Long-term Acclimation Research Articles

Species-specific oxidative stress responses and cellular energy allocation after coral shipping

Tropical corals cultured in or ex situ, are being increasingly shipped worldwide to supply wholesale aquarium companies and research institutions. Scleractinian corals are mostly traded for ornamental purposes, whereby octocorals are increasingly traded for biotechnological applications. During the shipping process corals can be affected by several stressors that in extreme cases can result in coral death. It is, therefore, important to understand the physiological effects of shipping on different coral species. For this, two octocoral species with biotechnological potential, Sinularia polydactyla and Sinularia asterolobata, were shipped from Indonesia to our research facilities in Europe, Portugal. Oxidative stress (catalase – CAT, glutathione S-transferase - GST, and total glutathione - tGSH), oxidative damage (lipid peroxidation - LPO), energy reserves (lipids, proteins, carbohydrates) and electron transport system (ETS) were measured at arrival and after three months of husbandry to evaluate stress and cellular damage after a common shipping practice. The CAT depletion, together with higher activity of the second line of oxidative defence (GST and tGSH) and energy consumption (measured as ETS activity), for S. polydactyla immediately after shipping, evidenced the detoxification effort to avoid cellular damage. However, colonies of this species did not recover from shipping and perished 24 h after arrival. S. asterolobata activated the antioxidative pathways (CAT, GST and tGSH) to control damage right after shipping. Nonetheless, after three months, levels of LPO were significantly higher. Post-stress regeneration and long-term acclimation to captivity conditions, are suggested explanations for these peroxidation values. This study highlights differences in species susceptibility to shipping stress and evidence the need to consider species-specific requirements to a successful shipping and reduce the loss of valuable biomass.

Arabidopsis REI-LIKE proteins activate ribosome biogenesis during cold acclimation

Arabidopsis REIL proteins are cytosolic ribosomal 60S-biogenesis factors. After shift to 10 °C, reil mutants deplete and slowly replenish non-translating eukaryotic ribosome complexes of root tissue, while controlling the balance of non-translating 40S- and 60S-subunits. Reil mutations respond by hyper-accumulation of non-translating subunits at steady-state temperature; after cold-shift, a KCl-sensitive 80S sub-fraction remains depleted. We infer that Arabidopsis may buffer fluctuating translation by pre-existing non-translating ribosomes before de novo synthesis meets temperature-induced demands. Reil1 reil2 double mutants accumulate 43S-preinitiation and pre-60S-maturation complexes and alter paralog composition of ribosomal proteins in non-translating complexes. With few exceptions, e.g. RPL3B and RPL24C, these changes are not under transcriptional control. Our study suggests requirement of de novo synthesis of eukaryotic ribosomes for long-term cold acclimation, feedback control of NUC2 and eIF3C2 transcription and links new proteins, AT1G03250, AT5G60530, to plant ribosome biogenesis. We propose that Arabidopsis requires biosynthesis of specialized ribosomes for cold acclimation.

A gene regulatory network for antenna size control in carbon dioxide-deprived Chlamydomonas reinhardtii cells.

In green microalgae, prolonged exposure to inorganic carbon depletion requires long-term acclimation responses, involving modulated gene expression and the adjustment of photosynthetic activity to the prevailing supply of carbon dioxide. Here, we describe a microalgal regulatory cycle that adjusts the light-harvesting capacity at photosystem II (PSII) to the prevailing supply of carbon dioxide in Chlamydomonas (Chlamydomonas reinhardtii). It engages low carbon dioxide response factor (LCRF), a member of the squamosa promoter-binding protein (SBP) family of transcription factors, and the previously characterized cytosolic translation repressor nucleic acid-binding protein 1 (NAB1). LCRF combines a DNA-binding SBP domain with a conserved domain for protein-protein interaction. LCRF transcription is rapidly induced by carbon dioxide depletion. LCRF activates NAB1 transcription by specifically binding to tetranucleotide motifs present in its promoter. Accumulation of the NAB1 protein enhances translational repression of its prime target mRNA, encoding the PSII-associated major light-harvesting protein LHCBM6. The resulting truncation of the PSII antenna size helps maintaining a low excitation during carbon dioxide limitation. Analyses of low carbon dioxide acclimation in nuclear insertion mutants devoid of a functional LCRF gene confirm the essentiality of this novel transcription factor for the regulatory circuit.

