Photorespiration and photoinhibition in the bracts of cotton under water stress

Gas exchange and chlorophyll fluorescence parameters of PSII were analyzed in the bracts and leaves of cotton plants after anthesis. Photosynthetic activity and photorespiration were measured in the leaves and bracts of cotton grown under either normal or reduced water-saving drip irrigation. The photosynthetic performance, amount of chlorophyll and Rubisco, and net photosynthesis were greater in the bracts than that in the leaves under water stress. The actual photochemical efficiency of PSII decreased in both the bracts and leaves after anthesis under reduced irrigation. However, the decrease was smaller in the bracts than in the leaves, indicating that the bracts experienced less severe photoinhibition compared to the leaves. The greater drought tolerance of bracts could be related to differences in relative water content, instantaneous water-use efficiency, and photorespiration rate. The ratio of photorespiration to net photosynthesis was much higher in the bracts than in leaves. Furthermore, water deficiency (due to the water-saving drip irrigation) had no significant effect on that ratio in the bracts. We hypothesized that photorespiration in the bracts alleviated photoinhibition and maintained photosynthetic activity.     

Peroxisomal APX knockdown triggers antioxidant mechanisms favourable for coping with high photorespiratory H2O2 induced by CAT deficiency in rice

The physiological role of peroxisomal ascorbate peroxidases (pAPX) is unknown; therefore, we utilized pAPX4 knockdown rice and catalase (CAT) inhibition to assess its role in CAT compensation under high photorespiration. pAPX4 knockdown induced co‐suppression in the expression of pAPX3. The rice mutants exhibited metabolic changes such as lower CAT and glycolate oxidase (GO) activities and reduced glyoxylate content; however, APX activity was not altered. CAT inhibition triggered different changes in the expression of CAT, APX and glutathione peroxidase (GPX) isoforms between non‐transformed (NT) and silenced plants. These responses were associated with alterations in APX, GPX and GO activities, suggesting redox homeostasis differences. The glutathione oxidation‐reduction states were modulated differently in mutants, and the ascorbate redox state was greatly affected in both genotypes. The pAPX suffered less oxidative stress and photosystem II (PSII) damage and displayed higher photosynthesis than the NT plants. The improved acclimation exhibited by the pAPX plants was indicated by lower H₂O₂ accumulation, which was associated with lower GO activity and glyoxylate content. The suppression of both pAPXs and/or its downstream metabolic and molecular effects may trigger favourable antioxidant and compensatory mechanisms to cope with CAT deficiency. This physiological acclimation may involve signalling by peroxisomal H₂O₂, which minimized the photorespiration.     

Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubisco and expression of photosynthetic genes in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Brassinosteroids (BRs) are a new group of plant growth substances that promote plant growth and productivity. We showed in this study that improved growth of cucumber (Cucumis sativus) plants after treatment with 24-epibrassinolide (EBR), an active BR, was associated with increased CO₂ assimilation and quantum yield of PSII (Φ_PSII). Treatment of brassinazole (Brz), a specific inhibitor for BR biosynthesis, reduced plant growth and at the same time decreased CO₂ assimilation and Φ_PSII. Thus, the growth-promoting activity of BRs can be, at least partly, attributed to enhanced plant photosynthesis. To understand how BRs enhance photosynthesis, we have analyzed the effects of EBR and Brz on a number of photosynthetic parameters and their affecting factors, including the contents and activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Northern and Western blotting demonstrated that EBR upregulated, while Brz downregulated, the expressions of rbcL, rbcS and other photosynthetic genes. In addition, EBR had a positive effect on the activation of Rubisco based on increased maximum Rubisco carboxylation rates (V _c,max), total Rubisco activity and, to a greater extent, initial Rubisco activity. The accumulation patterns of Rubisco activase (RCA) based on immunogold-labeling experiments suggested a role of RCA in BR-regulated activation state of Rubisco. Enhanced expression of genes encoding other Calvin cycle genes after EBR treatment may also play a positive role in RuBP regeneration (J _max), thereby increasing maximum carboxylation rate of Rubisco (V _c,max). Thus, BRs promote photosynthesis and growth by positively regulating synthesis and activation of a variety of photosynthetic enzymes including Rubisco in cucumber.     

