Photosynthesis In C3 Plants Research Articles

Review Aktivitas Fotosintesis pada Tanaman Sorgum (Sorghum bicolor) dalam Kondisi Cekaman Kekeringan

Sorghum (Sorghum bicolor L. Moench) is a cereal crop that has the potential to be developed in Indonesia as a food, feed and industrial crop. Sorghum is a C4 plant that has the advantage of efficiency in hot and dry environments. Drought stress is one of the most limiting environmental factors for crop productivity worldwide, and can be caused by water deficits in the soil and in the atmosphere. On the decreasing leaf water status, the rate of CO2 assimilation and the conductance of stomata decreased rapidly. The CO2 concentration mechanism is able to saturate C4 photosynthesis under the relatively low intercellular CO2 concentration. In addition, CO2 photorespiration is likely to be repaired before it exits the bundle sheat cells. The effects of non-stomatal factors include reduced activity of photosynthetic enzymes, inhibition of nitrate assimilation, induction of premature aging, and changes in leaf anatomy. Photosynthesis in C4 plants, including sorghum, involves, others, the PEPC and Rubisco enzymes. Drought can also trigger oxidative stress, which is an environmental condition that has increased Reactive Oxygen Species (ROS) due to an over reduction of the photosynthesis process.

Chloroplastic photoprotective strategies differ between bundle sheath and mesophyll cells in maize (Zea mays L.) Under drought.

Bundle sheath cells play a crucial role in photosynthesis in C4 plants, but the structure and function of photosystem II (PSII) in these cells is still controversial. Photoprotective roles of bundle sheath chloroplasts at the occurrence of environmental stresses have not been investigated so far. Non-photochemical quenching (NPQ) of chlorophyll a fluorescence is the photoprotective mechanism that responds to a changing energy balance in chloroplasts. In the present study, we found a much higher NPQ in bundle sheath chloroplasts than in mesophyll chloroplasts under a drought stress. This change was accompanied by a more rapid dephosphorylation of light-harvesting complex II (LHCII) subunits and a greater increase in PSII subunit S (PsbS) protein abundance than in mesophyll cell chloroplasts. Histochemical staining of reactive oxygen species (ROS) suggested that the high NPQ may be one of the main reasons for the lower accumulation of ROS in bundle sheath chloroplasts. This may maintain the stable functioning of bundle sheath cells under drought condition. These results indicate that the superior capacity for dissipation of excitation energy in bundle sheath chloroplasts may be an environmental adaptation unique to C4 plants.

How Light Reactions of Photosynthesis in C4 Plants Are Optimized and Protected under High Light Conditions.

Most C4 plants that naturally occur in tropical or subtropical climates, in high light environments, had to evolve a series of adaptations of photosynthesis that allowed them to grow under these conditions. In this review, we summarize mechanisms that ensure the balancing of energy distribution, counteract photoinhibition, and allow the dissipation of excess light energy. They secure effective electron transport in light reactions of photosynthesis, which will lead to the production of NADPH and ATP. Furthermore, a higher content of the cyclic electron transport components and an increase in ATP production are observed, which is necessary for the metabolism of C4 for effective assimilation of CO2. Most of the data are provided by studies of the genus Flaveria, where species belonging to different metabolic subtypes and intermediate forms between C3 and C4 are present. All described mechanisms that function in mesophyll and bundle sheath chloroplasts, into which photosynthetic reactions are divided, may differ in metabolic subtypes as a result of the different organization of thylakoid membranes, as well as the different demand for ATP and NADPH. This indicates that C4 plants have plasticity in the utilization of pathways in which efficient use and dissipation of excitation energy are realized.

A scheme for C4 evolution derived from a comparative analysis of the closely related C3, C3-C4 intermediate, C4-like, and C4 species in the genus Flaveria.