Long-term thermal acclimation drives adaptive physiological adjustments of a marine gastropod to reduce sensitivity to climate change

Ocean warming is predicted to challenge the persistence of a variety of marine organisms, especially when combined with ocean acidification. While temperature affects virtually all physiological processes, the extent to which thermal history mediates the adaptive capacity of marine organisms to climate change has been largely overlooked. Using populations of a marine gastropod (Turbo undulatus) with different thermal histories (cool vs. warm), we compared their physiological adjustments following exposure (8-week) to ocean acidification and warming. Compared to cool-acclimated counterparts, we found that warm-acclimated individuals had a higher thermal threshold (i.e. increased CTmax by 2 °C), which was unaffected by the exposure to ocean acidification and warming. Thermal history also strongly mediated physiological effects, where warm-acclimated individuals adjusted to warming by conserving energy, suggested by lower respiration and ingestion rates, energy budget (i.e. scope for growth) and O:N ratio. After exposure to warming, warm-acclimated individuals had higher metabolic rates and greater energy budget due to boosted ingestion rates, but such compensatory feeding disappeared when combined with ocean acidification. Overall, we suggest that thermal history can be a critical mediator of physiological performance under future climatic conditions. Given the relatively gradual rate of global warming, marine organisms may be better able to adaptively adjust their physiology to future climate than what short-term experiments currently convey.

Impacts of Acclimation in Warm-Low pH Conditions on the Physiology of the Sea Urchin Heliocidaris erythrogramma and Carryover Effects for Juvenile Offspring

Ocean warming (OW) and acidification (OA) affects nearly all aspects of marine organism physiology and it is important to consider both stressors when predicting responses to climate change. We investigated the effects of long-term exposure to OW and OA on the physiology of adults of the sea urchin,Heliocidaris erythrogramma, a species resident in the southeast Australia warming hotspot. The urchins were slowly introduced to stressor conditions in the laboratory over a 7-week adjustment period to three temperature (ambient, +2°C, +3°C) and two pH (ambient: pHT8.0; −0.4 units: pHT7.6) treatments. They were then maintained in a natural pattern of seasonal temperature and photoperiod change, and fixed pH, for 22 weeks. Survival was monitored through week 22 and metabolic rate was measured at 4 and 12 weeks of acclimation, feeding rate and ammonia excretion rate at 12 weeks and assimilation efficiency at 13 weeks. Acclimation to +3°C was deleterious regardless of pH. Mortality from week 6 indicated that recent marine heatwaves are likely to have been deleterious to this species. Acclimation to +2°C did not affect survival. Increased temperature decreased feeding and increased excretion rates, with no effect of acidification. While metabolic rate increased additively with temperature and low pH at week 4, there was no difference between treatments at week 12, indicating physiological acclimation in surviving urchins to stressful conditions. Regardless of treatment,H. erythrogrammahad a net positive energy budget indicating that the responses were not due to energy limitation. To test for the effect of parental acclimation on offspring responses, the offspring of acclimated urchins were reared to the juvenile stage in OW and OA conditions. Parental acclimation to warming, but not acidification altered juvenile physiology with an increase in metabolic rate. Our results show that incorporation of gradual seasonal environmental change in long-term acclimation can influence outcomes, an important consideration in predicting the consequences of changing climate for marine species.

Increased photosynthesis from a deep-shade to high-light regime occurs by enhanced CO2 diffusion into the leaf of Selaginella martensii

The current understanding of photosynthesis across land plant phylogeny strongly indicates that ancient vascular plants are mainly limited by strong constitutive CO2 diffusional constraints, particularly low stomatal and mesophyll conductance. Considering that the lycophyte Selaginella martensii can demonstrate long-term light acclimation, this study addresses the regulation extent of CO2 assimilation in this species cultivated under contrasting light regimes of deep shade, medium shade and high light. Comparative analyses of photosynthetic traits, CO2 conductance and leaf morpho-anatomy revealed acclimation plasticity similar to that of seed plants, though occurring in the context of an inherently low photosynthetic capacity typical of lycophytes. Specific modulations of the stomatal density and aperture, chloroplast surface exposed to mesophyll airspaces and cell wall thickness sustained a marked improvement in CO2 diffusion from deep shade to high light. However, the maximum carboxylation rate was comparatively less effectively upregulated, leading to a greater incidence of biochemical limitations of photosynthesis. Because of a low carboxylation capacity under any light regime, a lycophyte prevents potential photodamage to the chloroplast by not only exploiting the thermal dissipation of excess absorbed energy but also diverting a large fraction of photosynthetic electrons to sinks alternative to carboxylation.

