In Situ Accumulation of CaOx Crystals in C. quitensis Leaves and Its Relationship with Anatomy and Gas Exchange.

The accumulation of crystal calcium oxalate (CaOx) in plants is linked to a type of stress-induced photosynthesis termed 'alarm photosynthesis', serving as a carbon reservoir when carbon dioxide (CO2) exchange is constrained. Colobanthus quitensis is an extremophyte found from southern Mexico to Antarctica, which thrives in high-altitude Andean regions. Growing under common garden conditions, C. quitensis from different latitudinal provenances display significant variations in CaOx crystal accumulation. This raises the following questions: are these differences maintained under natural conditions? And is the CaOx accumulation related to mesophyll conductance (gm) and net photosynthesis (AN) performed in situ? It is hypothesized that in provenances with lower gm, C. quitensis will exhibit an increase in the use of CaOx crystals, resulting in reduced crystal leaf abundance. Plants from Central Chile (33°), Patagonia (51°), and Antarctica (62°) were measured in situ and sampled to determine gas exchange and CaOx crystal accumulation, respectively. Both AN and gm decrease towards higher latitudes, correlating with increases in leaf mass area and leaf density. The crystal accumulation decreases at higher latitudes, correlating positively with AN and gm. Thus, in provenances where environmental conditions induce more xeric traits, the CO2 availability for photosynthesis decreases, making the activation of alarm photosynthesis feasible as an internal source of CO2.

Ecophysiology of Antarctic Vascular Plants: An Update on the Extreme Environment Resistance Mechanisms and Their Importance in Facing Climate Change.

Antarctic flowering plants have become enigmatic because of their unique capability to colonize Antarctica. It has been shown that there is not a single trait that makes Colobanthus quitensis and Deschampsia antarctica so special, but rather a set of morphophysiological traits that coordinately confer resistance to one of the harshest environments on the Earth. However, both their capacity to inhabit Antarctica and their uniqueness remain not fully explained from a biological point of view. These aspects have become more relevant due to the climatic changes already impacting Antarctica. This review aims to compile and update the recent advances in the ecophysiology of Antarctic vascular plants, deepen understanding of the mechanisms behind their notable resistance to abiotic stresses, and contribute to understanding their potential responses to environmental changes. The uniqueness of Antarctic plants has prompted research that emphasizes the role of leaf anatomical traits and cell wall properties in controlling water loss and CO2 exchange, the role of Rubisco kinetics traits in facilitating efficient carbon assimilation, and the relevance of metabolomic pathways in elucidating key processes such as gas exchange, nutrient uptake, and photoprotection. Climate change is anticipated to have significant and contrasting effects on the morphophysiological processes of Antarctic species. However, more studies in different locations outside Antarctica and using the latitudinal gradient as a natural laboratory to predict the effects of climate change are needed. Finally, we raise several questions that should be addressed, both to unravel the uniqueness of Antarctic vascular species and to understand their potential responses to climate change.

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study.

Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses AFPs and is a promising candidate for cryopreservation applications. In this study, we investigated the cryoprotective properties of AFPs from D. antarctica extracts on Atlantic salmon spermatozoa. Apoplastic extracts were used to determine ice recrystallization inhibition (IRI), thermal hysteresis (TH) activities and ice crystal morphology. Spermatozoa were cryopreserved using a standard cryoprotectant medium (C+) and three alternative media supplemented with apoplastic extracts. Flow cytometry was employed to measure plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) postthaw. Results showed that a low concentration of AFPs (0.05 mg/mL) provided significant IRI activity. Apoplastic extracts from D. antarctica demonstrated a cryoprotective effect on salmon spermatozoa, with PMI comparable to the standard medium. Moreover, samples treated with apoplastic extracts exhibited a higher percentage of cells with high MMP. These findings represent the first and preliminary report that suggests that AFPs derived from apoplastic extracts of D. antarctica have the potential to serve as cryoprotectants and could allow the development of novel freezing media.

Leaf hydraulic properties of Antarctic plants: effects of growth temperature and its coordination with photosynthesis.

