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ispartof: FUNCTIONAL ECOLOGY vol:37 issue:7 pages:1948-1961 status: published

Little is known on the dynamics of under-ice phytoplankton communities. We investigated phytoplankton communities in the upper (0–20 m) and lower (30–35 m) layer of oligotrophic Lake Tovel, Brenta Dolomites (Italy) over 6 years during summer and under ice. Winter conditions were different from one year to another with respect to ice thickness and snow cover. Proxies for light transmission (Secchi disc transparency, light attenuation) were similar between seasons, even though the incident solar radiation was lower in winter. Algal richness and chlorophyll-a were not different between seasons while biomass was higher during summer. In four of the 6 years, Bacillariophyta dominated during summer and Miozoa (class Dinophyceae) under ice while in 2 years Bacillariophyta also dominated under ice. Generally, a shift to larger size classes from summer to under ice was observed for Bacillariophyta, Chlorophyta, and Ochrophyta (class Chrysophyceae) while Dinophyceae showed the opposite pattern. No strong links between phytoplankton community composition and abiotic factors (under-ice convective mixing, snow on ice, under-ice light) were found. We suggest that inter-species relationships and more precise indicators of under-ice light should be considered to better understand under-ice processes.

International audience; Aquatic plants are crucial for the proper functioning of aquatic environments. So far, mixotrophy, widespread in aquatic microbial communities and in certain terrestrial plants, has been studied rarely in aquatic plants other than carnivorous ones. New evidence, particularly in Lemna and other close genera, suggests that mixotrophy is much more widespread than previously thought. Three processes could be involved, namely biotrophic associations, capture of prey (e.g., in carnivorous plants), and absorbotrophy (uptake of dissolved organic carbon). The latter process seems to be the most probable hypothesis in regard to the physiology of aquatic plants, but further research at both organism and cellular levels is needed to understand and clarify the processes involved and their importance. This opinion opens a new window of opportunity in aquatic ecology, with possible unsuspected applications in water management.

Mycorrhizal fungi are central to the biology of land plants. However, to what extent mycorrhizal shifts - broad evolutionary transitions in root-associated fungal symbionts - are related to changes in plant trophic modes remains poorly understood. We built a comprehensive DNA dataset of Orchidaceae fungal symbionts and a dated plant molecular phylogeny to test the hypothesis that shifts in orchid trophic modes follow a stepwise pattern, from autotrophy over partial mycoheterotrophy (mixotrophy) to full mycoheterotrophy, and that these shifts are accompanied by switches in fungal symbionts. We estimate that at least 17 independent shifts from autotrophy towards full mycoheterotrophy occurred in orchids, mostly through an intermediate state of partial mycoheterotrophy. A wide range of fungal partners was inferred to occur in the roots of the common ancestor of this family, including 'rhizoctonias', ectomycorrhizal, and wood- or litter-decaying saprotrophic fungi. Phylogenetic hypothesis tests further show that associations with ectomycorrhizal or saprotrophic fungi were most likely a prerequisite for evolutionary shifts towards full mycoheterotrophy. We show that shifts in trophic mode often coincided with switches in fungal symbionts, suggesting that the loss of photosynthesis selects for different fungal communities in orchids. We conclude that changes in symbiotic associations and ecophysiological traits are tightly correlated throughout the diversification of orchids. ispartof: NEW PHYTOLOGIST vol:231 issue:2 pages:791-800 ispartof: location:England status: published

Summary Galdieria sulphuraria is a cosmopolitan microalga found in volcanic hot springs and calderas. It grows at low pH in photoautotrophic (use of light as a source of energy) or heterotrophic (respiration as a source of energy) conditions, using an unusually broad range of organic carbon sources. Previous data suggested that G. sulphuraria cannot grow mixotrophically (simultaneously exploiting light and organic carbon as energy sources), its photosynthetic machinery being repressed by organic carbon.Here, we show that G. sulphuraria SAG21.92 thrives in photoautotrophy, heterotrophy and mixotrophy. By comparing growth, biomass production, photosynthetic and respiratory performances in these three trophic modes, we show that addition of organic carbon to cultures (mixotrophy) relieves inorganic carbon limitation of photosynthesis thanks to increased CO2 supply through respiration. This synergistic effect is lost when inorganic carbon limitation is artificially overcome by saturating photosynthesis with added external CO2.Proteomic and metabolic profiling corroborates this conclusion suggesting that mixotrophy is an opportunistic mechanism to increase intracellular CO2 concentration under physiological conditions, boosting photosynthesis by enhancing the carboxylation activity of Ribulose‐1,5‐bisphosphate carboxylase‐oxygenase (Rubisco) and decreasing photorespiration.We discuss possible implications of these findings for the ecological success of Galdieria in extreme environments and for biotechnological applications.

WOS:000614340800002; The impact of changes in nitrogen (N) and phosphorus (P) availability on the trophic strategies of planktonic protists was evaluated in a disturbed Mediterranean lagoon (Biguglia lagoon, France) using short-term bioassays. Natural communities were collected in three periods, i.e., autumn, spring and summer, to address the influence of the different environmental conditions. The responses of autotrophic plankton communities to experimentally induced N and/or P limitations were assessed as changes in chlorophyll a (Chl a) concentrations and in the abundances of potentially mixotrophic protists taxa. We observed blooms (\textgreater 105 cells l−1) of nanoflagellates in autumn, and of phycocyanin-rich picocyanobacteria in summer. Communities showed a co-limitation by N and P at the three sampling periods, despite high N:P ratios in autumn and spring. The high abundances of potentially mixotrophic dinoflagellates during these periods suggest the involvement of alternative trophic pathways for their maintenance in the lagoon. After bioassay incubations using different nutrient enrichment treatments, we often observed reduced abundances of mixotrophic protists containing Chl a with a concomitant increased abundance of protists without Chl a. This indicates a loss of chloroplasts and photoautotrophic abilities in protists cells, possibly reflecting a shift towards heterotrophy that could be sustained by phagotrophy.

12 pages, 6 figures, 2 tables, supplementary data https://doi.org/10.1016/j.marenvres.2022.105558.-- Dataset: Duarte Ferreira, Guilherme; Figueira, Joana; Marques, Sónia Cotrim; Hansen, Per Juel; Calbet, Albert; 2022; The strengths and weaknesses of Live Fluorescently Labelled Algae (LFLA) to estimate herbivory in protozooplankton and mixoplankton [Dataset]; DIGITAL.CSIC; https://doi.org/10.20350/digitalCSIC/14483; http://hdl.handle.net/10261/257594, The Live Fluorescently Labelled Algae (LFLA) technique has been used numerous times to estimate microzooplankton herbivory. Yet, it is unknown how mixoplankton (i.e., single-cell organisms that can combine phototrophy and phagotrophy) affect the outcome of this technique. Hence, we conducted a broad-spectrum assessment of the strengths and weaknesses of the LFLA technique, using several mixoplanktonic and protozooplanktonic grazers. Species from different taxonomic groups and different feeding mechanisms were tested in short-term experiments (ca. 5 h) in the laboratory, at different prey concentrations and during light and dark periods of the day. Overall, our findings suggest that the LFLA technique, due to its short-term nature, is an effective tracker of diel ingestion and digestion rates, and can detect new mixoplanktonic predators. We recommend that, irrespective of the prey concentration, incubations to measure grazing rates with this technique should generally be concluded within 1 h (adaptable to the environmental temperature). Nevertheless, our results also call for caution whenever using LFLA in the field: feeding mechanisms other than direct engulfment (like peduncle feeding) may provide severely biased ingestion rates. Furthermore, size and species selectivity are very hard to circumvent. To reduce the effects of selectivity, we propose the combined use of two distinctly coloured fluorochromes (i.e., distinct emission spectra). With this modification, one could either label different size ranges of prey or account for species-specific interactions in the food web, This project has received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 766327. [...] This project is a contribution of the Marine Zooplankton Ecology Group from the Generalitat de Catalunya (2017 SGR 87) with the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). Thanks for financial support are also due to MARE (UIDB/04292/2020 + UIDP/04292/2020), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020

Current mixotrophic culture systems for Dunaliella salina have technical limitations to achieve high growth and productivity. The purpose of this study was to optimize the mixotrophic conditions imposed by glycerol, light, and salinity that lead to the highest biomass and β-carotene yields in D. salina. The combination of 12.5 mM glycerol, 3.0 M salinity, and 50 μmol photons m−2 s−1 light intensity enabled significant assimilation of glycerol by D. salina and consequently enhanced growth (2.1 × 106 cell mL−1) and β-carotene accumulation (4.43 pg cell−1). The saline and light shock induced the assimilation of glycerol by this microalga. At last stage of growth, the increase in light intensity (300 μmol photons m−2 s−1) caused the β-carotene to reach values higher than 30 pg cell−1 and tripled the β-carotene values obtained from photoautotrophic cultures using the same light intensity. Increasing the salt concentration from 1.5 to 3.0 M NaCl (non-isosmotic salinity) produced higher growth and microalgal β-carotene than the isosmotic salinity 3.0 M NaCl. The mixotrophic strategy developed in this work is evidenced in the metabolic capability of D. salina to use both photosynthesis and organic carbon, viz., glycerol that leads to higher biomass and β-carotene productivity than that of an either phototrophic or heterotrophic process alone. The findings provide insights into the key role of exogenous glycerol with a strategic combination of salinity and light, which evidenced unknown roles of this polyol other than that in osmoregulation, mainly on the growth, pigment accumulation, and carotenogenesis of D. salina.

