Overexpression of Water-Responsive Genes Promoted by Elevated CO2 Reduces ROS and Enhances Drought Tolerance in Coffea Species.

Drought is a major constraint to plant growth and productivity worldwide and will aggravate as water availability becomes scarcer. Although elevated air [CO2] might mitigate some of these effects in plants, the mechanisms underlying the involved responses are poorly understood in woody economically important crops such as Coffea. This study analyzed transcriptome changes in Coffea canephora cv. CL153 and C. arabica cv. Icatu exposed to moderate (MWD) or severe water deficits (SWD) and grown under ambient (aCO2) or elevated (eCO2) air [CO2]. We found that changes in expression levels and regulatory pathways were barely affected by MWD, while the SWD condition led to a down-regulation of most differentially expressed genes (DEGs). eCO2 attenuated the impacts of drought in the transcripts of both genotypes but mostly in Icatu, in agreement with physiological and metabolic studies. A predominance of protective and reactive oxygen species (ROS)-scavenging-related genes, directly or indirectly associated with ABA signaling pathways, was found in Coffea responses, including genes involved in water deprivation and desiccation, such as protein phosphatases in Icatu, and aspartic proteases and dehydrins in CL153, whose expression was validated by qRT-PCR. The existence of a complex post-transcriptional regulatory mechanism appears to occur in Coffea explaining some apparent discrepancies between transcriptomic, proteomic, and physiological data in these genotypes.

Xylem embolism spread is largely prevented by interconduit pit membranes until the majority of conduits are gas-filled.

Xylem embolism resistance varies across species influencing drought tolerance, yet little is known about the determinants of the embolism resistance of an individual conduit. Here we conducted an experiment using the optical vulnerability method to test whether individual conduits have a specific water potential threshold for embolism formation and whether pre-existing embolism in neighbouring conduits alters this threshold. Observations were made on a diverse sample of angiosperm and conifer species through a cycle of dehydration, rehydration and subsequent dehydration to death. Upon rehydration after the formation of embolism, no refilling was observed. When little pre-existing embolism was present, xylem conduits had a conserved, individual embolism-resistance threshold that varied across the population of conduits. The consequence of a variable conduit-specific embolism threshold is that a small degree of pre-existing embolism in the xylem results in apparently more resistant xylem in subsequent dehydrations, particularly in angiosperms with vessels. While our results suggest that pit membranes separating xylem conduits are critical for maintaining a conserved individual conduit threshold for embolism when little pre-existing embolism is present, as the percentage of embolized conduits increases, gas movement, local pressure differences and connectivity between conduits increasingly contribute to embolism spread.

Impaired auxin signaling increases vein and stomatal density but reduces hydraulic efficiency and ultimately net photosynthesis.

Auxins are known to regulate xylem development in plants, but their effects on water transport efficiency are poorly known. Here we used tomato plants with the diageotropica mutation (dgt), which has impaired function of a cyclophilin 1 cis-trans isomerase involved in auxin signaling, and the corresponding wild type (WT) to explore the mutation's effects on plant hydraulics and leaf gas exchange. The xylem of the dgt mutant showed a reduced hydraulically weighted vessel diameter (Dh) (24-43%) and conduit number (25-58%) in petioles and stems, resulting in lower theoretical hydraulic conductivities (Kt); on the other hand, no changes in root Dh and Kt were observed. The measured stem and leaf hydraulic conductances of the dgt mutant were lower (up to 81%), in agreement with the Kt values; however, despite dgt and WT plants showing similar root Dh and Kt, the measured root hydraulic conductance of the dgt mutant was 75% lower. The dgt mutation increased the vein and stomatal density, which could potentially increase photosynthesis. Nevertheless, even though it had the same photosynthetic capacity as WT plants, the dgt mutant showed a photosynthetic rate c. 25% lower, coupled with a stomatal conductance reduction of 52%. These results clearly demonstrate that increases in minor vein and stomatal density only result in higher leaf gas exchange when accompanied by higher hydraulic efficiency.

Specific leaf area is modulated by nitrogen via changes in primary metabolism and parenchymal thickness in pepper.

