Photosynthetic H2 production by green microalgae requires sustained low O2 while preserving photosynthetic electron supply to [FeFe]-hydrogenase. We developed a two-compartment bioreactor in which a hydrophobic PTFE membrane separates a concentrated Chlamydomonas reinhardtii pgr5 culture from an iron-based O2 absorbent, enabling continuous O2 removal without direct contact between cells and scavenger. After brief N2 flushing, dissolved O2 in the culture rapidly dropped to near zero and remained at microoxic levels for multi-day operation; adding silica-gel desiccant improved absorbent performance in the humid sealed reactor and prolonged low-O2 maintenance. Under high light (350 mu mol photons m- 2 s- 1) at 30 degrees C, the system supported sustained H2 production for 12 days, reaching 1350 +/- 80 mu L H2 mL- 1 under the optimal regime. Dark preincubation before high-light exposure reduced H2 yield and impaired photosystem II performance. These results identify membrane-assisted O2 removal and product-gas management as effective process-engineering strategies for prolonged algal H2 production.

The Gothenburg University Marine Algae Culture Collection (GUMACC) is a regionally specialized Nordic resource for marine microalgal research, biotechnology and education. The collection has a focus on Scandinavian coastal, brackish, and polar environments and is particularly rich in diatoms, including a large representation of Skeletonema marinoi. In this resource review we summarize the taxonomic and geographic composition of the collection, describe the public GUMACC database, and provide a complete strain inventory. The public database currently includes 80 strains and represents the historically established public core of the collection rather than the full currently maintained inventory of over 450 strains. We also review how GUMACC strains have supported research in taxonomy and systematics, ecophysiology, climate-change biology, genomics, environmental monitoring, biotechnology and bioresource development. Examples include description of new diatom taxa, studies of responses to temperature, salinity, carbon dioxide, nutrients and ultraviolet radiation, and the establishment of Skeletonema marinoi R05AC as a genetic model for chain-forming marine diatoms. Applied studies have explored lipid production for biodiesel, wastewater remediation, bioactive compounds, and diatom photonic materials. GUMACC also contributes to university teaching, school activities, museum exhibitions, and ocean-literacy initiatives. We discuss current challenges such as the reliance on serial subculturing, variable levels of strain documentation, incomplete public database coverage and the need for improved long-term preservation strategies. Finally, we identify future priorities for strengthening the collection. These include public database updating, expanded genomic and targeted transcriptomic characterization, integration with global biodiversity databases, and targeted expansion of Arctic and sub-Arctic strains relevant to climate resilience.

Phosphate (Pi) is essential for photosynthesis in the chloroplast of algae and plants. Pi homeostasis in the chloroplast is maintained by transporters from several families, whose identities in algae are largely unknown as compared with land plants. Here, we assess the role of the putative PHOSPHATE TRANSPORTER 4-9 from Chlamydomonas reinhardtii (CrPHT4-9) in maintaining chloroplast Pi homeostasis and modulating photosynthesis. Based on phylogenetic analyses and heterologous expression in a yeast (Saccharomyces cerevisiae) strain lacking Pi transporters, we demonstrate that CrPHT4-9 is a Pi transporter closely related to the chloroplast members of the PHT4 family in Arabidopsis (Arabidopsis thaliana). CrPHT4-9 is localized within the chloroplast, more specifically in the thylakoid membrane network and the tubules traversing the CO2-fixing pyrenoid. Two mutants lacking CrPHT4-9 (Crpht4-9) exhibit defective photoautotrophic growth, altered cell morphology and chloroplast ultrastructure under CO2-limiting conditions. In the Crpht4-9 mutants, we further show an increased proton motive force across the thylakoid membrane, enhanced energy- and state-transition-dependent non-photochemical quenching of chlorophyll a fluorescence, and diminished photosynthetic electron transport and ATP synthase activity. The Crpht4-9 mutants exhibit reduced affinity to inorganic carbon, indicating an impaired carbon-concentrating mechanism. These phenotypes are largely recovered by genetic complementation as well as by ample CO2 supply and, interestingly, by Pi deprivation. Therefore, we conclude that the thylakoid- and pyrenoid-localized CrPHT4-9 maintains Pi homeostasis within the chloroplast and is essential for photosynthesis and growth.