The traditional separation between primary producers (autotrophs) and consumers (heterotrophs) at the base of the marine food web is being increasingly replaced by the paradigm that mixoplankton, planktonic protists with the nutritional ability to use both phago(hetero)trophy and photo(auto)trophy to access energy are widespread globally. Thus, many ‘phytoplankton’ eat, while 50% of ‘protozooplankton’ also perform photosynthesis. Mixotrophy may enhance primary production, biomass transfer to higher trophic levels and the efficiency of the biological pump to sequester atmospheric CO2 into the deep ocean. Although this view is gaining traction, science lacks a tool to quantify the relative contributions of autotrophy and heterotrophy in planktonic protists. This hinders our understanding of their impacts on carbon cycling within marine pelagic ecosystems. It has been shown that the hydrogen (H) isotopic signature of lipids is uniquely sensitive to heterotrophy relative to autotrophy in plants and bacteria. Here, we explored whether it is also sensitive to the trophic status in protists. The new understanding of H isotope signature of lipid biomarkers suggests it offers great potential as a novel tool for quantifying the prevalence of mixotrophy in diverse marine microorganisms and thus for investigating the implications of the ‘mixoplankton’ paradigm.

Mixed microbial cultures have become a preferred choice of biocatalyst for chain elongation systems due to their ability to convert complex substrates into medium-chain carboxylates. However, the complexity of the effects of process parameters on the microbial metabolic networks is a drawback that makes the task of optimizing product selectivity challenging. Here, we studied the effects of small air contaminations on the microbial community dynamics and the product formation in anaerobic bioreactors fed with lactate, acetate and H2/CO2. Two stirred tank reactors and two bubble column reactors were operated with H2/CO2 gas recirculation for 139 and 116 days, respectively, at pH 6.0 and 32°C with a hydraulic retention time of 14 days. One reactor of each type had periods with air contamination (between 97 ± 28 and 474 ± 33 mL O2 L−1 d−1, lasting from 4 to 32 days), while the control reactors were kept anoxic. During air contamination, production of n-caproate and CH4 was strongly inhibited, whereas no clear effect on n-butyrate production was observed. In a period with detectable O2 concentrations that went up to 18%, facultative anaerobes of the genus Rummeliibacillus became predominant and only n-butyrate was produced. However, at low air contamination rates and with O2 below the detection level, Coriobacteriia and Actinobacteria gained a competitive advantage over Clostridia and Methanobacteria, and propionate production rates increased to 0.8–1.8 mmol L−1 d−1 depending on the reactor (control reactors 0.1–0.8 mmol L−1 d−1). Moreover, i-butyrate production was observed, but only when Methanobacteria abundances were low and, consequently, H2 availability was high. After air contamination stopped completely, production of n-caproate and CH4 recovered, with n-caproate production rates of 1.4–1.8 mmol L−1 d−1 (control 0.7–2.1 mmol L−1 d−1). The results underline the importance of keeping strictly anaerobic conditions in fermenters when consistent n-caproate production is the goal. Beyond that, micro-aeration should be further tested as a controllable process parameter to shape the reactor microbiome. When odd-chain carboxylates are desired, further studies can develop strategies for their targeted production by applying micro-aerobic conditions.

Background: Mixotrophy, combining phagotrophy and photoautotrophy in order to acquire nutrients and energy, is a widespread trophic mode in marine protist plankton. Acantharia (Radiolaria) are ubiquitous, but still uncultured oceanic protists. Many of them are mixoplanktic by endosymbiotic relations with microalgae. Here we aimed at quantitatively assessing phototrophy (inorganic nutrients) and phagotrophy (organic nutrients) of photosymbiotic Acantharia, to understand their physiology, and thereby improve integrations of mixotrophy into ecological models of oceanic ecosystems. Methods: Freshly collected Acantharia were incubated with stable isotopes of inorganic carbon and nitrogen to determine photosynthetic uptake rates. Grazing experiments (prey disappearance) were done with different algal cultures as potential food organisms to measure the contribution of prey ingestion to the acantharian metabolism. Fluorescently (and isotopically) labelled prey was used to verify prey uptake, qualitatively. Results/Conclusions: Carbon uptake rates were unaffected by the nitrogen source (i.e., nitrate or ammonium). Total carbon inorganic uptake rate was 1112±82 pgC h-1 Acantharia‑1, 22.3±1.6 pgC h-1 symbiont cell-1assuming 50 symbionts per Acantharia, at ~155-μmol photons m−2 s−1 irradiance. The Acantharia studied could use both inorganic ammonium and nitrate, but ammonium was taken up at a ~5 times higher rate. Prey ingestion of the haptophyte, Isochrysis galbana, was detected using labelled algae. Significant grazing by Acantharia could only be established on the dinoflagellate Effrenium voratum, with a grazing rate of 728 prey Acantharia‑1 hour-1 (i.e., ~56.3 ngC h-1, 46% of total holobiont carbon content) at a ratio of 1.06x104 prey predator-1. Daily photosynthetic carbon uptake rates made up ~14.5% of the total holobiont carbon content (0.9% hourly). The extent to which photosynthates are used and assimilated by the acantharian cell and/or if it is used for catabolic processes to obtain energy is still to be studied. Isotopic ratios further suggests seasonal differences in the usage of each trophic mode.

12 pages, 4 figures, 2 tables,1 appendix supplementary materials https://doi.org/10.1016/j.hal.2022.102292.-- Data availability: Data will be made available on request, Phaeocystis is a globally widespread marine phytoplankton genus, best known for its colony-forming species that can form large blooms and odorous foam during bloom decline. In the North Sea, Phaeocystis globosa typically becomes abundant towards the end of the spring bloom, when nutrients are depleted and the share of mixotrophic protists increases. Although mixotrophy is widespread across the eukaryotic tree of life and is also found amongst haptophytes, a mixotrophic nutrition has not yet been demonstrated in Phaeocystis. Here, we sampled two consecutive Phaeocystis globosa spring blooms in the coastal North Sea. In both years, bacterial cells were observed inside 0.6 – 2% of P. globosa cells using double CARD-FISH hybridizations in combination with laser scanning confocal microscopy. Incubation experiments manipulating light and nutrient availability showed a trend towards higher occurrence of intracellular bacteria under P-deplete conditions. Based on counts of bacteria inside P. globosa cells in combination with theoretical values of prey digestion times, maximum ingestion rates of up to 0.08 bacteria cell−1 h−1 were estimated. In addition, a gene-based predictive model was applied to the transcriptome assemblies of seven Phaeocystis strains and 24 other haptophytes to assess their trophic mode. This model predicted a phago-mixotrophic feeding strategy in several (but not all) strains of P. globosa, P. antarctica and other haptophytes that were previously assumed to be autotrophic. The observation of bacterial cells inside P. globosa and the gene-based model predictions strongly suggest that the phago-mixotrophic feeding strategy is widespread among members of the Phaeocystis genus and other haptophytes, and might contribute to their remarkable success to form nuisance blooms under nutrient-limiting conditions, This work was supported by a start-up budget provided to SW by IBED-UvA. With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)

Diatoms are unicellular photosynthetic protists which constitute one of the most successful microalgae contributing enormously to global primary productivity and nutrient cycles in marine and freshwater habitats. Though they possess the ability to biosynthesize high value compounds like eicosatetraenoic acid (EPA), fucoxanthin (Fx) and chrysolaminarin (Chrl) the major bottle neck in commercialization is their inability to attain high density growth. However, their unique potential of acquiring diverse carbon sources via varied mechanisms enables them to adapt and grow under phototrophic, mixotrophic as well as heterotrophic modes. Growth on organic carbon substrates promotes higher biomass, lipid, and carbohydrate productivity, which further triggers the yield of various biomolecules. Since, the current mass culture practices primarily employ open pond and tubular photobioreactors for phototrophic growth, they become cost intensive and economically non-viable. Therefore, in this review we attempt to explore and compare the mechanisms involved in organic carbon acquisition in diatoms and its implications on mixotrophic and heterotrophic growth and biomolecule production and validate how these strategies could pave a way for future exploration and establishment of sustainable diatom biorefineries for novel biomolecules.

