Importance of AOX pathway in optimizing photosynthesis under high light stress: role of pyruvate and malate in activating AOX

The present study shows the importance of alternative oxidase (AOX) pathway in optimizing photosynthesis under high light (HL). The responses of photosynthesis and respiration were monitored as O₂ evolution and O₂ uptake in mesophyll protoplasts of pea pre‐incubated under different light intensities. Under HL (3000 µmol m^?2 s^?1), mesophyll protoplasts showed remarkable decrease in the rates of NaHCO₃‐dependent O₂ evolution (indicator of photosynthetic carbon assimilation), while decrease in the rates of respiratory O₂ uptake were marginal. While the capacity of AOX pathway increased significantly by two fold under HL, the capacity of cytochrome oxidase (COX) pathway decreased by >50% compared with capacities under darkness and normal light (NL). Further, the total cellular levels of pyruvate and malate, which are assimilatory products of active photosynthesis and stimulators of AOX activity, were increased remarkably parallel to the increase in AOX protein under HL. Upon restriction of AOX pathway using salicylhydroxamic acid (SHAM), the observed decrease in NaHCO₃‐dependent O₂ evolution or p‐benzoquinone (BQ)‐dependent O₂ evolution [indicator of photosystem II (PSII) activity] and the increase in total cellular levels of pyruvate and malate were further aggravated/promoted under HL. The significance of raised malate and pyruvate levels in activation of AOX protein/AOX pathway, which in turn play an important role in dissipating excess chloroplastic reducing equivalents and sustenance of photosynthetic carbon assimilation to balance the effects of HL stress on photosynthesis, was depicted as a model.     

Photosynthesis,chlorophyll fluorescence,and antioxidant enzyme responses of invasive weed <Emphasis Type="Italic">Mikania micrantha</Emphasis> to <Emphasis Type="Italic">Bemisia tabaci</Emphasis> infestation

In a glasshouse, Bemisia tabaci infestation largely reduced response of photosynthesis to irradiance and CO₂ concentration of Mikania micrantha compared with the non-infested control (C) ones. The maximum irradiance-saturated photosynthetic rate (P _max) and saturation irradiance (SI) of the infested M. micrantha were only 21.3 % and 6.5 % of the C-plants, respectively. B. tabaci infestation led to the reduction of contents of chlorophyll and carotenoids in M. micrantha, which was accompanied with the decrease of actual photosystem 2 (PS2) efficiency (Φ_PS2), efficiency of excitation energy capture by open PS2 reaction centres (F_v′/F_m′), electron transport rate (ETR), and photochemical quenching (q_P). Moreover, superoxide dismutase and catalase activities significantly decreased while proline and glutathione contents significantly increased in infested M. micrantha. Hence B. tabaci infestation not only induced direct damage of photosynthetic apparatus but also altered the antioxidant enzymes activities in M. micrantha, which might as consequences accelerate senescence of this weed.     

Role of calcium ion in protection against heat and high irradiance stress-induced oxidative damage to photosynthesis of wheat leaves

Cross stress of heat and high irradiance (HI) resulted in the accumulation of active oxygen species and photo-oxidative damage to photosynthetic apparatus of wheat leaves during grain development. Pre-treatment with calcium ion protected the photosynthetic system from oxidative damage by reducing O^-. ₂ production, inhibiting lipid peroxidation, and retarding electrolyte leakage from cell. Therefore, high F_v/F_m [maximal photochemical efficiency of photosystem 2 (PS2) while all PS2 reaction centres are open], F_m/F₀ (another expression for the maximal photochemical efficiency of PS2), Φ_PS2 (actual quantum yield of PS2 under actinic irradiation), q_P (photochemical quenching coefficient), and P _N (net photosynthetic rate) were maintained, and lower q_NP (non-photochemical quenching coefficient) of the leaves was kept under heat and HI stress. EGTA (a chelant of calcium ion) and LaCl₃ (a blocker of Ca²⁺ channel in cytoplasmic membrane) had the opposite effect. Thus Ca ion may help protect the photosynthetic system of wheat leaves from oxidative damage induced by the cross stress of heat and HI.     

