Photosynthetic plasticity aggravates the susceptibility of magnesium-deficient leaf to high light in rapeseed plants: the importance of Rubisco and mesophyll conductance.

Plants grown under low magnesium (Mg) soils are highly susceptible to encountering light intensities that exceed the capacity of photosynthesis (A), leading to a depression of photosynthetic efficiency and eventually to photooxidation (i.e., leaf chlorosis). Yet, it remains unclear which processes play a key role in limiting the photosynthetic energy utilization of Mg-deficient leaves, and whether the plasticity of A in acclimation to irradiance could have cross-talk with Mg, hence accelerating or mitigating the photodamage. We investigated the light acclimation responses of rapeseed (Brassica napus) grown under low- and adequate-Mg conditions. Magnesium deficiency considerably decreased rapeseed growth and leaf A, to a greater extent under high than under low light, which is associated with higher level of superoxide anion radical and more severe leaf chlorosis. This difference was mainly attributable to a greater depression in dark reaction under high light, with a higher Rubisco fallover and a more limited mesophyll conductance to CO2 (gm ). Plants grown under high irradiance enhanced the content and activity of Rubisco and gm to optimally utilize more light energy absorbed. However, Mg deficiency could not fulfill the need to activate the higher level of Rubisco and Rubisco activase in leaves of high-light-grown plants, leading to lower Rubisco activation and carboxylation rate. Additionally, Mg-deficient leaves under high light invested more carbon per leaf area to construct a compact leaf structure with smaller intercellular airspaces, lower surface area of chloroplast exposed to intercellular airspaces, and CO2 diffusion conductance through cytosol. These caused a more severe decrease in within-leaf CO2 diffusion rate and substrate availability. Taken together, plant plasticity helps to improve photosynthetic energy utilization under high light but aggravates the photooxidative damage once the Mg nutrition becomes insufficient.

Leaf anatomy does not explain the large variability of mesophyll conductance across C3 crop species.

Increasing mesophyll conductance of CO2 (gm ) is a strategy to improve photosynthesis in C3 crops. However, the relative importance of different anatomical traits in determining gm in crops is unclear. Mesophyll conductance measurements were performed on 10 crops using the online carbon isotope discrimination method and the 'variable J' method in parallel. The influences of crucial leaf anatomical traits on gm were evaluated using a one-dimensional anatomical CO2 diffusion model. The gm values measured using two independent methods were compatible, although significant differences were observed in their absolute values. Quantitative analysis showed that cell wall thickness and chloroplast stroma thickness are the most important elements along the diffusion pathway. Unexpectedly, the large variability of gm across crops was not associated with any investigated leaf anatomical traits except chloroplast thickness. The gm values estimated using the anatomical model differed remarkably from the values measured in vivo in most species. However, when the species-specific effective porosity of the cell wall and the species-specific facilitation effect of CO2 diffusion across the membrane and chloroplast stoma were taken into account, the model could output gm values very similar to those measured in vivo. These results indicate that gm variation across crops is probably also driven by the effective porosity of the cell wall and effects of facilitation of CO2 transport across the membrane and chloroplast stroma in addition to the thicknesses of the elements.

Greater mesophyll conductance and leaf photosynthesis in the field through modified cell wall porosity and thickness via AtCGR3 expression in tobacco.

Mesophyll conductance (gm) describes the ease with which CO2 passes from the sub-stomatal cavities of the leaf to the primary carboxylase of photosynthesis, Rubisco. Increasing gm is suggested as a means to engineer increases in photosynthesis by increasing [CO2] at Rubisco, inhibiting oxygenation and accelerating carboxylation. Here, tobacco was transgenically up-regulated with Arabidopsis Cotton Golgi-related 3 (CGR3), a gene controlling methylesterification of pectin, as a strategy to increase CO2 diffusion across the cell wall and thereby increase gm. Across three independent events in tobacco strongly expressing AtCGR3, mesophyll cell wall thickness was decreased by 7%-13%, wall porosity increased by 75% and gm measured by carbon isotope discrimination increased by 28%. Importantly, field-grown plants showed an average 8% increase in leaf photosynthetic CO2 uptake. Up-regulating CGR3 provides a new strategy for increasing gm in dicotyledonous crops, leading to higher CO2 assimilation and a potential means to sustainable crop yield improvement.

