Steady-state Photosynthesis Research Articles

Photosynthesis and photoprotection in top leaves respond faster to irradiance fluctuations than bottom leaves in a tomato canopy.

Accounting for the dynamic responses of photosynthesis and photoprotection to naturally fluctuating irradiance can improve predictions of plant performance in the field, but the variation of these dynamics within crop canopies is poorly understood. We conducted a detailed study of dynamic and steady-state photosynthesis, photoprotection, leaf pigmentation, and stomatal anatomy in four leaf layers (100, 150, 200 and 250cm from the floor) of a fully-grown tomato canopy in the greenhouse. We found that leaves at the top of the canopy exhibited higher photosynthetic capacity and slightly faster photosynthetic induction compared to lower-canopy leaves, accompanied by higher stomatal conductance and a faster activation of carboxylation and linear electron transport capacities. In upper-canopy leaves, non-photochemical quenching showed faster induction and relaxation after in- and decreases in irradiance, allowing for more effective photoprotection in these leaves. Despite these observed differences in transient responses between leaf layers, steady-state rather than dynamic photosynthesis traits were more influential for predicting photosynthesis under fluctuating irradiance. Also, a model analysis revealed that time-averaged photosynthesis under fluctuating irradiance could be accurately predicted by one set of Rubisco activation/deactivation parameters across all four leaf layers, thereby greatly simplifying future modelling efforts of whole-canopy photosynthesis.

Effects of Red and Blue Light on the Growth, Photosynthesis, and Subsequent Growth under Fluctuating Light of Cucumber Seedlings.

The effects of red and blue light on growth and steady-state photosynthesis have been widely studied, but there are few studies focusing on dynamic photosynthesis and the effects of LED pre-treatment on cucumber seedlings' growth, so in this study, cucumber (Cucumis sativus L. cv. Jinyou 365) was chosen as the test material. White light (W), monochromatic red light (R), monochromatic blue light (B), and mixed red and blue lights with different red-to-blue ratios (9:1, 7:3, 5:5, 3:7, and 1:9) were set to explore the effects of red and blue light on cucumber seedlings' growth, steady-state photosynthesis, dynamic photosynthesis, and subsequent growth under fluctuating light. The results showed that compared with R and B, mixed red and blue light was more suitable for cucumber seedlings' growth, and the increased blue light ratios would decrease the biomass of cucumber seedlings under mixed red and blue light; cucumber seedlings under 90% red and 10% blue mixed light (9R1B) grew better than other treatments. For steady-state photosynthesis, blue light decreased the actual net photosynthetic rate but increased the maximum photosynthetic capacity by promoting stomatal development and opening; 9R1B exhibited higher actual net photosynthetic rate, but the maximum photosynthetic capacity was low. For dynamic photosynthesis, the induction rate of photosynthetic rate and stomatal conductance were also accelerated by blue light. For subsequent growth under fluctuating light, higher maximum photosynthetic capacity and photoinduction rate could not promote the growth of cucumber seedlings under subsequent fluctuating light, while seedlings pre-treated with 9R1B and B grew better under subsequent fluctuating light due to the high plant height and leaf area. Overall, cucumber seedlings treated with 9R1B exhibited the highest biomass and it grew better under subsequent fluctuating light due to the higher actual net photosynthetic rate, plant height, and leaf area.

Extra O2 evolution reveals an O2-independent alternative electron sink in photosynthesis of marine diatoms.

Following the principle of oxygenic photosynthesis, electron transport in the thylakoid membranes (i.e., light reaction) generates ATP and NADPH from light energy, which is subsequently utilized for CO2 fixation in the Calvin-Benson-Bassham cycle (i.e., dark reaction). However, light and dark reactions could discord when an alternative electron flow occurs with a rate comparable to the linear electron flow. Here, we quantitatively monitored O2 and total dissolved inorganic carbon (DIC) during photosynthesis in the pennate diatom Phaeodactylum tricornutum, and found that evolved O2 was larger than the consumption of DIC, which was consistent with 14CO2 measurements in literature. In our measurements, the stoichiometry of O2 evolution to DIC consumption was always around 1.5 during photosynthesis at different DIC concentrations. The same stoichiometry was observed in the cells grown under different CO2 concentrations and nitrogen sources except for the nitrogen-starved cells showing O2 evolution 2.5 times larger than DIC consumption. An inhibitor to nitrogen assimilation did not affect the extra O2 evolution. Further, the same physiological phenomenon was observed in the centric diatom Thalassiosira pseudonana. Based on the present dataset, we propose that the marine diatoms possess the metabolic pathway(s) functioning as the O2-independent electron sink under steady state photosynthesis that reaches nearly half of electron flux of the Calvin-Benson-Bassham cycle.