Green microalgae, such as Chlamydomonas reinhardtii , show great potential for producing green hydrogen using only water and sunlight, with no carbon emissions. However, sustainable hydrogen production requires addressing the hydrogenase sensitivity to oxygen and enhancing electron allocation to this enzyme. Previous methods for hydrogen photoproduction rely on a brief nitrogen flushing followed by a dark incubation phase to establish anoxia prior to exposure to high light. In this study, we present a straightforward protocol involving a mixotrophic growth phase followed by a semi‐autotrophic hydrogen production phase. During the hydrogen production phase, extended nitrogen flushing induced anoxia in the liquid algae culture, even though oxygen was still present in the headspace. Anoxia was maintained under light at moderate intensity (120 μmol m −2 s −1 ) and a controlled temperature of 30°C, with an efficient mixing system. Throughout the hydrogen production phase, we monitored dissolved oxygen levels in the culture alongside traditional oxygen measurements in the headspace. Using our protocol with the pgr5 mutant of C. reinhardtii , we achieved a maximum specific rate of 72 μmol H₂ mg −1 Chl h −1 and an average rate of 30–35 μmol H₂ mg −1 Chl h −1 over 10 hours of illumination. Additional nitrogen flushing steps extended anoxia, resulting in a total hydrogen yield of 220 ± 20 mL L −1 over 48 hours of illumination. This performance is attributed to maintaining the redox balance of the plastoquinone pool and minimizing photodamage to the photosystem II complex. Our protocol offers a significant advancement for scalable and sustainable green hydrogen production.

Climate warming increases the risk of harmful leaf temperatures in terrestrial plants, particularly in tropical tree species that have evolved in warm and thermally stable environments. We examined heat tolerance thresholds of photosynthetic light reactions in sun-exposed leaves of 12 tropical montane tree species with different strategies for growth and water use. Leaf chlorophyll a fluorescence, gas exchange, morphology and thylakoid membrane lipid composition were measured at three common gardens along an elevation and temperature gradient in Rwanda. Tree species with traits predisposing them to higher leaf temperatures, such as lower stomatal conductance and large leaves, had higher photosynthetic heat tolerance, but narrower thermal safety margins (TSMs). Photosynthetic heat tolerance partially acclimated to increased growth temperature, increasing by 0.31°C on average for every 1°C increase in growth temperature. Thus, TSMs were narrower for trees grown at the warmer sites. Heat tolerance and its acclimation were linked to the adjustment of thylakoid membrane lipid composition. Moreover, TSMs were larger in species with high leaf mass per area. Our results show that (i) leaf temperature is more important than heat tolerance in controlling interspecific variation in TSMs, and that (ii) tropical trees have limited ability to thermally acclimate to increasing temperatures.