International audience; Mycoheterotrophic plants have lost photosynthesis and obtain carbon through mycorrhizal fungi colonizing their roots. They are likely to have evolved from mixotrophic ancestors, which rely on both photosynthesis and fungal carbon for their development. Whereas our understanding of the ecological and genomic changes associated with the evolutionary shift to mycoheterotrophy is deepening, little information is known about the specific metabolic and physiological features driving this evolution. We investigated this issue in naturally occurring achlorophyllous variants of temperate mixotrophic orchids. We carried out an integrated transcriptomic and metabolomic analysis of the response to achlorophylly in the leaves of three mixotrophic species sampled in natura. Achlorophyllous leaves showed major impairment of their photosynthetic and mineral nutrition functions, strong accumulation of free amino acids, overexpression of enzymes and transporters related to sugars, amino acids and fatty acid catabolism, as well as induction of some autophagy-related and biotic stress genes. Such changes were reminiscent of these reported for variegated leaves and appeared to be symptomatic of a carbon starvation response. Rather than decisive metabolic innovations, we suggest that the evolution towards mycoheterotrophy in orchids is more likely to be reliant on the versatility of plant metabolism and an ability to exploit fungal organic resources, especially amino acids, to replace missing photosynthates.

The competition–defense tradeoff is a significant source of functional diversity in ecological communities. Here, we present a theoretical framework to describe the competition–defense tradeoff and apply it to a size‐based model of a unicellular plankton community. Specifically, we investigate how the emergent community structure depends on the shape of the trade‐off, and on whether the cost of defense is paid for by a lowered resource affinity or by an elevated metabolic rate. The inclusion of defense affects the size distribution and trophic strategies of the emerging community dependent on environmental conditions (eutrophic versus oligotrophic) and leads to increased diversity in size and trophic strategy under eutrophic conditions. Eutrophic conditions allow for better‐defended organisms than oligotrophic conditions. In most scenarios, competition–defense tradeoffs dampen trophic cascades in the seasonal cycle simulations, and increase the abundance of mixotrophs. We further demonstrate that it matters how the cost of defense is manifest (decreased affinity vs. increased metabolic rate), and that it has a significant effect on the resulting plankton community (overall biomass, size and feeding strategy diversity), particularly when the efficiency of the defense increases in direct proportion to the investment. Our results demonstrate that the structure of the ecosystem crucially depends on details of the defense tradeoff. This finding highlights the importance of a mechanistic understanding of defense tradeoffs, e.g. obtained through experimental measurements of specific defense mechanisms.

Chlorella is one of the most well-known microalgal genera, currently comprising approximately a hundred species of single-celled green algae according to the AlgaeBase. Strains of the genus Chlorella have the ability to metabolize both inorganic and organic carbon sources in various trophic modes and synthesize valuable metabolites that are widely used in many industries. The aim of this work was to investigate the impact of three trophic modes on the growth parameters, productivities of individual cell components, and biochemical composition of Chlorella sorokiniana, Chloroidium saccharofilum, and Chlorella vulgaris cells with special consideration of protein profiles detected by SDS-PAGE gel electrophoresis and two-dimensional gel electrophoresis with MALDI-TOF/TOF MS. Mixotrophic conditions with the use of an agro-industrial by-product stimulated the growth of all Chlorella species, which was confirmed by the highest specific growth rates and the shortest biomass doubling times. The mixotrophic cultivation of all Chlorella species yielded a high amount of protein-rich biomass with reduced contents of chlorophyll a, chlorophyll b, carotenoids, and carbohydrates. Additionally, this work provides the first information about the proteome of Chloroidium saccharofilum, Chlorella sorokiniana, and Chlorella vulgaris cells cultured in molasses supplementation conditions. The proteomic analysis of the three Chlorella species growing photoheterotrophically and mixotrophically showed increased accumulation of proteins involved in the cell energy metabolism and carbon uptake, photosynthesis process, and protein synthesis, as well as proteins involved in intracellular movements and chaperone proteins.

Green microalgae belonging to the genus Chlorella have long been used as health foods and additives for human consumption, as well as animal feed. These photosynthetic microorganisms have also found application in the bioremediation of wastewater and bio-mitigation of CO2. In the last decades, Chlorella microalgae have attracted considerable attention due to their great potential as feedstock for production of biofuels and high-value products. This review explores key aspects related to the cultivation of different Chlorella species under mixotrophic conditions. Common organic compounds and cheap substrates used for mixotrophic microalgal growth are presented. Bottlenecks and strategies to increase economic feasibility and sustainability of mixotrophic cultivation of Chlorella for future commercial production are also discussed. In this sense, the use of wastewater as a nutrient source for microalgae cultivation seems to be a promising alternative to improve the sustainability and economic feasibility of microalgae production systems. However, since wastewater presents a complex composition, methods to minimize the risk of contamination should be developed. Additionally, the screening of novel mixotrophic species of Chlorella capable of both assimilating a wide range of organic compounds from wastewater and accumulating high content of molecules of interest (e.g., lipids) would be highly recommended.

Background The production of microalgal biofuels, despite their sustainable and renowned potential, is not yet cost-effective compared to current conventional fuel technologies. However, the biorefinery concept increases the prospects of microalgal biomass as an economically viable feedstock suitable for the co-production of multiple biofuels along with value-added chemicals. To integrate biofuels production within the framework of a microalgae biorefinery, it is not only necessary to exploit multi-product platforms, but also to identify optimal microalgal cultivation strategies maximising the microalgal metabolites from which biofuels are obtained: starch and lipids. Whilst nutrient limitation is widely known for increasing starch and lipid formation, this cultivation strategy can greatly reduce microalgal growth. This work presents an optimisation framework combining predictive modelling and experimental methodologies to effectively simulate and predict microalgal growth dynamics and identify optimal cultivation strategies. Results Microalgal cultivation strategies for maximised starch and lipid formation were successfully established by developing a multi-parametric kinetic model suitable for the prediction of mixotrophic microalgal growth dynamics co-limited by nitrogen and phosphorus. The model’s high predictive capacity was experimentally validated against various datasets obtained from laboratory-scale cultures of Chlamydomonas reinhardtii CCAP 11/32C subject to different initial nutrient regimes. The identified model-based optimal cultivation strategies were further validated experimentally and yielded significant increases in starch (+ 270%) and lipid (+ 74%) production against a non-optimised strategy. Conclusions The optimised microalgal cultivation scenarios for maximised starch and lipids, as identified by the kinetic model presented here, highlight the benefits of exploiting modelling frameworks as optimisation tools that facilitate the development and commercialisation of microalgae-to-fuel technologies.

This study examined the effects of monochromatic illumination (blue, red, green and yellow) employing light-emitting diodes (LEDs), trophic conditions (photoautotrophic and mixotrophic), and nitrogen availability (high and low peptone concentration) on the growth and biochemical composition of Auxenochlorella protothecoides. The results revealed that mixotrophic conditions did not favor A. protothecoides, giving lower growth rates compared to heterotrophy (dark conditions). However, mixotrophy gave significantly higher growth rates compared to photoautotrophy. The best light wavelengths for mixotrophic cultivation were that of white and red. In all cases investigated in this study, high peptone concentration (4 g/L) resulted in decreased growth rates. Regarding the biochemical composition of A. protothecoides, the strongest effect, irrespective of trophic conditions, was caused by nitrogen availability (peptone concentration). Specifically, at nitrogen replete conditions (4 g/L peptone), biomass was rich in proteins (32–67%), whereas under deplete conditions (0.5 g/L peptone), A. protothecoides accumulated mainly carbohydrates (up to 56%). Mixotrophic conditions generally favored higher carbohydrate content, whereas photoautotrophic conditions favored higher protein content. The different illumination spectra did not have any clear effect on the biochemical composition (metabolites content), except that, in all trophic conditions, the use of the green spectrum resulted in higher chlorophyll b content. Chlorophyll a fluorescence studies revealed that the trophic conditions and the high peptone concentrations impacted the photosystem II (PSII) performance, and also affected plastoquinone re-oxidation kinetics and the heterogeneity of the PSII reaction centers.