MAIN CONCLUSION: Nitrogen promotes changes in SLA through metabolism and anatomical traits in Capsicum plants. Specific leaf area (SLA) is a key trait influencing light interception and light use efficiency that often impacts plant growth and production. SLA is a key trait explaining growth variations of plant species under different environments. Both light and nitrogen (N) supply are important determinants of SLA. To better understand the effect of irradiance level and N on SLA in Capsicum chinense, we evaluated primary metabolites and morphological traits of two commercial cultivars (Biquinho and Habanero) in response to changes in both parameters. Both genotypes showed increased SLA with shading, and a decrease in SLA in response to increased N supply, however, with Habanero showing a stable SLA in the range of N deficiency to sufficient N doses. Correlation analyses indicated that decreased SLA in response to higher N supply was mediated by altered amino acids, protein, and starch levels, influencing leaf density. Moreover, in the range of moderate N deficiency to N sufficiency, both genotypes exhibited differences in SLA response, with Biquinho and Habanero displaying alterations on palisade and spongy parenchyma, respectively. Altogether, the results suggest that SLA responses to N supply are modulated by the balance between certain metabolites content and genotype-dependent changes in the parenchyma cells influencing leaf thickness and density.

Limited plasticity in embolism resistance in response to light in leaves and stems in species with considerable vulnerability segmentation.

Xylem resistance to embolism is a key metric determining plant survival during drought. Yet, we have a limited understanding of the degree of plasticity in vulnerability to embolism. Here, we tested whether light availability influences embolism resistance in leaves and stems. The optical vulnerability method was used to assess stem and leaf resistance to embolism in Phellodendron amurense and Ilex verticillata acclimated to sun and shade microenvironments within the same canopy. In both species, we found considerable segmentation in xylem resistance to embolism between leaves and stems, but only minor acclimation in response to light availability. With the addition of a third species, Betula pubescens, which shows no vulnerability segmentation, we sought to investigate xylem anatomical traits that might correlate with strong vulnerability segmentation. We found a correlation between the area fraction of vessels in the xylem and embolism resistance across species and tissue types. Our results suggest that minimal acclimation of embolism resistance occurs in response to light environment in the same individual and that the degree of vulnerability segmentation between leaves and stems might be determined by the vessel lumen fraction of the xylem.

A Transcriptomic Approach to Understanding the Combined Impacts of Supra-Optimal Temperatures and CO2 Revealed Different Responses in the Polyploid Coffea arabica and Its Diploid Progenitor C. canephora.

Understanding the effect of extreme temperatures and elevated air (CO2) is crucial for mitigating the impacts of the coffee industry. In this work, leaf transcriptomic changes were evaluated in the diploid C. canephora and its polyploid C. arabica, grown at 25 °C and at two supra-optimal temperatures (37 °C, 42 °C), under ambient (aCO2) or elevated air CO2 (eCO2). Both species expressed fewer genes as temperature rose, although a high number of differentially expressed genes (DEGs) were observed, especially at 42 °C. An enrichment analysis revealed that the two species reacted differently to the high temperatures but with an overall up-regulation of the photosynthetic machinery until 37 °C. Although eCO2 helped to release stress, 42 °C had a severe impact on both species. A total of 667 photosynthetic and biochemical related-DEGs were altered with high temperatures and eCO2, which may be used as key probe genes in future studies. This was mostly felt in C. arabica, where genes related to ribulose-bisphosphate carboxylase (RuBisCO) activity, chlorophyll a-b binding, and the reaction centres of photosystems I and II were down-regulated, especially under 42°C, regardless of CO2. Transcriptomic changes showed that both species were strongly affected by the highest temperature, although they can endure higher temperatures (37 °C) than previously assumed.

Silicon nutrition mitigates the negative impacts of iron toxicity on rice photosynthesis and grain yield.

Excess iron (Fe) is commonly observed in wetland rice (Oryza sativa L.) plants, impairing crop growth and productivity. Some information suggests that silicon (Si) can reduce Fe content in leaves and roots of rice (vegetative phase), but nothing is known if Si could mitigate the effects of Fe toxicity on rice production and photosynthesis. Here, we assessed the role of Si in alleviating the well-known effects of Fe toxicity on nutritional imbalances, biomass accumulation, photosynthesis and grain yield using two rice cultivars having differential abilities to tolerate excess Fe. Plants were hydroponically grown under two Fe levels (25 µM or 5 mM) and the nutrient solutions were amended with Si (0 or 2 mM). Under excess Fe were detected (i) nutritional deficiencies, especially of calcium and magnesium in leaves; (ii) negligible changes in grain nutritional composition, independently of Si application; (iii) decreases in net photosynthetic rates, stomatal conductance and electron transport rate, in parallel to decreased grain yield components (total grain biomass, 1000-grain mass, percentage of filled grains, number of grains per plant and harvest index), especially in the Fe-sensitive cultivar. These impairments were partially reversed by the application of Si. Results also suggest that Si alleviated the negative impacts of Fe on spikelet sterility. In summary, we conclude that the use of Si can be recommended as an effective management strategy to reduce the negative impacts of Fe toxicity on rice photosynthetic performance and crop yield.