A comparative analysis of the genus Flaveria showed a C4 evolutionary process in which the anatomical and metabolic features of C4 photosynthesis were gradually acquired through C3-C4 intermediate stages. C4 photosynthesis has been acquired in multiple lineages of angiosperms during evolution to suppress photorespiration. Crops that perform C4 photosynthesis exhibit high rates of CO2 assimilation and high grain production even under high-temperature in semiarid environments; therefore, engineering C4 photosynthesis in C3 plants is of great importance in the application field. The genus Flaveria contains a large number of C3, C3-C4 intermediate, C4-like, and C4 species, making it a good model genus to study the evolution of C4 photosynthesis, and these studies indicate the direction for C4 engineering. C4 photosynthesis was acquired gradually through the C3-C4 intermediate stage. First, a two-celled C2 cycle called C2 photosynthesis was acquired by localizing glycine decarboxylase activity in the mitochondria of bundle sheath cells. With the development of two-cell metabolism, anatomical features also changed. Next, the replacement of the two-celled C2 cycle by the two-celled C4 cycle was induced by the acquisition of cell-selective expression in addition to the upregulation of enzymes in the C4 cycle during the C3-C4 intermediate stage. This was supported by an increase in cyclic electron transport activity in response to an increase in the ATP/NADPH demand for metabolism. Suppression of the C3 cycle in mesophyll cells was induced after the functional establishment of the C4 cycle, and optimization of electron transport by suppressing the activity of photosystem II also occurred during the final phase of C4 evolution.

Understanding Maize Response to Nitrogen Limitation in Different Light Conditions for the Improvement of Photosynthesis.

The photosynthetic capacity of leaves is determined by their content of nitrogen (N). Nitrogen involved in photosynthesis is divided between soluble proteins and thylakoid membrane proteins. In C4 plants, the photosynthetic apparatus is partitioned between two cell types: mesophyll cells and bundle sheath. The enzymes involved in the C4 carbon cycle and assimilation of nitrogen are localized in a cell-specific manner. Although intracellular distribution of enzymes of N and carbon assimilation is variable, little is known about the physiological consequences of this distribution caused by light changes. Light intensity and nitrogen concentration influence content of nitrates in leaves and can induce activity of the main enzymes involved in N metabolism, and changes that reduce the photosynthesis rate also reduce photosynthetic N use efficiency. In this review, we wish to highlight and discuss how/whether light intensity can improve photosynthesis in maize during nitrogen limitation. We described the general regulation of changes in the main photosynthetic and nitrogen metabolism enzymes, their quantity and localization, thylakoid protein abundance, intracellular transport of organic acids as well as specific features connected with C4 photosynthesis, and addressed the major open questions related to N metabolism and effects of light on photosynthesis in C4 plants.

Photosynthetic Linear Electron Flow Drives CO2 Assimilation in Maize Leaves.

Photosynthetic organisms commonly develop the strategy to keep the reaction center chlorophyll of photosystem I, P700, oxidized for preventing the generation of reactive oxygen species in excess light conditions. In photosynthesis of C4 plants, CO2 concentration is kept at higher levels around ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) by the cooperation of the mesophyll and bundle sheath cells, which enables them to assimilate CO2 at higher rates to survive under drought stress. However, the regulatory mechanism of photosynthetic electron transport for P700 oxidation is still poorly understood in C4 plants. Here, we assessed gas exchange, chlorophyll fluorescence, electrochromic shift, and near infrared absorbance in intact leaves of maize (a NADP-malic enzyme C4 subtype species) in comparison with mustard, a C3 plant. Instead of the alternative electron sink due to photorespiration in the C3 plant, photosynthetic linear electron flow was strongly suppressed between photosystems I and II, dependent on the difference of proton concentration across the thylakoid membrane (ΔpH) in response to the suppression of CO2 assimilation in maize. Linear relationships among CO2 assimilation rate, linear electron flow, P700 oxidation, ΔpH, and the oxidation rate of ferredoxin suggested that the increase of ΔpH for P700 oxidation was caused by the regulation of proton conductance of chloroplast ATP synthase but not by promoting cyclic electron flow. At the scale of intact leaves, the ratio of PSI to PSII was estimated almost 1:1 in both C3 and C4 plants. Overall, the photosynthetic electron transport was regulated for P700 oxidation in maize through the same strategies as in C3 plants only except for the capacity of photorespiration despite the structural and metabolic differences in photosynthesis between C3 and C4 plants.

Molecular Functional Characterisation of MechlPPDK Promoter in Transgenic Tobacco

Chloroplastic pyruvate phosphate dikinase (PPDK) (chlPPDK) is a key enzyme in the photosynthesis of C4 plants. PPDK is expressed in high abundance in C4 plants but only in trace amounts in C3 plants. The existing research reveals a higher expression of MechlPPDK in cultivated cassava varieties than that in wild cassava W14. However, knowledge about the transcriptional regulation of the MechlPPDK gene in cassava (Manihot esculenta Crantz) is insufficient. Therefore, we aim to identify the transcription profile of MechlPPDK and the core promoter region of MechlPPDK. We cloned the MechlPPDK coding sequence and its 5′-flanking sequence from the cassava variety Ku50. A series of deletion constructions fused to a uidA reporter gene were performed in the 5′-flanking sequence and stably transformed into tobacco. We found that P1 (from −590 bp to +114 bp) had the highest promoter activity among P1 to P3. The 5′-flanking sequence of MechlPPDK responded differently to varied abiotic stresses. Our results will further the understanding of the regulation of MechlPPDK expression.

