Untreated sewage outfalls do not promote Trichodesmium blooms in the coasts of the Canary Islands.

During the summer of 2017, recurrent extensive blooms of the diazotrophic cyanobacterium Trichodesmium invaded the beaches and coastal waters of the Canary Islands, causing great social alarm. Some local media and public sectors ascribed, without any strong scientific evidence, the origin and reactivation of these blooms to untreated sewage outfalls distributed along the coasts. In order to test whether sewage outfalls could have any influence on the metabolic activity of Trichodesmium, we performed 13C and 15N2 uptake experiments with colonies experiencing three different bloom development stages, incubated both with clear seawater and sewage water from an outfall south of Gran Canaria island. Our results showed that sewage outfalls did not promote any increase in dinitrogen (N2) fixation in Trichodesmium, supporting the hypothesis that decaying blooms were generated offshore and transported shoreward by local currents and winds, accumulating mostly leeward of the islands. The combination of unusually warm seawater temperatures, enhanced and sustained stratification of the upper water column and recurrent dust deposition events would have favored the development of the Trichodesmium blooms, which lasted for at least four months.

Multiscale Multiobjective Systems Analysis (MiMoSA): an advanced metabolic modeling framework for complex systems.

In natural environments, cells live in complex communities and experience a high degree of heterogeneity internally and in the environment. Even in 'ideal' laboratory environments, cells can experience a high degree of heterogeneity in their environments. Unfortunately, most of the metabolic modeling approaches that are currently used assume ideal conditions and that each cell is identical, limiting their application to pure cultures in well-mixed vessels. Here we describe our development of Multiscale Multiobjective Systems Analysis (MiMoSA), a metabolic modeling approach that can track individual cells in both space and time, track the diffusion of nutrients and light and the interaction of cells with each other and the environment. As a proof-of concept study, we used MiMoSA to model the growth of Trichodesmium erythraeum, a filamentous diazotrophic cyanobacterium which has cells with two distinct metabolic modes. The use of MiMoSA significantly improves our ability to predictively model metabolic changes and phenotype in more complex cell cultures.

Colonies of marine cyanobacteria Trichodesmium interact with associated bacteria to acquire iron from dust.

Iron (Fe) bioavailability limits phytoplankton growth in vast ocean regions. Iron-rich dust uplifted from deserts is transported in the atmosphere and deposited on the ocean surface. However, this dust is a poor source of iron for most phytoplankton since dust-bound Fe is poorly soluble in seawater and dust rapidly sinks out of the photic zone. An exception is Trichodesmium, a globally important, N2 fixing, colony forming, cyanobacterium, which efficiently captures and shuffles dust to its colony core. Trichodesmium and bacteria that reside within its colonies carry out diverse metabolic interactions. Here we show evidence for mutualistic interactions between Trichodesmium and associated bacteria for utilization of iron from dust, where bacteria promote dust dissolution by producing Fe-complexing molecules (siderophores) and Trichodesmium provides dust and optimal physical settings for dissolution and uptake. Our results demonstrate how intricate relationships between producers and consumers can influence productivity in the nutrient starved open ocean.

Reduced nitrogenase efficiency dominates response of the globally important nitrogen fixer Trichodesmium to ocean acidification.

The response of the prominent marine dinitrogen (N2)-fixing cyanobacteria Trichodesmium to ocean acidification (OA) is critical to understanding future oceanic biogeochemical cycles. Recent studies have reported conflicting findings on the effect of OA on growth and N2 fixation of Trichodesmium. Here, we quantitatively analyzed experimental data on how Trichodesmium reallocated intracellular iron and energy among key cellular processes in response to OA, and integrated the findings to construct an optimality-based cellular model. The model results indicate that Trichodesmium growth rate decreases under OA primarily due to reduced nitrogenase efficiency. The downregulation of the carbon dioxide (CO2)-concentrating mechanism under OA has little impact on Trichodesmium, and the energy demand of anti-stress responses to OA has a moderate negative effect. We predict that if anthropogenic CO2 emissions continue to rise, OA could reduce global N2 fixation potential of Trichodesmium by 27% in this century, with the largest decrease in iron-limiting regions.

CO2 modulation of the rates of photosynthesis and light-dependent O2 consumption in Trichodesmium.