The temporal variability of planktonic ciliates was studied in an extreme oligotrophic environment with special focus on trophic modes and size classes. Abundance, biomass, size classes, mixotrophy vs. heterotrophy, and species composition of planktonic ciliates were investigated focusing on temporal (samples collected on a monthly basis during 2019) and vertical (7 depth layers in the euphotic zone, from surface to 120 m) distribution at a coastal station in the oligotrophic Eastern Mediterranean. Abundance was in general very low (20 to 1150 cells L–1), except for September, which presented the highest abundance and biomass. Aloricate species dominated the ciliate community in all months and depths (% contribution from 77% in September to 99% in April). In general, oligotrichs presented maximum abundance at 2–10 m (except for June, July, and November: 100–120 m) whereas choreotrichs were more homogeneously distributed [and showed maxima at deep chlorophyll maximum (DCM)]. Small heterotrophs dominated the ciliate community at all depths and months, on average by 76% (they were 3 times more abundant than mixotrophs in terms of abundance and 2.5 times in terms of biomass). They were equally distributed both vertically and seasonally (and also in terms of size classes). In contrast, mixotrophs were found mainly at the surface layer to 20 m throughout the year, except for June and July (max at 100, 50 m). On average, 63% of integrated aloricate abundance was species 50 μm mixotrophic species were found only above the DCM. In contrast, during the mixing period of December to May, mixotrophic ciliates were very few and were dominated by small and medium-sized species. It seems that mixotrophic and heterotrophic ciliates, on one hand, and the four size classes, on the other, are very distinct groups characterized by different distributions both vertically and seasonally. This may have important consequences for the structure and function of the microbial food web of which they are part, as well as for the carbon flow to the higher trophic levels of this oligotrophic environment in which ciliates are the main grazers.

Allelopathy, a type of interference competition involving exuded chemicals, has been documented for several types of organisms including terrestrial plants, aquatic macrophytes, microbes, and planktonic algae. However, due to the dynamic nature of the aquatic environment it is unclear whether allelopathy is an evolutionarily stable competition mechanism in such a setting. In this research, we consider a cosmopolitan harmful algae species, Prymnesium parvum, for which multiple ecological roles of its produced deleterious chemicals have been suggested, including broadcast allelopathy (chemicals produced and exuded) and micropredation via cell–cell interactions (chemicals produced and held within the cell). To further investigate the ecological role of the deleterious chemicals, bioassays (with phytoplankton and zooplankton target organisms) were conducted using various fractions of a P. parvum culture grown under balanced N:P stoichiometry or imbalanced, P‐reduced stoichiometry. In addition, time‐series counts were generated with cell density enumerations and observations of behavior of a mixed species culture at intervals over a 24‐h period. Our results suggest that the apparent ecological role of the chemicals in a relatively high‐density population shifts depending on system stoichiometry in regard to nitrogen and phosphorus. We show that under balanced N:P conditions, deleterious chemical production is low and cell contact mediates mortality of prey. Differently, under imbalanced, P‐reduced conditions chemical effect is high and mass mortality occurs independent of cell contact. Thus, imbalanced N:P ratios shift the apparent ecological role of the deleterious chemicals from one of micropredation to one of broadcast allelopathy. These differences likely influence bloom dynamics and result in different ecological outcomes for systems affected by blooms. Further, we suggest that these differences indicate the importance of predatory mixotrophic feeding in the evolutionary maintenance of deleterious chemical production and the occurrence of allelopathic effects as a byproduct.

Mixotrophy, i.e., the capability of both phototrophy and phagotrophy within a single organism, is a prominent trophic mode in aquatic ecosystems. Mixotrophic strategies can be highly advantageous when feeding or photosynthesis alone does not sustain metabolic needs. In the current review, we discuss the functional types of mixotrophic marine protists (herein mixoplankton) within the context of evolution. Permanent plastids have been established in large due to gene transfer from prey and/or endosymbionts to the host cell. In some kleptoplastidic mixoplankton, prior gene transfers and active transcription of plastid related genes in the host can help maintain and extend retention of the current kleptoplast. In addition to kleptoplasts, the prey nucleus is also sometimes retained and actively transcribed to help maintain and even replicate the kleptoplasts. Endosymbiotic relations vary considerably in the extent to which hosts affect symbionts. For example, some endosymbionts are heavily modified to increase photosynthetic efficiency, or are controlled in their cell division. It can be proposed that many kleptoplasts and endosymbionts are in fact en route to becoming permanent plastids. Conditions such as increased temperature and limiting nutrients seem to favor phagotrophy in mixoplankton. However, responses of mixoplankton to changing environmental conditions like light irradiance, temperature, nutrient, and prey availability are variable and species-specific. Studying mixotrophs with temporary plastids could elucidate past and future evolutionary mechanisms and dynamics of processes such as phagotrophy and the establishment of (secondary) permanent plastids.

This study represents a comprehensive community-wide genome-centered metagenome analysis of biological soil crust (BSC) communities in arid environments, providing insights into the distribution of genes encoding different energy generation mechanisms, as well as survival strategies, among populations in an arid soil ecosystem. It reveals the metabolic potential of several uncultured and previously unsequenced microbial genera, families, and orders, as well as differences in the metabolic potential between the most abundant BSC populations and their cultured relatives, highlighting once more the danger of inferring function on the basis of taxonomy., Desert surface soils devoid of plant cover are populated by a variety of microorganisms, many with yet unresolved physiologies and lifestyles. Nevertheless, a common feature vital for these microorganisms inhabiting arid soils is their ability to survive long drought periods and reactivate rapidly in rare incidents of rain. Chemolithotrophic processes such as oxidation of atmospheric hydrogen and carbon monoxide are suggested to be a widespread energy source to support dormancy and resuscitation in desert soil microorganisms. Here, we assessed the distribution of chemolithotrophic, phototrophic, and desiccation-related metabolic potential among microbial populations in arid biological soil crusts (BSCs) from the Negev Desert, Israel, via population-resolved metagenomic analysis. While the potential to utilize light and atmospheric hydrogen as additional energy sources was widespread, carbon monoxide oxidation was less common than expected. The ability to utilize continuously available energy sources might decrease the dependency of mixotrophic populations on organic storage compounds and carbon provided by the BSC-founding cyanobacteria. Several populations from five different phyla besides the cyanobacteria encoded CO2 fixation potential, indicating further potential independence from photoautotrophs. However, we also found population genomes with a strictly heterotrophic genetic repertoire. The highly abundant Rubrobacteraceae (Actinobacteriota) genomes showed particular specialization for this extreme habitat, different from their closest cultured relatives. Besides the ability to use light and hydrogen as energy sources, they encoded extensive O2 stress protection and unique DNA repair potential. The uncovered differences in metabolic potential between individual, co-occurring microbial populations enable predictions of their ecological niches and generation of hypotheses on the dynamics and interactions among them. IMPORTANCE This study represents a comprehensive community-wide genome-centered metagenome analysis of biological soil crust (BSC) communities in arid environments, providing insights into the distribution of genes encoding different energy generation mechanisms, as well as survival strategies, among populations in an arid soil ecosystem. It reveals the metabolic potential of several uncultured and previously unsequenced microbial genera, families, and orders, as well as differences in the metabolic potential between the most abundant BSC populations and their cultured relatives, highlighting once more the danger of inferring function on the basis of taxonomy. Assigning functional potential to individual populations allows for the generation of hypotheses on trophic interactions and activity patterns in arid soil microbial communities and represents the basis for future resuscitation and activity studies of the system, e.g., involving metatranscriptomics.