Changes in photosynthetic capacity and polypeptide patterns during natural senescence and rejuvenation of <Emphasis Type="Italic">Cucurbita pepo</Emphasis> L. (zucchini) cotyledons

Natural senescence of Cucurbita pepo (zucchini) cotyledons was accompanied by a gradual degradation of reserve proteins (globulins) and an intensive decrease in the content of both large subunit (LSU) and small subunit (SSU) of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). The net photosynthetic rate, the primary photochemical activity of PSII, estimated by the variable fluorescence (F_v)/maximal fluorescence (F_m) ratio (F_v/F_m) and the actual quantum yield of PSII electron transport in the light-adapted state (ΦPSII) also progressively decreased during natural senescence. In contrast, the fraction of the absorbed light energy, which is not used for photochemistry (LNU) increased with progression of senescence. The decline in the photosynthetic rate started earlier in ontogenesis compared with the down-regulation of the functional activity of PSII, thus suggesting the existence of protective mechanisms which maintain higher efficiency of the photochemical electron transport reactions of photosynthesis compared with the dark reactions of the Calvin cycle during earlier stages of natural senescence. Decapitation of the epicotyl above the senescing cotyledons resulted in full recovery of the polypeptide profile in the rejuvenated cotyledons. In addition, the photosynthetic rate increased reaching values that exceeded those measured in juvenile cotyledons. The photochemical efficiency of PSII also gradually recovered, although it did not reach the maximum values measured in the presenescent cotyledons.     

Distribution and functional role of carbonic anhydrase Cah3 associated with thylakoid membranes in the chloroplast and pyrenoid of <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

Localization of lumenal carbonic anhydrase Cah3 in thylakoid membranes of Chlamydomonas reinhardtii was studied using wild-type algae and photosynthetic mutants with different composition of chlorophyll-protein complexes in the photosystems. In addition, the photosynthetic characteristics of wild-type C. reinhardtii and cia3 mutants lacking the activity of carbonic anhydrase Cah3 were examined. Western blot analysis revealed the lack of cross reaction with antibodies to Cah3 in the mutant lacking the photosystem II (PSII) reaction center, in contrast to the mutant deficient in light-harvesting complex of PSII. These data show that the lumenal Cah3 is associated with polypeptides on the donor side of PSII reaction center. Using immunoelectron microscopy and antibodies to Cah3 from C. reinhardtii, we showed for the first time that the major part of thylakoid Cah3 is localized in the pyrenoid where the bulk of Rubisco is located. The rate of photosynthetic oxygen evolution and PSII photochemical efficiency were lower in C. reinhardtii cia3 mutant than in the wild type, especially in the cells grown at limiting CO₂ concentrations. These observations show that Cah3 takes part in CO₂-concentrating mechanism of the chloroplast. The results support our hypothesis [1, 2] that the carboxylation reaction in microalgae proceeds in the pyrenoid, a specific Rubisco-containing part of the chloroplast, which acquires CO₂ from the lumen of intrapyrenoid thylakoids. We discuss significance of the pyrenoid as an autonomous metabolic microcompartment, in which Cah3 plays a key role in the production and concentration of CO₂ for Rubisco. These functions may promote the photosynthetic efficiency owing to the effective CO₂ supply for the Calvin cycle.     

An Excel tool for deriving key photosynthetic parameters from combined gas exchange and chlorophyll fluorescence: theory and practice

Combined photosynthetic gas exchange and modulated fluorometres are widely used to evaluate physiological characteristics associated with phenotypic and genotypic variation, whether in response to genetic manipulation or resource limitation in natural vegetation or crops. After describing relatively simple experimental procedures, we present the theoretical background to the derivation of photosynthetic parameters, and provide a freely available Excel‐based fitting tool (EFT) that will be of use to specialists and non‐specialists alike. We use data acquired in concurrent variable fluorescence–gas exchange experiments, where A/C_i and light–response curves have been measured under ambient and low oxygen. From these data, the EFT derives light respiration, initial PSII (photosystem II) photochemical yield, initial quantum yield for CO₂ fixation, fraction of incident light harvested by PSII, initial quantum yield for electron transport, electron transport rate, rate of photorespiration, stomatal limitation, Rubisco (ribulose 1·5‐bisphosphate carboxylase/oxygenase) rate of carboxylation and oxygenation, Rubisco specificity factor, mesophyll conductance to CO₂ diffusion, light and CO₂ compensation point, Rubisco apparent Michaelis–Menten constant, and Rubisco CO₂‐saturated carboxylation rate. As an example, a complete analysis of gas exchange data on tobacco plants is provided. We also discuss potential measurement problems and pitfalls, and suggest how such empirical data could subsequently be used to parameterize predictive photosynthetic models.     