As atmospheric CO2 concentrations increase, so too does the dissolved CO2 and HCO3- concentrations in the world's oceans. There are still many uncertainties regarding the biological response of key groups of organisms to these changing conditions, which is crucial for predicting future species distributions, primary productivity rates, and biogeochemical cycling. In this study, we established the relationship between gross photosynthetic O2 evolution and light-dependent O2 consumption in Trichodesmium erythraeum IMS101 acclimated to three targeted pCO2 concentrations (180 µmol mol-1=low-CO2, 380 µmol mol-1=mid-CO2, and 720 µmol mol-1=high-CO2). We found that biomass- (carbon) specific, light-saturated maximum net O2 evolution rates (PnC,max) and acclimated growth rates increased from low- to mid-CO2, but did not differ significantly between mid- and high-CO2. Dark respiration rates were five times higher than required to maintain cellular metabolism, suggesting that respiration provides a substantial proportion of the ATP and reductant for N2 fixation. Oxygen uptake increased linearly with gross O2 evolution across light intensities ranging from darkness to 1100 µmol photons m-2 s-1. The slope of this relationship decreased with increasing CO2, which we attribute to the increased energetic cost of operating the carbon-concentrating mechanism at lower CO2 concentrations. Our results indicate that net photosynthesis and growth of T. erythraeum IMS101 would have been severely CO2 limited at the last glacial maximum, but that the direct effect of future increases of CO2 may only cause marginal increases in growth.

Iron and phosphorus deprivation induce sociality in the marine bloom-forming cyanobacterium Trichodesmium.

Trichodesmium spp. are diazotrophic cyanobacteria that exist as single filaments (trichomes) and as macroscopic colonies of varying shapes formed by aggregating trichomes. The causes and dynamics of colony formation and disassociation are not yet elucidated. we demonstrate that limited availability of dissolved phosphorus (P) or iron (Fe) stimulated trichome mobility and induced colony formation in Trichodesmium erythraeum IMS101 cultures. The specific nutrient limitation differentially affected the rate of colony formation and morphology of the colonies. Fe starvation promoted rapid colony formation (10-48 h from depletion) while 5-7 days were required for colonies to form in P-depleted cultures. Video analyses confirmed that the probability of trichomes to cluster increased from 12 to 35% when transferred from nutrient replete to Fe-depleted conditions. Moreover, the probability for Fe-depleted aggregates to remain colonial increased to 50% from only 10% in nutrient replete cultures. These colonies were also characterized by stronger attachment forces between the trichomes. Enrichment of nutrient-depleted cultures with the limited nutrient-stimulated colony dissociation into single trichomes. We postulate that limited P and Fe availability enhance colony formation of Trichodesmium and primarily control the abundance and distribution of its different morphologies in the nutrient-limited surface ocean.

Benefit from decline: the primary transcriptome of Alteromonas macleodii str. Te101 during Trichodesmium demise.

Interactions between co-existing microorganisms deeply affect the physiology of the involved organisms and, ultimately, the function of the ecosystem as a whole. Copiotrophic Alteromonas are marine gammaproteobacteria that thrive during the late stages of phytoplankton blooms in the marine environment and in laboratory co-cultures with cyanobacteria such as Trichodesmium. The response of this heterotroph to the sometimes rapid and transient changes in nutrient supply when the phototroph crashes is not well understood. Here, we isolated and sequenced the strain Alteromonas macleodii str. Te101 from a laboratory culture of Trichodesmium erythraeum IMS101, yielding a chromosome of 4.63 Mb and a single plasmid of 237 kb. Increasing salinities to ≥43 ppt inhibited the growth of Trichodesmium but stimulated growth of the associated Alteromonas. We characterized the transcriptomic responses of both microorganisms and identified the complement of active transcriptional start sites in Alteromonas at single-nucleotide resolution. In replicate cultures, a similar set of genes became activated in Alteromonas when growth rates of Trichodesmium declined and mortality was high. The parallel activation of fliA, rpoS and of flagellar assembly and growth-related genes indicated that Alteromonas might have increased cell motility, growth, and multiple biosynthetic activities. Genes with the highest expression in the data set were three small RNAs (Aln1a-c) that were identified as analogs of the small RNAs CsrB-C in E. coli or RsmX-Z in pathogenic bacteria. Together with the carbon storage protein A (CsrA) homolog Te101_05290, these RNAs likely control the expression of numerous genes in responding to changes in the environment.

Phytoplankton responses to aluminum enrichment in the South China Sea.