Protective Effects of Exogenous Antioxidants and Phenolic Compounds on Photosynthesis of Wheat Leaves under High Irradiance and Oxidative Stress

Infiltration of methyl viologen (MV, source of O₂ ^–) and Na-diethyldithiocarbamate (DDC, inhibitor of SOD) into wheat leaves resulted in the accumulation of active oxygen species and photo-oxidative damage to photosynthetic apparatus under both moderate and high irradiance. Exogenous antioxidants, ascorbate (ASA) and mannitol, scavenged active oxygen efficiently, protected the photosynthetic system from MV and DDC induced oxidative damage, and maintained high F_v/F_m [maximal photochemical efficiency of photosystem 2 (PS2) while all PS2 reaction centres are open], F_m/F₀ (another expression for the maximal photochemical efficiency of PS2), _PS2 (actual quantum yield of PS2 under actinic irradiation), q_P (photochemical quenching coefficient), P _N (net photosynthetic rate), and lowered q_NP (non-photochemical quenching coefficient) of the leaves kept under high irradiance and oxidative stress. Phenolic compounds used in these experiments, catechol (Cat), resorcinol (Res), and tannic acid (Tan), had similar anti-oxidative activity and protective effect on photosynthetic apparatus as ASA and mannitol. The anti-oxidative activity and the protective effect of phenolic compounds increased with increase in their concentration from 100 to 300 g m^–3. The number and the position of hydroxyl group in phenolic molecules seemed to influence their antioxidative activity.     

Effect of Cd on growth, photosynthetic gas exchange, and chlorophyll fluorescence of wild and Cd-sensitive mutant rice

Growth, photosynthetic gas exchange, and chlorophyll fluorescence characteristics were investigated in wild type (WT) and Cd-sensitive mutant rice (Oryza sativa L.) plants using 50 μM Cd treatment for 12 d followed by a 3-d recovery. Under Cd stress, net dry mass and pigment contents were significantly lower in the mutant plants than in the WT. The mutant had lower net photosynthetic rate (P _N), transpiration rate (E), and stomatal conductance (g _s) than WT rice, however, it had higher intercellular CO₂ concentration (C _i), indicating that non-stomatal factors accounted for the inhibition of P _N. Maximal photochemical efficiency of photosystem 2 (F_v/F_m), effective quantum yield of PS2 (Φ_PS2), and photochemical quenching (q_P) decreased much in the mutant under Cd stress. Cd content in roots and leaves of the mutant was significantly higher than those in the WT. Hence Cd toxicity was associated with the marked increases in Cd contents of plant tissue. After the recovery for 3 d, the WT rice had higher capacity to recover from Cd injury than the mutant.     

Exogenous application of trehalose induced H2O2 production and stomatal closure in Vicia faba

Trehalose can reduce stomatal aperture by a hydrogen-peroxide-dependent pathway in Vicia faba L. (cv. Daqingpi) resulting in significantly lower values of net photosynthetic rate (P_N), stomatal conductance (g_s), and transpiration rate (E). At 8 and 24 h, the lower P_N in trehalose-treated plants was accompanied by significant decrease in intercellular CO₂ concentration (c_i) suggesting that the reduction of P_N was caused by stomatal limitation. At 48 and 72 h, trehalose decreased apparent carboxylation efficiency (P_N/c_i) and did not decrease c_i and g_s compared with controls; therefore the reduction in photosynthesis was caused by non-stomatal limitation. Trehalose treatment resulted in significantly higher effective photochemical efficiency of PS II (Φ_PSII) and did not affect maximum photochemical efficiency of PS II (F_v/F_m). At 24, 48, and 72 h, trehalose decreased non-photochemical quenching (NPQ) and increased photochemical quenching (qP). Our results suggest that trehalose did not damage photosynthetic reaction centers.     

High photosynthetic efficiency of a rice (Oryza sativa L.) xantha mutant

Comparative analysis revealed that a xantha rice mutant (cv. Huangyu B) had higher ratios of chlorophyll (Chl) a/b and carotenoids/Chl, and higher photosynthetic efficiency than its wild type parent (cv. II32 B). Unexpectedly, the mutant had higher net photosynthetic rate (P _N) than II32 B. This might have resulted from its lower non-photochemical quenching (q_N) but higher maximal photochemical efficiency (F_V/F_M), higher excitation energy capture efficiency of photosystem 2 (PS2) reaction centres (F_V′/F_M′), higher photochemical quenching (q_P), higher effective PS2 quantum yield (Φ_PS2), and higher non-cyclic electron transport rate (ETR). This is the first report of a chlorophyll mutant that has higher photosynthetic efficiency and main Chl fluorescence parameters than its wild type. This mutant could become a unique material both for the basic research on photosynthesis and for the development of high yielding rice cultivars.     