One of the well-documented effects of regional warming in Antarctica is the impact on flora. Warmer conditions modify several leaf anatomical traits of Antarctic vascular plants, increasing photosynthesis and growth. Given that CO2 and water vapor partially share their diffusion pathways through the leaf, changes in leaf anatomy could also affect the hydraulic traits of Antarctic plants. We evaluated the effects of growth temperature on several anatomical and hydraulic parameters of Antarctic plants and assessed the trait co-variation between these parameters and photosynthetic performance. Warmer conditions promoted an increase in leaf and whole plant hydraulic conductivity, correlating with adjustments in carbon assimilation. These adjustments were consistent with changes in leaf vasculature, where Antarctic species displayed different strategies. At higher temperature, Colobanthus quitensis decreased the number of leaf xylem vessels, but increased their diameter. In contrast, in Deschampsia antarctica the diameter did not change, but the number of vessels increased. Despite this contrasting behavior, some traits such as a small leaf diameter of vessels and a high cell wall rigidity were maintained in both species, suggesting a water-conservation response associated with the ability of Antarctic plants to cope with harsh environments.

Pre-Harvest Salicylic Acid Application Affects Fruit Quality and Yield under Deficit Irrigation in Aristotelia chilensis (Mol.) Plants.

Salicylic acid (SA) application is a promising agronomic tool. However, studies under field conditions are required, to confirm the potential benefits of SA. Thus, SA application was evaluated under field conditions for its effect on abscisic acid levels, antioxidant related-parameters, fruit quality, and yield in Aristotelia chilensis subjected to different levels of irrigation. During two growing seasons, three-year-old plants under field conditions were subjected to full irrigation (FI: 100% of reference evapotranspiration (ETo), and deficit irrigation (DI: 60% ETo). During each growth season, a single application of 0.5 mM SA was performed at fruit color change by spraying fruits and leaves of both irrigation treatments. The results showed that DI plants experienced moderate water stress (-1.3 MPa), which increased ABA levels and oxidative stress in the leaves. The SA application facilitated the recovery of all physiological parameters under the DI condition, increasing fruit fresh weight by 44%, with a 27% increase in fruit dry weight, a 1 mm increase in equatorial diameter, a 27% improvement in yield per plant and a 27% increase in total yield, with lesser oxidative stress and tissue ABA levels in leaves. Also, SA application significantly increased (by about 10%) the values of fruit trait variables such as soluble solids, total phenols, and antioxidant activity, with the exceptions of titratable acidity and total anthocyanins, which did not vary. The results demonstrated that SA application might be used as an agronomic strategy to improve fruit yield and quality, representing a saving of 40% regarding water use.

Transcriptome Analysis of Diurnal and Nocturnal-Warmed Plants, the Molecular Mechanism Underlying Cold Deacclimation Response in Deschampsia antarctica.

Warming in the Antarctic Peninsula is one of the fastest on earth, and is predicted to become more asymmetric in the near future. Warming has already favored the growth and reproduction of Antarctic plant species, leading to a decrease in their freezing tolerance (deacclimation). Evidence regarding the effects of diurnal and nocturnal warming on freezing tolerance-related gene expression in D. antarctica is negligible. We hypothesized that freezing tolerance-related gene (such as CBF-regulon) expression is reduced mainly by nocturnal warming rather than diurnal temperature changes in D. antarctica. The present work aimed to determine the effects of diurnal and nocturnal warming on cold deacclimation and its associated gene expression in D. antarctica, under laboratory conditions. Fully cold-acclimated plants (8 °C/0 °C), with 16h/8h thermoperiod and photoperiod duration, were assigned to four treatments for 14 days: one control (8 °C/0 °C) and three with different warming conditions (diurnal (14 °C/0 °C), nocturnal (8 °C/6 °C), and diurnal-nocturnal (14 °C/6 °C). RNA-seq was performed and differential gene expression was analyzed. Nocturnal warming significantly down-regulated the CBF transcription factors expression and associated cold stress response genes and up-regulated photosynthetic and growth promotion genes. Consequently, nocturnal warming has a greater effect than diurnal warming on the cold deacclimation process in D. antarctica. The eco-physiological implications are discussed.

The Potential of CRISPR/Cas Technology to Enhance Crop Performance on Adverse Soil Conditions.