Compared to extensive studies reporting the aluminum (Al) toxicity to terrestrial plants and freshwater organisms, very little is known about how marine phytoplankton responds to Al in the field. Here we report the marine phytoplankton responses to Al enrichment in the South China Sea (SCS) using on-deck bottle incubation experiments during eight cruises from May 2010 to November 2013. Generally, Al addition alone enhanced the growth of diatom and Trichodesmium, and nitrogen fixation, but it inhibited the growth of dinoflagellates and Synechococcus. Nevertheless, Al addition alone did not influence the chlorophyll a concentration of the entire phytoplankton assemblages. By adding nitrate and phosphate simultaneously, Al enrichment led to substantial increases in chlorophyll a concentration (especially that of the picophytoplankton<3µm), and cell abundances of diatom and photosynthetic picoeukaryotes. These results indicate varied responses of phytoplankton in different size fractions and taxonomic groups to Al enrichment. Further, by simultaneously adding different macronutrients and/or sufficient trace metals including iron, we found that the phytoplankton responses to Al enrichment were relevant to nutrients coexisting in the environment. Al enrichment may give some phytoplankton a competitive edge over using nutrients, especially the limited ones. The possible influences of Al on the competitors and grazers (predators) of some phytoplankton might indirectly contribute to the positive responses of the phytoplankton to Al enrichment. Our results indicate that Al may influence marine carbon cycle by impacting phytoplankton growth and structure in natural seawater.

The complex effects of ocean acidification on the prominent N2-fixing cyanobacterium Trichodesmium.

Acidification of seawater caused by anthropogenic carbon dioxide (CO2) is anticipated to influence the growth of dinitrogen (N2)-fixing phytoplankton, which contribute a large fraction of primary production in the tropical and subtropical ocean. We found that growth and N2-fixation of the ubiquitous cyanobacterium Trichodesmium decreased under acidified conditions, notwithstanding a beneficial effect of high CO2 Acidification resulted in low cytosolic pH and reduced N2-fixation rates despite elevated nitrogenase concentrations. Low cytosolic pH required increased proton pumping across the thylakoid membrane and elevated adenosine triphosphate production. These requirements were not satisfied under field or experimental iron-limiting conditions, which greatly amplified the negative effect of acidification.

The use of genome-scale metabolic network reconstruction to predict fluxes and equilibrium composition of N-fixing versus C-fixing cells in a diazotrophic cyanobacterium, Trichodesmium erythraeum.

BACKGROUND: Computational, genome based predictions of organism phenotypes has enhanced the ability to investigate the biological phenomena that help organisms survive and respond to their environments. In this study, we have created the first genome-scale metabolic network reconstruction of the nitrogen fixing cyanobacterium T. erythraeum and used genome-scale modeling approaches to investigate carbon and nitrogen fluxes as well as growth and equilibrium population composition. RESULTS: We created a genome-scale reconstruction of T. erythraeum with 971 reactions, 986 metabolites, and 647 unique genes. We then used data from previous studies as well as our own laboratory data to establish a biomass equation and two distinct submodels that correspond to the two cell types formed by T. erythraeum. We then use flux balance analysis and flux variability analysis to generate predictions for how metabolism is distributed to account for the unique productivity of T. erythraeum. Finally, we used in situ data to constrain the model, infer time dependent population compositions and metabolite production using dynamic Flux Balance Analysis. We find that our model predicts equilibrium compositions similar to laboratory measurements, approximately 15.5% diazotrophs for our model versus 10-20% diazotrophs reported in literature. We also found that equilibrium was the most efficient mode of growth and that equilibrium was stoichiometrically mediated. Moreover, the model predicts that nitrogen leakage is an essential condition of optimality for T. erythraeum; cells leak approximately 29.4% total fixed nitrogen when growing at the optimal growth rate, which agrees with values observed in situ. CONCLUSION: The genome-metabolic network reconstruction allows us to use constraints based modeling approaches to predict growth and optimal cellular composition in T. erythraeum colonies. Our predictions match both in situ and laboratory data, indicating that stoichiometry of metabolic reactions plays a large role in the differentiation and composition of different cell types. In order to realize the full potential of the model, advance modeling techniques which account for interactions between colonies, the environment and surrounding species need to be developed.

Molecular and physiological evidence of genetic assimilation to high CO2 in the marine nitrogen fixer Trichodesmium.