Sulfur starvation in rice: the effect on photosynthesis, carbohydrate metabolism, and oxidative stress protective pathways

Sulfur-deficient plants generate a lower yield and have a reduced nutritional value. The process of sulfur acquisition and assimilation play an integral role in plant metabolism, and response to sulfur deficiency involves a large number of plant constituents. Rice (Oryza sativa) is the second most consumed cereal grain, and the effects of sulfur deprivation in rice were analyzed by measuring changes in photosynthesis, carbohydrate metabolism, and antioxidants. The photosynthetic apparatus was severely affected under sulfur deficiency. The Chl content was reduced by 49% because of a general reduction of PSII and PSI and the associated light-harvesting antenna. The PSII efficiency was 31% lower at growth light, and the ability of PSI to photoreduce NADP+ was decreased by 61%. The Rubisco content was also significantly reduced in the sulfur-deprived plants. The imbalances between PSII and PSI, and between photosynthesis and carbon fixation led to a general over-reduction of the photosynthetic electron carriers (higher 1-q(P)). Chromatographic analysis showed that the level of monosaccharides was lower and starch content higher in the sulfur-deprived plants. In contrast, no changes in metabolite levels were found in the tricarboxylic acid or Calvin cycle. The level of the thiol-containing antioxidant, GSH, was 70% lower and the redox state was significantly more oxidized. These changes in GSH status led to an upregulation of the cytosolic isoforms of GSH reductase and monodehydroascorbate reductase. In addition, alternative antioxidants like flavonoids and anthocyanins were increased in the sulfur-deprived plants.     

A role for brassinosteroids in the regulation of photosynthesis in Cucumis sativus

The effects of 24-epibrassinolide (EBR) spray application on gas-exchange, chlorophyll fluorescence characteristics, Rubisco activity, and carbohydrate metabolism were investigated in cucumber (Cucumis sativus L. cv. Jinchun No. 3) plants grown in a greenhouse. EBR significantly increased the light-saturated net CO(2) assimilation rate (A(sat)) from 3 h to 7d after spraying, with 0.1 mg l(-1) EBR proving most effective. Increased A(sat) in EBR-treated leaves was accompanied by increases in the maximum carboxylation rate of Rubisco (V(c,max)) and in the maximum rate of RuBP regeneration (J(max)). EBR-treated leaves also had a higher quantum yield of PSII electron transport (phi(PSII)) than the controls, which was mainly due to a significant increase in the photochemical quenching (q(P)), with no change in the efficiency of energy capture by open PSII reaction centres (F'(v)/F'(m)). EBR did not influence photorespiration. In addition, significant increases in the initial activity of Rubisco and in the sucrose, soluble sugars, and starch contents were observed followed by substantial increases in sucrose phosphate synthase (SPS), sucrose synthase (SS), and acid invertase (AI) activities after EBR treatment. It was concluded that EBR increases the capacity of CO(2) assimilation in the Calvin cycle, which was mainly attributed to an increase in the initial activity of Rubisco.     