Harnessing photosynthetic C18O16O discrimination dynamics under leaf water nonsteady state to estimate mesophyll conductance: a new, regression-based method.

Mesophyll conductance (gm) is a crucial plant trait that can significantly limit photosynthesis. Measurement of photosynthetic C18O16O discrimination (Δ18O) has proved to be the only viable means of resolving gm in both C3 and C4 plants. However, the currently available methods to exploit Δ18O for gm estimation are error prone due to their inadequacy in constraining the degree of oxygen isotope exchange (θ) during mesophyll CO2 hydration. Here, we capitalized on experimental manipulation of leaf water isotopic dynamics to establish a novel, nonsteady state, regression-based approach for simultaneous determination of gm and θ from online Δ18O measurements. We demonstrated the methodological and theoretical robustness of this new Δ18O-gm estimation approach and showed through measurements on several C3 and C4 species that this approach can serve as a benchmark method against which to identify previously-unrecognized biases of the existing Δ18O-gm methods. Our results highlight the unique value of this nonsteady state-based approach for contributing to ongoing efforts toward quantitative understanding of mesophyll conductance for crop yield improvement and carbon cycle modeling.

Uncoupling of stomatal conductance and photosynthesis at high temperatures: mechanistic insights from online stable isotope techniques.

The strong covariation of temperature and vapour pressure deficit (VPD) in nature limits our understanding of the direct effects of temperature on leaf gas exchange. Stable isotopes in CO2 and H2 O vapour provide mechanistic insight into physiological and biochemical processes during leaf gas exchange. We conducted combined leaf gas exchange and online isotope discrimination measurements on four common European tree species across a leaf temperature range of 5-40°C, while maintaining a constant leaf-to-air VPD (0.8 kPa) without soil water limitation. Above the optimum temperature for photosynthesis (30°C) under the controlled environmental conditions, stomatal conductance (gs ) and net photosynthesis rate (An ) decoupled across all tested species, with gs increasing but An decreasing. During this decoupling, mesophyll conductance (cell wall, plasma membrane and chloroplast membrane conductance) consistently and significantly decreased among species; however, this reduction did not lead to reductions in CO2 concentration at the chloroplast surface and stroma. We question the conventional understanding that diffusional limitations of CO2 contribute to the reduction in photosynthesis at high temperatures. We suggest that stomata and mesophyll membranes could work strategically to facilitate transpiration cooling and CO2 supply, thus alleviating heat stress on leaf photosynthetic function, albeit at the cost of reduced water-use efficiency.

Shorting the metaphorical circuit: vascular partitioning and stomatal patchiness can create apparent unsaturation and CO2 gradient inversion in the Ohmic analogy for leaf gas exchange.

Analyses of leaf gas exchange rely on an Ohmic analogy that arrays single stomatal, internal air space, and mesophyll conductances in series. Such models underlie inferences of mesophyll conductance and the relative humidity of leaf airspaces, reported to fall as low as 80%. An unresolved question is whether such series models are biased with respect to real leaves, whose internal air spaces are chambered at various scales by vasculature. To test whether unsaturation could emerge from modeling artifacts, we compared series model estimates with true parameter values for a chambered leaf with varying distributions and magnitudes of leaf surface conductance ('patchiness'). Distributions of surface conductance can create large biases in gas exchange calculations. Both apparent unsaturation and internal CO2 gradient inversion can be produced by the evolution of broader distributions of stomatal apertures consistent with a decrease in surface conductance, as might occur under increasing vapor pressure deficit. In gas exchange experiments, the behaviors of derived quantities defined by simple series models are highly sensitive to the true partitioning of flux and stomatal apertures across leaf surfaces. New methods are needed to disentangle model artifacts from real biological responses.

High photosynthesis rates in Brassiceae species are mediated by leaf anatomy enabling high biochemical capacity, rapid CO2 diffusion and efficient light use.

