction and relaxation method (FIRe). Photosynthesis and N2-fixation increased with CO2 and this effect was even more pronounced under high light. Moreover, our data suggest a shift in the distribution of metabolic energy between photosynthesis, carbon acquisition and N2-fixation in Trichodesmium. The observed stimulation in photosynthesis and N2-fixation may enhance the productivity in N-limited oligotrophic regions

Seawater carbonate chemistry and combined mechanistic effects of CO2 and light on the N2-fixing cyanobacterium Trichodesmium IMS101, 2010

The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO2 partial pressure (pCO2) with higher N2 fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO2(150 and 900 µatm) and light (50 and 200 µmol photons m-2 s-1) on TrichodesmiumIMS101. We expand on a complementary study that demonstrated that while elevated pCO2 enhanced N2 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 pCO2 and light controlled the operation of the CO2-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO2 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 N2 fixation and growth at elevated pCO2 and light. We suggest that changes in the redox state of the photosynthetic electron transportchain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enableTrichodesmium to flourish in future surface oceans characterized by elevated pCO2, higher temperatures, and high light

Combined Effects of CO2 and Light on the N2-Fixing Cyanobacterium Trichodesmium IMS101: Physiological Responses1[OA]

Recent studies on the diazotrophic cyanobacterium Trichodesmium erythraeum (IMS101) showed that increasing CO2 partial pressure (pCO2) enhances N2 fixation and growth. Significant uncertainties remain as to the degree of the sensitivity to pCO2, 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 pCO2 (150 and 900 μatm) and irradiance (50 and 200 μmol photons m−2 s−1). Growth rates as well as cellular carbon and nitrogen contents increased with increasing pCO2 and light levels in the cultures. The pCO2-dependent stimulation in organic carbon and nitrogen production was highest under low light. High pCO2 stimulated rates of N2 fixation and prolonged the duration, while high light affected maximum rates only. Gross photosynthesis increased with light but did not change with pCO2. HCO3− was identified as the predominant carbon source taken up in all treatments. Inorganic carbon uptake increased with light, but only gross CO2 uptake was enhanced under high pCO2. A comparison between carbon fluxes in vivo and those derived from 13C fractionation indicates high internal carbon cycling, especially in the low-pCO2 treatment under high light. Light-dependent oxygen uptake was only detected under low pCO2 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 pCO2 effect on N2 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 pCO2. 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