杂交酸模叶片线粒体交替氧化酶呼吸途径在光破坏防御中的作用

以杂交酸模(Rumex K-1)为试材,研究了不同光强下线粒体交替氧化酶呼吸途径(AOX途径)对酸模叶片光破坏的防御作用.结果表明:在200 μmol·m-2·s-1弱光下,用水杨基羟肟酸抑制AOX途径后,Rumex K-1叶片的PSⅡ实际光化学效率、光合线性电子传递速率以及光合放氧速率均显著下降,非还原性QB反应中心显著升高,加重了叶片的光抑制,而活性氧清除机制上调,避免了活性氧的过量积累,部分缓解了Rumex K-1叶片的光抑制;在800 μmol·m-2·s-1强光下,AOX途径受抑,导致Rumex K-1叶片发生严重的光抑制,而此时活性氧清除机制的上调不足以缓解活性氧过量的积累.无论在强光还是弱光下,AOX途径在Rumex K-1叶片的光破坏防御过程中都起着重要作用,而且在强光下,AOX途径对叶片的光破坏防御作用是叶绿体内其他光破坏防御途径所不能代替的.     

Photosynthetic responses of radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> var. <Emphasis Type="Italic">longipinnatus</Emphasis>) plants to infection by turnip mosaic virus

Plant growth, chlorophyll (Chl) content, photosynthetic gas exchange, ribulose-1,5-bisphosphate carboxylase (RuBPCO) enzyme activity, and Chl fluorescence in radish (Raphanus sativus var. longipinnatus) plants were examined after turnip mosaic virus (TuMV) infection. Plant fresh mass, dry mass, Chl content, net photosynthetic rate (P _N), transpiration rate (E), stomatal conductance (g _s), and RuBPCO activity were significantly lower in infected plants after 5 weeks of virus infection as compared to healthy plants. The 5-week virus infection did not induce significant differences in intercellular CO₂ concentration (C _i, photochemical efficiency of photosystem 2, PS2 (F_v/F_m), excitation capture efficiency of open PS2 reaction centres (F_v'/F_m'), effective quantum efficiency of photosystem 2 (ΔF/F_m'), and photochemical quenching (q_P), but non-photochemical quenching (q_N) and alternative electron sink (AES) were significantly enhanced. Thus the decreased plant biomass of TuMV-infected plants might be associated with the decreased photosynthetic activity mainly due to reduced RuBPCO activity.     

Enhancement of susceptivity to photoinhibition and photooxidation in rice chlorophyll <Emphasis Type="Italic">b</Emphasis>-less mutants

Two rice chlorophyll (Chl) b-less mutants (VG28-1, VG30-5) and the respective wild type (WT) plant (cv. Zhonghua No. 11) were analyzed for the changes in Chl fluorescence parameters, xanthophyll cycle pool, and its de-epoxidation state under exposure to strong irradiance, SI (1 700 μmol m⁻² s⁻¹). We also examined alterations in the chloroplast ultrastructure of the mutants induced by methyl viologen (MV) photooxidation. During HI (0–3.5 h), the photoinactivation of photosystem 2 (PS2) appeared earlier and more severely in Chl b-less mutants than in the WT. The decreases in maximal photochemical efficiency of PS2 in the dark (F_v/F_m), quantum efficiency of PS2 electron transport (Φ_PS2), photochemical quenching (q_P), as well as rate of photochemistry (P_rate), and the increases in de-epoxidation state (DES) and rate of thermal dissipation of excitation energy (D_rate) were significantly greater in Chl b-mutants compared with the WT plant. A relatively larger xanthophyll pool and 78–83 % conversion of violaxanthin into antheraxanthin and zeaxanthin in the mutants after 3.5 h of HI was accompanied with a high ratio of inactive/total PS2 (0.55–0.73) and high 1–q_P (0.57–0.68) which showed that the activities of the xanthophyll cycle were probably insufficient to protect the photosynthetic apparatus against photoinhibition. No apparent difference of chloroplast ultrastructure was found between Chl b-less mutants and WT plants grown under low, LI (180 μmol m⁻² s⁻¹) and high, HI (700 μmol m⁻² s⁻¹) irradiance. However, swollen chloroplasts and slight dilation of thylakoids occurred in both mutants and the WT grown under LI followed by MV treatment. These typical symptoms of photooxidative damage were aggravated as plants were exposed to HI. Distorted and loose scattered thylakoids were observed in particular in the Chl b-less mutants. A greater extent of photoinhibition and photooxidation in these mutants indicated that the susceptibility to HI and oxidative stresses was enhanced in the photosynthetic apparatus without Chl b most likely as a consequence of a smaller antenna size.     