Certain species in the Brassicaceae family exhibit high photosynthesis rates, potentially providing a valuable route toward improving agricultural productivity. However, factors contributing to their high photosynthesis rates are still unknown. We compared Hirschfeldia incana, Brassica nigra, Brassica rapa and Arabidopsis thaliana, grown under two contrasting light intensities. Hirschfeldia incana matched B. nigra and B. rapa in achieving very high photosynthesis rates under high growth-light condition, outperforming A. thaliana. Photosynthesis was relatively more limited by maximum photosynthesis capacity in H. incana and B. rapa and by mesophyll conductance in A. thaliana and B. nigra. Leaf traits such as greater exposed mesophyll specific surface enabled by thicker leaf or high-density small palisade cells contributed to the variation in mesophyll conductance among the species. The species exhibited contrasting leaf construction strategies and acclimation responses to low light intensity. High-light plants distributed Chl deeper in leaf tissue, ensuring even distribution of photosynthesis capacity, unlike low-light plants. Leaf anatomy of H. incana, B. nigra and B. rapa facilitated effective CO2 diffusion, efficient light use and provided ample volume for their high maximum photosynthetic capacity, indicating that a combination of adaptations is required to increase CO2-assimilation rates in plants.

Temperature responses of leaf respiration in light and darkness are similar and modulated by leaf development.

Our ability to predict temperature responses of leaf respiration in light and darkness (RL and RDk ) is essential to models of global carbon dynamics. While many models rely on constant thermal sensitivity (characterized by Q10 ), uncertainty remains as to whether Q10 of RL and RDk are actually similar. We measured short-term temperature responses of RL and RDk in immature and mature leaves of two evergreen tree species, Castanopsis carlesii and Ormosia henry in an open field. RL was estimated by the Kok method, the Yin method and a newly developed Kok-iterCc method. When estimated by the Yin and Kok-iterCc methods, RL and RDk had similar Q10 (c. 2.5). The Kok method overestimated both Q10 and the light inhibition of respiration. RL /RDk was not affected by leaf temperature. Acclimation of respiration in summer was associated with a decline in basal respiration but not in Q10 in both species, which was related to changes in leaf nitrogen content between seasons. Q10 of RL and RDk in mature leaves were 40% higher than in immature leaves. Our results suggest similar Q10 values can be used to model RL and RDk while leaf development-associated changes in Q10 require special consideration in future respiration models.

The interplay of short-term mesophyll and stomatal conductance responses under variable environmental conditions.

Understanding the short-term responses of mesophyll conductance (gm) and stomatal conductance (gsc) to environmental changes remains a challenging yet central aspect of plant physiology. This review synthesises our current knowledge of these short-term responses, which underpin CO2 diffusion within leaves. Recent methodological advances in measuring gm using online isotopic discrimination and chlorophyll fluorescence have improved our confidence in detecting short-term gm responses, but results need to be carefully evaluated. Environmental factors like vapour pressure deficit and CO2 concentration indirectly impact gm through gsc changes, highlighting some of the complex interactions between the two parameters. Evidence suggests that short-term responses of gm are not, or at least not fully, mechanistically linked to changes in gsc, cautioning against using gsc as a reliable proxy for gm. The overarching challenge lies in unravelling the mechanistic basis of short-term gm responses, which will contribute to the development of accurate models bridging laboratory insights with broader ecological implications. Addressing these gaps in understanding is crucial for refining predictions of gm behaviour under changing environmental conditions.

The long and tortuous path towards improving photosynthesis by engineering elevated mesophyll conductance.

The growing demand for global food production is likely to be a defining issue facing humanity over the next 50 years. To tackle this challenge, there is a desire to bioengineer crops with higher photosynthetic efficiencies, to increase yields. Recently, there has been a growing interest in engineering leaves with higher mesophyll conductance (gm), which would allow CO2 to move more efficiently from the substomatal cavities to the chloroplast stroma. However, if crop yield gains are to be realised through this approach, it is essential that the methodological limitations associated with estimating gm are fully appreciated. In this review, we summarise these limitations, and outline the uncertainties and assumptions that can affect the final estimation of gm. Furthermore, we critically assess the predicted quantitative effect that elevating gm will have on assimilation rates in crop species. We highlight the need for more theoretical modelling to determine whether altering gm is truly a viable route to improve crop performance. Finally, we offer suggestions to guide future research on gm, which will help mitigate the uncertainty inherently associated with estimating this parameter.

Caught between two states: The compromise in acclimation of photosynthesis, transpiration and mesophyll conductance to different amplitudes of fluctuating irradiance.