Variation in Photosynthetic Efficiency under Fluctuating Light between Rose Cultivars and its Potential for Improving Dynamic Photosynthesis

Photosynthetic efficiency under both steady-state and fluctuating light can significantly affect plant growth under naturally fluctuating light conditions. However, the difference in photosynthetic performance between different rose genotypes is little known. This study compared the photosynthetic performance under steady-state and fluctuating light in two modern rose cultivars (Rose hybrida), "Orange Reeva" and "Gelato", and an old Chinese rose plant Rosa chinensis cultivar, "Slater's crimson China". The light and CO2 response curves indicated that they showed similar photosynthetic capacity under steady state. The light-saturated steady-state photosynthesis in these three rose genotypes was mainly limited by biochemistry (60%) rather than diffusional conductance. Under fluctuating light conditions (alternated between 100 and 1500 μmol photons m-2 m-1 every 5 min), stomatal conductance gradually decreased in these three rose genotypes, while mesophyll conductance (gm) was maintained stable in Orange Reeva and Gelato but decreased by 23% in R. chinensis, resulting in a stronger loss of CO2 assimilation under high-light phases in R. chinensis (25%) than in Orange Reeva and Gelato (13%). As a result, the variation in photosynthetic efficiency under fluctuating light among rose cultivars was tightly related to gm. These results highlight the importance of gm in dynamic photosynthesis and provide new traits for improving photosynthetic efficiency in rose cultivars.

Carbon assimilation in upper subtidal macroalgae is determined by an inverse correlation between Rubisco carboxylation efficiency and CO2 concentrating mechanism effectiveness.

Seaweeds have a wide ecophysiological and phylogenetic diversity with species expressing different Rubisco forms that frequently coexist with biophysical CO2 concentrating mechanisms (CCMs), an adaptation that overcomes the low CO2 availability and gas diffusion in seawater. Here, we assess the possible coevolution between the Rubisco catalysis and the type and effectiveness of CCMs present in six upper subtidal macroalgal species belonging to three phylogenetic groups of seaweeds. A wide diversity in the Rubisco kinetic traits was found across the analyzed species, although the specificity factor was the only parameter explained by the expressed Rubisco form. Differences in the catalytic trade-offs were found between Rubisco forms, indicating that ID Rubiscos could be better adapted to the intracellular O2 :CO2 ratio found in marine organisms during steady-state photosynthesis. The biophysical components of the CCMs also differed among macroalgal species, resulting in different effectiveness to concentrate CO2 around Rubisco active sites. Interestingly, an inverse relationship was found between the effectiveness of CCMs and the invitro Rubisco carboxylation efficiency, which possibly led to a similar carboxylation potential across the analyzed macroalgal species. Our results demonstrate a coevolution between Rubisco kinetics and CCMs across phylogenetically distant marine macroalgal species sharing the same environment.

Activation of CO2 assimilation during photosynthetic induction is slower in C4 than in C3 photosynthesis in three phylogenetically controlled experiments.