Surfing the Hyperbola Equations of the Steady-State Farquhar\u2013von Caemmerer\u2013Berry C3 Leaf Photosynthesis Model: What Can a Theoretical Analysis of Their Oblique Asymptotes and Transition Points Tell Us?

The asymptotes and transition points of the net CO2 assimilation (A/Ci) rate curves of the steady-state Farquhar–von Caemmerer–Berry (FvCB) model for leaf photosynthesis of C3 plants are examined in a theoretical study, which begins from the exploration of the standard equations of hyperbolae after rotating the coordinate system. The analysis of the A/Ci quadratic equations of the three limitation states of the FvCB model—abbreviated as Ac, Aj and Ap—allows us to conclude that their oblique asymptotes have a common slope that depends only on the mesophyll conductance to CO2 diffusion (gm). The limiting values for the transition points between any two states of the three limitation states c, j and p do not depend on gm, and the results are therefore valid for rectangular and non-rectangular hyperbola equations of the FvCB model. The analysis of the variation of the slopes of the asymptotes with gm casts doubts about the fulfilment of the steady-state conditions, particularly, when the net CO2 assimilation rate is inhibited at high CO2 concentrations. The application of the theoretical analysis to extended steady-state FvCB models, where the hyperbola equations of Ac, Aj and Ap are modified to accommodate nitrogen assimilation and amino acids export via the photorespiratory pathway, is also discussed.

Sedimentary organic matter and δ13CKerogen variation on the southern Iberian palaeomargin (Betic Cordillera, SE Spain) during the latest Pliensbachian–Early Toarcian

The Early Toarcian Oceanic Anoxic Event (T-OAE) is characterised by major palaeoenvironmental and palaeoceanographical changes (ocean anoxia, global warming, ocean acidification, among other), and a severe perturbation of the global carbon cycle. Although widespread oceanic anoxia was a significant control on the occurrence of organic-rich facies during the T-OAE in several locations of the northern Tethyan margin, the effects of anoxia, oceanic productivity, and dilution are poorly understood in other worldwide locations, such as the southern Tethyan and Iberian margins.In this study, we report new geochemical [total organic carbon (TOC), total sulphur (TS), total carbonate content (CaCO3), δ13C from kerogen isolates (δ13CKerogen)], palynofacies and thermal maturation data from the Upper Pliensbachian–Lower Toarcian successions in La Cerradura and Fuente Vidriera, External Subbetic domain, Betic Cordillera (SE Spain). The obtained results indicate that these successions have low TOC contents (~ 0.3 wt.%), with TOC reaching up to 0.46 wt.% around the Polymorphum–Serpentinum ammonite zone boundary in La Cerradura. δ13CKerogen negative carbon isotope excursions (CIE) are observed at the base of the Polymorphum Zone, the Pliensbachian–Toarcian Event (Pl-Toa), and base of the Serpentinum Zone (T-OAE). Overall, the low maturity kerogen assemblages are dominated by terrestrial OM and with a minor marine component. Small increments in terrestrial OM, non-opaque phytoclasts, and palynomorphs are observed just before and during the T-OAE in the studied area.During the Early Toarcian, in the External Subbetic domain, accumulation and preservation of OM was low and with a significant terrestrial contribution, similarly to other locations around the southern margins of Iberia and Tethys. The dominance of the opaque phytoclast subgroup, the occurrence of Classopollis, and the relatively more positive δ13C of terrestrial kerogen suggest that OM was mostly sourced from an area characterised by a semi-arid climate (i.e. with pronounced arid periods) during most of the Polymorphum Zone. The T-OAE negative CIE is accompanied by an increase in terrestrial OM, with a slight increase in the non-opaque phytoclast subgroup and terrestrial palynomorph subgroup. The increase of non-opaque phytoclast and terrestrial palynomorphs are in line with the posited enhancements of the hydrological cycle and export of terrestrial OM into marine environments during the T-OAE. This study suggests a link between 13C fractionation during photosynthesis in C3 plants and climate during the Early Toarcian. This link suggests coupling of the water and terrestrial carbon cycles before and during the T-OAE.