PHOTOPROTECTIIVE STRATEGIES OF UNICELULAR RED ALGA RHODELLA VIOLACEA DURING ACCLIMATION TO STRONG LIGHT

Red algae contain in their photosynthetic machinery water-soluble antenna complexes - phycobilisomes (PBSs) attached to thylakoid membranes to transfer excitation energy to photosystems. Strong light absorbed by the PBSs triggers a fast formation of transthylakoid ΔpH that follows the non-photochemical quenching of chlorophyll (Chl) fluorescence. The ΔpH build-up seems to be essential for photoprotecting the photosynthetic apparatus in the absence of xanthophyll cycle common to higher plants. However, the photoprotective mechanisms of red algae are not studied in details yet. 
 We present here our research of the Chl fluorescence quenching in unicellular red algae Rhodella violacea and its correlation with the ΔpH gradient being formed. The relation of this phenomenon to photoprotection of photosystem 2 (PS 2) in the normal and high light-acclimated Rhodella cells is also examined. 
 Under the photoinhibitory conditions (white light of 2000-3000 μE/m2s), the ΔpH-dependent Chl fluorescence quenching was found to delay the kinetics of PS 2 photoinhibition. The uncouplers like nigericin and NH4Cl are known to break down ΔpH gradient, lead to the dissipation of Chl fluorescence quenching followed by enhancing the PS 2 photoinhibition rate. The same effect showed far-red (FR) light consuming transthylakoid ΔpH. ATPase inhibitor, DCCD, having no impact on ΔpH didn’t influence PS 2 photoinhibition as well this implies the photoprotection to be fulfilled by the proton gradient rather than by ATP synthesis. 
 Long-term acclimation of Rhodella cells to higher irradiances (500-1000 μE/m2s) results in a partial loss of the periphery phycoerythrin-containing subunits by PBSs. The light-acclimated cultures display a higher resistance to the photoinhibitory light than the non-acclimated ones. This could be explained by diminishing the energy transfer from the reduced PBSs to PS 2 as well as light screening by the secondary carotenoids synthesized during light exposure. 
 Data on low-temperature (77K) fluorescence allow to evaluate the molecular mechanisms of light-induced Chl fluorescence suppression in Rhodella cells and its recovery in darkness.

Chloroplast Electron Chain, ROS Production, and Redox Homeostasis Are Modulated by COS-OGA Elicitation in Tomato (Solanum lycopersicum) Leaves.

The stimulation of plant innate immunity by elicitors is an emerging technique in agriculture that contributes more and more to residue-free crop protection. Here, we used RNA-sequencing to study gene transcription in tomato leaves treated three times with the chitooligosaccharides–oligogalacturonides (COS-OGA) elicitor FytoSave® that induces plants to fend off against biotrophic pathogens. Results showed a clear upregulation of sequences that code for chloroplast proteins of the electron transport chain, especially Photosystem I (PSI) and ferredoxin. Concomitantly, stomatal conductance decreased by half, reduced nicotinamide adenine dinucleotide phosphate [NAD(P)H] content and reactive oxygen species production doubled, but fresh and dry weights were unaffected. Chlorophyll, β-carotene, violaxanthin, and neoxanthin contents decreased consistently upon repeated elicitations. Fluorescence measurements indicated a transient decrease of the effective PSII quantum yield and a non-photochemical quenching increase but only after the first spraying. Taken together, this suggests that plant defense induction by COS-OGA induces a long-term acclimation mechanism and increases the role of the electron transport chain of the chloroplast to supply electrons needed to mount defenses targeted to the apoplast without compromising biomass accumulation.

Dying by drying: Timing of physiological stress thresholds related to tree death is not significantly altered by highly elevated CO2.