Transcriptomic Leaf Profiling Reveals Differential Responses of the Two Most Traded Coffee Species to Elevated [CO2].

As atmospheric [CO2] continues to rise to unprecedented levels, understanding its impact on plants is imperative to improve crop performance and sustainability under future climate conditions. In this context, transcriptional changes promoted by elevated CO2 (eCO2) were studied in genotypes from the two major traded coffee species: the allopolyploid Coffea arabica (Icatu) and its diploid parent, C. canephora (CL153). While Icatu expressed more genes than CL153, a higher number of differentially expressed genes were found in CL153 as a response to eCO2. Although many genes were found to be commonly expressed by the two genotypes under eCO2, unique genes and pathways differed between them, with CL153 showing more enriched GO terms and metabolic pathways than Icatu. Divergent functional categories and significantly enriched pathways were found in these genotypes, which altogether supports contrasting responses to eCO2. A considerable number of genes linked to coffee physiological and biochemical responses were found to be affected by eCO2 with the significant upregulation of photosynthetic, antioxidant, and lipidic genes. This supports the absence of photosynthesis down-regulation and, therefore, the maintenance of increased photosynthetic potential promoted by eCO2 in these coffee genotypes.

Impaired Malate and Fumarate Accumulation Due to the Mutation of the Tonoplast Dicarboxylate Transporter Has Little Effects on Stomatal Behavior.

Malate is a central metabolite involved in a multiplicity of plant metabolic pathways, being associated with mitochondrial metabolism and playing significant roles in stomatal movements. Vacuolar malate transport has been characterized at the molecular level and is performed by at least one carrier protein and two channels in Arabidopsis (Arabidopsis thaliana) vacuoles. The absence of the Arabidopsis tonoplast Dicarboxylate Transporter (tDT) in the tdt knockout mutant was associated previously with an impaired accumulation of malate and fumarate in leaves. Here, we investigated the consequences of this lower accumulation on stomatal behavior and photosynthetic capacity as well as its putative metabolic impacts. Neither the stomatal conductance nor the kinetic responses to dark, light, or high CO2 were highly affected in tdt plants. In addition, we did not observe any impact on stomatal aperture following incubation with abscisic acid, malate, or citrate. Furthermore, an effect on photosynthetic capacity was not observed in the mutant lines. However, leaf mitochondrial metabolism was affected in the tdt plants. Levels of the intermediates of the tricarboxylic acid cycle were altered, and increases in both light and dark respiration were observed. We conclude that manipulation of the tonoplastic organic acid transporter impacted mitochondrial metabolism, while the overall stomatal and photosynthetic capacity were unaffected.

Coordinated plasticity maintains hydraulic safety in sunflower leaves.

The xylem cavitation threshold water potential establishes a hydraulic limit on the ability of woody species to survive in water-limiting environments, but herbs may be more plastic in terms of their ability to adapt to drying conditions. Here, we examined the capacity of sunflower (Helianthus annuus L.) leaves to adapt to reduced water availability by modifying the sensitivity of xylem and stomata to soil water deficit. We found that sunflower plants grown under water-limited conditions significantly adjusted leaf osmotic potential, which was linked to a prolongation of stomatal opening as soil dried and a reduced sensitivity of photosynthesis to water-stress-induced damage. At the same time, the vulnerability of midrib xylem to water-stress-induced cavitation was observed to be highly responsive to growth conditions, with water-limited plants producing conduits with thicker cell walls which were more resistant to xylem cavitation. Coordinated plasticity in osmotic potential and xylem vulnerability enabled water-limited sunflowers to safely extract water from the soil, while protecting leaf xylem against embolism. High plasticity in sunflower xylem contrasts with data from woody plants and may suggest an alternative strategy in herbs.

The genetic architecture of photosynthesis and plant growth-related traits in tomato.