From the southwestern termination of the Agulhas current, anticyclonic eddies are emitted and drift across the South Atlantic Ocean. This study is based on a FORSA (Following Ocean Rings in the South Atlantic) oceanographic cruise, from Cape Town (South AFRica) to Arraial do Cabo (Brazil) in June 2015, during which three eddies of different ages (E1, 7 months; E3, 11 months; E5, 24 months) from the Agulhas current were sampled for microplankton identification and determination of functional traits. The stations where sampling occurred at each eddy included a control outside the eddy and three stations inside the eddy—border, midway (between the border and center), and center (identified through satellite images of sea level anomaly—SLA). Functional traits were determined based on microscopic observations and consultation of the literature. An evident decay in the Agulhas eddies toward the west was observed, and each eddy proved to be different. E1 represented a younger and more robust structure. At the same time, the other eddies, E3 and E5, were more alike with similar physical, chemical, and ecological characteristics and almost the same indices values of functional diversity, demonstrating that although their species compositions were different, the strategies used by the species were the same. The most crucial ecological trait for microplankton was nutrition mode. The microplankton contained mainly mixotrophic dinoflagellates and cyanobacteria adapted to oligotrophic conditions. The functional strategy of microplankton did not differ among the eddies and stations, with mixotrophy being the most striking trait. Therefore, the older eddies’ microplankton community fits the neutrality theory, whereby species perform similar ecological functions, and the younger eddy fits in the niche complementarity. Even with the species composition being different in each eddy and/or within the same eddy, the functional strategy was the same, with scarce resources and species selected that best use any source of nutrients or use evolutionary advantages to live in an oligotrophic environment.

Many species of the ciliate genus Strombidium can acquire functional chloroplasts from a wide range of algal prey and are thus classified as generalist non-constitutive mixotrophs. Little, however, is known about the influence of irradiance and prey availability on their ability to exploit the photosynthetic potential of the chloroplasts, and how this may explain their spatial and temporal distribution in nature. In this study, inorganic carbon uptake, growth, and ingestion rates were measured for S. cf. basimorphum under three different irradiances (10, 40, and 120 μmol photons m–2 s–1) when acclimated to three different prey densities (5 × 103, 1 × 104, and 4 × 104 cells mL–1), as well as when allowed to deplete the prey. After prey depletion, cultures survived without prey longest (∼6 days) at the medium irradiance treatment (40 μmol photons m–2 s–1), while ciliate density, inorganic carbon uptake rates, and cellular chl-a content declined fastest at the highest irradiance treatment. This indicates that the ciliates may be unable to maintain the chloroplasts functionally without replacement at high irradiances. Ingestion rates were not shown to be significantly influenced by irradiance. The maximum gross growth efficiency (GGE) in this study (1.1) was measured in cultures exposed to the medium test irradiance and lowest prey density treatment (5 × 103 cells mL–1). The relative contribution of inorganic carbon uptake to the ciliate carbon budget was also highest in this treatment (42%). A secondary GGE peak (0.99) occurred when cultures were exposed to the highest test irradiance and the medium prey density. These and other results suggest that S. cf. basimorphum, and other generalist non-constitutive mixotrophs, can flexibly exploit many different environmental conditions across the globe.

Microalgae are a promising source of polyunsaturated fatty acids as well as bioactive antioxidant compounds such as carotenoids, phenolics and tocopherols. However, the accumulation of these biomolecules is often promoted by conflicting growth conditions. In this study, a phased bioprocessing strategy was developed to simultaneously enhance the lipid and antioxidant amounts by tailoring nitrogen content in the cultivation medium and applying light stress. This approach increased the overall contents of total fatty acids, carotenoids, phenolics, and α-tocopherol in Chlorella vulgaris by 2.2-, 2.2-, 1.5-, and 2.1-fold, respectively. Additionally, the bioaccessibility of the lipids and bioactives from the obtained biomasses improved after pulsed electric field (5 μs, 20 kV cm−1, 31.8 kJ kg−1sus) treatment (up to +12%) and high-pressure homogenization (100 MPa, 5-6 passes) (+41-76%). This work represents a step towards the generation of more efficient algae biorefineries, thus expanding the alternative resources available for essential nutrients., Bioresource Technology, 347, ISSN:1873-2976, ISSN:0960-8524

The extremophilic red alga Galdieria partita is a facultative heterotroph that occupies mostly low-light microhabitats. However, the exceptional detection of abundant populations of G. partita in sunlight-exposed soil raises the possibility that exogenous organic carbon sources protect cells from photo-oxidative damage. The present study aimed to identify the photoprotective process activated by exogenous glucose under photo-oxidative stress. We demonstrated that exogenous glucose mitigated the photo-oxidative damage of cells exposed to 300 μmol photons m–2 s–1 photosynthetic active radiation. Photosynthesis carbon assimilation scarcely contributed to the cell growth in the presence of glucose, but the photosynthetic apparatus was nevertheless maintained and protected by glucose in a concentration-dependent manner. Supplementation of glucose increased expression of the L-gulonolactone oxidase gene essential for ascorbic acid biosynthesis, whereas no enhanced expression of the genes involved in carotenoid or tocopherol biosynthesis was observed. Under the photo-oxidative stress condition, the ascorbic acid content was strongly enhanced by exogenous glucose. We propose that the biosynthesis of ascorbic acid is one of the major photoprotective processes induced by exogenous glucose. The elucidation of how ascorbic acid is involved in scavenging reactive oxygen species provides key insights into the photoprotective mechanism in red algae.

Memoria de tesis doctoral presentada por Laura Rubinat Ripoll para obtener el título de Docteur en la Spécialité Écologie por la Sorbonne Université, realizada bajo la dirección del Dr. Ramiro Logares del Institut de Ciències del Mar (ICM-CSIC), del Dr. Colomban de Vargas y el Dr. Olivier Jaillon.-- 181 pages, figures, tables, [EN] Primary production is the synthesis of organic matter out of inorganic molecules, and in most ecosystems is achieved through photosynthesis. Eukaryotic microbial phototrophs and mixotrophs are main contributors to primary production in the global oligotrophic ocean, supporting processes of energy and biomass trophic transfer at a planetary scale. Despite the ecological value of these organisms, their wide taxonomic and functional diversity remains largely unknown. In this thesis, we use different types of molecular data obtained from the Tara Oceans circumglobal expedition to assess the composition and trophic diversity of picoeukaryotic primary producers in the oligotrophic ocean. In the first part of the thesis, we compare metagenomic and metabarcoding data to assess the evolutionary diversity and relative abundance of picophototrophic organisms. We identified phototrophic bacteria as three-fold more abundant and significantly less phylogenetically diverse than phototrophic picoeukayrotes. Prymnesiophyceae, Mamiellophyceae, Pelagophyceae and Dictiochophyceae appeared as the dominant groups of phototrophic picoeukaryotes in terms of relative richness and abundance. In the fourth chapter, we describe a predictive model to quantify the abundance of trophic groups in metagenomic samples. This taxonomy-free approach revealed the dominance of photo-trophic organisms across all ocean basins, while the contribution of phagomixo- and phago-trophs oscillated around 25% of the relative abundance in most samples. In the last study included in this thesis we assess the distribution of a collection of single-cell amplified genomes across all Tara Oceans samples. Our results argue that single-cell sequencing technique has the potential to recover the genome of dominant protists in the global oligotrophic ocean with a relatively low sampling effort. Overall, this work describes a number of approaches based on molecular data for the assessment of primary producers distribution and diversity in marine environments, [FR] La production primaire est la synthèse de matière organique à partir de molécules inorganiques et, dans la plupart des écosystèmes, elle est réalisée à travers de la photosynthèse. Les microbiens eucaryotes phototrophes et mixotrophes sont les principaux contributeurs à la production primaire dans l’océan oligotrophe global, soutenant les processus de transfert trophique d’énergie et de biomasse à l’échelle planétaire. [...]

Basic understanding of the photosynthetic physiology of the oleaginous green microalga Ettlia oleoabundans is still very limited, including the modulation of the photosynthetic membrane upon metabolism conversion from autotrophy to mixotrophy. It was previously reported that, upon glucose supply in the culture medium, E. oleoabundans preserves photosystem II (PSII) from degradation by virtue of a higher packing of thylakoid complexes. In this work, it was investigated whether in the mixotrophic exponential growth phase the PSII activity is merely preserved or even enhanced. Modulated fluorescence parameters were then recorded under short-term treatments with increasing irradiance values of white light. It was found that the mixotrophic microalga down-regulated the chlororespiratory electron recycling from photosystem I (PSI), but enhanced the linear electron flow from PSII to PSI. Ability to keep PSII more open than in autotrophic growth conditions indicated that the respiration of the glucose taken up from the medium fed the carbon fixing reactions with CO2. The overall electron poise was indeed well regulated, with a lesser need for thermal dissipation of excess absorbed energy. It is proposed that the significant, though small, increase in PSII maximum quantum yield in mixotrophic cells just reflects an improved light energy use and an increased photochemical capacity as compared to the autotrophic cells.

