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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.

Ciliophora is a phylum of unicellular eukaryotes that are common and have pivotal roles in aquatic environments. Sea ice is a marine habitat, which is composed of a matrix of solid ice and pockets of saline water in which Ciliophora thrive. Here, we used phylogenetic placement to identify Ciliophora 18S ribosomal RNA reads obtained from wintertime water and sea ice, and assigned functions to the reads based on this taxonomic information. Based on our results, sea-ice Ciliophora assemblages are poorer in taxonomic and functional richness than under-ice water and water-column assemblages. Ciliophora diversity stayed stable throughout the ice-covered season both in sea ice and in water, although the assemblages changed during the course of our sampling. Under-ice water and the water column were distinctly predominated by planktonic orders Choreotrichida and Oligotrichida, which led to signifcantly lower taxonomic and functional evenness in water than in sea ice. In addition to planktonic Ciliophora, assemblages in sea ice included a set of moderately abundant surface-oriented species. Omnivory (feeding on bacteria and unicellular eukaryotes) was the most common feeding type but was not as predominant in sea ice as in water. Sea ice included cytotrophic (feeding on unicellular eukaryotes), bacterivorous and parasitic Ciliophora in addition to the predominant omnivorous Ciliophora. Potentially mixotrophic Ciliophora predominated the water column and heterotrophic Ciliophora sea ice. Our results highlight sea ice as an environment that creates a set of variable habitats, which may be threatened by the diminishing extent of sea ice due to changing climate. DNA metabarcoding · Phylogenetic placement · Mixotrophy · Predator–prey interactions · Winter ecology

Diatoms are microalgae well known for their high variability and high primary productivity, being responsible for about 20% of the annual global carbon fixation. Moreover, they are interesting as potential feedstocks for the production of biofuels and high-value lipids and carotenoids. Diatoms exhibit trophic flexibility and, under certain conditions, they can grow mixotrophically by combing photosynthesis and respiration. So far, only a few species of diatoms have been tested for their mixotrophic metabolism; in some cases, they produced more biomass and with higher lipid content when grown under this condition. Phaeodactylum tricornutum is the most studied diatom species for its mixotrophic metabolism due to available genome sequence and molecular tools. However, studies in additional species are needed to better understand the conservation of this process in diatoms and its potential in industrial applications. Here, we describe the photosynthetic and respiratory pathways involved in mixotrophy and provide an overview of the trophic variability in diatoms. This review also highlights promising areas of industrial applications for diatoms when cultivated under mixotrophy.

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.

Background Mixotrophy can confer a higher growth rate than the sum of photoautotrophy and heterotrophy in many microalgal species. Thus, it has been applied to biodiesel production and wastewater utilization. However, its carbon and energy metabolic mechanism is currently poorly understood. Results To elucidate underlying carbon and energy metabolic mechanism of mixotrophy, Chromochloris zofingiensis was employed in the present study. Photosynthesis and glucose metabolism were found to operate in a dynamic balance during mixotrophic cultivation, the enhancement of one led to the lowering of the other. Furthermore, compared with photoautotrophy, non-photochemical quenching and photorespiration, considered by many as energy dissipation processes, were significantly reduced under mixotrophy. Comparative transcriptome analysis suggested that the intermediates of glycolysis could directly enter the chloroplast and replace RuBisCO-fixed CO2 to provide carbon sources for chloroplast organic carbon metabolism under mixotrophy. Therefore, the photosynthesis rate-limiting enzyme, RuBisCO, was skipped, allowing for more efficient utilization of photoreaction-derived energy. Besides, compared with heterotrophy, photoreaction-derived ATP reduced the need for TCA-derived ATP, so the glucose decomposition was reduced, which led to higher biomass yield on glucose. Based on these results, a mixotrophic metabolic mechanism was identified. Conclusions Our results demonstrate that the intermediates of glycolysis could directly enter the chloroplast and replace RuBisCO-fixed CO2 to provide carbon for photosynthesis in mixotrophy. Therefore, the photosynthesis rate-limiting enzyme, RuBisCO, was skipped in mixotrophy, which could reduce energy waste of photosynthesis while promote cell growth. This finding provides a foundation for future studies on mixotrophic biomass production and photosynthetic metabolism.

Premise Difficulties in comparing extremely divergent features in fully mycoheterotrophic plants with those in closely related chlorophyllous plants have complicated attempts to reveal the evolutionary patterns and processes of fully mycoheterotrophic plants. Albino mutants of partially mycoheterotrophic plants, generally observed in Orchidaceae, have provided an ideal model for investigating the evolution of mycoheterotrophy within similar genetic backgrounds. In 2018, we found a putative albino population of Pyrola (Ericaceae). Here we aimed to reveal the identity of the albino pyroloid and confirm its fully mycoheterotrophic status. Methods To reveal the putative albino pyroloid's identity, we examined its morphology and sequenced its chloroplast DNA. In addition, we assessed the trophic status of the putative albino pyroloid by analyzing chlorophyll fluorescence, chlorophyll concentration, and natural 13 C and 15 N abundances. Results We identified albino individuals as P. japonica-otherwise a partially mycoheterotrophic species. We confirmed their albino status by their considerably lower chlorophyll fluorescence and concentrations than those of sympatrically occurring chlorophyllous plants. 13 C abundance in the albino individuals was significantly higher than in the green individuals of P. japonica. Conclusions This first report of albino mutants from partially mycoheterotrophic species in angiosperms other than orchids will play a valuable role in further studies focused on mycoheterotrophy. For instance, their δ13 C and δ15 N values represent a reference for fully mycoheterotrophic plants in Pyrola. Our findings also indicate the strong dependence of some leafy Pyrola species on fungal C during their entire life cycle.