Despite their importance for the global carbon cycle and crop production, species with C4 photosynthesis are still somewhat understudied relative to C3 species. Although the benefits of the C4 carbon concentrating mechanism are readily observable under optimal steady state conditions, it is less clear how the presence of C4 affects activation of CO2 assimilation during photosynthetic induction. In this study we aimed to characterise differences between C4 and C3 photosynthetic induction responses by analysing steady state photosynthesis and photosynthetic induction in three phylogenetically linked pairs of C3 and C4 species from Alloteropsis, Flaveria, and Cleome genera. Experiments were conducted both at 21% and 2% O2 to evaluate the role of photorespiration during photosynthetic induction. Our results confirm C4 species have slower activation of CO2 assimilation during photosynthetic induction than C3 species, but the apparent mechanism behind these differences varied between genera. Incomplete suppression of photorespiration was found to impact photosynthetic induction significantly in C4 Flaveria bidentis, whereas in the Cleome and Alloteropsis C4 species, delayed activation of the C3 cycle appeared to limit induction and a potentially supporting role for photorespiration was also identified. The sheer variation in photosynthetic induction responses observed in our limited sample of species highlights the importance of controlling for evolutionary distance when comparing C3 and C4 photosynthetic pathways.

The Dynamic Changes of Alternative Electron Flows upon Transition from Low to High Light in the Fern Cyrtomium fortune and the Gymnosperm Nageia nagi

In photosynthetic organisms except angiosperms, an alternative electron sink that is mediated by flavodiiron proteins (FLVs) plays the major role in preventing PSI photoinhibition while cyclic electron flow (CEF) is also essential for normal growth under fluctuating light. However, the dynamic changes of FLVs and CEF has not yet been well clarified. In this study, we measured the P700 signal, chlorophyll fluorescence, and electrochromic shift spectra in the fern Cyrtomium fortune and the gymnosperm Nageia nagi. We found that both species could not build up a sufficient proton gradient (∆pH) within the first 30 s after light abruptly increased. During this period, FLVs-dependent alternative electron flow was functional to avoid PSI over-reduction. This functional time of FLVs was much longer than previously thought. By comparison, CEF was highly activated within the first 10 s after transition from low to high light, which favored energy balancing rather than the regulation of a PSI redox state. When FLVs were inactivated during steady-state photosynthesis, CEF was re-activated to favor photoprotection and to sustain photosynthesis. These results provide new insight into how FLVs and CEF interact to regulate photosynthesis in non-angiosperms.

A System Dynamics Approach to Model Photosynthesis at Leaf Level Under Fluctuating Light.

Photosynthesis has been mainly studied under steady-state conditions even though this assumption results inadequate for assessing the biochemical responses to rapid variations occurring in natural environments. The combination of mathematical models with available data may enhance the understanding of the dynamic responses of plants to fluctuating environments and can be used to make predictions on how photosynthesis would respond to non-steady-state conditions. In this study, we present a leaf level System Dynamics photosynthesis model based and validated on an experiment performed on two soybean varieties, namely, the wild type Eiko and the chlorophyll-deficient mutant MinnGold, grown in constant and fluctuating light conditions. This mutant is known to have similar steady-state photosynthesis compared to the green wild type, but it is found to have less biomass at harvest. It has been hypothesized that this might be due to an unoptimized response to non-steady-state conditions; therefore, this mutant seems appropriate to investigate dynamic photosynthesis. The model explained well the photosynthetic responses of these two varieties to fluctuating and constant light conditions and allowed to make relevant conclusions on the different dynamic responses of the two varieties. Deviations between data and model simulations are mostly evident in the non-photochemical quenching (NPQ) dynamics due to the oversimplified combination of PsbS- and zeaxanthin-dependent kinetics, failing in finely capturing the NPQ responses at different timescales. Nevertheless, due to its simplicity, the model can provide the basis of an upscaled dynamic model at a plant level.

Short-term elevated temperature and CO2 promote photosynthetic induction in the C3 plant Glycine max, but not in the C4 plant Amaranthus tricolor.