Most investigations of biogeochemically important microbes have focused on plastic (short-term) phenotypic responses in the absence of genetic change, whereas few have investigated adaptive (long-term) responses. However, no studies to date have investigated the molecular progression underlying the transition from plasticity to adaptation under elevated CO2 for a marine nitrogen-fixer. To address this gap, we cultured the globally important cyanobacterium Trichodesmium at both low and high CO2 for 4.5 y, followed by reciprocal transplantation experiments to test for adaptation. Intriguingly, fitness actually increased in all high-CO2 adapted cell lines in the ancestral environment upon reciprocal transplantation. By leveraging coordinated phenotypic and transcriptomic profiles, we identified expression changes and pathway enrichments that rapidly responded to elevated CO2 and were maintained upon adaptation, providing strong evidence for genetic assimilation. These candidate genes and pathways included those involved in photosystems, transcriptional regulation, cell signaling, carbon/nitrogen storage, and energy metabolism. Conversely, significant changes in specific sigma factor expression were only observed upon adaptation. These data reveal genetic assimilation as a potentially adaptive response of Trichodesmium and importantly elucidate underlying metabolic pathways paralleling the fixation of the plastic phenotype upon adaptation, thereby contributing to the few available data demonstrating genetic assimilation in microbial photoautotrophs. These molecular insights are thus critical for identifying pathways under selection as drivers in plasticity and adaptation.

Bloom of Trichodesmium (Oscillatoriales, Phormidiaceae) and seasonality of potentially harmful phytoplankton in San Pedro Bay, Leyte, Philippines / Proliferación de Trichodesmium (Oscillatoriales: Euphorbiaceae) y estacionalidad de fitoplancton potencialmente dañino en Bahía San Pedro, Leyte, Filipinas

AbstractSince 1983, San Pedro Bay in the Philippines had been reported to be the site of episodic Pyrodinium bahamense var. compressum blooms that caused paralytic shellfish poisoning in its nearby coastal communities. This bay is also subjected to numerous storms; the strongest was super typhoon Haiyan in November 8, 2013. For the first time, the seasonal dynamics of potentially toxic and harmful phytoplankton in this bay is elucidated. This is also the first record of a bloom of the cyanobacteria, Trichodesmium erythraeum that reached 70 000 colonies/L in April 2013 in this area. There were other 19 potentially toxic and harmful phytoplankton encountered during the sampling period. These consisted of a haptophyte, Phaeocystis globosa, the diatom Pseudonitzschia and 17 dinoflagellates. Seven of these harmful algae had densities high enough to be traced through time. Normally, diatoms abound during the dry season. But Pseudo-nitzschia increased in abundance during the wet season of 2012 and 2013. The dinoflagellates and Phaeocystis globosa behaved as expected and exhibited a relative increase in cell density during the rainy season of both years too. High nutrient availability during this season must have influenced the behavior of the phytoplankton despite differences in temperature and light intensity among seasons. Other notable but rare harmful species found only in plankton net tows during the study were Pyrodinium bahamense var. compressum, Alexandrium tamiyavanichii, Cochlodiniumpolykrikoides, and Noctiluca scintillans. Rev. Biol. Trop. 64 (2): 897-911. Epub 2016 June 01.

ResumenDesde 1983, la Bahía de San Pedro en Filipinas ha sido reportada como un sitio de proliferación de Pyrodinium bahamense var. compressum que causó intoxicación paralítica en sus comunidades costeras cercanas. Esta bahía también está sometida a numerosas tormentas; entre las más fuertes se presentó un súper tifón en Haiyan, el 8 de noviembre 2013. Por primera vez, se explica la dinámica estacional del fitoplancton potencialmente tóxico y dañino en esta bahía. Este es también el primer registro en esta área de una proliferación de cianobacterias (Trichodesmium erythraeum) que alcanzó 70 000 colonias/L en abril 2013. Durante el periodo de muestreo se presentaron otras 19 proliferaciones de fitoplancton potencialmente tóxicas y dañinas. Estos consistían en una haptófita, Phaeocystis globosa, la diatomea Pseudo-nitzschia y 17 dinoflagelados. Siete de estas algas nocivas tenían densidades suficientemente altas como para ser rastreadas a través del tiempo. Normalmente, las diatomeas abundan durante la estación seca. Pero Pseudo-nitzschia aumentó en abundancia durante la temporada de lluvias de 2012 y 2013. Los dinoflagelados y Phaeocystis globosa se comportaron como se esperaba y exhibieron un aumento relativo de la densidad celular durante la temporada de lluvias en los dos años. La alta disponibilidad de nutrientes durante esta temporada debe haber influido en el comportamiento del fitoplancton a pesar de las diferencias en la temperatura y la intensidad de la luz entre estaciones. Otras especies nocivas notables, pero raras que se encontraron sólo en las redes de arrastre de plancton durante el estudio fueron: Pyrodinium bahamense var. compressum, Alexandriumtamiyavanichii, Cochlodinium polykrikoides y Noctiluca scintillans.