Although Haematococcus lacustris has been developed for astaxanthin production for decades, the production cost is still high. In order to modify the production processes, we proposed a novel strategy of cultivation, featured by sequential indoor continuous mixotrophic cultivation for the production of green cells followed by outdoor phototrophic induction for astaxanthin accumulation. The continuous mixotrophic cultivation was first optimized indoor, and then the seed culture of mixotrophic cultivation was inoculated into outdoor open raceway ponds for photoinduction. The results showed that mixotrophically grown cultures could efficiently grow without losing their photosynthetic efficiency and yielded higher biomass concentration (0.655 g L−1) and astaxanthin content (2.2% DW), compared to phototrophically grown seed culture controls. This novel strategy might be a promising alternative to the current approaches to advance the production technology of astaxanthin from microalgae.

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.

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.

The trophic ecology of mixotrophic, zooxanthellate jellyfishes potentially spans a wide spectrum between autotrophy and heterotrophy. However, their degree of trophic plasticity along this spectrum is not well known. To better characterize their trophic ecology, we sampled the zooxanthellate medusa Mastigias papua in contrasting environments and sizes in Palau (Micronesia). We characterized their trophic ecology using isotopic (bulk δ13C and δ15N), elemental (C:N ratios), and fatty acid compositions. The different trophic indicators were correlated or anti-correlated as expected (Pearson’s correlation coefficient, rP > 0.5 or < -0.5 in 91.1% of cases, p < 0.05), indicating good agreement. The sampled M. papua were ordered in a trophic spectrum between autotrophy and heterotrophy (supported by decreasing δ13C, C:N, proportion of neutral lipid fatty acids (NLFA:TLFA), n-3:n-6 and increasing δ15N, eicosapentaenoic acid to docosahexaenoic acid ratio (EPA:DHA)). This trophic spectrum was mostly driven by sampling location with little influence of medusa size. Moreover, previous observations have shown that in a given location, the trophic ecology of M. papua can change over time. Thus, the positions on the trophic spectrum of the populations sampled here are not fixed, suggesting high trophic plasticity in M. papua. The heterotrophic end of the trophic spectrum was occupied by non-symbiotic M. papua, whereas the literature indicates that the autotrophic end of the spectrum corresponds to dominant autotrophy, where more than 100% of the carbon requirement is obtained by photosynthesis. Such high trophic plasticity has critical implications for the trophic ecology and blooming ability of zooxanthellate jellyfishes.

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.

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.

Partial mycoheterotrophy, the ability of plants to obtain carbon from fungi throughout their life cycle in combination with photosynthesis, appears to be more common within the Plant Kingdom than previously anticipated. Recent studies using stable isotope analyses have indicated that isotope signatures in partially mycoheterotrophic plants vary widely among species, but the relative contributions of family- or species-specific characteristics and the identity of the fungal symbionts to the observed differences remain unclear. Here, we investigated in detail mycorrhizal communities and isotopic signatures in four co-occurring terrestrial orchids (Platanthera chlorantha, Epipactis helleborine, E. neglecta and the mycoheterotrophic Neottia nidus-avis). All investigated species were mycorrhizal generalists (i.e., associated with a large number of fungi simultaneously), but mycorrhizal communities differed significantly between species. Mycorrhizal communities associating with the two Epipactis species consisted of a wide range of fungi belonging to different families, whereas P. chlorantha and N. nidus-avis associated mainly with Ceratobasidiaceae and Sebacinaceae species, respectively. Isotopic signatures differed significantly between both Epipactis species, with E. helleborine showing near autotrophic behavior and E. neglecta showing significant enrichment in both carbon and nitrogen. No significant differences in photosynthesis and stomatal conductance were observed between the two partially mycoheterotrophic orchids, despite significant differences in isotopic signatures. Our results demonstrate that partially mycoheterotrophic orchids of the genus Epipactis formed mycorrhizas with a wide diversity of fungi from different fungal families, but variation in mycorrhizal community composition was not related to isotope signatures and thus transfer of C and N to the plant. We conclude that the observed differences in isotope signatures between E. helleborine and E. neglecta cannot solely be explained by differences in mycorrhizal communities, but most likely reflect a combination of inherent physiological differences and differences in mycorrhizal communities.