Enhanced photosystem 2 thermostability during leaf growth of Elm (Ulmus pumila) seedlings

We examined photosynthetic activities and thermostability of photosystem 2 (PS2) in leaves of elm (Ulmus pumila) seedlings from initiation to full expansion. During leaf development, net photosynthetic rate (P _N) increased gradually and reached the maximum when leaves were fully developed. In parallel with the increase of P _N, chlorophyll (Chl) content was significantly elevated. Chl a fluorescence measurements showed that the maximum quantum yield of PS2 (ϕ_PS2), the efficiency a trapped exciton, moved an electron into the electron transport chain further than Q_A ⁻ (Ψ_o), and the quantum yield of electron transport beyond Q_A (ϕ_Eo) increased gradually. These results were independently confirmed by our low irradiance experiments. When subjected to progressive heat stress, the young leaves exhibited considerably lower ϕ_PS2 and higher minimal fluorescence (F₀) than the mature leaves, revealing the highly sensitive nature of PS2 under heat in the newly initiating leaves. Further analysis showed that PS2 structure in the newly initiating leaves was strongly altered under heat, as evidenced by the increased fluorescence signals at the position of the K step. We therefore demonstrated an inhibition in the oxygen-evolving complex (OEC) in the young leaves. This resulted in decrease in amount of the functional PS2 reaction centres and relative increase in the PS2 reaction centres with inhibited electron transport at the acceptor side under heat. We suggest that the enhanced thermostability of PS2 during leaf development is associated with improved OEC stability.     

Effects of simulated acid rain on the photosynthetic characteristics of Phaseolus vulgaris L.

A single treatment with a low pH solution of bean plants led to serious changes in the net photosynthetic rate (P _N) as well as in various parameters of photosystem 2 (PS2) activity. A considerable suppression of P _N was established already in the first hours (3 and 5) following the acid treatment (pH 2.4-1.8). The period of strong inhibition of CO₂ uptake and photochemical activity was followed by the period of recovery (24-72 h). At a single spraying, pH values exceeding 2.0 did not lead to irreversible damages of the photosynthetic apparatus. The damages resulting from treatments with pH 2.0 and 1.8 were on the threshold of irreversible ones and were the cause of faster ageing. This revised version was published online in June 2006 with corrections to the Cover Date.     

Photosynthesis of lichen symbiotic alga Trebouxia erici as affected by irradiance and osmotic stress

The relation between oxygen evolution rate (OER) and quantum yield of photochemical reactions in photosystem 2 (Φ_PS2) was examined in lichen symbiotic alga Trebouxia erici Ahmadjian (strain UTEX 911) exposed to different irradiances and osmotic stress (2 M sucrose for 60 h). Linear relationship was found between OER and Φ_PS2 in control cell suspension within irradiance range of 0 – 500 μmol m⁻² s⁻¹. Under osmotic stress, OER and Φ_PS2 were significantly reduced. Relation between OER and Φ_PS2 was curvilinear due to strong osmotically-induced inhibition of OER at high irradiance. The highest used irradiance (500 μmol m⁻² s⁻¹) was photoinhibitory for osmotically-stressed T. erici because non-photochemical quenching (NPQ) increased substantially. Energy-dependent quenching represented major part of NPQ increase. Osmotic stress led also to the reduction of capacity of photochemical processes in PS 2 (F_V/F_M) and increase in F₀/F_M. These changes indicated negative effects of osmoticum on structure and function of photosynthetic apparatus.     