Drought-induced tree mortality is expected to occur more frequently under predicted climate change. However, the extent of a possibly mitigating effect of simultaneously rising atmospheric [CO2 ] on stress thresholds leading to tree death is not fully understood, yet. Here, we studied the drought response, the time until critical stress thresholds were reached and mortality occurrence of Pinus halepensis (Miller). In order to observe a large potential benefit from eCO2 , the seedlings were grown with ample of water and nutrient supply under either highly elevated [CO2 ] (eCO2 , c. 936 ppm) or ambient (aCO2 , c. 407 ppm) during 2 years. The subsequent exposure to a fast or a slow lethal drought was monitored using whole-tree gas exchange chambers, measured leaf water potential and non-structural carbohydrates. Using logistic regressions to derive probabilities for physiological parameters to reach critical drought stress thresholds, indicated a longer period for halving needle starch storage under eCO2 than aCO2 . Stomatal closure, turgor loss, the duration until the daily tree C balance turned negative, leaf water potential at thresholds and time-of-death were unaffected by eCO2 . Overall, our study provides for the first-time insights into the chronological interplay of physiological drought thresholds under long-term acclimation to elevated [CO2 ].

Mechanisms underlying photosynthetic acclimation to high temperature are different between Vitis vinifera cv. Syrah and Grenache

Photosynthesis acclimation to high temperature differs among and within species. Grapevine intra-specific variation in photosynthetic acclimation to elevated temperature has been scarcely assessed. Our objectives were to (i) evaluate the mechanisms underlying long-term acclimation of photosynthesis to elevated temperature in grapevine, and (ii) determine whether these responses are similar among two varieties. A warming experiment with well irrigated Grenache and Syrah field-grown plants was performed during two growing seasons comparing plants exposed at ambient temperature (control) with plants in open-top chambers (heating) that increased mean air temperature between 1.5 and 3.6°C. Photosynthetic acclimation was assessed through the response of net assimilation (An), Rubisco carboxylation rate (Vcmax) and electron transport rate (Jmax), at leaf temperatures from 20 to 40°C. Our results evidenced different mechanisms for photosynthetic acclimation to elevated temperature. Compared with control, Grenache heated increased An, maintaining higher Vcmax and Jmax at temperatures above 35°C. By contrast, Syrah heated and control presented similar values of An, Vcmax and Jmax, evidencing an adjustment of photosynthesis without increasing C assimilation. Both varieties increased the optimum temperature for An, but to a lesser extent when growth temperature was higher. Our study provides evidence that grapevine varieties present different acclimation mechanisms to expected warming.

Developmental acclimation of the thylakoid proteome to light intensity in Arabidopsis.

Photosynthetic acclimation, the ability to adjust the composition of the thylakoid membrane to optimise the efficiency of electron transfer to the prevailing light conditions, is crucial to plant fitness in the field. While much is known about photosynthetic acclimation in Arabidopsis, to date there has been no study that combines both quantitative label-free proteomics and photosynthetic analysis by gas exchange, chlorophyll fluorescence and P700 absorption spectroscopy. Using these methods we investigated how the levels of 402 thylakoid proteins, including many regulatory proteins not previously quantified, varied upon long-term (weeks) acclimation of Arabidopsis to low (LL), moderate (ML) and high (HL) growth light intensity and correlated these with key photosynthetic parameters. We show that changes in the relative abundance of cytb6 f, ATP synthase, FNR2, TIC62 and PGR6 positively correlate with changes in estimated PSII electron transfer rate and CO2 assimilation. Improved photosynthetic capacity in HL grown plants is paralleled by increased cyclic electron transport, which positively correlated with NDH, PGRL1, FNR1, FNR2 and TIC62, although not PGR5 abundance. The photoprotective acclimation strategy was also contrasting, with LL plants favouring slowly reversible non-photochemical quenching (qI), which positively correlated with LCNP, while HL plants favoured rapidly reversible quenching (qE), which positively correlated with PSBS. The long-term adjustment of thylakoid membrane grana diameter positively correlated with LHCII levels, while grana stacking negatively correlated with CURT1 and RIQ protein abundance. The data provide insights into how Arabidopsis tunes photosynthetic electron transfer and its regulation during developmental acclimation to light intensity.