Worldwide food security is under threat in the actual scenery of global climate change because the major staple food crops are not adapted to hostile climatic and soil conditions. Significant efforts have been performed to maintain the actual yield of crops, using traditional breeding and innovative molecular techniques to assist them. However, additional strategies are necessary to achieve the future food demand. Clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) technology, as well as its variants, have emerged as alternatives to transgenic plant breeding. This novelty has helped to accelerate the necessary modifications in major crops to confront the impact of abiotic stress on agriculture systems. This review summarizes the current advances in CRISPR/Cas applications in crops to deal with the main hostile soil conditions, such as drought, flooding and waterlogging, salinity, heavy metals, and nutrient deficiencies. In addition, the potential of extremophytes as a reservoir of new molecular mechanisms for abiotic stress tolerance, as well as their orthologue identification and edition in crops, is shown. Moreover, the future challenges and prospects related to CRISPR/Cas technology issues, legal regulations, and customer acceptance will be discussed.

How Does Diurnal and Nocturnal Warming Affect the Freezing Resistance of Antarctic Vascular Plants?

The Antarctic Peninsula has rapidly warmed up in past decades, and global warming has exhibited an asymmetric trend; therefore, it is interesting to understand whether nocturnal or diurnal warming is the most relevant for plant cold deacclimation. This study aimed to evaluate the effect of diurnal and nocturnal warming on Antarctic vascular plant's freezing resistance under laboratory conditions. This was studied by measuring the lethal temperature for 50% of tissue (LT50), ice nucleation temperature (INT), and freezing point (FP) on Deschampsia antarctica and Colobanthus quitensis plants. Additionally, soluble carbohydrates content and dehydrin levels were analyzed during nocturnal and diurnal temperatures increase. Nocturnal warming led to a 7 °C increase in the LT50 of D. antarctica and reduced dehydrin-like peptide expression. Meanwhile, C. quitensis warmed plants reduce their LT50 to about 3.6 °C. Both species reduce their sucrose content by more than 28% in warming treatments. Therefore, nocturnal warming leads to cold deacclimation in both plant species, while C. quitensis plants are also cold-deacclimated upon warm days. This suggests that even when the remaining freezing resistance of both species allows them to tolerate summer freezing events, C. quitensis can reach its boundaries of freezing vulnerability in the near future if warming in the Antarctic Peninsula progress.

Vegetation drives the response of the active fraction of the rhizosphere microbial communities to soil warming in Antarctic vascular plants.

In the Antarctic Peninsula, increases in mean annual temperature are associated with the coverage and population density of the two Antarctic vascular plant species-Deschampsia antarctica and Colobanthus quitensis-potentially modifying critical soil processes. In this study, we characterized the diversity and community composition of active microorganisms inhabiting the vascular plant rhizosphere in two sites with contrasting vegetation cover in King George Island, Western Antarctic Peninsula. We assessed the interplay between soil physicochemical properties and microbial diversity and composition, evaluating the effect of an in situ experimental warming on the microbial communities of the rhizosphere from D. antarctica and C. quitensis. Bacteria and Eukarya showed different responses to warming in both sites, and the effect was more noticeable in microbial eukaryotes from the low vegetation site. Furthermore, important changes were found in the relative abundance of Tepidisphaerales (Bacteria) and Ciliophora (Eukarya) between warming and control treatments. Our results showed that rhizosphere eukaryal communities are more sensitive to in situ warming than bacterial communities. Overall, our results indicate that vegetation drives the response of the active fraction of the microbial communities from the rhizosphere of Antarctic vascular plants to soil warming.

Respiratory and Photosynthetic Responses of Antarctic Vascular Plants Are Differentially Affected by CO2 Enrichment and Nocturnal Warming.

Projected rises in atmospheric CO2 concentration and minimum night-time temperatures may have important effects on plant carbon metabolism altering the carbon balance of the only two vascular plant species in the Antarctic Peninsula. We assessed the effect of nocturnal warming (8/5 °C vs. 8/8 °C day/night) and CO2 concentrations (400 ppm and 750 ppm) on gas exchange, non-structural carbohydrates, two respiratory-related enzymes, and mitochondrial size and number in two species of vascular plants. In Colobanthus quitensis, light-saturated photosynthesis measured at 400 ppm was reduced when plants were grown in the elevated CO2 or in the nocturnal warming treatments. Growth in elevated CO2 reduced stomatal conductance but nocturnal warming did not. The short-term sensitivity of respiration, relative protein abundance, and mitochondrial traits were not responsive to either treatment in this species. Moreover, some acclimation to nocturnal warming at ambient CO2 was observed. Altogether, these responses in C. quitensis led to an increase in the respiration-assimilation ratio in plants grown in elevated CO2. The response of Deschampsia antarctica to the experimental treatments was quite distinct. Photosynthesis was not affected by either treatment; however, respiration acclimated to temperature in the elevated CO2 treatment. The observed short-term changes in thermal sensitivity indicate type I acclimation of respiration. Growth in elevated CO2 and nocturnal warming resulted in a reduction in mitochondrial numbers and an increase in mitochondrial size in D. antarctica. Overall, our results suggest that with climate change D. antarctica could be more successful than C. quitensis, due to its ability to make metabolic adjustments to maintain its carbon balance.