Rubisco mutagenesis provides new insight into limitations on photosynthesis and growth in Synechocystis PCC6803

Orthophosphate (Pi) stimulates the activation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) while paradoxically inhibiting its catalysis. Of three Pi-binding sites, the roles of the 5P- and latch sites have been documented, whereas that of the 1P-site remained unclear. Conserved residues at the 1P-site of Rubisco from the cyanobacterium Synechocystis PCC6803 were substituted and the kinetic properties of the enzyme derivatives and effects on cell photosynthesis and growth were examined. While Pi-stimulated Rubisco activation diminished for enzyme mutants T65A/S and G404A, inhibition of catalysis by Pi remained unchanged. Together with previous studies, the results suggest that all three Pi-binding sites are involved in stimulation of Rubisco activation, whereas only the 5P-site is involved in inhibition of catalysis. While all the mutations reduced the catalytic turnover of Rubisco (K(cat)) between 6- and 20-fold, the photosynthesis and growth rates under saturating irradiance and inorganic carbon (Ci) concentrations were only reduced 40-50% (in the T65A/S mutants) or not at all (G404A mutant). Analysis of the mutant cells revealed a 3-fold increase in Rubisco content that partially compensated for the reduced K(cat) so that the carboxylation rate per chlorophyll was one-third of that in the wild type. Correlation between the kinetic properties of Rubisco and the photosynthetic rate (P(max)) under saturating irradiance and Ci concentrations indicate that a >60% reduction in K(cat) can be tolerated before P(max) in Synechocystsis PCC6803 is affected. These results indicate that the limitation of Rubisco activity on the rate of photosynthesis in Synechocystis is low. Determination of Calvin cycle metabolites revealed that unlike in higher plants, cyanobacterial photosynthesis is constrained by phosphoglycerate reduction probably due to limitation of ATP or NADPH.     

Transgenic approaches to manipulate the environmental responses of the C3 carbon fixation cycle

The limitation to photosynthetic CO2 assimilation in C3 plants in hot, dry environments is dominated by ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco) because CO2 availability is restricted and photorespiration is stimulated. Using a combination of genetic engineering and transgenic technology, three approaches to reduce photorespiration have been taken; two of these focused on increasing the carboxylation efficiency of Rubisco either by reducing the oxygenase reaction directly or by manipulating the Rubisco enzyme by concentrating CO2 in the region of Rubisco through the introduction of enzymes of the C4 pathway. The third approach attempted to reduce photorespiration directly by manipulation of enzymes in this pathway. The progress in each of these areas is discussed, and the most promising approaches are highlighted. Under saturating CO2 conditions, Rubisco did not limit photosynthesis, and limitation shifted to ribulose bisphosphate (RuBP) regeneration capacity of the C3 cycle. Transgenic analysis was used to identify the specific enzymes that may be targets for improving carbon fixation, and the way this may be exploited in the high CO2 future is considered.     

大气CO2浓度升高对干旱条件下冬小麦叶片光合适应的影响

大气CO₂浓度升高是全球气候变化的主要特征,但大气CO₂浓度长期升高条件下冬小麦叶片发生光合适应的机制尚不十分清楚。本研究以盆栽冬小麦‘郑麦9023’为试验材料,在人工气候控制室内设置2个CO₂浓度(400和600 μmol·mol^-1)、2个水分条件(田间持水量的80%±5%和55%±5%),测定拔节期和抽穗期的光合特征曲线、叶绿素荧光动力学参数、叶氮含量和收获后的籽粒产量等指标,探讨干旱条件下库源关系改变对叶片光合适应的影响。结果表明: 在小麦拔节期,干旱条件下CO₂浓度升高处理的小麦PSⅡ实际光化学效率没有显著增加,但通过提升最大电子传递速率和电子向光化学方向的传递比例,增强了Rubisco的羧化速率,从而提高了最大净光合速率;在抽穗期,功能叶最大电子传递速率和电子向光化学方向的传递比例虽然较高,但PSⅡ实际光化学转换效率降低,Rubisco羧化速率和丙糖磷酸利用效率下降,以致最大净光合速率降低。干旱条件下,CO₂浓度升高增加了小麦单茎生物量、单穗粒数和穗粒重,降低了不孕小穗数,提高了籽粒产量。土壤干旱条件下,CO₂浓度升高对收获期小麦单茎籽粒产量的促进作用可能主要来自于生长前期的光合产物积累。生长后期光合适应发生的主要原因是功能叶PSⅡ实际光化学转换效率和丙糖磷酸利用效率的降低,而不是最大电子传递速率、光化学方向的电子传递比例和新叶库强的变化。     

Photosynthetic characteristics and tolerance to photo-oxidation of transgenic rice expressing C4 photosynthesis enzymes