Polyhydroxyalkanoates (PHAs) are the biopolymer of choice if we look for a substitute of petroleum-based non-biodegradable plastics. Microbial production of PHAs as carbon reserves has been studied for decades and PHAs are gaining attention for a wide range of applications in various fields. Still, their uneconomical production is the major concern largely attributed to high cost of organic substrates for PHA producing heterotrophic bacteria. Therefore, microalgae/cyanobacteria, being photoautotrophic, prove to have an edge over heterotrophic bacteria. They have minimal metabolic requirements, such as inorganic nutrients (CO2, N, P, etc.) and light, and they can survive under adverse environmental conditions. PHA production under photoautotrophic conditions has been reported from cyanobacteria, the only candidate among prokaryotes, and few of the eukaryotic microalgae. However, an efficient cultivation system is still required for photoautotrophic PHA production to overcome the limitations associated with (1) stringent management of closed photobioreactors and (2) optimization of monoculture in open pond culture. Thus, a hybrid system is a necessity, involving the participation of microalgae/cyanobacteria and bacteria, i.e., both photoautotrophic and heterotrophic components having mutual interactive benefits for each other under different cultivation regime, e.g., mixotrophic, successive two modules, consortium based, etc. Along with this, further strategies like optimization of culture conditions (N, P, light exposure, CO2 dynamics, etc.), bioengineering, efficient downstream processes, and the application of mathematical/network modeling of metabolic pathways to improve PHA production are the key areas discussed here. Conclusively, this review aims to critically analyze cyanobacteria as PHA producers and proposes economically sustainable production of PHA from microbial autotrophs and heterotrophs in “hybrid biological system.”

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.

The mixotrophic cultivation of microalgae often couples high biomass production with lipid accumulation, and thus it is considered a useful approach to obtain high quantities of added value microalgal biomass. In this study, the marine diatom Thalassiosira pseudonana was supplied with glycerol to determine the potential of the mixotrophic alga for the accumulation of lipids. In preliminarily experiments, T. pseudonana was cultivated in the presence of increasing concentrations of glucose, acetate and glycerol in order to select the best organic carbon source for growth promotion, and glycerol was selected for further experiments. The best glycerol dose for both biomass productivity and lipid production was 2.5 g L−1. To overcome the costs of the pure glycerol supply, the crude glycerol derived from a biodiesel production plant was also tested, as a low-cost alternative. Beside growth and photosynthetic parameters useful to monitor the algal conditions, lipid amount and profile were compared between autotrophic and pure/crude glycerol-treated algae. Interestingly, an almost doubled lipid content was accumulated in mixotrophic algae compared to the autotrophic ones. Moreover, an evident increase in a relatively short-chain fatty acid (C14:0, myristic acid), potentially useful for bio-jet fuels, and in a long-chain omega3-polyunsaturated fatty acid (C18:4; stearidonic acid), interesting for nutritional purposes, was highlighted in mixotrophic samples, especially when cultivated with crude glycerol. Differently, the fatty acid profile obtained from the autotrophic cultures suggested a preferable employment of the algal biomass for biodiesel production, owing to the prevalence of palmitic and palmitoleic acids. This study highlights that the mixotrophic cultivation of T. pseudonana drives the algal metabolism towards the production of different lipids, suitable for diversified applications. Moreover, the use of biodiesel-derived crude glycerol could make mixotrophy economically advantageous, in particular considering a bio-refinery approach.

Despite being an autotrophic organism, Galdieria phlegrea (Galdieriaceae) has the ability to use glycerol thereby switching to heterotrophy in the dark and mixotrophy in the presence of light. To examine cellular changes during the switch to mixotrophic metabolism and finally to photoautotrophic metabolism, heterotrophic cells of G. phlegrea were exposed to light and split into two subcultures. Cells exposed only to light but cultivated in medium containing glycerol grew with a recovery time of at least 3 days. In a parallel culture, the simultaneous removal of glycerol from the culture medium and light exposure allowed Galdieria cells to rapidly recover their growth rate due to their ability to rapidly absorb ammonium from the medium. However, contrary to expectation, a higher content of total soluble protein was observed in light-exposed cells cultivated in medium containing glycerol compared to cells cultivated without glycerol. In addition, the level of Rubisco in cells exposed to light and cultivated without glycerol was higher than those in cells cultivated in medium containing glycerol, indicating full photosynthetic functionality after only 3 days of light treatment. The greater chlorophyll a content confirms that the photosynthetic activity of cells cultivated without glycerol recovers earlier than that of heterotrophic cells exposed to light but still cultivated in medium containing glycerol.

Many marine ciliate species retain functional chloroplasts from their photosynthetic prey. In some species, the functionality of the acquired plastids is connected to the simultaneous retention of prey nuclei. To date, this has never been documented in plastidic Strombidium species. The functionality of the sequestered chloroplasts in Strombidium species is thought to be independent from any nuclear control and only maintained via frequent replacement of chloroplasts from newly ingested prey. Chloroplasts sequestered from the cryptophyte prey Teleaulax amphioxeia have been shown to keep their functionality for several days in the ciliate Strombidium cf. basimorphum. To investigate the potential retention of prey genetic material in this ciliate, we applied a molecular marker specific for this cryptophyte prey. Here, we demonstrate that the genetic material from prey nuclei, nucleomorphs, and ribosomes is detectable inside the ciliate for at least 5 days after prey ingestion. Moreover, single-cell transcriptomics revealed the presence of transcripts of prey nuclear origin in the ciliate after 4 days of prey starvation. These new findings might lead to the reconsideration of the mechanisms regulating chloroplasts retention in Strombidium ciliates. The development and application of molecular tools appear promising to improve our understanding on chloroplasts retention in planktonic protists.