The continuous increases of atmospheric temperature and CO2 concentration will impact global photosynthesis. However, there are few studies considering the interaction of elevated temperature (eT) and elevated CO2 (eCO2 ) on dynamic photosynthesis, particularly for C4 species. We examine dynamic photosynthesis under four different temperature and [CO2 ] treatments: (1) 400ppm×28°C (CT); (2) 400ppm×33°C (CT+); (3) 800ppm×28°C (C+T); and (4) 800ppm×33°C (C+T+). In Glycine max L., the time required to reach 50% (T 50%A ) and 90% (T 90%A ) of full photosynthetic induction was smaller under the CT+, C+T, and C+T+ treatments than those under the CT treatment. In Amaranthus tricolor L., however, neither T 50%A nor T 90%A was not significantly affected by eT or eCO2 . In comparison with the CT treatment, the achieved carbon gain was increased by 58.3% (CT+), 112% (C+T), and 136.6% (C+T+) in G. max and was increased by 17.1% (CT+), 2.6% (C+T) and 56.9% (C+T+) in A. tricolor . The increases of achieved carbon gain in G. max were attributable to both improved photosynthetic induction efficiency (IE) and enhanced steady-state photosynthesis, whereas those in A. tricolor were attributable to enhanced steady-state photosynthesis.

Evolution of a biochemical model of steady-state photosynthesis.

On the occasion of the 40th anniversary of the publication of the landmark model by Farquhar, von Caemmerer & Berry on steady‐state C3 photosynthesis (known as the “FvCB model”), we review three major further developments of the model. These include: (1) limitation by triose phosphate utilization, (2) alternative electron transport pathways, and (3) photorespiration‐associated nitrogen and C1 metabolisms. We discussed the relation of the third extension with the two other extensions, and some equivalent extensions to model C4 photosynthesis. In addition, the FvCB model has been coupled with CO2‐diffusion models. We review how these extensions and integration have broadened the use of the FvCB model in understanding photosynthesis, especially with regard to bioenergetic stoichiometries associated with photosynthetic quantum yields. Based on the new insights, we present caveats in applying the FvCB model. Further research needs are highlighted.

The role of Cytochrome b6f in the control of steady-state photosynthesis: a conceptual and quantitative model

Here, we present a conceptual and quantitative model to describe the role of the Cytochrome hbox {b}_{6}hbox {f} complex in controlling steady-state electron transport in hbox {C}_{3} leaves. The model is based on new experimental methods to diagnose the maximum activity of Cyt hbox {b}_{6}hbox {f} in vivo, and to identify conditions under which photosynthetic control of Cyt hbox {b}_{6}hbox {f} is active or relaxed. With these approaches, we demonstrate that Cyt hbox {b}_{6}hbox {f} controls the trade-off between the speed and efficiency of electron transport under limiting light, and functions as a metabolic switch that transfers control to carbon metabolism under saturating light. We also present evidence that the onset of photosynthetic control of Cyt hbox {b}_{6}hbox {f} occurs within milliseconds of exposure to saturating light, much more quickly than the induction of non-photochemical quenching. We propose that photosynthetic control is the primary means of photoprotection and functions to manage excitation pressure, whereas non-photochemical quenching functions to manage excitation balance. We use these findings to extend the Farquhar et al. (Planta 149:78–90, 1980) model of hbox {C}_{3} photosynthesis to include a mechanistic description of the electron transport system. This framework relates the light captured by PS I and PS II to the energy and mass fluxes linking the photoacts with Cyt hbox {b}_{6}hbox {f}, the ATP synthase, and Rubisco. It enables quantitative interpretation of pulse-amplitude modulated fluorometry and gas-exchange measurements, providing a new basis for analyzing how the electron transport system coordinates the supply of Fd, NADPH, and ATP with the dynamic demands of carbon metabolism, how efficient use of light is achieved under limiting light, and how photoprotection is achieved under saturating light. The model is designed to support forward as well as inverse applications. It can either be used in a stand-alone mode at the leaf-level or coupled to other models that resolve finer-scale or coarser-scale phenomena.

Dynamics of photosynthetic induction and relaxation within the canopy of rice and two wild relatives.

Wild rice species are a source of genetic material for improving cultivated rice (Oryza sativa) and a means to understand its evolutionary history. Renewed interest in non‐steady‐state photosynthesis in crops has taken place due its potential in improving sustainable productivity. Variation was characterized for photosynthetic induction and relaxation at two leaf canopy levels in three rice species. The wild rice accessions had 16%–40% higher rates of leaf CO2 uptake (A) during photosynthetic induction relative to the O. sativa accession. However, O. sativa had an overall higher photosynthetic capacity when compared to accessions of its wild progenitors. Additionally, O. sativa had a faster stomatal closing response, resulting in higher intrinsic water‐use efficiency during high‐to‐low light transitions. Leaf position in the canopy had a significant effect on non‐steady‐state photosynthesis, but not steady‐state photosynthesis. The results show potential to utilize wild material to refine plant models and improve non‐steady‐state photosynthesis in cultivated rice for increased productivity.