A Cytosolic Bypass and G6P Shunt in Plants Lacking Peroxisomal Hydroxypyruvate Reductase.

The oxygenation of ribulose 1,5-bisphosphate by Rubisco is the first step in photorespiration and reduces the efficiency of photosynthesis in C3 plants. Our recent data indicate that mutants in photorespiration have increased rates of photosynthetic cyclic electron flow around photosystem I. We investigated mutant lines lacking peroxisomal hydroxypyruvate reductase to determine if there are connections between 2-phosphoglycolate accumulation and cyclic electron flow in Arabidopsis (Arabidopsis thaliana). We found that 2-phosphoglycolate is a competitive inhibitor of triose phosphate isomerase, an enzyme in the Calvin-Benson cycle that converts glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. This block in metabolism could be overcome if glyceraldehyde 3-phosphate is exported to the cytosol, where cytosolic triose phosphate isomerase could convert it to dihydroxyacetone phosphate. We found evidence that carbon is reimported as glucose-6-phosphate, forming a cytosolic bypass around the block of stromal triose phosphate isomerase. However, this also stimulates a glucose-6-phosphate shunt, which consumes ATP, which can be compensated by higher rates of cyclic electron flow.

Comparative transcriptomics method to infer gene coexpression networks and its applications to maize and rice leaf transcriptomes

Time-series transcriptomes of a biological process obtained under different conditions are useful for identifying the regulators of the process and their regulatory networks. However, such data are 3D (gene expression, time, and condition), and there is currently no method that can deal with their full complexity. Here, we developed a method that avoids time-point alignment and normalization between conditions. We applied it to analyze time-series transcriptomes of developing maize leaves under light-dark cycles and under total darkness and obtained eight time-ordered gene coexpression networks (TO-GCNs), which can be used to predict upstream regulators of any genes in the GCNs. One of the eight TO-GCNs is light-independent and likely includes all genes involved in the development of Kranz anatomy, which is a structure crucial for the high efficiency of photosynthesis in C4 plants. Using this TO-GCN, we predicted and experimentally validated a regulatory cascade upstream of SHORTROOT1, a key Kranz anatomy regulator. Moreover, we applied the method to compare transcriptomes from maize and rice leaf segments and identified regulators of maize C4 enzyme genes and RUBISCO SMALL SUBUNIT2 Our study provides not only a powerful method but also novel insights into the regulatory networks underlying Kranz anatomy development and C4 photosynthesis.

Antagonism or synergism? Combined effects of enhanced UV-B radiation and acid rain on photosynthesis in seedlings of two C4 plants

Enhanced ultraviolet-B (UV-B) radiation and acid precipitation (AP), abiotic stress factors that may have a strong impact on plants because of their detrimental effects on photosynthetic components. Therefore, understanding the mechanisms of UV-B radiation and acid rain on plant photosynthesis is a crucial question in plant ecology. However, research regarding their combined effects on photosynthesis of C4 plants is still scarce. Here we monitored the response of net photosynthetic rate, chlorophyll content and enzyme activities of NADP-ME C4 photosynthetic pathway monocotyledon plant waxy corn (Zea mays L. certain Kulesh), and NAD-ME C4 photosynthetic pathway dicotyledon plant edible amaranth (Amaranthus mangostanus L.) to enhanced UV-B and acid rain and their interaction. We grew them in the green house under 3 levels of UV-B radiation (0, 2.88, 5.76 kJ·m−2·d−1 for Z.mays and 0, 2.88, 4.32 kJ·m−2·d−1 for A.mangostanus) and 3 levels of acid rain (pH 6.5, pH 4.5, pH 3.5) in a full three-way randomized block design. The key results were as follows: Enhanced UV-B radiation and acid rain and their interaction all decreased the net photosynthesis (Pn), PEPCase and RuBPCase activity of both species, compared with the control A0B0. UV-B decreased Chla, Chlb and Car in Amaranth seedlings, however, increased them in Maize seedlings, compared with those of the control. As a whole, acid rain had negative effects on chlorophyll content, however, positive effects on Car in two plants seedlings. Both UV-B radiation and (or) acid rain did not affect Chla/Chlb and Car/Chl. In conclusion, the negative effect of UV-B radiation was greater than that of acid rain. Whereas, more synergism was observed in monocotyledonous C4 plants, Maize seedlings, and more antagonism was observed in dicotyledonous C4 plants, Amaranth seedlings, which could explain why the Maize was affected less by the combined UV-B irradiation and acid rain.