As the number of mixotrophic protists has been increasingly documented, “mixoplankton”, a third category separated from the traditional categorization of plankton into “phytoplankton” and “zooplankton”, has become a new paradigm and research hotspot in aquatic plankton ecology. While species of dinoflagellates are a dominant group among all recorded members of mixoplankton, the trophic modes of Karlodinium, a genus constituted of cosmopolitan toxic species, were reviewed due to their representative features as mixoplankton and harmful algal blooms (HABs)-causing dinoflagellates. Among at least 15 reported species in the genus, three have been intensively studied for their trophic modes, and all found to be phagotrophic. Their phagotrophy exhibits multiple characteristics: (1) omnivority, i.e., they can ingest a variety of preys in many forms; (2) flexibility in phagotrophic mechanisms, i.e., they can ingest small preys by direct engulfment and much bigger preys by myzocytosis using a peduncle; (3) cannibalism, i.e., species including at least K. veneficum can ingest the dead cells of their own species. However, for some recently described and barely studied species, their tropical modes still need to be investigated further regarding all of the above-mentioned aspects. Mixotrophy of Karlodinium plays a significant role in the population dynamics and the formation of HABs in many ways, which thus deserves further investigation in the aspects of physiological ecology, environmental triggers (e.g., levels of inorganic nutrients and/or presence of preys), energetics, molecular (genes and gene expression regulations) and biochemical (e.g., relevant enzymes and signal molecules) bases, origins, and evaluation of the advantages of being a phagotroph.

Pico-/nanoeukaryotes (P/NEs) comprise both primary producers and bacterial predators, playing important biogeochemical and ecological roles in the marine microbial loop. Besides the difference in size, these small-sized fractions can be distinguished from microplankton by certain functional and ecological traits. Nevertheless, little information is available regarding patterns of their taxonomic and functional diversity and community composition along environmental gradients in coastal marine ecosystems. In this study, we applied high-throughput sequencing of 18S rRNA gene to assess the taxonomic species richness and community composition of P/NEs in surface waters of Bohai Sea and North Yellow Sea, northern China spanning a 600-km distance during summer and winter of 2011. The richness of operational taxonomic units (OTUs) formed a U-shaped relationship with concentration of chlorophyll a (Chl-a, a proxy of primary productivity), but a stronger, negative relationship with concentration of dissolved oxygen (DO). These two factors also significantly co-varied with the OTU-based community composition of P/NEs. The effect of geographic distance on community composition of P/NEs was negligible. Among the three functional groups defined by trophic traits, heterotrophs had the highest OTU richness, which exhibited a U-shaped relationship with both DO and Chl-a. The community of P/NEs was dominated by heterotrophs and mixotrophs in terms of read numbers, which showed a trade-off along the gradient of phosphate, but no significant changes along DO and Chl-a gradients, indicating functional redundancy. Similarly, the proportion of phototrophs was significantly and positively correlated with the concentration of silicate. Our results indicate that taxonomic and functional composition of P/NEs are decoupled on a regional scale, and limiting nutrients are important factors in modulating functional composition of these microorganisms in the studied area. These findings contribute toward gaining a better understanding of how diversity of small eukaryotes and their functions are structured in coastal oceans and the effect of environmental changes on the structuring process.

Mixotrophic organisms are increasingly recognized as important components of ecosystems, but the factors controlling their nutrition pathways (in particular their autotrophy-heterotrophy balance) are little known. Both autotrophy and heterotrophy are expected to respond to density-dependent mechanisms but not necessarily in the same direction and/or strength. We hypothesize that the autotrophy-heterotrophy balance of mixotrophic organisms might therefore be a function of population densities. To investigate this relationship, we sampled mixotrophic jellyfish holobionts (host, Mastigias papua etpisoni; symbiont, Cladocopium sp.) in a marine lake (Palau, Micronesia) on six occasions (from 2010 to 2018). Over this period, population densities varied similar to 100 fold. We characterized the nutrition of the holobionts using the delta C-13 and delta N-15 signatures as well as C:N ratios. delta C-13 values increased and delta N-15 values decreased with increasing population densities (respectively, R-2 = 0.86 and 0.70, P < 0.05). Although less distinct, C:N ratios increased with increasing population densities (R-2 = 0.59, 0.1 > P > 0.05). This indicates that the autotrophy-heterotrophy balance tends toward autotrophy when population densities increase. We propose that the availability of zooplanktonic prey is the main driver of this pattern. These results demonstrate that the autotrophy-heterotrophy balance of mixotrophic jellyfishes can be tightly regulated by density-dependent mechanisms.