Leaf N content regulates the speed of photosynthetic induction under fluctuating light among canola genotypes (Brassica napus L.).

Nitrogen is an essential element for plant growth, and the relationship between leaf N content and photosynthesis has been widely studied in different species under steady-state light. However, under natural conditions, the light intensity at the leaf level is always changing, inherently heterogeneous in time and space. Therefore, the effect of leaf N content on photosynthesis under dynamic light conditions needs further study. At present, the effects of leaf N content on leaf non-steady-state photosynthesis have not been reported in canola (Brassica napus L.). To clarify the relationship between leaf N content and the speed of the response leaf gas exchange to variations in light intensity, eight genotypes of canola varying in leaf N content were used to study the temporal response of gas exchange to a step increase in irradiance. We found there were significant differences in non-steady-state photosynthesis, physiological characteristics, and anatomical traits across genotypes (the maximum amplitude was about fivefold), despite the lack of contrast under normal, steady-state photosynthesis. In addition, initial stomatal conductance to water vapor in the darkness and leaf N content per leaf area were negatively correlated with the time required to achieve 50% and 100% of the maximum photosynthetic rate. Contrarily, the time required to reach 50% of the maximum stomatal conductance was positively correlated with the time required to achieve 90% of the maximum photosynthetic rate across genotypes. It is concluded that the genotypes of canola with higher N content per leaf area show a faster induction of photosynthesis to fluctuating light conditions.

Identification of quantitative trait loci for dynamic and steady-state photosynthetic traits in a barley mapping population.

Enhancing the photosynthetic induction response to fluctuating light has been suggested as a key target for improvement in crop breeding programmes, with the potential to substantially increase whole-canopy carbon assimilation and contribute to crop yield potential. Rubisco activation may be the main physiological process that will allow us to achieve such a goal. In this study, we assessed the phenotype of Rubisco activation rate in a doubled haploid (DH) barley mapping population [131 lines from a Yerong/Franklin (Y/F) cross] after a switch from moderate to saturating light. Rates of Rubisco activation were found to be highly variable across the mapping population, with a median activation rate of 0.1 min−1 in the slowest genotype and 0.74 min−1 in the fastest genotype. A unique quantitative trait locus (QTL) for Rubisco activation rate was identified on chromosome 7H. This is the first report on the identification of a QTL for Rubisco activation rate in planta and the discovery opens the door to marker-assisted breeding to improve whole-canopy photosynthesis of barley. This also suggests that genetic factors other than the previously characterized Rubisco activase (RCA) isoforms on chromosome 4H control Rubisco activity. Further strength is given to this finding as this QTL co-localized with QTLs identified for steady-state photosynthesis and stomatal conductance. Several other distinct QTLs were identified for these steady-state traits, with a common overlapping QTL on chromosome 2H, and distinct QTLs for photosynthesis and stomatal conductance identified on chromosomes 4H and 5H, respectively. Future work should aim to validate these QTLs under field conditions so that they can be used to aid plant breeding efforts.

Rice Cultivar Takanari Has Higher Photosynthetic Performance Under Fluctuating Light Than Koshihikari, Especially Under Limited Nitrogen Supply and Elevated CO2.