Functional Characterization of Cytosolic Pyruvate Phosphate Dikinase Gene (MecyPPDK) and Promoter (MecyPPDKP) of Cassava in Response to Abiotic Stress in Transgenic Tobacco

Pyruvate phosphate dikinase (PPDK) is a key enzyme in the photosynthesis of C4 plants; its expression significantly affects the accumulation of photosynthetic products in C4 plants. Pyruvate phosphate dikinase presents two subtypes, namely, chloroplastic PPDK and cytosolic PPDK. Cytosolic PPDK is ubiquitous in C3 and C4 plants. This subtype is also highly expressed in metabolically active storage organs. Cytosolic PPDK in cassava (Manihot esculenta Crantz, MecyPPDK) is considerately expressed mainly in functional leaves. However, research on the function of PPDK in cassava has not yet been reported. In this study, we cloned the 5′ flanking sequence of MecyPPDK fused to the uidA reporter gene and stably transfected tobacco (Nicotiana benthamiana Domin). Slight β‐glucuronidase activity was observed in transformed tobacco. This activity was upregulated remarkably under low temperature, drought, waterlogging, and abscisic acid treatments. Results suggest that MecyPPDK facilitates the survival of cassava plants in response to abiotic stress. Our results will provide further understanding regarding the regulation of MecyPPDK expression and a novel stress‐inducible promoter in transgenic tobacco.

Uncovering C4-like photosynthesis in C3 vascular cells.

In C4 plants, the vascularization of the leaf is extended to include a ring of photosynthetic bundle sheath cells, which have essential and specific functions. In contrast to the substantial knowledge of photosynthesis in C4 plants, relatively little is known about photosynthesis in C3 plant veins, which differs substantially from that in C3 mesophyll cells. In this review we highlight the specific photosynthetic machinery present in C3 vascular cells, which likely evolved prior to the divergence between C3 and C4 plants. The associated primary processes of carbon recapture, nitrogen transport, and antioxidant metabolism are discussed. This review of the basal C4 photosynthesis in C3 plants is significant in the context of promoting the potential for biotechnological development of C4-transgenic rice crops.

Small-molecule inhibition of pyruvate phosphate dikinase targeting the nucleotide binding site.

Pyruvate phosphate dikinase (PPDK) is an essential enzyme of C4 photosynthesis in plants, catalyzing the ATP-driven conversion of pyruvate to phosphoenolpyruvate (PEP). It is further used by some bacteria and unicellular protists in the reverse, ATP-forming direction. Many weed species use C4 photosynthesis in contrast to world’s major crops, which are C3 plants. Hence inhibitors of PPDK may be used as C4-specific herbicides. By screening a library of 80 commercially available kinase inhibitors, we identified compounds derived from bisindolylmaleimide (bisindolylmaleimide IV, IC50 = 0.76 ± 0.13 μM) and indirubin (indirubin-3’-monoxime, IC50 = 4.2 ± 0.9 μM) that showed high inhibitory potency towards PPDK and are among the most effective PPDK inhibitors described today. Physiological studies on leaf tissues of a C4 model plant confirmed in vivo inhibition of C4-driven photosynthesis by these substances. Moreover, comparative docking studies of non-inhibitory bisindolylmaleimide derivatives suggest that the selectivity towards PPDK may be increased by addition of functional groups to the core structure.

Photosynthesis: Mining grasses for a better Rubisco.

Photosynthesis in C3 plants is limited by the kinetics of the CO2-fixing enzyme Rubisco. Natural variation in Rubisco can be exploited to provide new avenues for adapting photosynthetic performance to a changing climate while reducing the environmental footprint of agriculture.

Towards engineering carboxysomes into C3 plants.