The photosynthetic characteristics of four transgenic rice lines over-expressing rice NADP-malic enzyme (ME), and maize phosphoenolpyruvate carboxylase (PC), pyruvate,orthophosphate dikinase (PK), and PC+PK (CK) were investigated using outdoor-grown plants. Relative to untransformed wild-type (WT) rice, PC transgenic rice exhibited high PC activity (25-fold increase) and enhanced activity of carbonic anhydrase (more than two-fold increase), while the activity of ribulose-bisphosphate carboxylase/oxygenase (Rubisco) and its kinetic property were not significantly altered. The PC transgenic plants also showed a higher light intensity for saturation of photosynthesis, higher photosynthetic CO₂ uptake rate and carboxylation efficiency, and slightly reduced CO₂ compensation point. In addition, chlorophyll a fluorescence analysis indicates that PC transgenic plants are more tolerant to photo-oxidative stress, due to a higher capacity to quench excess light energy via photochemical and non-photochemical means. Furthermore, PC and CK transgenic rice produced 22–24% more grains than WT plants. Taken together, these results suggest that expression of maize C₄ photosynthesis enzymes in rice, a C₃ plant, can improve its photosynthetic capacity with enhanced tolerance to photo-oxidation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Acclimation of rice photosynthesis to irradiance under field conditions

Acclimation to irradiance was measured in terms of light-saturated photosynthetic carbon assimilation rates (P(max)), Rubisco, and pigment content in mature field-grown rice (Oryza sativa) plants in tropical conditions. Measurements were made at different positions within the canopy alongside irradiance and daylight spectra. These data were compared with a second experiment in which acclimation to irradiance was assessed in uppermost leaves within whole-plant shading regimes (10% low light [LL], 40% medium light [ML], and 100% high light [HL] of full natural sunlight). Two varieties, japonica (tropical; new plant type [NPT]) and indica (IR72) were compared. Values for Rubisco amount, chlorophyll a/b, and P(max) all declined from the top to the base of the canopy. In the artificial shading experiment, acclimation of P(max) (measured at 350 microL L(-1) CO(2)) occurred between LL and ML for IR72 with no difference observed between ML and HL. The Rubisco amount increased between ML and HL in IR72. A different pattern was seen for NPT with higher P(max) (measured at 350 microL L(-1) CO(2)) at LL than IR72 and some acclimation of this parameter between ML and HL. Rubisco levels were higher in NPT than IR72 contrasting with P(max). Comparison of data from both experiments suggests a leaf aging effect between the uppermost two leaf positions, which was not a result of irradiance acclimation. Results are discussed in terms of: (a) acclimation of photosynthesis and radiation use efficiency at high irradiance in rice, and (b) factors controlling photosynthetic rates of leaves within the canopy.     

遮荫条件下草莓的光合持性变化

对宝交早生和硕丰两个草莓品种遮荫处理后测定其光合特性变化结果表明,遮荫处理使两个草莓品种叶片光合速率显著降低,分别下降了20％和47％,而表观量子效率分别提高了13％和8％,叶片中叶绿素含量升高而可溶性蛋白含量显著降低,光系统II电子传递活笥（PSII活性）分别下降了22．5％和53．7％,1,5二磷酸核酮糖羧化酶（Rubisco)活性分别低了19．6％和35．3％,进一步讨论了草莓光饱和速率下降的生理基础。     

Characteristics of photosynthesis and functions of the water-water cycle in rice (Oryza sativa) leaves in response to potassium deficiency