High levels of heterogeneity in diazotroph diversity and activity within a putative hotspot for marine nitrogen fixation.

Australia's tropical waters represent predicted 'hotspots' for nitrogen (N2) fixation based on empirical and modelled data. However, the identity, activity and ecology of diazotrophs within this region are virtually unknown. By coupling DNA and cDNA sequencing of nitrogenase genes (nifH) with size-fractionated N2 fixation rate measurements, we elucidated diazotroph dynamics across the shelf region of the Arafura and Timor Seas (ATS) and oceanic Coral Sea during Austral spring and winter. During spring, Trichodesmium dominated ATS assemblages, comprising 60% of nifH DNA sequences, while Candidatus Atelocyanobacterium thalassa (UCYN-A) comprised 42% in the Coral Sea. In contrast, during winter the relative abundance of heterotrophic unicellular diazotrophs (δ-proteobacteria and γ-24774A11) increased in both regions, concomitant with a marked decline in UCYN-A sequences, whereby this clade effectively disappeared in the Coral Sea. Conservative estimates of N2 fixation rates ranged from <1 to 91 nmol l(-1) day(-1), and size fractionation indicated that unicellular organisms dominated N2 fixation during both spring and winter, but average unicellular rates were up to 10-fold higher in winter than in spring. Relative abundances of UCYN-A1 and γ-24774A11 nifH transcripts negatively correlated to silicate and phosphate, suggesting an affinity for oligotrophy. Our results indicate that Australia's tropical waters are indeed hotspots for N2 fixation and that regional physicochemical characteristics drive differential contributions of cyanobacterial and heterotrophic phylotypes to N2 fixation.

Effect of light on the growth of non-nitrogen-fixing and nitrogen-fixing phytoplankton in an aquatic system.

We discuss a mathematical model of growth of two types of phytoplankton, non-nitrogen-fixing and nitrogen-fixing, that both require light in order to grow. We use general functional responses to represent the inhibitory effect their biomass has on the exposure to light. We give conditions for the existence and local stability of all of the possible steady-states (die out, single species survival, and coexistence). We derive conditions for global stability of the die out and single-species steady-states and for persistence of both species when the coexistence steady-state exists. Numerical investigation illustrates the qualitative dynamics demonstrating that even under constant environmental conditions, both stable intrinsic oscillatory behavior and a period doubling route to chaotic dynamics are possible. We also show that competitor-mediated coexistence can occur due to the positive feedback resulting from recycling by the nitrogen-fixing phytoplankton. To show the impact of seasonal change in water depth, we also allow the water depth to vary in an annual cycle and discuss echo blooms in this context.

Bloom of Trichodesmium (Oscillatoriales, Phormidiaceae) and seasonality of potentially harmful phytoplankton in San Pedro Bay, Leyte, Philippines.

Since 1983, San Pedro Bay in the Philippines had been reported to be the site of episodic Pyrodinium bahamense var. compressum blooms that caused paralytic shellfish poisoning in its nearby coastal communities. This bay is also subjected to numerous storms; the strongest was super typhoon Haiyan in November 8, 2013. For the first time, the seasonal dynamics of potentially toxic and harmful phytoplankton in this bay is elucidated. This is also the first record of a bloom of the cyanobacteria, Trichodesmium erythraeum that reached 70 000 colonies/L in April 2013 in this area. There were other 19 potentially toxic and harmful phytoplankton encountered during the sampling period. These consisted of a haptophyte, Phaeocystis globosa, the diatom Pseudo-nitzschia and 17 dinoflagellates. Seven of these harmful algae had densities high enough to be traced through time. Normally, diatoms abound during the dry season. But Pseudo-nitzschia increased in abundance during the wet season of 2012 and 2013. The dinoflagellates and Phaeocystis globosa behaved as expected and exhibited a relative increase in cell density during the rainy season of both years too. High nutrient availability during this season must have influenced the behavior of the phytoplankton despite differences in temperature and light intensity among seasons. Other notable but rare harmful species found only in plankton net tows during the study were Pyrodinium bahamense var. compressum, Alexandrium tamiyavanichii, Cochlodinium polykrikoides, and Noctiluca scintillans.