To identify genomic regions involved in the regulation of fundamental physiological processes such as photosynthesis and respiration, a population of Solanum pennellii introgression lines was analyzed. We determined phenotypes for physiological, metabolic, and growth related traits, including gas exchange and chlorophyll fluorescence parameters. Data analysis allowed the identification of 208 physiological and metabolic quantitative trait loci with 33 of these being associated to smaller intervals of the genomic regions, termed BINs. Eight BINs were identified that were associated with higher assimilation rates than the recurrent parent M82. Two and 10 genomic regions were related to shoot and root dry matter accumulation, respectively. Nine genomic regions were associated with starch levels, whereas 12 BINs were associated with the levels of other metabolites. Additionally, a comprehensive and detailed annotation of the genomic regions spanning these quantitative trait loci allowed us to identify 87 candidate genes that putatively control the investigated traits. We confirmed 8 of these at the level of variance in gene expression. Taken together, our results allowed the identification of candidate genes that most likely regulate photosynthesis, primary metabolism, and plant growth and as such provide new avenues for crop improvement.

Enhanced Photosynthesis and Growth in atquac1 Knockout Mutants Are Due to Altered Organic Acid Accumulation and an Increase in Both Stomatal and Mesophyll Conductance.

Stomata control the exchange of CO2 and water vapor in land plants. Thus, whereas a constant supply of CO2 is required to maintain adequate rates of photosynthesis, the accompanying water losses must be tightly regulated to prevent dehydration and undesired metabolic changes. Accordingly, the uptake or release of ions and metabolites from guard cells is necessary to achieve normal stomatal function. The AtQUAC1, an R-type anion channel responsible for the release of malate from guard cells, is essential for efficient stomatal closure. Here, we demonstrate that mutant plants lacking AtQUAC1 accumulated higher levels of malate and fumarate. These mutant plants not only display slower stomatal closure in response to increased CO2 concentration and dark but are also characterized by improved mesophyll conductance. These responses were accompanied by increases in both photosynthesis and respiration rates, without affecting the activity of photosynthetic and respiratory enzymes and the expression of other transporter genes in guard cells, which ultimately led to improved growth. Collectively, our results highlight that the transport of organic acids plays a key role in plant cell metabolism and demonstrate that AtQUAC1 reduce diffusive limitations to photosynthesis, which, at least partially, explain the observed increments in growth under well-watered conditions.

Photosynthetic and metabolic acclimation to repeated drought events play key roles in drought tolerance in coffee.

Over the last decades, most information on the mechanisms underlying tolerance to drought has been gained by considering this stress as a single event that happens just once in the life of a plant, in contrast to what occurs under natural conditions where recurrent drought episodes are the rule. Here we explored mechanisms of drought tolerance in coffee (Coffea canephora) plants from a broader perspective, integrating key aspects of plant physiology and biochemistry. We show that plants exposed to multiple drought events displayed higher photosynthetic rates, which were largely accounted for by biochemical rather than diffusive or hydraulic factors, than those submitted to drought for the first time. Indeed, these plants displayed higher activities of RuBisCO and other enzymes associated with carbon and antioxidant metabolism. Acclimation to multiple drought events involved the expression of trainable genes related to drought tolerance and was also associated with a deep metabolite reprogramming with concordant alterations in central metabolic processes such as respiration and photorespiration. Our results demonstrate that plants exposed to multiple drought cycles can develop a differential acclimation that potentiates their defence mechanisms, allowing them to be kept in an 'alert state' to successfully cope with further drought events.

Stomatal dynamics are limited by leaf hydraulics in ferns and conifers: results from simultaneous measurements of liquid and vapour fluxes in leaves.

Stomatal responsiveness to vapour pressure deficit (VPD) results in continuous regulation of daytime gas-exchange directly influencing leaf water status and carbon gain. Current models can reasonably predict steady-state stomatal conductance (gs ) to changes in VPD but the gs dynamics between steady-states are poorly known. Here, we used a diverse sample of conifers and ferns to show that leaf hydraulic architecture, in particular leaf capacitance, has a major role in determining the gs response time to perturbations in VPD. By using simultaneous measurements of liquid and vapour fluxes into and out of leaves, the in situ fluctuations in leaf water balance were calculated and appeared to be closely tracked by changes in gs thus supporting a passive model of stomatal control. Indeed, good agreement was found between observed and predicted gs when using a hydropassive model based on hydraulic traits. We contend that a simple passive hydraulic control of stomata in response to changes in leaf water status provides for efficient stomatal responses to VPD in ferns and conifers, leading to closure rates as fast or faster than those seen in most angiosperms.

Sustained enhancement of photosynthesis in coffee trees grown under free-air CO2 enrichment conditions: disentangling the contributions of stomatal, mesophyll, and biochemical limitations.