Plants rely on water and light for photosynthesis, but water droplets on leaves can focus light into high-intensity spots, risking photodamage. Excessive light can impair growth or induce cell death, making it essential for plants to detect and respond to light fluctuations. While Ca2+ signaling has been linked to high light (HL) acclimation, the subcellular dynamics remain unclear. Here, we investigate Ca2+ responses to HL exposure in Arabidopsis thaliana. Using a glass bead to simulate light-focusing by water droplets, a biphasic increase of Ca2+ concentration was detected in the chloroplast stroma by the genetically encoded calcium indicator YC3.6 and confirmed using a newly established stroma-localized R-GECO1 (NTRC-R-GECO1). The stromal response was largely independent of light wavelength and unaffected in phot1 phot2 and cry1 cry2 mutants. Chemical inhibition of photosynthetic electron transport, microscopy-based Fv/Fm experiments, and measurement of the reactive oxygen species (ROS)-redox balance with roGFP-based reporters and Singlet Oxygen Sensor Green (SOSG) chemical dye suggested that photodamage and singlet oxygen contribute to the stromal Ca2+ response. While blue and white light also triggered a Ca2+ response in the cytosol and nucleus, pharmacological inhibition with cyclopiazonic acid (CPA) and loss-of-function mutants of the Ca2+ transporters BIVALENT CATION TRANSPORTER 2 (BICAT2) and endoplasmic reticulum (ER)-type Ca2+-ATPase (ECA) suggested that the HL response depends on a Ca2+ exchange between the ER and chloroplast stroma. The response was primarily light dependent but accelerated by increasing external temperature. This study implicates a novel Ca2+-mediated acclimation mechanism to HL stress, a process of growing relevance in the context of climate change.

Mixotrophy, the concurrent use of inorganic and organic carbon in the presence of light for microalgal growth, holds ecological and industrial significance. However, it is poorly explored in diatoms, especially in ecologically relevant species like Skeletonema marinoi. This study strategically employed mixotrophic metabolism to optimize the growth of a strain of Skeletonema marinoi (Sm142), which was found potentially important for biomass production on the west coast of Sweden in winter conditions. The aim of this study was to discern the most effective organic carbon sources by closely monitoring microalgal growth through the assessment of optical density, chlorophyll a fluorescence, and biomass concentration. The impact of various carbon sources on the physiology of Sm142 was investigated using photosynthetic and respiratory parameters. The findings revealed that glycerol exhibited the highest potential for enhancing the biomass concentration of Sm142 in a multi-cultivator under the specified experimental conditions, thanks to the increase in respiration activity. Furthermore, the stimulatory effect of glycerol was confirmed at a larger scale using environmental photobioreactors simulating the winter conditions on the west coast of Sweden; it was found comparable to the stimulation by CO2-enriched air versus normal air. These results were the first evidence of the ability of Skeletonema marinoi to perform mixotrophic metabolism during the winter and could explain the ecological success of this diatom on the Swedish west coast. These findings also highlight the importance of both organic and inorganic carbon sources for enhancing biomass productivity in harsh winter conditions.

In many eukaryotic algae, CO2 fixation by Rubisco is enhanced by a CO2-concentrating mechanism, which utilizes a Rubisco-rich organelle called the pyrenoid. The pyrenoid is traversed by a network of thylakoid membranes called pyrenoid tubules, which are proposed to deliver CO2. In the model alga Chlamydomonas (Chlamydomonas reinhardtii), the pyrenoid tubules have been proposed to be tethered to the Rubisco matrix by a bestrophin-like transmembrane protein, BST4. Here, we show that BST4 forms a complex that localizes to the pyrenoid tubules. A Chlamydomonas mutant impaired in the accumulation of BST4 (bst4) formed normal pyrenoid tubules, and heterologous expression of BST4 in Arabidopsis (Arabidopsis thaliana) did not lead to the incorporation of thylakoids into a reconstituted Rubisco condensate. Chlamydomonas bst4 mutants did not show impaired growth under continuous light at air level CO2 but were impaired in their growth under fluctuating light. By quantifying the non-photochemical quenching (NPQ) of chlorophyll fluorescence, we propose that bst4 has a transiently lower thylakoid lumenal pH during dark-to-light transition compared to control strains. We conclude that BST4 is not a tethering protein but is most likely a pyrenoid tubule ion channel involved in the ion homeostasis of the lumen with particular importance during light fluctuations.