A transporter Slr1512 involved in bicarbonate and pH-dependent acclimation mechanism to high light stress in Synechocystis sp. PCC 6803

High light (HL) exposure leads to photoinhibition and excess accumulation of toxic reactive oxygen species (ROS) in photosynthetic organisms, negatively impacting the global primary production. In this study, by screening a mutant library, a gene related with bicarbonate transport, slr1512, was found involved in HL acclimation in model cyanobacterium Synechocystis sp. PCC 6803. Comparative growth analysis showed that the slr1512 knockout mutant dramatically enhanced the tolerance of Synechocystis towards long-term HL stress (200 μmol photons m−2 s−1) than the wild type, achieving an enhanced growth by ~1.95-folds after 10 d. The phenotype differences between Δslr1512 and the wild type were analyzed via absorption spectrum and chlorophyll a content measurement. In addition, the accessible bicarbonate controlled by slr1512 and decreased PSII activity were demonstrated, and they were found to be the key factors affecting the tolerance of Synechocystis against HL stress. Further analysis confirmed that intracellular bicarbonate can significantly affect the activity of photosystem II, leading to the altered accumulation of toxic ROS under HL. Finally, a comparative transcriptomics was applied to determine the differential responses to HL between Δslr1512 and the wild type. This work provides useful insights to long-term acclimation mechanisms towards HL and valuable information to guide the future tolerance engineering of cyanobacteria against HL.

Transcriptional Response of Osmolyte Synthetic Pathways and Membrane Transporters in a Euryhaline Diatom During Long-term Acclimation to a Salinity Gradient.

How diatoms respond to fluctuations in osmotic pressure is important from both ecological and applied perspectives. It is well known that osmotic stress affects photosynthesis and can result in the accumulation of compounds desirable in pharmaceutical and alternative fuel industries. Gene expression responses to osmotic stress have been studied in short-term trials, but it is unclear whether the same mechanisms are recruited during long-term acclimation. We used RNA-seq to study the genome-wide transcription patterns in the euryhaline diatom, Cyclotella cryptica, following long-term acclimation to salinity that spanned the natural range of fresh to oceanic water. Long-term acclimated C. cryptica exhibited induced synthesis or repressed degradation of the osmolytes glycine betaine, taurine and dimethylsulfoniopropionate (DMSP). Although changes in proline concentration is one of the main responses in short-term osmotic stress, we did not detect a transcriptional change in proline biosynthetic pathways in our long-term experiment. Expression of membrane transporters showed a general tendency for increased import of potassium and export of sodium, consistent with the electrochemical gradients and dependence on co-transported molecules. Our results show substantial between-genotype differences in growth and gene expression reaction norms and suggest that the regulation of proline synthesis important in short-term osmotic stress might not be maintained in long-term acclimation. Further examination using time-course gene expression experiments, metabolomics and genetic validation of gene functions would reinforce patterns inferred from RNA-seq data.

The influence of a stepwise pH increase on volatile fatty acids production and phosphorus release during Al-waste activated sludge fermentation

In this study, the performance of volatile fatty acids (VFAs) production and phosphorus (P) release during Al-waste activated sludge (Al-WAS) anaerobic fermentation with stepwise pH increases from 8 to 11 was investigated via a long-term acclimation strategy. As results, VFAs concentration increased with increasing pH and the maximum yield of VFAs was 358.03 mg-COD/g VS at pH 11, which was much higher than at pH 8. P was also released during the process, and the P concentration increased gradually from 26 mg/L at pH 8 to 186 mg/L at pH 11, accounting for 35.8% of the total P in the Al-WAS. The P distribution results demonstrated the dissolution of non-apatite inorganic phosphorus (NAIP) and organic P in the sludge contributed to release P into the liquid at pH 8, 9, and 10, while inorganic P release originated from the dissolution of NAIP at pH 11.

The Molecular Function of Plant mTERFs as Key Regulators of Organellar Gene Expression.