Ciliates represent an important trophic link between nanoplankton and mesoplankton. Many species acquire functional chloroplasts from photosynthetic prey, being thus mixotrophs. Little is known about which algae they exploit,and of the relevance of inorganic carbon assimilation to theirmetabolism. To get insights into these aspects, laboratory cultures of three mixotrophic Strombidium spp. were established and 35 photosynthetic algal species were tested as prey.The relative contributions of ingestion and photosynthesis to total carbon uptake were determined, and responses to prey starvation were studied.Ciliate growth was supported by algal species in the 2–12 μm size range, with cryptophytes and chlorophytes being the best prey types. Inorganic carbon incorporation was only quantitatively important when prey concentration was low (3–100 μgCL−1), when it led to increased gross growth efficiencies. Chla specific inorganic carbon uptake rates were reduced by 60–90% compared to that of the photosynthetic prey. Inorganic carbon uptake alone could not sustain survival of cultures and ciliate populations declined by 25–30% during 5 days of starvation. The results suggest thatmixotrophy in Strombidium spp. may substantially bolster the efficiency of trophic transfer when biomass of small primary producers is low. Ciliates represent an important trophic link between nanoplankton and mesoplankton. Many species acquire functional chloroplasts from photosynthetic prey, being thus mixotrophs. Little is known about which algae they exploit, and of the relevance of inorganic carbon assimilation to theirmetabolism. To get insights into these aspects, laboratory cultures of three mixotrophic Strombidium spp. were established and 35 photosynthetic algal species were tested as prey. The relative contributions of ingestion and photosynthesis to total carbon uptake were determined, and responses to prey starvation were studied.Ciliate growth was supported by algal species in the 2–12 μm size range, with cryptophytes and chlorophytes being the best prey types. Inorganic carbon incorporation was only quantitatively important when prey concentration was low (3–100 μgCL−1), when it led to increased gross growth efficiencies. Chla specific inorganic carbon uptake rates were reduced by 60–90% compared to that of the photosynthetic prey. Inorganic carbon uptake alone could not sustain survival of cultures and ciliate populations declined by 25–30% during 5 days of starvation. The results suggest thatmixotrophy in Strombidium spp. may substantially bolster the efficiency of trophic transfer when biomass of small primary producers is low.

Chloroflexales (Chloroflexi) are typical members of the anoxygenic photosynthesizing component of microbial mats and have mostly been characterized from communities associated to hot springs. Here, we report the assembly of five metagenome-assembled genomes (MAGs) of a novel lineage of Chloroflexales found in mesophilic lithifying microbial mats (microbialites) in Lake Alchichica (Mexico). Genomic and phylogenetic analyses revealed that the bins shared 92% of their genes, and these genes were nearly identical despite being assembled from samples collected along a depth gradient (1–15 m depth). We tentatively name this lineage Candidatus Lithoflexus mexicanus. Metabolic predictions based on the MAGs suggest that these chlorosome-lacking mixotrophs share features in central carbon metabolism, electron transport, and adaptations to life under oxic and anoxic conditions, with members of two related lineages, Chloroflexineae and Roseiflexineae. Contrasting with the other diverse microbialite community members, which display much lower genomic conservation along the depth gradient, Ca. L. mexicanus MAGs exhibit remarkable similarity. This might reflect a particular flexibility to acclimate to varying light conditions with depth or the capacity to occupy a very specific spatial ecological niche in microbialites from different depths. Alternatively, Ca. L. mexicanus may also have the ability to modulate its gene expression as a function of the local environmental conditions during diel cycles in microbialites along the depth gradient.

Many marine microbial eukaryotes combine photosynthetic with phagotrophic nutrition, but incomplete understanding of such mixotrophic protists, their functional diversity, and underlying physiological mechanisms limits the assessment and modeling of their roles in present and future ocean ecosystems. We developed an experimental system to study responses of mixotrophic protists to availability of living prey and light, and used it to characterize contrasting physiological strategies in two stramenopiles in the genus Ochromonas. We show that oceanic isolate CCMP1393 is an obligate mixotroph, requiring both light and prey as complementary resources. Interdependence of photosynthesis and heterotrophy in CCMP1393 comprises a significant role of mitochondrial respiration in photosynthetic electron transport. In contrast, coastal isolate CCMP2951 is a facultative mixotroph that can substitute photosynthesis by phagotrophy and hence grow purely heterotrophically in darkness. In contrast to CCMP1393, CCMP2951 also exhibits a marked photoprotection response that integrates non-photochemical quenching and mitochondrial respiration as electron sink for photosynthetically produced reducing equivalents. Facultative mixotrophs similar to CCMP2951 might be well adapted to variable environments, while obligate mixotrophs similar to CCMP1393 appear capable of resource efficient growth in oligotrophic ocean environments. Thus, the responses of these phylogenetically close protists to the availability of different resources reveals niche differentiation that influences impacts in food webs and leads to opposing carbon cycle roles.

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.

The uptake of dissolved organic compounds, that is, osmotrophy, has been shown to be an efficient nutritional strategy for algae. However, this mode of nutrition may affect the biochemical composition, for example, the fatty acid (FA) contents, of algal cells. This study focused on the osmotrophic assimilation of glucose and leucine by selected seven algal strains belonging to chlorophytes, chrysophytes, cryptophytes, dinoflagellates and euglenoids. Our laboratory experiments with stable isotope labeling showed that osmotrophy occurred in four of the selected seven strains. However, only three of these produced long chain omega-3 FAs eicosapentaenoic acid (EPA; 20:5ω3) and docosahexaenoic acid (DHA; 22:6ω3). High glucose content (5 mg L−1) affected negatively on the total FAs ofMallomonas kalinaeand the total omega-3 FAs ofCryptomonassp. Further, glucose assimilation explained 35% (negative effect) and leucine assimilation 48% (positive effect) of the variation of EPA, DHA and the FAs related to their synthesis inCryptomonassp. Moderate glucose concentration (2 mg L−1) was found to enhance the growth ofCryptomonas ozolinii, whereas low leucine (20 µg L−1) enhanced the growth ofM. kalinae. However, no systematic effect of osmotrophy on growth rates was detected. Our study shows that osmotrophic assimilation of algae is species and compound specific, and that the effects of the assimilated compounds on algal metabolism also varies depending on the species.