While dynamic regulation of photosynthesis in fluctuating light is increasingly recognized as an important driver of carbon uptake, acclimation to realistic irradiance fluctuations is still largely unexplored. We subjected Arabidopsis thaliana (L.) wild-type and jac1 mutants to irradiance fluctuations with distinct amplitudes and average irradiance. We examined how irradiance fluctuations affected leaf structure, pigments and physiology. A wider amplitude of fluctuations produced a stronger acclimation response. Large reductions of leaf mass per area under fluctuating irradiance framed our interpretation of changes in photosynthetic capacity and mesophyll conductance as measured by three separate methods, in that photosynthetic investment increased markedly on a mass basis, but only a little on an area basis. Moreover, thermal imagery showed that leaf transpiration was four times higher under fluctuating irradiance. Leaves growing under fluctuating irradiance, although thinner, maintained their photosynthetic capacity, as measured through light- and CO2-response curves; suggesting their photosynthesis may be more cost-efficient than those under steady light, but overall may incur increased maintenance costs. This is especially relevant for plant performance globally because naturally fluctuating irradiance creates conflicting acclimation cues for photosynthesis and transpiration that may hinder progress towards ensuring food security under climate-related extremes of water stress.

The importance of species-specific and temperature-sensitive parameterisation of A/Ci models: A case study using cotton (Gossypium hirsutum L.) and the automated 'OptiFitACi' R-package.

Leaf gas exchange measurements are an important tool for inferring a plant's photosynthetic biochemistry. In most cases, the responses of photosynthetic CO2 assimilation to variable intercellular CO2 concentrations (A/Ci response curves) are used to model the maximum (potential) rate of carboxylation by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, Vcmax) and the rate of photosynthetic electron transport at a given incident photosynthetically active radiation flux density (PAR; JPAR). The standard Farquhar-von Caemmerer-Berry model is often used with default parameters of Rubisco kinetic values and mesophyll conductance to CO2 (gm) derived from tobacco that may be inapplicable across species. To study the significance of using such parameters for other species, here we measured the temperature responses of key in vitro Rubisco catalytic properties and gm in cotton (Gossypium hirsutum cv. Sicot 71) and derived Vcmax and J2000 (JPAR at 2000 µmol m-2 s-1 PAR) from cotton A/Ci curves incrementally measured at 15°C-40°C using cotton and other species-specific sets of input parameters with our new automated fitting R package 'OptiFitACi'. Notably, parameterisation by a set of tobacco parameters produced unrealistic J2000:Vcmax ratio of <1 at 25°C, two- to three-fold higher estimates of Vcmax above 15°C, up to 2.3-fold higher estimates of J2000 and more variable estimates of Vcmax and J2000, for our cotton data compared to model parameterisation with cotton-derived values. We determined that errors arise when using a gm,25 of 2.3 mol m-2 s-1 MPa-1 or less and Rubisco CO2-affinities in 21% O2 (KC 21%O2) at 25°C outside the range of 46-63 Pa to model A/Ci responses in cotton. We show how the A/Ci modelling capabilities of 'OptiFitACi' serves as a robust, user-friendly, and flexible extension of 'plantecophys' by providing simplified temperature-sensitivity and species-specificity parameterisation capabilities to reduce variability when modelling Vcmax and J2000.

Carboxylation capacity is the main limitation of carbon assimilation in High Arctic shrubs.

Increases in shrub height, biomass and canopy cover are key whole-plant features of warming-induced vegetation change in tundra. We investigated leaf functional traits underlying photosynthetic capacity of Arctic shrub species, particularly its main limiting processes such as mesophyll conductance. In this nutrient-limited ecosystem, we expect leaf nitrogen concentration to be the main limiting factor for photosynthesis. We measured the net photosynthetic rate at saturated light (Asat) in three Salix species throughout a glacial valley in High-Arctic tundra and used a causal approach to test relationships between leaf stomatal and mesophyll conductances (gsc, gm), carboxylation capacity (Vcmax), nitrogen and phosphorus concentration (Narea, Parea) and leaf mass ratio (LMA). Arctic Salix species showed no difference in Asat compared to a global data set, while being characterized by higher Narea, Parea and LMA. Vcmax, gsc and gm independently increased Asat, with Vcmax as its main limitation. We highlighted a nitrogen-influenced pathway for increasing photosynthesis in the two prostrate mesic habitat species. In contrast, the erect wetland habitat Salix richardsonii mainly increased Asat with increasing gsc. Overall, our study revealed high photosynthetic capacities of Arctic Salix species but contrasting regulatory pathways that may influence shrub ability to respond to environmental changes in High Arctic tundra.

Leaf anatomical alterations reduce cotton's mesophyll conductance under dynamic drought stress conditions.