Plants in the field experience dynamic changes of sunlight rather than steady-state irradiation. Therefore, increasing the photosynthetic rate of an individual leaf under fluctuating light is essential for improving crop productivity. The high-yielding indica rice (Oryza sativa L.) cultivar Takanari is considered a potential donor of photosynthesis genes because of its higher steady-state photosynthesis at both atmospheric and elevated CO2 concentrations than those of several Japanese commercial cultivars, including Koshihikari. Photosynthetic induction after a sudden increase in light intensity is faster in Takanari than in Koshihikari, but whether the daily carbon gain of Takanari outperforms that of Koshihikari under fluctuating light in the field is unclear. Here we report that Takanari has higher non-steady-state photosynthesis, especially under low nitrogen (N) supply, than Koshihikari. In a pot experiment, Takanari had greater leaf carbon gain during the initial 10 min after a sudden increase in irradiation and higher daily CO2 assimilation under simulated natural fluctuating light, at both atmospheric (400 ppm) and elevated (800 ppm) CO2 concentrations. The electron transport rate during a day under field conditions with low N supply was also higher in Takanari than in Koshihikari. Although the advantages of Takanari were diminished under high N supply, photosynthetic N use efficiency was consistently higher in Takanari than in Koshihikari, under both low and high N supply. This study demonstrates that Takanari is a promising donor parent to use in breeding programs aimed at increasing CO2 assimilation in a wide range of environments, including future higher CO2 concentrations.

Artifleck: The Study of Artifactual Responses to Light Flecks with Inappropriate Leaves.

Methods in sunfleck research commonly employ the use of experimental leaves which were constructed in homogeneous light. These experimental organs may behave unnaturally when they are challenged with fluctuating light. Photosynthetic responses to heterogeneous light and leaf macronutrient relations were determined for Cycas micronesica, Glycine max, and Zea mays leaves that were grown in homogeneous shade, heterogeneous shade, or full sun. The speed of priming where one light fleck increased the photosynthesis during a subsequent light fleck was greatest for the leaves grown in heterogeneous shade. The rate of induction and the ultimate steady-state photosynthesis were greater for the leaves that were grown in heterogeneous shade versus the leaves grown in homogeneous shade. The leaf mass per area, macronutrient concentration, and macronutrient stoichiometry were also influenced by the shade treatments. The amplitude and direction in which the three developmental light treatments influenced the response variables were not universal among the three model species. The results indicate that the historical practice of using experimental leaves which were constructed under homogeneous light to study leaf responses to fluctuating light may produce artifacts that generate dubious interpretations.

Declining root water transport drives stomatal closure in olive under moderate water stress.

Efficient water transport from soil to leaves sustains stomatal opening and steady-state photosynthesis. The aboveground portion of this pathway is well-described, yet the roots and their connection with the soil are still poorly understood due to technical limitations. Here we used a novel rehydration technique to investigate changes in the hydraulic pathway between roots and soil and within the plant body as individual olive plants were subjected to a range of water stresses. Whole root hydraulic resistance (including the radial pathway from xylem to the soil-root interface) constituted 81% of the whole-plant resistance in unstressed plants, increasing to >95% under a moderate level of water stress. The decline in this whole root hydraulic conductance occurred in parallel with stomatal closure and contributed significantly to the reduction in canopy conductance according to a hydraulic model. Our results demonstrate that losses in root hydraulic conductance, mainly due to a disconnection from the soil during moderate water stress in olive plants, are profound and sufficient to induce stomatal closure before cavitation occurs. Future studies will determine whether this core regulatory role of root hydraulics exists more generally among diverse plant species.

Red/blue light ratio strongly affects steady-state photosynthesis, but hardly affects photosynthetic induction in tomato (Solanum lycopersicum).

Plants are often subjected to rapidly alternating light intensity and quality. While both short- and long-term changes in red and blue light affect leaf photosynthesis, their impact on dynamic photosynthesis is not well documented. It was tested how dynamic and steady-state photosynthetic traits were affected by red/blue ratios, either during growth or during measurements, in tomato leaves. Four red/blue ratios were used: monochromatic red (R100 ), monochromatic blue (B100 ), a red/blue light ratio of 9:1 (R90 B10 ) and a red/blue light ratio of 7:3 (R70 B30 ). R100 grown leaves showed decreased photosynthetic capacity (maximum rates of light-saturated photosynthesis, carboxylation, electron transport and triose phosphate use), leaf thickness and nitrogen concentrations. Acclimation to various red/blue ratios had limited effects on photosynthetic induction in dark-adapted leaves. B100 -grown leaves had a approximately 15% larger initial NPQ transient than the other treatments, which may be beneficial for photoprotection under fluctuating light. B100 -grown leaves also showed faster stomatal closure when exposed to low light intensity, which likely resulted from smaller stomata and higher stomatal density. When measured under different red/blue ratios, stomatal opening rate and photosynthetic induction rate were hardly accelerated by increased fractions of blue light in both growth chamber-grown leaves and greenhouse-grown leaves. However, steady-state photosynthesis rate 30 min after photosynthetic induction was strongly reduced in leaves exposed to B100 during the measurement. We conclude that varying red/blue light ratios during growth and measurement strongly affects steady-state photosynthesis, but has limited effects on photosynthetic induction rate.