Photosynthetic performance and acclimation of <Emphasis Type="Italic">Incarvillea delavayi</Emphasis> to water stress

The photosynthetic performance and related leaf traits of Incarvillea delavayi Bur. et Franch were studied at different water regimes to assess its capacity for photosynthetic acclimation to water stress. The initial response of I. delavayi to water stress was the closure of stomata, which resulted in down-regulation of photosynthesis. The stomatal limitation (S_L) represented the main component to photosynthetic limitations but non-stomatal limitation (NS_L) increased quickly with the increasing water stress, and had similar magnitude to SL under severe water stress (soil moisture 25–30 % of field capacity). Chlorophyll (Chl) a fluorescence parameters characterizing photosystem (PS) 2 photochemical efficiency (Φ_PS2), electron transport rate (J) and photochemical quenching (qP) decreased with the increasing water stress, indicating impaired photosynthetic apparatus. However, the water-stressed plants had a increased mesophyll CO₂ diffusional conductance, Chl a/b ratio, leaf nitrogen partitioning in RuBPCO and bioenergetics in later grown parts, indicating that I. delavay had a substantial physiological plasticity and showed a good tolerance to water stress.     

Effects of rhizobia inoculation and nitrogen fertilization on photosynthetic physiology of soybean

Plant growth, contents of photosynthetic pigments, photosynthetic gas exchange, and chlorophyll (Chl) fluorescence in soybean [Glycine max (L.) Merr. cv. Heinong37] were investigated after it was inoculated with Sinorhizobium fredii USDA191 or treated with 5 mM (NH₄)₂SO₄ (N5) and 30 mM (NH₄)₂SO₄ (N30), respectively. In the plants following N5 fertilization, not only plant biomass, leaf area, and Chl content, but also net photosynthetic rate (P _N), stomatal conductance (g _s), carboxylation efficiency (CE), maximum photochemical efficiency (F_v/F_m) of photosystem 2 (PS2), and quantum yield of PS2 (Φ_PS2) were markedly improved as compared with the control plants. There were also positive effects on plant growth and plant photosynthesis after rhizobia inoculation, but the effects were much less than those of N5 fertilization. For N30 plants there were no significant positive effects on plant growth and photosynthetic capacity. Plant biomass, P _N, and g _s were similar to those of N-limited (control) plants. Φ_PS2 and photochemical quenching (q_P) were obviously declined while content of carotenoids and non-photochemical quenching (q_N) were significantly enhanced in N30 treated plants. This indicated that excess N supply may cause some negative effects on soybean plants.     

交替呼吸途径对CuCl2胁迫下菜豆叶片光系统Ⅱ的保护作用

以“农普”12号菜豆幼苗为材料,采用1mmol·L^-1的水杨基氧肟酸（SHAM）抑制交替呼吸途径活性,探讨了在CuCl₂胁迫下交替呼吸途径对菜豆幼苗叶片光系统Ⅱ的保护作用。结果表明,随着CuCl₂胁迫浓度的增加,菜豆幼苗叶片潜在最大光化学效率F_v/F_m、光适应下叶片的最大光化学效率F_v'/F_m'、PSⅡ的实际光化学效率Y（Ⅱ）以及光化学猝灭系数qP、叶绿素含量均呈下降趋势,而非光学猝灭系数NPQ和交替呼吸途径的容量水平则呈上升趋势。较之在CuCl₂处理下的菜豆幼苗,用交替呼吸途径抑制剂预处理后的菜豆再置于CuCl₂的胁迫下,则会导致F_v/F_m、F_v'/F_m'、Y（Ⅱ）、qP以及叶绿素含量的进一步下降和NPQ的进一步上升。上述观察表明,在CuCl₂胁迫下交替呼吸途径可以缓解PSⅡ光化学效率的下降、维持PSⅡ反应中心的开放程度、减少天线色素的热耗散以及缓解叶绿素含量的降低,从而保护菜豆叶片光系统Ⅱ免受CuCl₂胁迫的伤害。     