Drought stress significantly affects cotton's net photosynthetic rate (A) by restraining stomatal (gs ) and mesophyll conductance (gm ) as well as perturbing its biochemical process, resulting in yield reductions. Despite the significant progress in dissecting effects of drought on photosynthesis, the variability observed in cotton's gm , and the mechanisms contributing to that variability under dynamic drought stress conditions are poorly understood. For that reason, a controlled-environment experiment with two cotton genotypes (Dexiamian 1, Yuzaomian 9110), three water levels (soil relative water content: control [75 ± 5]%, moderate drought [60 ± 5]%, severe drought [45 ± 5]%), and two drought durations (10 and 31 days) were conducted. The results indicated that the cotton boll biomass was significantly decreased under 10-day severe drought and 31-day moderate and severe drought. Decreases in gs were later accompanied by decreases in gm and further combined with reductions in electron transport rate, as drought stress progressed in duration and severity, ultimately resulting in significant reductions in A of subtending leaf. Stomatal and mesophyll conductance constraints were the primary factors limiting photosynthesis, while biochemical constraints decreased, as drought stress progressed. Considering gm , its decline was ascribed to increases in the diffusion resistance of CO2 through cytoplasm (rcyt ), under short- or long-term drought, as well as to increases in leaf dry mass (LMA), and decreases in the chloroplast surface area exposed to intercellular air space (Sc /S), under long-term drought. It was concluded that A could be enhanced, under dynamic drought stress conditions, by increasing gm through increasing Sc /S and reducing LMA and rcyt .

Interspecific variation in the temperature response of mesophyll conductance is related to leaf anatomy.

Although mesophyll conductance (gm ) is known to be sensitive to temperature (T), the mechanisms underlying the temperature response of gm are not fully understood. In particular, it has yet to be established whether interspecific variation in gm -T relationships is associated with mesophyll anatomy and vein traits. In the present study, we measured the short-term response of gm in eight crop species, and leaf water potential (Ψleaf ) in five crop species over a temperature range of 15-35°C. The considered structural parameters are surface areas of mesophyll cells and chloroplasts facing intercellular airspaces per unit leaf area (Sm and Sc ), cell wall thickness (Tcw ), and vein length per area (VLA). We detected large interspecific variations in the temperature responses of gm and Ψleaf . The activation energy for gm (Ea,gm ) was found to be positively correlated with Sc , although it showed no correlation with Tcw . In contrast, VLA was positively correlated with the slope of the linear model of Ψleaf -T (a), whereas Ea,gm was marginally correlated with VLA and a. A two-component model was subsequently used to model gm -T relationships, and the mechanisms underlying the temperature response of gm are discussed. The data presented here indicate that leaf anatomy is a major determinant of the interspecific variation in gm -T relationships.

Genotype-dependent changes of cell wall composition influence physiological traits of a long and a non-long shelf-life tomato genotypes under distinct water regimes.

Water shortage strongly affects plants' physiological performance. Since tomato (Solanum lycopersicum) non-long shelf-life (nLSL) and long shelf-life (LSL) genotypes differently face water deprivation, we subjected a nLSL and a LSL genotype to four treatments: control (well watering), short-term water deficit stress at 40% field capacity (FC) (ST 40% FC), short-term water deficit stress at 30% FC (ST 30% FC), and short-term water deficit stress at 30% FC followed by recovery (ST 30% FC-Rec). Treatments promoted genotype-dependent elastic adjustments accompanied by distinct photosynthetic responses. While the nLSL genotype largely modified mesophyll conductance (gm ) across treatments, it was kept within a narrow range in the LSL genotype. However, similar gm values were achieved under ST 30% FC conditions. Particularly, modifications in the relative abundance of cell wall components and in sub-cellular anatomic parameters such as the chloroplast surface area exposed to intercellular air space per leaf area (Sc /S) and the cell wall thickness (Tcw ) regulated gm in the LSL genotype. Instead, only changes in foliar structure at the supra-cellular level influenced gm in the nLSL genotype. Even though further experiments testing a larger range of genotypes and treatments would be valuable to support our conclusions, we show that even genotypes of the same species can present different elastic, anatomical, and cell wall composition-mediated mechanisms to regulate gm when subjected to distinct water regimes.

CO2 mesophyll conductance regulated by light: a review.