Methanothermobacter Met2 is a metagenome-assembled genome (MAG) that encodes a putative mixotrophic methanogen constituting the major populations in thermophilic fixed-bed anaerobic digesters. In order to characterize its physiology, the present work isolated an archaeon (strain Met2-1) that represents Met2-type methanogens by using a combination of enrichments under a nitrogen atmosphere, colony formation on solid media and limiting dilution under high partial pressures of hydrogen. Strain Met2-1 utilizes hydrogen and carbon dioxide for methanogenesis, while the growth is observed only when culture media are additionally supplemented with acetate. It does not grow on acetate in the absence of hydrogen. The results demonstrate that Methanothermobacter sp. strain Met2-1 is a novel methanogen that exhibits obligate mixotrophy.

Resource competition theory predicts that when two species compete for a single, finite resource, the better competitor should exclude the other. However, in some cases, weaker competitors can persist through intraguild predation, that is, by eating their stronger competitor. Mixotrophs, species that meet their carbon demand by combining photosynthesis and phagotrophic heterotrophy, may function as intraguild predators when they consume the phototrophs with which they compete for light. Thus, theory predicts that mixotrophy may allow for coexistence of two species on a single limiting resource. We tested this prediction by developing a new mathematical model for a unicellular mixotroph and phytoplankter that compete for light, and comparing the model's predictions with a laboratory experimental system. We find that, like other intraguild predators, mixotrophs can persist when an ecosystem is sufficiently productive (i.e., the supply of the limiting resource, light, is relatively high), or when species interactions are strong (i.e., attack rates and conversion efficiencies are high). Both our mathematical and laboratory models show that, depending upon the environment and species traits, a variety of equilibrium outcomes, ranging from competitive exclusion to coexistence, are possible.

PREMISE OF THE STUDY The evolution of full mycoheterotrophy is one of the most interesting topics within plant evolution. The leafless orchid Cymbidium macrorhizon is often assumed to be fully mycoheterotrophic even though it has a green stem and fruit capsule. Here, we assessed the trophic status of this species by analyzing the chlorophyll content and the natural 13 C and 15 N abundance in the sprouting and the fruiting season. METHODS The chlorophyll content was measured in five sprouting and five fruiting individuals of C. macrorhizon that were co-occurring. In addition, their 13 C and 15 N isotopic signatures were compared with those of neighboring autotrophic and partially mycoheterotrophic reference plants. KEY RESULTS Fruiting individuals of C. macrorhizon were found to contain a remarkable amount of chlorophyll compared to their sprouting counterparts. In addition, the natural abundance of 13 C in the tissues of the fruiting plants was slightly depleted relative to the sprouting ones. Linear two-source mixing model analysis revealed that fruiting C. macrorhizon plants obtained approximately 73.7 ± 2.0% of their total carbon from their mycorrhizal fungi when the sprouting individuals were used as the 100% carbon gain standard. CONCLUSIONS Our results indicated that despite its leafless status, fruiting plants of C. macrorhizon were capable of fixing significant quantities of carbon. Considering the autotrophic carbon gain increases during the fruiting season, its photosynthetic ability may contribute to fruit and seed production. These results indicate that C. macrorhizon should, therefore, be considered a partially mycoheterotrophic species rather than fully mycoheterotrophic, at least during the fruiting stage.

Microzooplankton community structure, distribution, growth and herbivory were examined in the eastern Fram Strait and Arctic Ocean shelf affected by the Atlantic water inflow in May (during the spring bloom, chlorophyll up to 9 µg l-1) and August (post-bloom, 0.13 – 1.7 µg l-1) 2014. Microzooplankton were collected along two longitudinal transects at 79°N and 79.4 °N from the slope to the ice edge and crossed the Western Spitsbergen Current (WSC). In May, integrated microzooplankton biomass in the upper 100 m ranged from 0.16 g C m-2 above the slope to 2.3 g C m-2 within WSC (0.71 g C m-2 on average). Mixotrophic oligotrich ciliates from the genus Strombidium dominated in the spring and formed a surface bloom (79 x 103 cells L-1, 206 µg C L-1). This is the largest microzooplankton biomass recorded in the Arctic so far. The heterotrophic dinoflagellates Gyrodinium and Protoperidinium were abundant at the diatom-dominated stations in the marginal ice zone. In the summer, a more diverse community included a large proportion of heterotrophic and mixotrophic dinoflagellates, tintinnids, and other ciliates. Microzooplankton biomass increased to the average of 1.27 g C m-2. At the ice-covered and open water stations in the Yermak shelf and deep basin, microzooplankton grew at 0.04 to 0.38 d-1; their species-specific growth rates were up to 1.79 d-1. Microzooplankton herbivory on average removed 72% (36 to >100%) of daily primary production with the exception of Phaeocystis pouchetii colonies. These results indicate that the microbial food web plays a central role in the carbon cycle in this Atlantic-influenced polar system.

Eutrophication of inland waters is expected to increase the frequency and severity of harmful algal blooms (HABs). Toxin-production associated with HABs has negative effects on human health and aquatic ecosystem functioning. Despite evidence that flagellates can ingest toxin-producing cyanobacteria, interactions between members of the microbial loop are underestimated in our understanding of the food web and algal bloom dynamics. Physical and allelopathic interactions between a mixotrophic flagellate (Cryptomonas sp.) and two strains of a cyanobacteria (Microcystis aeruginosa) were investigated in a full-factorial experiment in culture. The maximum population growth rate of the mixotroph (0.25 day−1) occurred during incubation with filtrate from toxic M. aeruginosa. Cryptomonas was able to ingest toxic and non-toxic M. aeruginosa at maximal rates of 0.5 and 0.3 cells day−1, respectively. The results establish that although Cryptomonas does not derive benefits from co-incubation with M. aeruginosa, it may obtain nutritional supplement from filtrate. We also provide evidence of a reduction in cyanotoxin concentration (microcystin-LR) when toxic M. aeruginosa is incubated with the mixotroph. Our work has implications for “trophic upgrading” within the microbial food web, where cyanobacterivory by nanoflagellates may improve food quality for higher trophic levels and detoxify secondary compounds.

We studied quagmires around 17 soft-water lakes in southern and southeastern Estonia. Vegetation analysis was carried out at the level of moss and field layer synusiae and plant communities. The aims of the current study were to elucidate the main factors determining the species richness of these quagmires, in order to ascertain what types of synusiae and plant communities form their vegetation, and what are their indicator species. Increasing the pH of peat-water increased the number of bryophyte species and the total number of species. The number of bryophyte species was positively related to through-flowing lakes and neighbouring forest vegetation. In total nine societies of bryophyte synusiae, 14 societies of vascular plants and eight community types were distinguished. Six community types represented minerotrophic quaking fen, and two types were classified as mixotrophic quaking bog. Our results show clearly a relative independency of synusiae; similar moss synusiae can associate with synusiae of various vascular plant societies and vice versa.

Some jellyfish host zooxanthellae in their tissues (mostly from the family Symbiodiniaceae; Dinophyceae) and supplement their heterotrophic nutrition with their symbiont's photosynthates. The mixotrophy of zooxanthellate jellyfishes (as holobionts) renders the study of their nutrition, growth, and population dynamics complicated. Here, we used an experimental approach to assess how carbon and nitrogen stable isotopes (delta C-13 and delta N-15) as well as the elemental composition (C:N ratios) of zooxanthellate jellyfishes are affected by variations in nutrition sources: i.e. predation (heterotrophic) versus photosynthesis (autotrophic). Our laboratory experiment, conducted on the zooxanthellate jellyfish Cassiopea sp. medusae (including symbionts) in the presence or absence of light and prey during 24 days, showed conclusive results. Presence of light decreased delta N-15, increased delta C-13 and C:N ratios, whereas presence of prey increased delta N-15, and decreased delta C-13 and C:N ratios. The medusae incubated with both light and prey had intermediate delta N-15, delta C-13 and C:N ratios. Variations in zooxanthellate jellyfishes' nutrition sources (autotrophy vs. heterotrophy) are thus reflected by their isotopic and elemental composition. By disentangling the effects of autotrophy and of heterotrophy on zooxanthellate jellyfish isotopic and elemental compositions, these results would help to interpret the values of delta C-13, delta N-15 and C:N ratios that can be observed on these organisms in fieldwork studies.