Microalgae are photosynthetic microorganisms that have been extensively studied for their ability to produce biofuels, including hydrogen (H2). While microalgae immobilization techniques have been extensively researched for wastewater treatment, their application for H2 photoproduction is a relatively new area of study. In this review, we evaluate microalgae immobilization as a promising technology to improve the commercial maturity of green H2 production by microalgae, as it offers multiple advantages over traditional suspended growth systems. It facilitates cell harvesting, reduces contamination risk, and improves control of cultivation conditions, biomass retention, and efficiency of H2 production. However, the effectiveness of immobilization largely depends on the technical parameters, including the type of immobilization method used, the choice of immobilization materials, and the operational conditions. Here we provide a comparative review on different parameters and conditions suggested in the literature for microalgae immobilization, and propose a protocol optimized to achieve an efficient H2 production.

Plants require water and light for photosynthesis, but light, when focused by water droplets on leaves, can create high light intensity spots that are harmful to plants. As excessive light intensity can reduce growth or even induce cell death, it is vital for plants to detect and react to changes in light exposure and acclimate to high light stress. Ca2+ signaling was previously implicated in high light acclimation. However, the dynamics of free Ca2+ concentration in the chloroplast, the primary site of photosynthesis, or in the nucleus and in the cytoplasm, where transcription and translation for long-term acclimation occurs, remain unknown. Here we studied the dynamics and mechanism of the Ca2+ response to high light exposure. Focusing light through a glass bead to mimic water droplets triggered an increase of the free Ca2+ concentration in the chloroplast stroma of Arabidopsis thaliana . This finding was corroborated using established and newly developed genetically encoded calcium indicators, which revealed a biphasic increase in the stromal free Ca2+ concentration when exposed to varying intensities and qualities of light. Among photosynthetic by-products, reactive oxygen and lipophilic species in particular, have been implicated in high light stress acclimation. A H2O2 signature was induced, albeit with different dynamics than the Ca2+ response, while chemical inhibition of the photosynthetic electron transport points towards singlet oxygen as a potential culprit of the high light-induced increase in stromal free Ca2+ concentration. The observed dynamics differed from those of a heat-shock induced Ca2+ signature, although temperature had a positive effect on the Ca2+ response. Based on Ca2+ inhibitor treatments and the free Ca2+ concentration dynamics, we suggest that the high light-induced stromal Ca2+ is derived from the endoplasmic reticulum rather than from the cytoplasm. In conclusion, inspired by the burning glass effect of water droplets on leaves, we uncovered a Ca2+ response that implicates a novel mechanism for plants to acclimate to high light stress—a process that will become increasingly relevant in a changing climate. ### Competing Interest Statement The authors have declared no competing interest.

In many eukaryotic algae, CO2 fixation by Rubisco is enhanced by a CO2-concentrating mechanism, which utilizes a Rubisco-rich organelle called the pyrenoid. The pyrenoid is traversed by a network of thylakoid-membranes called pyrenoid tubules, proposed to deliver CO2. In the model alga Chlamydomonas reinhardtii (Chlamydomonas), the pyrenoid tubules have been proposed to be tethered to the Rubisco matrix by a bestrophin-like transmembrane protein, BST4. Here, we show that BST4 forms a complex that localizes to the pyrenoid tubules. A Chlamydomonas mutant impaired in the accumulation of BST4 (bst4) formed normal pyrenoid tubules and heterologous expression of BST4 in Arabidopsis thaliana did not lead to the incorporation of thylakoids into a reconstituted Rubisco condensate. Chlamydomonas bst4 mutant did not show impaired growth at air level CO2. By quantifying the non-photochemical quenching (NPQ) of chlorophyll fluorescence, we show that bst4 displays a transiently lower thylakoid lumenal pH during dark to light transition compared to control strains. When acclimated to high light, bst4 had sustained higher NPQ and elevated levels of light-induced H2O2 production. We conclude that BST4 is not a tethering protein, but rather is an ion channel involved in lumenal pH regulation possibly by mediating bicarbonate transport across the pyrenoid tubules.