A Thermotolerant Variant of Rubisco Activase From a Wild Relative Improves Growth and Seed Yield in Rice Under Heat Stress.

Genes encoding thermostable variants of the photosynthesis heat-labile protein Rubisco activase (Rca) from a wild relative Oryza australiensis were overexpressed in domesticated rice (Oryza sativa). Proteomics was used to quantify the abundance of O. australiensis Rca (Rca-Oa) in the resulting plants. Plants were grown to maturity in growth rooms and from early tillering until immediately prior to anthesis, they were exposed to daytime maximum temperatures of 28, 40, and 45°C and constant night temperatures of 22°C. Non-destructive measurements of leaf elongation and photosynthesis were used to compare the null segregant with a transfected line in which 19% of its total Rca content was the recombinant O. australiensis Rca (T-Oa-19). Height, fresh mass, panicle number, seed set, and seed number were measured at final harvest. Traits at maturity after heat stress at 45°C correlated strongly with recombinant protein abundance. Seed number was far the most responsive trait to an increase in Rca-Oa abundance, improving by up to 150%. Leaf elongation rates (LER) and tiller number were significantly greater in the transformed plants in the first two weeks of exposure to 45°C but tiller numbers later became equal in the two genotypes. Gas exchange measurements showed that T-Oa-19 had faster light induction of photosynthesis but not significantly higher CO2 assimilation rates, indicating that the carbon gain that resulted in large yield improvement after growth at 45°C was not strongly correlated with an instantaneous measurement of steady-state photosynthesis. When plants were grown at 40°C daytime maximum, there was no improvement in the final biomass, panicle or seed number when compared with 28°C, indicating that the threshold for heat damage and beneficial effects of the thermostable Rca recombinant protein was between 40 and 45°C, which corresponded to leaf temperatures in the range 38–42°C. The results suggest that the thermotolerant form of Rca from O. australiensis was sufficient to enhance carbohydrate accumulation and storage by rice over the life of the plant, dramatically improving yields after exposure to heat throughout the vegetative phase.

Genetic characterization of aflap1 null mutation in Arabidopsis npq4 and pgr5 plants suggests that the regulatory role of FLAP1 involves the control of proton homeostasis in chloroplasts.

Precise control of the proton concentration gradient across thylakoid membranes (ΔpH) is essential for photosynthesis and its regulation because the gradient contributes to the generation of the proton motive force used for ATP synthesis and also for the fast and reversible induction of non-photochemical quenching (NPQ) to avoid photoinhibition and photodamage. However, the regulatory mechanism(s) controlling ΔpH in response to fluctuating light has not been fully elucidated. We previously described a new NPQ-regulatory chloroplastic protein, Fluctuating-Light-Acclimation Protein1 (FLAP1), which is important for plant growth and modulation of ΔpH under fluctuating light conditions. For this report, we further characterized FLAP1 activity by individually crossing an Arabidopsis flap1 mutant with npq4 and pgr5 plants; npq4 is defective in PsbS-dependent NPQ, and pgr5 is defective in induction of steady-state proton motive force (pmf) and energy-dependent quenching (qE). Both npq4 and npq4 flap1 exhibited similar NPQ kinetics and other photosynthetic parameters under constant or fluctuating actinic light. Conversely, pgr5 flap1 had recovered NPQ, photosystem II quantum yield and growth under fluctuating light, each of which was impaired in pgr5. Together with other data, we propose that FLAP1 activity controls proton homeostasis under steady-state photosynthesis to manipulate luminal acidification levels appropriately to balance photoprotection and photochemical processes.