Coffee (Coffea spp.), a globally traded commodity, is a slow-growing tropical tree species that displays an improved photosynthetic performance when grown under elevated atmospheric CO2 concentrations ([CO2]). To investigate the mechanisms underlying this response, two commercial coffee cultivars (Catuaí and Obatã) were grown using the first free-air CO2 enrichment (FACE) facility in Latin America. Measurements were conducted in two contrasting growth seasons, which were characterized by the high (February) and low (August) sink demand. Elevated [CO2] led to increases in net photosynthetic rates (A) in parallel with decreased photorespiration rates, with no photochemical limitations to A. The stimulation of A by elevated CO2 supply was more prominent in August (56% on average) than in February (40% on average). Overall, the stomatal and mesophyll conductances, as well as the leaf nitrogen and phosphorus concentrations, were unresponsive to the treatments. Photosynthesis was strongly limited by diffusional constraints, particularly at the stomata level, and this pattern was little, if at all, affected by elevated [CO2]. Relative to February, starch pools (but not soluble sugars) increased remarkably (>500%) in August, with no detectable alteration in the maximum carboxylation capacity estimated on a chloroplast [CO2] basis. Upregulation of A by elevated [CO2] took place with no signs of photosynthetic downregulation, even during the period of low sink demand, when acclimation would be expected to be greatest.

Long-term elevated air [CO2 ] strengthens photosynthetic functioning and mitigates the impact of supra-optimal temperatures in tropical Coffea arabica and C. canephora species.

The tropical coffee crop has been predicted to be threatened by future climate changes and global warming. However, the real biological effects of such changes remain unknown. Therefore, this work aims to link the physiological and biochemical responses of photosynthesis to elevated air [CO2 ] and temperature in cultivated genotypes of Coffea arabica L. (cv. Icatu and IPR108) and Coffea canephora cv. Conilon CL153. Plants were grown for ca. 10 months at 25/20°C (day/night) and 380 or 700 µl CO2 l(-1) and then subjected to temperature increase (0.5°C day(-1) ) to 42/34°C. Leaf impacts related to stomatal traits, gas exchanges, C isotope composition, fluorescence parameters, thylakoid electron transport and enzyme activities were assessed at 25/20, 31/25, 37/30 and 42/34°C. The results showed that (1) both species were remarkably heat tolerant up to 37/30°C, but at 42/34°C a threshold for irreversible nonstomatal deleterious effects was reached. Impairments were greater in C. arabica (especially in Icatu) and under normal [CO2 ]. Photosystems and thylakoid electron transport were shown to be quite heat tolerant, contrasting to the enzymes related to energy metabolism, including RuBisCO, which were the most sensitive components. (2) Significant stomatal trait modifications were promoted almost exclusively by temperature and were species dependent. Elevated [CO2 ], (3) strongly mitigated the impact of temperature on both species, particularly at 42/34°C, modifying the response to supra-optimal temperatures, (4) promoted higher water-use efficiency under moderately higher temperature (31/25°C) and (5) did not provoke photosynthetic downregulation. Instead, enhancements in [CO2 ] strengthened photosynthetic photochemical efficiency, energy use and biochemical functioning at all temperatures. Our novel findings demonstrate a relevant heat resilience of coffee species and that elevated [CO2 ] remarkably mitigated the impact of heat on coffee physiology, therefore playing a key role in this crop sustainability under future climate change scenarios.

Antisense inhibition of the iron-sulphur subunit of succinate dehydrogenase enhances photosynthesis and growth in tomato via an organic acid-mediated effect on stomatal aperture.

Transgenic tomato (Solanum lycopersicum) plants expressing a fragment of the Sl SDH2-2 gene encoding the iron sulfur subunit of the succinate dehydrogenase protein complex in the antisense orientation under the control of the 35S promoter exhibit an enhanced rate of photosynthesis. The rate of the tricarboxylic acid (TCA) cycle was reduced in these transformants, and there were changes in the levels of metabolites associated with the TCA cycle. Furthermore, in comparison to wild-type plants, carbon dioxide assimilation was enhanced by up to 25% in the transgenic plants under ambient conditions, and mature plants were characterized by an increased biomass. Analysis of additional photosynthetic parameters revealed that the rate of transpiration and stomatal conductance were markedly elevated in the transgenic plants. The transformants displayed a strongly enhanced assimilation rate under both ambient and suboptimal environmental conditions, as well as an elevated maximal stomatal aperture. By contrast, when the Sl SDH2-2 gene was repressed by antisense RNA in a guard cell-specific manner, changes in neither stomatal aperture nor photosynthesis were observed. The data obtained are discussed in the context of the role of TCA cycle intermediates both generally with respect to photosynthetic metabolism and specifically with respect to their role in the regulation of stomatal aperture.