There is an increasing interest in algae-based biomass produced outdoors in natural and industrial settings for biotechnological applications. To predict the yield and biochemical composition of the biomass, it is important to understand how the transcriptome of species and strains of interest is affected by seasonal changes. Here we studied the effects of Nordic winter and summer on the transcriptome of two phytoplankton species, namely the diatom Skeletonema marinoi (Sm) and the eustigmatophyte Nannochloropsis granulata (Ng), recently identified as potentially important for biomass production on the west coast of Sweden. Cultures were grown in photobioreactors in simulated Nordic summer and winter, and the gene expression in two phases was quantified by Illumina RNA-sequencing. Five paired comparisons were made among the four conditions. Sm was overall more responsive to seasons since 70 % of the total transcriptome (14,783 genes) showed differential expression in at least one comparison as compared to 1.6 % (1403 genes) for Ng. For both species, we observed larger differences between the seasons than between the phases of the same season. In summer phase 1, Sm cells focused on photosynthesis and polysaccharide biosynthesis. Nitrate assimilation and recycling of intracellular nitrogen for protein biosynthesis were more active in summer phase 2 and throughout winter. Lipid catabolism was upregulated in winter relative to summer to supply carbon for respiration. Ng favored lipid accumulation in summer, while in winter activated different lipid remodeling pathways as compared to Sm. To cope with winter, Ng upregulated breakdown and transport of carbohydrates for energy production. Taken together, our transcriptome data reveal insights into adaptive seasonal responses of Sm and Ng important for biotechnological applications on the west coast of Sweden, but more work is required to decipher the molecular mechanisms behind these responses.

Magnesium (Mg2+) is essential for photosynthesis in the chloroplasts of land plants and algae. Being the central ion of chlorophyll, cofactor and activator of many photosynthetic enzymes including RuBisCO, magnesium-deficient plants may suffer from leaf chlorosis symptoms and retarded growth. Therefore, the chloroplast Mg2+ concentration is tightly controlled by magnesium transport proteins. Recently, three different transporters from two distinct families have been identified in the chloroplast inner envelope of the model plant Arabidopsis thaliana: MGT10, MGR8, and MGR9. Here, we assess the individual roles of these three proteins in maintaining chloroplast Mg2+ homeostasis and regulating photosynthesis, and if their role is conserved in the model green alga Chlamydomonas reinhardtii. Phylogenetic analysis and heterologous expression revealed that the CorC-like MGR8 and MGR9 transport Mg2+ by a different mechanism than the CorA-like MGT10. MGR8 and MGT10 genes are highest expressed in leaves, indicating a function in chloroplast Mg2+ transport. MGR9 is important for chloroplast function and plant adaptation in conditions of deficiency or excess of Mg2+. Transmission electron microscopy indicated that MGT10 plays a differential role in thylakoid stacking than MGR8 and MGR9. Furthermore, we report that MGR8, MGR9, and MGT10 are involved in building up the pH gradient across the thylakoid membrane and activating photoprotection in conditions of excess light, however the mechanism has not been resolved yet. While there are no chloroplast MGR-like transporters in Chlamydomonas, we show that MRS4 is a homolog of MGT10, that is required for photosynthesis and cell growth. Taken together, our findings reveal that the studied Mg2+ transporters play essential but differential roles in maintaining chloroplast Mg2+ homeostasis.