The mechanisms of photoprotection of photosynthesis and dissipation of excitation energy in rice leaves in response to potassium (K) deficiency were investigated. Net photosynthetic rate and the activity of ribulose-1,5-bisphosphate carboxylase/oxygenase decreased under K deficiency. Compared with the control, non-photochemical quenching of Chl fluorescence increased in K-deficient plant, whereas the efficiency of excitation transfer (F'(v)/F'(m)) and the photochemical quenching coefficient (q(P)) decreased. Thus, thermal dissipation of excitation energy increased as more excess electrons were accumulated in the photosynthetic chain. The electron transport rate through PSII (J(f)) was more sensitive to O2 concentration, and the fraction of electron transport rate required to sustain CO2 assimilation and photorespiration (J(g)/J(f)) was significantly decreased under K deficiency compared with the control. Furthermore, the alternative electron transport (J(a)/J(f)) was increased, indicating that a considerable amount of electrons had been transported to O2 during the water-water cycle in the K-deficient leaves. Although the fraction of electron transport to photorespiration (J(o)/J(f)) was also increased in the K-deficient leaves, it was less sensitive than that of the water-water cycle. With the generation of reactive oxygen species level, the activities of superoxide dismutase and ascorbate peroxidase, two of the key enzymes involved in scavenging of active oxygen species in the water-water cycle, also increased in K-deficient rice. Therefore, it is likely that a series of photoprotective mechanisms were initiated in rice plants in response to K deficiency and the water-water cycle might be critical for protecting photosynthetic apparatus under K deficiency in rice.     

Engineering Rubisco activase from thermophilic cyanobacteria into high-temperature sensitive plants

In the past decade, various strategies to improve photosynthesis and crop yield, such as leaf morphology, light interception and use efficiency, biochemistry of light reactions, stomatal conductance, carboxylation efficiency, and source to sink regulation, have been discussed at length. Leaf morphology and physiology are tightly coupled to light capturing efficiency, gas exchange capacity, and temperature regulation. However, apart from the photoprotective mechanism of photosystem-II (PSII), i.e. non-photochemical quenching, very low genetic variation in the components of light reactions has been observed in plants. In the last decade, biochemistry-based enhancement of carboxylation efficiency that improves photosynthesis in plants was one of the potential strategies for improving plant biomass production. Enhancement of activation of the ubiquitous enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) by Rubisco activase may be another potential strategy for improving a photosynthesis-driven increase in crop yield. Rubisco activase modifies the conformation of the active center in Rubisco by removing tightly bound inhibitors, thereby contributing to enzyme activation and rapid carboxylation. Thermophilic cyanobacteria are oxygenic photosynthetic bacteria that thrive in high-temperature environments. This critical review discusses the prospects for and the potential of engineering Rubisco activase from thermophilic cyanobacteria into temperature-sensitive plants, to increase the threshold temperature and survival of these plants in arid regions.     

Strategies for engineering C4 photosynthesis

C₃ photosynthesis is an inefficient process, because the enzyme that lies at the heart of the Benson–Calvin cycle, ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco) is itself a very inefficient enzyme. The oxygenase activity of Rubisco is an unavoidable side reaction that is a consequence of its reaction mechanism. The product of oxygenation, glycollate 2-P, has to be retrieved by photorespiration, a process which results in the loss of a quarter of the carbon that was originally present in glycollate 2-P. Photorespiration therefore reduces carbon gain. Purely in terms of carbon economy, there is, therefore, a strong selection pressure on plants to reduce the rate of photorespiration so as to increase carbon gain, but it also improves water- and nitrogen-use efficiency. Possibilities for the manipulation of plants to decrease the amount of photorespiration include the introduction of improved Rubisco from other species, reconfiguring photorespiration, or introducing carbon-concentrating mechanisms, such as inorganic carbon transporters, carboxysomes or pyrenoids, or engineering a full C₄ Kranz pathway using the existing evolutionary progression in C₃–C₄ intermediates as a blueprint. Possible routes and progress to suppressing photorespiration by introducing C₄ photosynthesis in C₃ crop plants will be discussed, including whether single cell C₄ photosynthesis is feasible, how the evolution of C₃–C₄ intermediates can be used as a blueprint for engineering C₄ photosynthesis, which pathway for the C₄ cycle might be introduced and the extent to which processes and structures in C₃ plant might require optimisation.     