Through the merger of previously independent lineages, symbiosis promotes the acquisition of new traits and exploitation of inaccessible ecological niches [1, 2], driving evolutionary innovation and important ecosystem functions [3–6]. The transient nature of establishment makes study of symbiotic origins difficult, but experimental comparison of independent origins could reveal the degree of convergence in the underpinning mechanisms [7, 8]. We compared the metabolic mechanisms of two independent origins of Paramecium bursaria-Chlorella photosymbiosis [9–11] using a reciprocal metabolomic pulse-chase method. This showed convergent patterns of nutrient exchange and utilization for host-derived nitrogen in the Chlorella genotypes [12, 13] and symbiont-derived carbon in the P. bursaria genotypes [14, 15]. Consistent with a convergent primary nutrient exchange, partner-switched host-symbiont pairings were functional. Direct competition of hosts containing native or recombined symbionts against isogenic symbiont-free hosts showed that the fitness benefits of symbiosis for hosts increased with irradiance but varied by genotype. Global metabolism varied more between the Chlorella than the P. bursaria genotypes and suggested divergent mechanisms of light management. Specifically, the algal symbiont genotypes either produced photo-protective carotenoid pigments at high irradiance or more chlorophyll, resulting in corresponding differences in photosynthetic efficiency and non-photochemical quenching among host-symbiont pairings. These data suggest that the multiple origins of P. bursaria-Chlorella symbiosis use a convergent nutrient exchange, whereas other photosynthetic traits linked to functioning of photosymbiosis have diverged. Although convergence enables partner switching among diverse strains, phenotypic mismatches resulting from divergence of secondary symbiotic traits could mediate host-symbiont specificity in nature. Sørensen et al. compare multiple independent evolutionary origins of Paramecium-Chlorella symbiosis to reveal the underpinning metabolic mechanisms. Although the independent origins use a convergent nutrient exchange, they have diverged in traits linked to photosynthesis, which could mediate host-symbiont specificity in nature.

Cryptophyte algae are globally distributed photosynthetic flagellates found in freshwater, estuarine, and neritic ecosystems. While cryptophytes can be highly abundant and are consumed by a wide variety of protistan predators, few studies have sought to quantify in situ grazing rates on their populations. Here we show that autumnal grazing rates on in situ communities of cryptophyte algae in Chesapeake Bay are high throughout the system, while growth rates, particularly in the lower bay, were low. Analysis of the genetic diversity of cryptophyte populations within dilution experiments suggests that microzooplankton may be selectively grazing the fastest-growing members of the population, which were generally Teleaulax spp. We also demonstrate that potential grazing rates of ciliates and dinoflagellates on fluorescently labeled (FL) Rhodomonas salina, Storeatula major, and Teleaulax amphioxeia can be high (up to 149 prey predator−1 d−1), and that a Gyrodinium sp. and Mesodinium rubrum could be selective grazers. Potential grazing was highest for heterotrophic dinoflagellates, but due to its abundance, M. rubrum also had a high overall impact. This study reveals that cryptophyte algae in Chesapeake Bay can experience extremely high grazing pressure from phagotrophic protists, and that this grazing likely shapes their community diversity.

Despite its ecological importance, essential aspects of microbial N2O reduction—such as the effect of O2 availability on the N2O sink capacity of a community—remain unclear. We studied N2O vs. aerobic respiration in a chemostat culture to explore (i) the extent to which simultaneous respiration of N2O and O2 can occur, (ii) the mechanism governing the competition for N2O and O2, and (iii) how the N2O-reducing capacity of a community is affected by dynamic oxic/anoxic shifts such as those that may occur during nitrogen removal in wastewater treatment systems. Despite its prolonged growth and enrichment with N2O as the sole electron acceptor, the culture readily switched to aerobic respiration upon exposure to O2. When supplied simultaneously, N2O reduction to N2 was only detected when the O2 concentration was limiting the respiration rate. The biomass yields per electron accepted during growth on N2O are in agreement with our current knowledge of electron transport chain biochemistry in model denitrifiers like Paracoccus denitrificans. The culture’s affinity constant (KS) for O2 was found to be two orders of magnitude lower than the value for N2O, explaining the preferential use of O2 over N2O under most environmentally relevant conditions. Electronic supplementary material The online version of this article (10.1007/s00253-018-9247-3) contains supplementary material, which is available to authorized users.

In this study, Tisochrysis lutea was cultivated in mixotrophic and heterotrophic cultures with glycerol as a carbon source and with glucose and acetate for comparison; autotrophic cultivation was the control group without a carbon source. It was found that T. lutea used glycerol and did not use glucose and acetate under mixotrophy. Mixotrophy slightly elevated the docosahexaenoic acid (DHA) and total fatty acids (TFA) content in the dry-weight and enhanced the DHA and TFA production in medium (41.3 and 31.9% respectively) at the end of a 16-day cultivation, while heterotrophy reduced the DHA content and TFA production. Under the mixotrophy, the glycerol contribution to the DHA production (16.19 mg/L) and the TFA production (97.8 mg/L) was not very high and the DHA yield [2.63% chemical oxygen demand (COD)] and TFA yield (13.1% COD) were also very low. Furthermore, T. lutea using glycerol had a period of adaptation, indicating that T. lutea was not an ideal microalga for organic carbon utilization. Electronic supplementary material The online version of this article (10.1186/s13568-018-0580-9) contains supplementary material, which is available to authorized users.

Identifying putative mixotrophic protist species in the environment is important for understanding their behavior, with the recovery of these species in culture essential for determining the triggers of feeding, grazing rates, and overall impact on bacterial standing stocks. In this project, mixotroph abundances determined using tracer ingestion in water and sea ice samples collected in the Ross Sea, Antarctica during the summer of 2011 were compared with data from the spring (Ross Sea) and fall (Arctic) to examine the impacts of bacterivory/mixotrophy. Mixotrophic nanoplankton (MNAN) were usually less abundant than heterotrophs, but consumed more of the bacterial standing stock per day due to relatively higher ingestion rates (1–7 bacteria mixotroph−1 h−1 vs. 0.1–4 bacteria heterotroph−1 h−1). Yet, even with these high rates observed in the Antarctic summer, mixotrophs appeared to have a smaller contribution to bacterivory than in the Antarctic spring. Additionally, putative mixotroph taxa were identified through incubation experiments accomplished with bromodeoxyuridine-labeled bacteria as food, immunoprecipitation (IP) of labeled DNA, and amplification and high throughput sequencing of the eukaryotic ribosomal V9 region. Putative mixotroph OTUs were identified in the IP samples by taxonomic similarity to known phototroph taxa. OTUs that had increased abundance in IP samples compared to the non-IP samples from both surface and chlorophyll maximum (CM) depths were considered to represent active mixotrophy and include ones taxonomically similar to Dictyocha, Gymnodinium, Pentapharsodinium, and Symbiodinium. These OTUs represent target taxa for isolation and laboratory experiments on triggers for mixotrophy, to be combined with qPCR to estimate their abundance, seasonal distribution and potential impact.

The exploitation of petroleum as energy and material source opened unprecedented possibilities for the development of our human societies, but only now we realize that the use of fossil resources comes at devastatingly high environmental costs. Consequently, our efforts to tap other carbon sources are steadily increasing. Industrial microbiology has the potential to use carbon dioxide directly as carbon source, thereby converting a foe into a friend. This thematic issue of FEMS Microbiology Letters sheds some light on recent developments for the understanding of microbial pathways for carbon dioxide fixation and on strategies for their industrial exploitation.

Among green microalgae, Scenedesmus sp. is known for its potential in wastewater remediation and lipid production, especially under starvation. Moreover, it is often characterised by a mixotrophic metabolism. In this work, we cultivated S. acutus PVUW12 in the presence of a liquid fraction of scotta (LFS), a cheese whey by-product, as source of nutrients. Subsequently, cultures were starved to evaluate lipid production. Cells were analysed to obtain information about growth, nutrient consumption during LFS cultivation, morphology and photosynthetic efficiency. We found that the alga boosted its growth when cultured in presence of LFS. Production of stromatic starch grains, polyphosphate granules, cell wall enlargement and reduction of the photosynthetic efficiency were also induced. Massive lipid accumulation was observed only during starvation, which also induced a strong slowdown of growth, loss of polyphosphate grains and further decrease in photosynthetic efficiency. This study demonstrates that S. acutus PVUW12 can be involved in a two-step cultivation, first by promoting growth using a by-product from cheese industry and second by transferring the microalgae on starvation to induce lipid accumulation for bioenergetics purposes.