MAIN CONCLUSION: Light quality and intensity regulate plant mesophyll conductance, which has played an essential role in photosynthesis by controlling leaf structural and biochemical properties. Mesophyll conductance (gm), a crucial physiological factor influencing the photosynthetic rate of leaves, is used to describe the resistance of CO2 from the sub-stomatal cavity into the chloroplast up to the carboxylation site. Leaf structural and biochemical components, as well as external environmental factors such as light, temperature, and water, all impact gm. As an essential factor of plant photosynthesis, light affects plant growth and development and plays a vital role in regulating gm as well as determining photosynthesis and yield. This review aimed to summarize the mechanisms of gm response to light. Both structural and biochemical perspectives were combined to reveal the effects of light quality and intensity on the gm, providing a guide for selecting the optimal conditions for intensifying photosynthesis in plants.

Unpredicted, rapid and unintended structural and functional changes occurred during early domestication of Silphium integrifolium, a perennial oilseed.

MAIN CONCLUSION: Selection for increased yield changed structure, physiology and overall resource-use strategy from conservative towards acquisitive leaves. Alternative criteria can be considered, to increase yield with less potentially negative traits. We compared the morphology, anatomy and physiology of wild and semi-domesticated (SD) accessions of Silphium integrifolium (Asteraceae), in multi-year experiments. We hypothesized that several cycles of selection for seed-yield would result in acquisitive leaves, including changes predicted by the leaf economic spectrum. Early-selection indirectly resulted in leaf structural and functional changes. Leaf anatomy changed, increasing mesophyll conductance and the size of xylem vessels and mesophyll cells increased. Leaves of SD plants were larger, heavier, with lower stomatal conductance, lower internal CO2 concentration, and lower resin concentration than those of wild types. Despite increased water use efficiency, SD plants transpired 25% more because their increase in leaf area. Unintended and undesired changes in functional plant traits could quickly become fixed during domestication, shortening the lifespan and increasing resource consumption of the crop as well as having consequences in the provision and regulation of ecosystem services.

Assessing the CO2 concentration at the surface of photosynthetic mesophyll cells.

We present a robust estimation of the CO2 concentration at the surface of photosynthetic mesophyll cells (cw ), applicable under reasonable assumptions of assimilation distribution within the leaf. We used Capsicum annuum, Helianthus annuus and Gossypium hirsutumas model plants for our experiments. We introduce calculations to estimate cw using independent adaxial and abaxial gas exchange measurements, and accounting for the mesophyll airspace resistances. The cw was lower than adaxial and abaxial estimated intercellular CO2 concentrations (ci ). Differences between cw and the ci of each surface were usually larger than 10 µmol mol-1 . Differences between adaxial and abaxial ci ranged from a few µmol mol-1 to almost 50 µmol mol-1 , where the largest differences were found at high air saturation deficits (ASD). Differences between adaxial and abaxial ci and the ci estimated by mixing both fluxes ranged from -30 to +20 µmol mol-1 , where the largest differences were found under high ASD or high ambient CO2 concentrations. Accounting for cw improves the information that can be extracted from gas exchange experiments, allowing a more detailed description of the CO2 and water vapor gradients within the leaf.

Is chloroplast size optimal for photosynthetic efficiency?

Improving photosynthetic efficiency has recently emerged as a promising way to increase crop production in a sustainable manner. While chloroplast size may affect photosynthetic efficiency in several ways, we aimed to explore whether chloroplast size manipulation can be a viable approach to improving photosynthetic performance. Several tobacco (Nicotiana tabacum) lines with contrasting chloroplast sizes were generated via manipulation of chloroplast division genes to assess photosynthetic performance under steady-state and fluctuating light. A selection of lines was included in a field trial to explore productivity. Lines with enlarged chloroplasts underperformed in most of the measured traits. Lines with smaller and more numerous chloroplasts showed a similar efficiency compared with wild-type (WT) tobacco. Chloroplast size only weakly affected light absorptance and light profiles within the leaf. Increasing chloroplast size decreased mesophyll conductance (gm ) but decreased chloroplast size did not increase gm . Increasing chloroplast size reduced chloroplast movements and enhanced non-photochemical quenching. The chloroplast smaller than WT appeared to be no better than WT for photosynthetic efficiency and productivity under field conditions. The results indicate that chloroplast size manipulations are therefore unlikely to lead to higher photosynthetic efficiency or growth.