Salicylic Acid Improves Antioxidant Defense System and Photosynthetic Performance in Aristotelia chilensis Plants Subjected to Moderate Drought Stress.

Salicylic acid (SA) has been shown to ameliorate drought stress. However, physiological and biochemical mechanisms involved in drought stress tolerance induced by SA in plants have not been well understood. Thus, this study aimed to study the role of SA application on enzymatic and non-enzymatic antioxidants, photosynthetic performance, and plant growth in A. chilensis plants subjected to moderate drought stress. One-year-old A. chilensis plants were subjected to 100% and 60% of field capacity. When plants reached moderate drought stress (average of stem water potential of -1.0 MPa, considered as moderate drought stress), a single SA application was performed on plants. Then, physiological and biochemical features were determined at different times during 14 days. Our study showed that SA application increased 13.5% plant growth and recovered 41.9% AN and 40.7% gs in drought-stressed plants on day 3 compared to drought-stressed plants without SA application. Interestingly, SOD and APX activities were increased 85% and 60%, respectively, in drought-stressed SA-treated plants on day 3. Likewise, SA improved 30% total phenolic content and 60% antioxidant capacity in drought-stressed A. chilensis plants. Our study provides insight into the SA mechanism to tolerate moderate drought stress in A. chilensis plants.

Calcium Oxalate Crystals in Leaves of the Extremophile Plant Colobanthus quitensis (Kunth) Bartl. (Caryophyllaceae).

The presence of calcium oxalate (CaOx) crystals has been widely reported in the plant kingdom. These structures play a central role in various physiological functions, including calcium regulation, metal detoxification, and photosynthesis. However, precise knowledge about their possible roles and functions in plants is still limited. Therefore, the present work aims to study the ecotypic variability of Colobanthus quitensis, an extremophile species, concerning CaOx crystal accumulation. The CaOx crystals were studied in leaves of C. quitensis collected from different provenances within a latitudinal gradient (From Andes mountains in central Chile to Antarctica) and grown under common garden conditions. Polarized light microscopy, digital image analysis, and electron microscopy were used to characterize CaOx crystals. The presence of CaOx crystals was confirmed in the four provenances of C. quitensis, with significant differences in the accumulation among them. The Andean populations presented the highest accumulation of crystals and the Antarctic population the lowest. Electron microscopy showed that CaOx crystals in C. quitensis are classified as druses based on their morphology. The differences found could be linked to processes of ecotypic differentiation and plant adaptation to harsh environments.

A high-throughput method for measuring critical thermal limits of leaves by chlorophyll imaging fluorescence.

Plant thermal tolerance is a crucial research area as the climate warms and extreme weather events become more frequent. Leaves exposed to temperature extremes have inhibited photosynthesis and will accumulate damage to PSII if tolerance thresholds are exceeded. Temperature-dependent changes in basal chlorophyll fluorescence (T-F0) can be used to identify the critical temperature at which PSII is inhibited. We developed and tested a high-throughput method for measuring the critical temperatures for PSII at low (CTMIN) and high (CTMAX) temperatures using a Maxi-Imaging fluorimeter and a thermoelectric Peltier plate heating/cooling system. We examined how experimental conditions of wet vs dry surfaces for leaves and heating/cooling rate, affect CTMIN and CTMAX across four species. CTMAX estimates were not different whether measured on wet or dry surfaces, but leaves were apparently less cold tolerant when on wet surfaces. Heating/cooling rate had a strong effect on both CTMAX and CTMIN that was species-specific. We discuss potential mechanisms for these results and recommend settings for researchers to use when measuring T-F0. The approach that we demonstrated here allows the high-throughput measurement of a valuable ecophysiological parameter that estimates the critical temperature thresholds of leaf photosynthetic performance in response to thermal extremes.

Decomposition of Calcium Oxalate Crystals in Colobanthus quitensis under CO2 Limiting Conditions.