Trait-based ecology allows much of the complexity of ecosystems to be projected onto a low-dimensional trait space. We conjecture that three key traits capture the main aspects of diversity of unicellular planktonic organisms: cell size, trophic strategies (relative investment in photosynthesis and phagotrophy) and vacuolation. The three selected traits are representative of two groups: mixotrophic protists, which in addition to phototrophy, use phagotrophic grazing as a food source, and diatoms that use fluid-filled vacuoles to increase their physical size relative to carbon biomass. We construct a trait-based model related to the three traits and determine the optimal trait values of cells of a given carbon size in a given environment. We also perform fully dynamic simulations to study the self-assembled trait distribution at steady state and throughout a seasonal cycle. Diatoms’ strategy is shown to be advantageous among small cells and under early spring conditions, in which highly vacuolated diatoms are simulated, while mixotrophy is dominant among larger cells when nutrients are limiting (summer conditions). The novelty of this approach lies in the mechanistic understanding of plankton physiology that is successfully used to capture how community trait structure emerges from trade-offs that constrain the different strategy among unicellular plankton.

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.

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.

In this study, Arthrospira platensis was grown in the presence of different glycerol concentrations (0.5&ndash, 9 g/L) under three light intensities (5, 10 and 15 Klux) in semi-continuous mode and under non-axenic conditions. The aim of this study was to investigate the growth performance, the biomass biochemical composition and any interactions between A. platensis and bacteria that would potentially grow as well on glycerol. The results here show that glycerol did not have any positive effect on biomass production of A. platensis. In contrast, it was observed that by increasing glycerol concentration the growth performance of A. platensis was restricted, while a gradual increase of bacteria population was observed, which apparently outcompeted and repressed A. platensis growth. Chlorophyll fluorescence measurements (Quantum Yields) revealed that glycerol was not an inhibiting factor per se of photosynthesis. On the other hand, cyanobacterial biomass grown on glycerol displayed a higher content in proteins and lipids. Especially, protein productivity was enhanced around 15&ndash, 35% with the addition of glycerol compared to the control. In distinction, carbohydrate and photosynthetic pigments (phycocyanin and chlorophyll-&alpha, ) content decreased with the increase of glycerol concentration. The results here suggest that A. platensis did not utilize glycerol for biomass production but most probably as metabolic energy carrier towards synthesis of proteins and lipids, which are more energy consuming metabolites compared to carbohydrates. The study revealed that the addition of glycerol at amounts of 0.5&ndash, 1.5 g/L could be a strategy to improve protein productivity by A. platensis.

Some green orchids obtain carbon from their mycorrhizal fungi, as well as from photosynthesis. These partially mycoheterotrophic orchids sometimes produce fully achlorophyllous, leaf-bearing (albino) variants. Comparing green and albino individuals of these orchids will help to uncover the molecular mechanisms associated with mycoheterotrophy. We compared green and albino Epipactis helleborine by molecular barcoding of mycorrhizal fungi, nutrient sources based on 15N and 13C abundances, and gene expression in their mycorrhizae by RNA-seq and cDNA de novo assembly. Molecular identification of mycorrhizal fungi showed that green and albino E. helleborine harbored similar mycobionts, mainly Wilcoxina. Stable isotope analyses indicated that albino E. helleborine plants were fully mycoheterotrophic, whereas green individuals were partially mycoheterotrophic. Gene expression analyses showed that genes involved in antioxidant metabolism were up-regulated in the albino variants, which indicates that these plants experience greater oxidative stress than the green variants, possibly due to a more frequent lysis of intracellular pelotons. It was also found that some genes involved in the transport of some metabolites, including carbon sources from plant to fungus, is higher in albino than in green variants. This result may indicate a bidirectional carbon flow even in the mycoheterotrophic symbiosis. The genes related to mycorrhizal symbiosis in autotrophic orchids and arbuscular mycorrhizal plants were also up-regulated in the albino variants, indicating the existence of common molecular mechanisms among the different mycorrhizal types. This article is protected by copyright. All rights reserved.

Mixotrophic species use both organic and mineral carbon sources. Some mixotrophic plants combine photosynthesis and a nutrition called mycoheterotrophy, where carbon is obtained from fungi forming mycorrhizal symbiosis with their roots. These species can lose photosynthetic abilities and evolve full mycoheterotrophy. Besides morphological changes, the latter transition is associated with a deep alteration of the plastid genome. Photosynthesis-related genes are lost first, followed by housekeeping genes, eventually resulting in a highly reduced genome. Whether relaxation of selective constraints already occurs for the plastid genome of mixotrophic species, which remain photosynthetic, is unclear. This is partly due to the difficulty of comparing plastid genomes of autotrophic, mixotrophic, and mycoheterotrophic species in a narrow phylogenetic framework. We address this question in the orchid tribe Neottieae, where this large assortment of nutrition types occurs. We sequenced 13 new plastid genomes, including 9 mixotrophic species and covering all 6 Neottieae genera. We investigated selective pressure on plastid genes in each nutrition type and conducted a phylogenetic inference of the group. Surprisingly, photosynthesis-related genes did not experience selection relaxation in mixotrophic species compared with autotrophic relatives. Conversely, we observed evidence for selection intensification for some plastid genes. Photosynthesis is thus still under purifying selection, maybe because of its role in fruit formation and thus reproductive success. Phylogenetic analysis resolved most relationships, but short branches at the base of the tree suggest an evolutionary radiation at the beginning of Neottieae history, which, we hypothesize, may be linked to mixotrophy emergence.