Photosynthesis in C3 plants is limited by features of the carbon-fixing enzyme Rubisco, which exhibits a low turnover rate and can react with O2 instead of CO2 , leading to photorespiration. In cyanobacteria, bacterial microcompartments, known as carboxysomes, improve the efficiency of photosynthesis by concentrating CO2 near the enzyme Rubisco. Cyanobacterial Rubisco enzymes are faster than those of C3 plants, though they have lower specificity toward CO2 than the land plant enzyme. Replacement of land plant Rubisco by faster bacterial variants with lower CO2 specificity will improve photosynthesis only if a microcompartment capable of concentrating CO2 can also be installed into the chloroplast. We review current information about cyanobacterial microcompartments and carbon-concentrating mechanisms, plant transformation strategies, replacement of Rubisco in a model C3 plant with cyanobacterial Rubisco and progress toward synthesizing a carboxysome in chloroplasts.

Physiological evidence for plasticity in glycolate/glycerate transport during photorespiration.

Photorespiration recycles fixed carbon following the oxygenation reaction of Ribulose, 1–5, carboxylase oxygenase (Rubisco). The recycling of photorespiratory C2 to C3 intermediates is not perfectly efficient and reduces photosynthesis in C3 plants. Recently, a plastidic glycolate/glycerate transporter (PLGG1) in photorespiration was identified in Arabidopsis thaliana, but it is not known how critical this transporter is for maintaining photorespiratory efficiency. We examined a mutant deficient in PLGG1 (plgg1-1) using modeling, gas exchange, and Rubisco biochemistry. Under low light (under 65 μmol m−2 s−1 PAR), there was no difference in the quantum efficiency of CO2 assimilation or in the photorespiratory CO2 compensation point of plgg1-1, indicating that photorespiration proceeded with wild-type efficiency under sub-saturating light irradiances. Under saturating light irradiance (1200 μmol m−2 s−1 PAR), plgg1-1 showed decreased CO2 assimilation that was explained by decreases in the maximum rate of Rubisco carboxylation and photosynthetic linear electron transport. Decreased rates of Rubisco carboxylation resulted from probable decreases in the Rubisco activation state. These results suggest that glycolate/glycerate transport during photorespiration can proceed in moderate rates through an alternative transport process with wild-type efficiencies. These findings also suggest that decreases in net CO2 assimilation that occur due to disruption to photorespiration can occur by decreases in Rubisco activity and not necessarily decreases in the recycling efficiency of photorespiration.Electronic supplementary materialThe online version of this article (doi:10.1007/s11120-016-0277-3) contains supplementary material, which is available to authorized users.

Metabolic Network Constrains Gene Regulation of C4 Photosynthesis: The Case of Maize.

Engineering C3 plants to increase their efficiency of carbon fixation as well as of nitrogen and water use simultaneously may be facilitated by understanding the mechanisms that underpin the C4 syndrome. Existing experimental studies have indicated that the emergence of the C4 syndrome requires co-ordination between several levels of cellular organization, from gene regulation to metabolism, across two co-operating cell systems—mesophyll and bundle sheath cells. Yet, determining the extent to which the structure of the C4 plant metabolic network may constrain gene expression remains unclear, although it will provide an important consideration in engineering C4 photosynthesis in C3 plants. Here, we utilize flux coupling analysis with the second-generation maize metabolic models to investigate the correspondence between metabolic network structure and transcriptomic phenotypes along the maize leaf gradient. The examined scenarios with publically available data from independent experiments indicate that the transcriptomic programs of the two cell types are co-ordinated, quantitatively and qualitatively, due to the presence of coupled metabolic reactions in specific metabolic pathways. Taken together, our study demonstrates that precise quantitative coupling will have to be achieved in order to ensure a successfully engineered transition from C3 to C4 crops.

Rapid responses of mesophyll conductance to changes of CO2 concentration, temperature and irradiance are affected by N supplements in rice

Photosynthesis in C3 plants is significantly limited by mesophyll conductance (gm ), which can vary with leaf anatomical traits and nitrogen (N) supplements. Several studies have investigated the response of gm to N supplements; however, none examined the implications of N supplements on the response of gm to rapid environmental changes. Here we investigated the effect of N supplement on gm and the response of gm to change of CO2 , temperature and irradiance in rice. High N supplement (HN) increased mesophyll cell wall surface area and chloroplast surface area exposed to intercellular airspace per leaf area, and reduced cell wall thickness. These changes resulted in increased gm . The gm of leaves with HN was more sensitive to changes in CO2 concentration, temperature and irradiance. The difference in leaf structural features between low N supplement and HN indicates that a rapid change in gm is related to the regulation of diffusion through biological membranes rather than leaf structural features. These results will contribute to an understanding of the determinants of gm response to rapid changes in environmental factors.