Brassinosteroids accelerate recovery of photosynthetic apparatus from cold stress by balancing the electron partitioning,carboxylation and redox homeostasis in cucumber

The aim of this study was to examine the role of brassinosteroids (BRs) in protecting the photosynthetic apparatus from cold‐induced damage in cucumber (Cucumis sativus) plants. Recovery at both high light (HL) and low light (LL) after a cooling at 10/7°C induced irreversible inhibition of CO₂ assimilation, photoinhibition at photosystem I (PSI) and inhibition of enzyme activities of Calvin cycle and ascorbate (AsA)‐reduced glutathione (GSH) cycle, followed by accumulation of H₂O₂ and malondialdehyde. However, cold‐induced photoinhibition at PSII was fully recovered at LL but not at HL. Meanwhile, recovery at HL increased electron flux to O₂‐dependent alternative pathway [Ja(O₂‐dependent)]. Foliar application of 24‐epibrassinolide (EBR) accelerated recovery from photoinhibition of PSII but not of PSI. EBR also significantly increased CO₂ assimilation, activity of Calvin cycle enzymes and electron flux to carbon reduction [Je(PCR)], with a concomitant decrease in Ja(O₂‐dependent); meanwhile EBR increased the activity of enzymes in AsA‐GSH cycle and cellular redox states. However, the positive effect of EBR on plant recovery was observed only at HL, but not LL. These results indicate that BR accelerates the recovery of photosynthetic apparatus at HL by activation of enzymes in Calvin cycle and increasing the antioxidant capacity, which in turn mitigate the photooxidative stress and the inhibition of plant growth during the recovery.     

Metabolome analysis of photosynthesis and the related primary metabolites in the leaves of transgenic rice plants with increased or decreased Rubisco content

Because the comprehensive effects on metabolism by genetic manipulation of leaf Rubisco content are unknown, metabolome analysis was carried out on transgenic rice plants with increased or decreased Rubisco content using the capillary electrophoresis-time-of-flight mass spectrometry (CE-TOFMS) technique. In RBCS-sense plants, an increase in Rubisco content did not improve light-saturated photosynthesis. Glyceraldehyde 3-phosphate and sedoheputulose 7-phosphate levels increased, but ribulose bisphosphate (RuBP), ATP and ADP levels were not affected. It is considered from these results that RuBP regeneration independent of ATP supply became a bottleneck for photosynthesis. In RBCS-antisense plants, a decline in Rubisco content decreased photosynthesis with a substantial accumulation of RuBP. ATP and ADP levels also increased and were associated with increases in the diphosphate and triphosphate compounds of other nucleosides. These results imply that a decline in Rubisco content slowed down the Calvin cycle and that the resultant excess energy of ATP was transferred to other nucleoside diphosphates and triphosphates. The levels of amino acids tended to decline in RBCS-sense plants and increase in RBCS-antisense plants, probably reflecting the demand for Rubisco synthesis. Starch and carbohydrate levels decreased only in RBCS-antisense plants. Thus, genetic manipulation of Rubisco contents widely affected C and N metabolism in rice.     

Interruption of the Calvin cycle inhibits the repair of Photosystem II from photodamage

In photosynthetic organisms, impairment of the activities of enzymes in the Calvin cycle enhances the extent of photoinactivation of Photosystem II (PSII). We investigated the molecular mechanism responsible for this phenomenon in the unicellular green alga Chlamydomonas reinhardtii. When the Calvin cycle was interrupted by glycolaldehyde, which is known to inhibit phosphoribulokinase, the extent of photoinactivation of PSII was enhanced. The effect of glycolaldehyde was very similar to that of chloramphenicol, which inhibits protein synthesis de novo in chloroplasts. The interruption of the Calvin cycle by the introduction of a missense mutation into the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) also enhanced the extent of photoinactivation of PSII. In such mutant 10-6C cells, neither glycolaldehyde nor chloramphenicol has any additional effect on photoinactivation. When wild-type cells were incubated under weak light after photodamage to PSII, the activity of PSII recovered gradually and reached a level close to the initial level. However, recovery was inhibited in wild-type cells by glycolaldehyde and was also inhibited in 10-6C cells. Radioactive labelling and Northern blotting demonstrated that the interruption of the Calvin cycle suppressed the synthesis de novo of chloroplast proteins, such as the D1 and D2 proteins, but did not affect the levels of psbA and psbD mRNAs. Our results suggest that the photoinactivation of PSII that is associated with the interruption of the Calvin cycle is attributable primarily to the inhibition of the protein synthesis-dependent repair of PSII at the level of translation in chloroplasts.