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.

Endosymbioses necessitate functional cooperation of cellular compartments to avoid pathway redundancy and streamline the control of biological processes. To gain insight into the metabolic compartmentation in chromerids, phototrophic relatives to apicomplexan parasites, we prepared a reference set of proteins probably localized to mitochondria, cytosol and the plastid, taking advantage of available genomic and transcriptomic data. Training of prediction algorithms with the reference set now allows a genome-wide analysis of protein localization in C. velia and V. brassicaformis. We confirm that the chromerid plastids house enzymatic pathways needed for their maintenance and photosynthetic activity, but for carbon and nitrogen allocation, metabolite exchange is necessary with the cytosol and mitochondria. This indeed suggests that the regulatory mechanisms operate in the cytosol to control carbon metabolism based on the availability of both light and nutrients. We discuss that this arrangement is largely shared with apicomplexans and dinoflagellates, possibly stemming from a common ancestral metabolic architecture, and supports the mixotrophy of the chromerid algae.

International audience; Background and AimsPyroloids, forest sub-shrubs of the Ericaceae family, are an important model for their mixotrophic nutrition, which mixes carbon from photosynthesis and from their mycorrhizal fungi. They have medical uses but are difficult to cultivate ex situ; in particular, their dust seeds contain undifferentiated, few-celled embryos, whose germination is normally fully supported by fungal partners. Their germination and early ontogenesis thus remain elusive.MethodsAn optimized in vitro cultivation system of five representatives from the subfamily Pyroloideae was developed to study the strength of seed dormancy and the effect of different media and conditions (including light, gibberellins and soluble saccharides) on germination. The obtained plants were analysed for morphological, anatomical and histochemical development.Key ResultsThanks to this novel cultivation method, which breaks dormancy and achieved up to 100 % germination, leafy shoots were obtained in vitro for representatives of all pyroloid genera (Moneses, Orthilia, Pyrola and Chimaphila). In all cases, the first post-germination stage is an undifferentiated structure, from which a root meristem later emerges, well before formation of an adventive shoot.ConclusionsThis cultivation method can be used for further research or for ex situ conservation of pyroloid species. After strong seed dormancy is broken, the tiny globular embryo of pyroloids germinates into an intermediary zone, which is functionally convergent with the protocorm of other plants with dust seeds such as orchids. Like the orchid protocorm, this intermediary zone produces a single meristem: however, unlike orchids, which produce a shoot meristem, pyroloids first generate a root meristem.

Mixotrophy by planktonic algae can dominate picoplankton predation in oligotrophic environments. We compared the grazing activity and prey preference of different mixotrophic taxa from natural communities of two oligotrophic shallow lakes (Escondido and Morenito) by performing eight laboratory grazing experiments using three different fluorescent prey (bacteria, picocyanobacteria, and picoeukaryotic algae). Eight taxa of mixotrophic flagellates (MF) corresponding to five different Classes, and two size categories of heterotrophic flagellates were distinguished. Chesson index showed that the ten identified protists preferred picocyanobacteria than bacteria, while picoeukaryotic algae were not ingested by any flagellate. In both lakes, MFs were responsible for ca. 80% of the picoplankton ingestion due to flagellates, and more than 50% of the total grazing impact when considering all flagellates, ciliates, and rotifers. Pseudopedinella spp. (Dictyochophyceae), Chrysochromulina parva (Haptophyceae), and Dinobryon divergens (Chrysophyceae) were the most important predators, while Plagioselmis lacustris (Cryptophyceae) and Chrysophyceae > 5 µm showed the lowest grazing rates. Overall, our findings support the idea of MF being the main picoplankton predators in oligotrophic systems. Besides, we also conclude that herbivory represents a key process in the mixotrophic carbon cycling in oligotrophic environments. Fil: Gerea, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Queimaliños, Claudia Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Unrein, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina

The in vitro rooting phase of Echinacea angustifolia DC. was studied through an unconventional approach, where micropropagated shoots from adult plants were grown under autotrophic conditions. The system employed artificial supports (cellulose plugs) soaked with a liquid sugar-less MS medium that contained 1 mg L−1 indole-3-butyric acid (microfloating). The culture vessels were provided with passive ventilation (PV) or forced ventilation (FV) by bubbling air into the liquid medium. Using this microfloating system, the E. angustifolia plantlets reached about 65% rooting capacity. The cultures were analyzed for their photosynthetic rates (PN), carbohydrate content and phytochemical profile. When the ventilation of the vessels was forced, the net photosynthetic rate was enhanced improving explant quality and survival during acclimatization, but the content of caffeic acid derivatives (CADs) was negatively affected. On the contrary, at the end of the multiplication phase in mixotrophic conditions, the photosynthesis was impaired and the production of CADs was improved. The mixotrophy or autotrophy status experienced by the E. angustifolia microplants during the micropropagation process can direct the cultures towards either the accumulation of active compounds or the production of good quality transplants suitable to undergo the acclimatization process.