Silicon nutrition alleviates the negative impacts of arsenic on the photosynthetic apparatus of rice leaves: an analysis of the key limitations of photosynthesis.

Silicon (Si) plays important roles in alleviating various abiotic stresses. In rice (Oryza sativa), arsenic (As) is believed to share the Si transport pathway for entry into roots, and Si has been demonstrated to decrease As concentrations. However, the physiological mechanisms through which Si might alleviate As toxicity in plants remain poorly elucidated. We combined detailed gas exchange measurements with chlorophyll fluorescence analysis to examine the effects of Si nutrition on photosynthetic performance in rice plants [a wild-type (WT) cultivar and its lsi1 mutant defective in Si uptake] challenged with As (arsenite). As treatment impaired carbon fixation (particularly in the WT genotype) that was unrelated to photochemical or biochemical limitations but, rather, was largely associated with decreased leaf conductance at the stomata and mesophyll levels. Indeed, regardless of the genotypes, in the plants challenged with As, photosynthetic rates correlated strongly with both stomatal (r(2) = 0.90) and mesophyll (r(2) = 0.95) conductances, and these conductances were, in turn, linearly correlated with each other. The As-related impairments to carbon fixation could be considerably reverted by Si in a time- and genotype-dependent manner. In conclusion, we identified Si nutrition as an important target in an attempt to not only decrease As concentrations but also to ameliorate the photosynthetic performance of rice plants challenged with As.

Improving the estimation of mesophyll conductance to CO2: on the role of electron transport rate correction and respiration.

Mesophyll conductance (gm) can markedly limit photosynthetic CO2 assimilation and is required to estimate the parameters of the Farquhar-von Caemmerer-Berry (FvCB) model properly. The variable J (electron transport rate) is the most frequently used method for estimating gm, and the correct determination of J is one of its requirements. Recent evidence has shown that calibrating J can lead to some errors in estimating gm, but to what extent the parameterization of the FvCB model is affected by calibrations is not well known. In addition to determining the FvCB parameters, variants of the J calibration method were tested to address whether varying CO2 or light levels, possible alternative electron sinks, or contrasting leaf structural properties might play a role in determining differences in αß, the product of the leaf absorptance (α) and the photosystem II optical cross-section (ß). It was shown that differences in αß were mainly attributed to the use of A/C(i) or A/PPFD curves to calibrate J. The different αß values greatly influenced g(m), leading to a high number of unrealistic values in addition to affecting the estimates of the FvCB model parameters. A new approach was devised to retrieve leaf respiration in the light from combined A/C(i) and A/C(c) curves and a framework to understand the high variation in observed gm values. Overall, a background is provided to decrease the noise in gm, facilitating data reporting and allowing better retrieval of the information presented in A/C(i) and A/C(c) curves.

Physiological and biochemical abilities of robusta coffee leaves for acclimation to cope with temporal changes in light availability.

The effects of varying intensities of light on plants depend on when they occur, even if the total amount of light received is kept constant. We designed an experiment using two clones of robusta coffee (Coffea canephora) intercropped with shelter trees in such a way that allowed us to compare coffee bushes shaded in the morning (SM) with those shaded in the afternoon (SA), and then confronting both with bushes receiving full sunlight over the course of the day (FS). The SM bushes displayed better gas-exchange performance than their SA and FS counterparts, in which the capacity for CO2 fixation was mainly constrained by stomatal (SA bushes) and biochemical (FS bushes) factors. Physiological traits associated with light capture were more responsive to temporal fluctuations of light rather than to the amount of light received, although this behavior could be a clone-specific response. The activity of key antioxidant enzymes differed minimally when comparing the SM and SA clones, but was much larger in FS clones. No signs of photoinhibition or cell damage were found regardless of the light treatments. Acclimations to varying light supplies had no apparent additional cost for constructing and maintaining the leaves regardless of the light supply. Both the SM and SA individuals displayed higher return in terms of revenue streams (e.g. higher mass-based light-saturated photosynthetic rates, photosynthetic nitrogen use efficiencies and long-term water use efficiencies) than their FS counterparts. In conclusion, shading may improve the physiological performance of coffee bushes growing in harsh, tropical environments.