Calcium oxalate (CaOx) crystals are widespread among plant species. Their functions are not yet completely understood; however, they can provide tolerance against multiple environmental stress factors. Recent evidence suggested that CaOx crystals function as carbon reservoirs since its decomposition provides CO2 that may be used as carbon source for photosynthesis. This might be advantageous in plants with reduced mesophyll conductance, such as the Antarctic plant Colobanthus quitensis, which have shown CO2 diffusion limitations. In this study, we evaluate the effect of two CO2 concentrations in the CaOx crystals decomposition and chlorophyll fluorescence of C. quitensis. Plants were exposed to airflows with 400 ppm and 11.5 ppm CO2 and the number and relative size of crystals, electron transport rate (ETR), and oxalate oxidase (OxO) activity were monitored along time (10 h). Here we showed that leaf crystal area decreases over time in plants with 11.5 ppm CO2, which was accompanied by increased OxO activity and only a slight decrease in the ETR. These results suggested a relation between CO2 limiting conditions and the CaOx crystals decomposition in C. quitensis. Hence, crystal decomposition could be a complementary endogenous mechanism for CO2 supply in plants facing the Antarctic stressful habitat.

Exploratory Study of Fatty Acid Profile in Two Filmy Ferns with Contrasting Desiccation Tolerance Reveal the Production of Very Long Chain Polyunsaturated Omega-3 Fatty Acids.

Lipids are fundamental components of cell membranes and play a significant role in their integrity and fluidity. Alteration in lipid composition of membranes has been reported to be a major response to abiotic environmental stresses. This work was focused on the characterization of frond lipid composition and membrane integrity during a desiccation-rehydration cycle of two filmy fern species with contrasting desiccation tolerance: Hymenophyllum caudiculatum (less tolerant) and Hymenophyllum plicatum (more tolerant). The relative water content decreased without differences between species when both filmy ferns were subjected to desiccation. However, H. plicatum reached a higher relative water content than H. caudiculatum after rehydration. Fatty acids profiles showed the presence of a very long chain polyunsaturated fatty acid during the desiccation-rehydration cycle, with eicosatrienoic acid being the most abundant. Additionally, propidium iodide permeation staining and confocal microscopy demonstrated that, following the desiccation-rehydration cycle, H. plicatum exhibited a greater membrane integrity than H. caudiculatum. The lack of some very long chain fatty acids such as C22:1n9 and C24:1n9 in this species contrasting with H. plicatum may be associated with its lower membrane stability during the desiccation-rehydration cycle. This report provides the first insight into the fatty acid composition and dynamics of the membrane integrity of filmy ferns during a desiccation-rehydration cycle. This could potentially play a role in determining the different levels of desiccation tolerance and microhabitat preferences exhibited by Hymenophyllaceae species.

Decoding Gene Networks Modules That Explain the Recovery of Hymenoglossum cruentum Cav. After Extreme Desiccation.

Hymenoglossum cruentum (Hymenophyllaceae) is a poikilohydric, homoiochlorophyllous desiccation-tolerant (DT) epiphyte fern. It can undergo fast and frequent dehydration-rehydration cycles. This fern is highly abundant at high-humidity/low-light microenvironments within the canopy, although rapid changes in humidity and light intensity are frequent. The objective of this research is to identify genes associated to desiccation-rehydration cycle in the transcriptome of H. cruentum to better understand the genetic dynamics behind its desiccation tolerance mechanism. H. cruentum plants were subjected to a 7 days long desiccation-rehydration process and then used to identify key expressed genes associated to its capacity to dehydrate and rehydrate. The relative water content (RWC) and maximum quantum efficiency (F v/F m) of H. cruentum fronds decayed to 6% and 0.04, respectively, at the end of the desiccation stage. After re-watering, the fern showed a rapid recovery of RWC and F v/F m (ca. 73% and 0.8, respectively). Based on clustering and network analysis, our results reveal key genes, such as UBA/TS-N, DYNLL, and LHC, orchestrating intracellular motility and photosynthetic metabolism; strong balance between avoiding cell death and defense (CAT3, AP2/ERF) when dehydrated, and detoxifying pathways and stabilization of photosystems (GST, CAB2, and ELIP9) during rehydration. Here we provide novel insights into the genetic dynamics behind the desiccation tolerance mechanism of H. cruentum.