The protein family of mTERFs (mitochondrial transcription termination factors) was initially studied in mammalian and insect mitochondria before the first Arabidopsis mTERF mutant was characterized. More than 10 years of research on the function of plant mTERFs in the flowering plants Arabidopsis thaliana, Zea mays and the green microalga Chlamydomonas reinhardtii has since highlighted that mTERFs are key regulators of organellar gene expression (OGE) in mitochondria and in chloroplasts. Additional functions to be fulfilled by plant mTERFs (e.g. splicing) and the fact that the expression of two organellar genomes had to be facilitated have led to a massive expansion of the plant mTERF portfolio compared to that found in mammals. Plant mTERFs are implicated in all steps of OGE ranging from the modulation of transcription to the maturation of tRNAs and hence translation. Furthermore, being regulators of OGE, mTERFs are required for a successful long-term acclimation to abiotic stress, retrograde signaling and interorganellar communication. Here, I review the recent progress in the elucidation of molecular mTERF functions.

Temporary addition of carbon fibers facilitates methanogenic degradation of ethanol during anaerobic treatment

Syntrophic methanogenesis can be improved by the addition of conductive materials. In this study, conductive carbon fibers (CFs) were applied to efficiently enrich syntrophic microorganisms with potential direct interspecies electron transfer (DIET) ability and promote methanogenic activity. With ethanol as the substrate, CFs shortened the acclimation time remarkably. The maximum methane production rate and the ethanol degradation rate of suspended biomass were increased by 40% and 68%, respectively, even when CFs were subsequently removed. However, with acetate and propionate as the mixed substrate, CFs decreased the methanogenic activity. In the reactor fed with ethanol, CFs increased the relative abundance of Geobacter, Desulfovibrio, and methanogens by 57%, 39%, and 63%, respectively. Methanosaeta possessed most methane production genes and might involve in DIET. Furthermore, CFs increased the relative abundance of ethanol-degradation genes assigned to Geobacter, Desulfovibrio and Pelobacter, suggesting the promoted ethanol-degradation. The triggered electron transport system activity and acetoclastic methanogenesis also explained the accelerated effects on ethanol-degradation by long-term acclimation with CFs. Notably, the dominance of Geobacter and Methanosaeta combined with the increased electron transfer constant in the CFs-amended ethanol reactor indicated the potential role of DIET after the removal of CFs, which deserved further clarification.

An Integrated and Multibiomarker approach to delineate oxidative stress status of Bellamya bengalensis under the interactions of elevated temperature and chlorpyrifos contamination

Synergistic effects of warming on bioconcentration and receptiveness of pollutants are still poorly unravelled in conjunction with cellular and molecular responses. The present study addressed the impact of an environmental relevant dose of chlorpyrifos (organophosphate pesticide), under control (25 °C) and elevated levels of temperature (30 °C, 35 °C) in Bellamya bengalensis, a freshwater gastropod for 60 days across various endpoints. Multiple levels of biomarkers were measured: growth conditions (organ to flesh weight ratio, condition index), oxidative stress status (SOD, CAT, GST, LPO) and DNA damage (Comet assay-3rd, 30th and 60th days only) after acute (24, 48 and 72 h) and long-term exposures (10th, 20th, 30th, 40th, 50th and 60th days). An integrated biomarker response (IBR) strategy was adapted to amalgamate results generated from various biomarkers to assess organism’s vulnerability to pesticide pollution and how it may shift with warming climate. Significant changes were observed in growth conditions under longer exposure periods. Acute as well as long-term exposures enhanced the antioxidant and detoxification enzyme activity. DNA damage was extensive under longer exposure to stress howbeit was also significantly escalated under acute severe warming. Antioxidant and detoxification mechanisms fell short in counteracting cellular level damage. The IBR results indicated long-term acclimation of B. bengalensis to elevated temperatures and pesticide contamination lead to an improved tolerance level howbeit, acute stress was more detrimental. This study provided evidence for the efficiency of employing an integrated biomarker approach for B. bengalensis in future bio-monitoring studies.