La mixotrofía es una estrategia de alimentación por medio de la cual un organismo combina la autotrofia y la heterotrofia. La dinámica estacional de cinco ciliados mixótrofos comúnmente presentes en las aguas superficiales de la zona interna y media del estuario de Bahía Blanca, Argentina, rico en nutrientes, somero y con alta turbidez, se estudió mensualmente desde enero a diciembre de 2009. Se registraron los valores de temperatura, salinidad, turbidez y clorofila a, así como también la abundancia y biomasa de Strombidium capitatum, Strombidium acutum, Cyrtostrombidium sp., Lohmanniella oviformis y Tontonia appendiculariformis. La abundancia más alta de los mixótrofos se registró en el invierno en la zona interna, mientras que en la zona media del estuario la presencia de estos ciliados fue casi nula. La contribución más alta de clorofila proveniente de los mixótrofos a la clorofila a total fue 6% en la zona interna y 23% en la zona media (ambas estimaciones registradas en otoño). La baja abundancia de mixótrofos en la zona media del estuario durante el invierno, como usualmente es observado en otros sistemas costeros, podría ser explicada por una presión de pastoreo elevada en esta zona (control de tipo “top-down”) por parte de mesozooplancton (e.g., copépodos), en comparación con la zona interna. La floración secundaria del fitoplancton observada durante los últimos veranos, dominada por diatomeas de pequeño tamaño y fitoflagelados del nanoplancton, podría haber sustentado la alta abundancia de ciliados mixótrofos que se registró en la zona media en otoño y verano. Mixotrophy is a feeding strategy by which some organisms combine autrotrophic and heterotrophic activities. The seasonal dynamics of the five most common mixotrophic ciliates were studied monthly in surface layers of the inner and middle zone of the Bahía Blanca estuary, a nutrient-rich, shallow and highly turbid environment in Argentina, from January to December 2009. Temperature, salinity, turbidity and chlorophyll a were recorded, as well as the abundance and biomass of Strombidium capitatum, Strombidium acutum, Cyrtostrombidium sp., Lohmanniella oviformis and Tontonia appendiculariformis. The highest mixotrophic ciliate abundance was recorded during the austral winter (June-July) in the inner zone, meanwhile in the middle zone of the estuary the presence of these ciliates was almost null. The highest chlorophyll contribution derived from mixotrophic ciliates to total chlorophyll a ranged from 6% in the inner zone to 23% in the middle zone, both registered in autumn. The low abundance of mixotrophs in the middle zone of the estuary during the winter, as is usually observed in other coastal ecosystems, could be explained through a higher grazing pressure in this zone (top-down control) by mesozooplankton (e.g., copepods) in comparison to the inner zone. The secondary bloom of phytoplankton consistently observed during the last summers dominated by small sized diatoms and nanoplankton phytoflagelates, could have sustained the high abundance of mixotrophic ciliates registered in the middle zone in autumn and summer.

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.

Results from a dynamic mathematical model are presented simulating the growth of the harmful algal bloom (HAB) mixotrophic dinoflagellate Karlodinium veneficum and its algal prey, Rhodomonas salina. The model describes carbon-nitrogen-phosphorus-based interactions within the mixotroph, interlinking autotrophic and phagotrophic nutrition. The model was tuned to experimental data from these species grown under autotrophic conditions and in mixed batch cultures in which nitrogen:phosphorus stoichiometry (input molar N:P of 4, 16, and 32) of both predator and prey varied. A good fit was attained to all experimentally derived carbon biomass data. The potential effects of temperature and nutrient changes on promoting growth of prey and thus K. veneficum bloom formation were explored using this simulation platform. The simulated biomass of K. veneficum was highest when they were functioning as mixotrophs and when they consumed prey under elevated N:P conditions. The scenarios under low N:P responded differently, with simulations showing larger deviation between mixotrophic and autotrophic growth, depending on temperature. When inorganic nutrients were in balanced proportions, lower biomass of the mixotroph was attained at all temperatures in the simulations, suggesting that natural systems might be more resilient against Karlodinium HAB development in warming conditions if nutrients were available in balanced proportions. These simulations underscore the need for models of HAB dynamics to include consideration of prey; modeling HAB as autotrophs is insufficient. The simulations also imply that warmer, wetter springs that may bring more N with lower N:P, such as predicted under climate change scenarios for Chesapeake Bay, may be more conducive to development of these HABs. Prey availability may also increase with temperature due to differential growth temperature responses of K. veneficum and its prey.

Polar marine ecosystems are characterized by low water temperatures, sea ice cover, and extreme annual variation in solar irradiance and primary productivity. A review of the available information from the Arctic suggests that mixotrophy (i.e. the combination of photosynthetic and phagotrophic modes of nutrition in one cell) is wide spread among plankton. In the central Arctic Ocean (AO) in summer, mixotrophic flagellates such as Micromonas and Dinobryon can account for most of bacterivory. Planktonic ciliates with acquired phototrophy form the bulk of microzooplankton biomass in both the ultra-oligotrophic deep basins of AO and its productive shelf seas. With the exception of the diatom bloom in the marginal ice zone, mixotrophic ciliates often dominate total chlorophyll in the mixed layer in summer taking advantage of the 24-h insolation. Their relatively high growth rates at low temperatures indicate that they are important component of primary and secondary production. The key Arctic copepod species preferentially feed on chloroplast-bearing ciliates, which form an important link in the planktonic food web. The limited year round data indicate that are available show that mixotrophic plankton persist in the water column during the long polar winter when irradiance is low or absent and ice cover reduces further light penetration. These observations suggest that at high latitudes an alternative food web based on mixotrophy may dominate the pelagic lower food web during much of the year.