Novel effects of methyl viologen on photosystem II function in spinach leaves

Methyl viologen (MV) is a well-known electron mediator that works on the acceptor side of photosystem I. We investigated the little-known, MV-induced inhibition of linear electron flow through photosystem II (PS II) in spinach-leaf discs. Even a low [MV] decreased the (1) average, light-adapted photochemical efficiency of PS II traps, (2) oxidation state of the primary quinone acceptor Q_A in PS II during illumination, (3) photochemical efficiency of light-adapted open PS II traps, (4) fraction of absorbed light energy dissipated constitutively in a light-independent manner or as chlorophyll (Chl) a fluorescence emission, (5) Chl a fluorescence yield corresponding to dark-adapted open reaction-center traps (F _o) and closed reaction-center traps (F _m), and (6) half-time for re-oxidation of Q_A⁻ in PS II after a single-turnover flash. These effects suggest that the presence of MV accelerates various “downhill” electron-transfer steps in PS II. Therefore, when using the MV to quantify cyclic electron flow, the inhibitory effect of MV on PS II should be taken into account.     

Photosynthesis and chlorophyll fluorescence response to low sink demand of tubers and roots in <Emphasis Type="Italic">Dahlia pinnata</Emphasis> source leaves

Photosynthetic rate (P_N) and chlorophyll (Chl) fluorescence induction of source leaves in response to a low sink demand created by girdling the branch (GB) between the root-tuber-system and the leaves were studied in Dahlia pinnata L. cv. Rigolet during the stage of rapid tuber growth in the greenhouse. GB resulted in significantly lower values of P_N, stomatal conductance (g_s), and transpiration rate (E), but in higher leaf temperature (T_l) compared with those of controls. With exception of maximum quantum yield of photosystem 2 (PS 2) photochemistry (F_v/F_m) and maximum ratio of quantum yields of photochemical and concurrent non-photochemical processes in PS 2 (F_v/F₀), no significant differences were observed in Chl fluorescence parameters between girdled and control leaves on days 1 and 2 after GB, indicating no apparent damage in the photosynthetic apparatus. However, longer girdling duration resulted in higher non-photochemical Chl fluorescence quenching (NPQ), but lower F_v/F₀, actual efficiency of energy conversion in PS 2 under steady-state conditions (Φ_PS2), and photochemical quenching coefficient (qP) in comparison with controls from 10:00 to 16:00 or 15:00 on days 4 and 5, respectively, indicating reversible injury in the photosynthetic apparatus.     

The hydroperoxide lyase branch of the oxylipin pathway protects against photoinhibition of photosynthesis

Main conclusion

This study describes a new role for hydroperoxide lyase branch of oxylipin biosynthesis pathway in protecting photosynthetic apparatus under high light conditions.

Lipid-derived signaling molecules, oxylipins, produced by a multi-branch pathway are central in regulation of a wide range of functions. The two most known branches, allene oxide synthase (AOS) and 13-hydroperoxide lyase (HPL) pathways, are best recognized as producers of defense compounds against biotic challenges. In the present work, we examine the role of these two oxylipin branches in plant tolerance to the abiotic stress, namely excessive light. Towards this goal, we have analyzed variable chlorophyll fluorescence parameters of intact leaves of Arabidopsis thaliana genotypes with altered oxylipin profile, followed by examining the impact of exogenous application of selected oxylipins on functional activity of photosynthetic apparatus in intact leaves and isolated thylakoid membranes. Our findings unequivocally bridge the function of oxylipins to photosynthetic processes. Specifically, HPL overexpressing lines display enhanced adaptability in response to high light treatment as evidenced by lower rate constant of photosystem 2 (PS2) photoinhibition and higher rate constant of PS2 recovery after photoinhibition. In addition, exogenous application of linolenic acid, 13-hydroperoxy linolenic acid, 12-oxophytodienoic acid, and methyl jasmonate individually, suppresses photochemical activity of PS2 in intact plants and isolated thylakoid membranes, while application of HPL-branch metabolites—does not. Collectively these data implicate function of HPL branch of oxylipin biosynthesis pathway in guarding PS2 under high light conditions, potentially exerted through tight regulation of free linolenic acid and 13-hydroperoxy linolenic acid levels, as well as competition with production of metabolites by AOS-branch of the oxylipin pathway.