Effect of in vitro cold acclimation of Deschampsia antarctica on the accumulation of proteins with antifreeze activity.

Deschampsia antarctica has managed to colonize the maritime Antarctic. One of the main factors associated with its tolerance to low temperatures is the presence of apoplastic proteins with antifreeze activity. This work focuses on the effect of cold acclimation of D. antarctica on the accumulation of apoplastic proteins with antifreeze activity. Antifreeze proteins present in apoplastic extracts were purified by ice affinity purification, and their identity was determined by protein sequencing. D. antarctica plants were subjected to 22 days of cold acclimation at 4 °C. The highest content of apoplastic proteins with antifreeze activity was obtained at between 12 and 16 days of acclimation. Protein sequencing allowed their identification with >95% probability. Percentage coverage was 74% with D. antarctica ice recrystallization inhibition protein 1 (DaIRIP1) and 55% with DaIRIP3. Cold acclimation of D. antarctica improved the yield of apoplastic proteins, and resulted in an increase in the antifreeze activity of apoplastic extracts. An in silico analysis suggested that the fluctuations presented by the three-dimensional structures of DaIRIPs help to explain the presence of certain DaIRIPs in apoplastic extracts under the cold acclimation conditions evaluated.

A comparative gene co-expression analysis using self-organizing maps on two congener filmy ferns identifies specific desiccation tolerance mechanisms associated to their microhabitat preference.

BACKGROUND: Filmy-ferns (Hymenophyllaceae) are poikilohydric, homoiochlorophyllous desiccation-tolerant (DT) epiphytes. They can colonize lower and upper canopy environments of humid forest. Filmy-ferns desiccate rapidly (hours), contrasting with DT angiosperms (days/weeks). It has been proposed that desiccation tolerance in filmy-ferns would be associated mainly with constitutive features rather than induced responses during dehydration. However, we hypothesize that the inter-specific differences in vertical distribution would be associated with different dynamics of gene expression within the dehydration or rehydration phases. A comparative transcriptomic analysis with an artificial neural network was done on Hymenophyllum caudiculatum (restricted to lower canopy) and Hymenophyllum dentatum (reach upper canopy) during a desiccation/rehydration cycle. RESULTS: Raw reads were assembled into 69,599 transcripts for H. dentatum and 34,726 transcripts for H. caudiculatum. Few transcripts showed significant changes in differential expression (DE). H. caudiculatum had ca. twice DE genes than H. dentatum and higher proportion of increased-and-decreased abundance of genes occurs during dehydration. In contrast, the abundance of genes in H. dentatum decreased significantly when transitioning from dehydration to rehydration. According to the artificial neural network results, H. caudiculatum enhanced osmotic responses and phenylpropanoid related pathways, whilst H. dentatum enhanced its defense system responses and protection against high light stress. CONCLUSIONS: Our findings provide a deeper understanding of the mechanisms underlying the desiccation tolerance responses of two filmy ferns and the relationship between the species-specific response and the microhabitats these ferns occupy in nature.

How do vascular plants perform photosynthesis in extreme environments? An integrative ecophysiological and biochemical story.

In this work, we review the physiological and molecular mechanisms that allow vascular plants to perform photosynthesis in extreme environments, such as deserts, polar and alpine ecosystems. Specifically, we discuss the morpho/anatomical, photochemical and metabolic adaptive processes that enable a positive carbon balance in photosynthetic tissues under extreme temperatures and/or severe water-limiting conditions in C3 species. Nevertheless, only a few studies have described the in situ functioning of photoprotection in plants from extreme environments, given the intrinsic difficulties of fieldwork in remote places. However, they cover a substantial geographical and functional range, which allowed us to describe some general trends. In general, photoprotection relies on the same mechanisms as those operating in the remaining plant species, ranging from enhanced morphological photoprotection to increased scavenging of oxidative products such as reactive oxygen species. Much less information is available about the main physiological and biochemical drivers of photosynthesis: stomatal conductance (gs ), mesophyll conductance (gm ) and carbon fixation, mostly driven by RuBisCO carboxylation. Extreme environments shape adaptations in structures, such as cell wall and membrane composition, the concentration and activation state of Calvin-Benson cycle enzymes, and RuBisCO evolution, optimizing kinetic traits to ensure functionality. Altogether, these species display a combination of rearrangements, from the whole-plant level to the molecular scale, to sustain a positive carbon balance in some of the most hostile environments on Earth.