The overall goal of this study was to provide solutions to innovative microalgae-based technology for wastewater remediation in a cold-water recirculating marine aquaculture system (RAS). This is based on the novel concept of integrated aquaculture systems in which fish nutrient-rich rearing water will be used for microalgae cultivation. The produced biomass can be used as fish feed, while the cleaned water can be reused, to create a highly eco-sustainable circular economy. Here, we tested three microalgae species Nannochloropis granulata (Ng), Phaeodactylum tricornutum (Pt), and Chlorella sp (Csp) for their ability to remove nitrogen and phosphate from the RAS wastewater and simultaneously produce high-value biomass, i.e., containing amino acids (AA), carotenoids, and polyunsaturated fatty acids (PUFAs). A high yield and value of biomass were achieved for all species in a two-phase cultivation strategy: i) a first phase using a medium optimized for best growth (f/2 14x, control); ii) a second "stress" phase using the RAS wastewater to enhance the production of high-value metabolites. Ng and Pt performed best in terms of biomass yield (i.e., 5-6 g of dry weight, DW.L-1) and efficient cleaning of the RAS wastewater from nitrite, nitrate, and phosphate (i.e., 100% removal). Csp produced about 3 g L-1 of DW and reduced efficiently only nitrate, and phosphate (i.e., about 76% and 100% removal, respectively). The biomass of all strains was rich in protein (30-40 % of DW) containing all the essential AA except Methionine. The biomass of all three species was also rich in PUFAs. Finally, all tested species are excellent sources of antioxidant carotenoids, including fucoxanthin (Pt), lutein (Ng and Csp) and β-carotene (Csp). All tested species in our novel two-phase cultivation strategy thus showed great potential to treat marine RAS wastewater and provide sustainable alternatives to animal and plant proteins with extra added values.

Fluctuating light intensity challenges fluent photosynthetic electron transport in plants, inducing photoprotection while diminishing carbon assimilation and growth, and also influencing photosynthetic signaling for regulation of gene expression. Here, we employed in vivo chlorophyll-a fluorescence and P700 difference absorption measurements to demonstrate the enhancement of photoprotective energy dissipation of both photosystems in wild-type Arabidopsis thaliana after 6 h exposure to fluctuating light as compared with constant light conditions. This acclimation response to fluctuating light was hampered in a triple mutant lacking the thylakoid ion transport proteins KEA3, VCCN1, and CLCe, leading to photoinhibition of photosystem I. Transcriptome analysis revealed upregulation of genes involved in biotic stress and defense responses in both genotypes after exposure to fluctuating as compared with constant light, yet these responses were demonstrated to be largely upregulated in triple mutant already under constant light conditions compared with wild type. The current study illustrates the rapid acclimation of plants to fluctuating light, including photosynthetic, transcriptomic, and metabolic adjustments, and highlights the connection among thylakoid ion transport, photosynthetic energy balance, and cell signaling.

As the most abundant cation in archaeal, bacterial, and eukaryotic cells, potassium (K + ) is an essential element for life. While much is known about the machinery of transcellular and paracellular K transport – channels, pumps, co-transporters, and tight-junction proteins - many quantitative aspects of K homeostasis in biological systems remain poorly constrained. Here we present measurements of the stable isotope ratios of potassium ( 41 K/ 39 K) in three biological systems (algae, fish, and mammals). When considered in the context of our current understanding of potential mechanisms of K isotope fractionation and K + transport in these biological systems, our results provide evidence that the fractionation of K isotopes depends on transport pathway and transmembrane transport machinery. Specifically, we find that passive transport of K + down its electrochemical potential through channels and pores in tight-junctions at favors 39 K, a result which we attribute to a kinetic isotope effect associated with dehydration and/or size selectivity at the channel/pore entrance. In contrast, we find that transport of K + against its electrochemical gradient via pumps and co-transporters is associated with less/no isotopic fractionation, a result that we attribute to small equilibrium isotope effects that are expressed in pumps/co-transporters due to their slower turnover rate and the relatively long residence time of K + in the ion pocket. These results indicate that stable K isotopes may be able to provide quantitative constraints on transporter-specific K + fluxes (e.g. the fraction of K efflux from a tissue by channels vs. co-transporters) and how these fluxes change under different physiological states. In addition, precise determination of K isotope effects associated with K + transport through channels, pumps, and co-transporters may provide unique constraints on the mechanisms of K transport that could be tested with steered molecular dynamic simulations.

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO2. The addition of either glycerol or CO2-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.