Screening, acclimation and ammonia tolerance of microalgae grown in food waste digestate

Anaerobic digestate of food waste as a waste product of anaerobic digestion contains a significant amount of nutrients making its direct disposal prohibitive due to environmental regulations. However, the nutrients in this waste are a valuable feedstock for waste-to-product endeavours such as microalgae cultivation coupled to the treatment of the digestate. A limitation to this path is the high toxic concentration of ammonia nitrogen in the digestate which limits microalgae growth, leading to the requirement for significant dilution before use. This study focused on the bioprospecting and sourcing of species capable of sustained growth in very high concentrations of ammonia nitrogen. Ten local strains of microalgae were isolated, comprising mainly of unicellular species, a colonial species, and a filamentous species. Three unicellular species were chosen (Chlorella sp., MUR 271; Scenedesmus obliquus (Tetradesmus obliquus), MUR 272; and Oocystis sp., MUR 273) and screened alongside previously isolated strains (Scenedesmus quadricauda, MUR 268; Chlorella sp., MUR 269; and Scenedesmus dimorphus (Tetradesmus dimorphus) MUR 270) which had undergone long-term acclimation in digestate. The most tolerant of the newly isolated strains was MUR 273 (Oocystis sp.), capable of proliferation in up to 600 mg L−1 NH3-N concentration in digestate. The maximum specific growth rate, μmax, of MUR 273 was 0.36 ± 0.01 day−1 at 150 mg L−1 NH3-N. The results indicate that MUR 273 displayed tolerance levels similar to that obtained with MUR 268 which had previously undergone long-term acclimation in digestate and could potentially be used in the treatment and valorization of the anaerobic digestate of food waste with significantly less dilution.

Field and acclimated metabolomes of a resurrection plant suggest strong environmental regulation in the extreme end of the species’ range

Plant secondary metabolites are significant mediators of the interactions of plants with their environment. The desiccation tolerant “resurrection plant” Myrothamnus flabellifolia Welw. is distributed throughout southern Africa and is vicariantly separated between arid Namibia and the comparatively mesic South Africa where it is subjected to contrasting climate regimes. Our earlier study indicated that these populations were metabolomically and phylogenetically distinct, leading us to hypothesise that the metabolic signature of the arid-adapted plants from Namibia was environmentally determined. To test this, we profiled the metabolomes of plants from both regions that had been sampled directly in the field (non-acclimated) with those of plants from the same regions that had been long-term acclimated in a greenhouse (acclimated). Long-term acclimation resulted in a convergence of the metabolic profiles of the plants collected from the different regions. However, while the South African plants demonstrated little difference between their acclimated and non-acclimated metabolomes, the metabolomes of the acclimated Namibian samples were distinct from those of the plants sampled in the field in Namibia. Further scrutiny of the metabolite differences between the acclimated and non-acclimated Namibian samples revealed several metabolites that were significantly more abundant in the field samples, and we propose that these might contribute to survival in this region. To further test the metabolomic response from a temporal perspective, we subjected the acclimated plants to short-term high-temperature treatment in climate chambers. However, this treatment had no significant impact on the metabolomes of the plants. Our findings suggest that long-term, rather than short-term, environmental conditions warrant large-scale reprogramming of the metabolome in M. flabellifolia, and that the climate conditions in the more arid region of the species’ range necessitate the production of particular phenolic metabolites.

A complex metabolic rearrangement towards the accumulation of glycerol and sugars consequence of a proteome remodeling is required for the survival of Chlamydomonas reinhardtii growing under osmotic stress

During its life, freshwater microalgae need to grow under varying water availabilities such as seasonal dryness of puddles or tidal variations. A vast knowledge of the physiological and molecular responses to osmotic stress is available for land plants, however these responses have been scarcely studied in microalgae. In this work we described the acclimation of Chlamydomonas reinhardtii to a mild osmotic stress during a 24 h time-course experiment. The microalgae stress-responses were reconstructed from the generated proteometabolomic datasets using a systems biology approach. Protein and metabolite dynamics showed a Chlamydomonas tolerance to osmotic stress which seems to rely on three strategies: the accumulation of osmolyte/osmoprotective sugars and glycerol, the increase of ROS scavenging mechanisms, and the alteration of cell physiology. These responses involved specific proteome and metabolome changes, probably tuned by several signaling proteins (MAPK, PP2Cs, PKL1, GUN4) and metabolites as 2-hydroxyglutarate which accumulated under stress. Moreover, the function of the plant orthologs of these proteins suggested the potential activation of brassinosteroid- and ABA-mediated pathways in a plant-like way and the activation of a HOG-like osmorelay resembling a yeast-like response. These pathways were also probably regulating a long-term acclimation mediated by epigenetic mechanisms, being promising targets for further osmotic response characterization or exploitation towards engineering strains accumulating glycerol or other industry-demanding biomolecules.