Leles, S.G. ... et. al.-- 6 pages, 3 figures, supplementary material https://royalsocietypublishing.org/doi/suppl/10.1098/rspb.2017.0664, This first comprehensive analysis of the global biogeography of marine protistan plankton with acquired phototrophy shows these mixotrophic organisms to be ubiquitous and abundant; however, their biogeography differs markedly between different functional groups. These mixotrophs, lacking a constitutive capacity for photosynthesis (i.e. non-constitutive mixotrophs, NCMs), acquire their phototrophic potential through either integration of prey-plastids or through endosymbiotic associations with photosynthetic microbes. Analysis of field data reveals that 40–60% of plankton traditionally labelled as (non-phototrophic) microzooplankton are actually NCMs, employing acquired phototrophy in addition to phagotrophy. Specialist NCMs acquire chloroplasts or endosymbionts from specific prey, while generalist NCMs obtain chloroplasts from a variety of prey. These contrasting functional types of NCMs exhibit distinct seasonal and spatial global distribution patterns. Mixotrophs reliant on ‘stolen’ chloroplasts, controlled by prey diversity and abundance, dominate in high-biomass areas. Mixotrophs harbouring intact symbionts are present in all waters and dominate particularly in oligotrophic open ocean systems. The contrasting temporal and spatial patterns of distribution of different mixotroph functional types across the oceanic provinces, as revealed in this study, challenges traditional interpretations of marine food web structures. Mixotrophs with acquired phototrophy (NCMs) warrant greater recognition in marine research

Pyroleae species are perennial understory shrubs, many of which are partial mycoheterotrophs. Most fungi colonizing Pyroleae roots are ectomycorrhizal (ECM) and share common mycobionts with their Pyroleae hosts. However, such mycobiont sharing has neither been examined in depth before nor has the interspecific variation in sharing among Pyroleae species. Here, we examined root-associated fungal communities in three co-existing Pyroleae species, including Pyrola alpina, Pyrola incarnata, and Orthilia secunda, with reference to co-existing ECM fungi on the surrounding trees in the same soil blocks in subalpine coniferous forests. We identified 42, 75, and 18 fungal molecular operational taxonomic units in P. alpina, P. incarnata, and O. secunda roots, respectively. Mycobiont sharing with surrounding trees, which was defined as the occurrence of the same mycobiont between Pyroleae and surrounding trees in each soil block, was most frequent among P. incarnata (31 of 44 plants). In P. alpina, sharing was confirmed in 12 of 37 plants, and the fungal community was similar to that of P. incarnata. Mycobiont sharing was least common in O. secunda, found in only 5 of 32 plants. Root-associated fungi of O. secunda were dominated by Wilcoxina species, which were absent from the surrounding ECM roots in the same soil blocks. These results indicate that mycobiont sharing with surrounding trees does not equally occur among Pyroleae plants, some of which may develop independent mycorrhizal associations with ECM fungi, as suggested in O. secunda at our research sites. Electronic supplementary material The online version of this article (doi:10.1007/s00572-017-0788-6) contains supplementary material, which is available to authorized users.

We have previously shown that Prochlorococcus sp. SS120 strain takes up glucose by using a multiphasic transporter encoded by the Pro1404 gene. Here, we studied the glucose uptake kinetics in multiple Prochlorococcus strains from different ecotypes, observing diverse values for the Ks constants (15–126.60 nM) and the uptake rates (0.48–6.36 pmol min-1 mg prot-1). Multiphasic kinetics was observed in all studied strains, except for TAK9803-2. Pro1404 gene expression studies during the 21st Atlantic Meridional Transect cruise showed positive correlation with glucose concentrations in the ocean. This suggests that the Pro1404 transporter has been subjected to diversification along the Prochlorococcus evolution, in a process probably driven by the glucose availabilities at the different niches it inhabits. The glucose uptake mechanism seems to be a primary transporter. Glucose addition induced detectable transcriptomic and proteomic changes in Prochlorococcus SS120, but photosynthetic efficiency was unaffected. Our studies indicate that glucose is actively taken up by Prochlorococcus, but its uptake does not significantly alter the trophic ways of this cyanobacterium, which continues performing photosynthesis. Therefore Prochlorococcus seems to remain acting as a fundamentally phototrophic organism, capable of using glucose as an extra resource of carbon and energy when available in the environment.

Autotrophic archaeal and bacterial ammonia-oxidisers (AOA and AOB) drive soil nitrification. Ammonia limitation, mixotrophy, and pH have been suggested as factors providing niche specialisation and differentiation between soil AOA and AOB. However, current data from genomes, cultures, field studies, and microcosms suggest that no single factor discriminates between AOA and AOB. In addition, there appears to be sufficient physiological diversity within each group for growth and activity in all soils investigated, with the exception of acidic soils (pH

noxia, and ciliates might play a very important role in the plankton community budget there. We analysed changes in the composition and biomass of the ciliate assemblage and other microbial loop components throughout the oxycline just at the end of stratification in a warm-monomictic lake, Lake Alchichica, Mexico (four samplings: 2006-2008, 2010); the results were compared with those obtained from another lake from the region, La Preciosa, sampled in 2010. Bacteria, autotrophic picoplankton (APP) and flagellates were analysed using epifluorescence microscopy. Ciliates were evaluated either in DAPI stained samples (looking for pigmented organelles and/or ingested phototrophs) or in quantitative protargol stain (QPS) permanent preparations, where they were identified at the genus or species level. The end of the stratification period in Lake Alchichica was characterized by almost uniform heterotrophic picoplankton (HPP) numbers (106 cells mL-1) throughout the water column. Meanwhile, APP showed epilimnetic and/or metalimnetic maxima of 105 cells mL-1 followed by an order of magnitude drop in the hypolimnion. A very important peak (105 cells mL-1) of the autotrophic or mixotrophic flagellate Pyramimonas sp. was observed repeatedly above and within the oxycline of Lake Alchichica. Ciliate biomass maxima were found around the oxycline and in the above-bottom layer. The top of the oxycline was dominated by Euplotes spp. and Spirostomum teres fine- to coarse-filter feeders (feeding upon APP, nanodiatoms and algae). Raptorial haptorids (in particular, Phialina sp.) were the second most important group, generally occupying the layer below euplotids, followed by Holophrya and Prorodon facultative anaerobic prostomes. Sometimes, strictly anaerobic Caenomorpha sp. was found to be important in the anoxic hypolimnion. Minute picoplankton feeding species (both APP and heterotrophic bacteria feeders) were important throughout the water column: in the epilimnion, vorticellids (2006-2008) or scuticociliates (2010) dominated. Typically, the scuticociliate maximum was located in the oxycline and/or above the bottom. Some microaerophilic species were isolated; thus, their identification could be carried out. However, the apparent polymorphic ciliate life cycles were not described completely, and the species composition was only estimated: two dominant species (SC 1 – Cristigera-like and SC 2 – Cyclidim-like) covered nearly the total scuticociliate biomass. Strictly anaerobic scuticociliates were not isolated but observed in the deepest layers of the lake (bacteria symbiotic Isocyclidium globosum and Cristigera sp.). Significant statistical relation within the ciliate distribution and environmental variables was not confirmed due to unique species composition in the respective years. However, general trends in the distribution of ciliates on a species level were observed. Scuticociliates, including two important tentatively identified species, did not present unambiguous ecological position, and the study of their live cycle should be the next step in investigations.

WOS:000372014200055; International audience; Mixotrophic plankton, which combine the uptake of inorganic resources and the ingestion of living prey, are ubiquitous in marine ecosystems, but their integrated biogeochemical impacts remain unclear. We address this issue by removing the strict distinction between phytoplankton and zooplankton from a global model of the marine plankton food web. This simplification allows the emergence of a realistic trophic network with increased fidelity to empirical estimates of plankton community structure and elemental stoichiometry, relative to a system in which autotrophy and heterotrophy are mutually exclusive. Mixotrophy enhances the transfer of bio-mass to larger sizes classes further up the food chain, leading to an approximately threefold increase in global mean organism size and an similar to 35% increase in sinking carbon flux.