Warming climate increases the risk for harmful leaf temperatures in terrestrial plants, causing heat stress and loss of productivity. The heat sensitivity may be particularly high in equatorial tropical tree species adapted to a thermally stable climate. Thermal thresholds of the photosynthetic system of sun-exposed leaves were investigated in three tropical montane tree species native to Rwanda with different growth and water use strategies (Harungana montana, Syzygium guineense and Entandrophragma exselsum). Measurements of chlorophyll fluorescence, leaf gas exchange, morphology, chemistry and temperature were made at three common gardens along an elevation/temperature gradient. Heat tolerance acclimated to maximum leaf temperature (Tleaf ) across the species. At the warmest sites, the thermal threshold for normal function of photosystem II was exceeded in the species with the highest Tleaf despite their higher heat tolerance. This was not the case in the species with the highest transpiration rates and lowest Tleaf . The results point to two differently effective strategies for managing thermal stress: tolerance through physiological adjustment of leaf osmolality and thylakoid membrane lipid composition, or avoidance through morphological adaptation and transpiratory cooling. More severe photosynthetic heat stress in low-transpiring montane climax species may result in a competitive disadvantage compared to high-transpiring pioneer species with more efficient leaf cooling.

Coping with changes in light intensity is challenging for plants, but well-designed mechanisms allow them to acclimate to most unpredicted situations. The thylakoid K+/H+ antiporter KEA3 and the voltage-dependent Cl− channel VCCN1 play important roles in light acclimation by fine-tuning electron transport and photoprotection. Good evidence exists that the thylakoid Cl− channel ClCe is involved in the regulation of photosynthesis and state transitions in conditions of low light. However, a detailed mechanistic understanding of this effect is lacking. Here we report that the ClCe loss-of-function in Arabidopsis thaliana results in lower levels of phosphorylated light-harvesting complex II (LHCII) proteins as well as lower levels of the photosystem I-LHCII complexes relative to wild type (WT) in low light conditions. The phosphorylation of the photosystem II core D1/D2 proteins was less affected either in low or high light conditions. In low light conditions, the steady-state levels of ATP synthase conductivity and of the total proton flux available for ATP synthesis were lower in ClCe loss-of-function mutants, but comparable to WT at standard and high light intensity. As a long-term acclimation strategy, expression of the ClCe gene was upregulated in WT plants grown in light-limiting conditions, but not in WT plants grown in standard light even when exposed for up to 8 h to low light. Taken together, these results suggest a role of ClCe in the regulation of the ATP synthase activity which under low light conditions impacts LHCII protein phosphorylation and state transitions.

At Nordic latitudes, year-round outdoor cultivation of microalgae is debatable due to seasonal variations in productivity. Shall the same species/strains be used throughout the year, or shall seasonal-adapted ones be used? To elucidate this, a laboratory study was performed where two out of 167 marine microalgal strains were selected for intended cultivation at the west coast of Sweden. The two local strains belong to Nannochloropsis granulata (Ng) and Skeletonema marinoi (Sm142). They were cultivated in photobioreactors and compared in conditions simulating variations in light and temperature of a year divided into three growth seasons (spring, summer and winter). The strains grew similarly well in summer (and also in spring), but Ng produced more biomass (0.225 vs. 0.066 g DW L-1 day(-1)) which was more energy rich (25.0 vs. 16.6 MJ kg(-1) DW). In winter, Sm142 grew faster and produced more biomass (0.017 vs. 0.007 g DW L-1 day(-1)), having similar energy to the other seasons. The higher energy of the Ng biomass is attributed to a higher lipid content (40 vs. 16% in summer). The biomass of both strains was richest in proteins (65%) in spring. In all seasons, Sm142 was more effective in removing phosphorus from the cultivation medium (6.58 vs. 4.14 mg L-1 day(-1) in summer), whereas Ng was more effective in removing nitrogen only in summer (55.0 vs. 30.8 mg L-1 day(-1)). Our results suggest that, depending on the purpose, either the same or different local species can be cultivated, and are relevant when designing outdoor studies.