Analysis of the photosynthetic response induced by variation potential in geranium

Electrical signals (action and variation potentials) caused by environmental stimuli induce a number of physiological responses in plants including changes in photosynthesis; however, mechanisms of these changes remain unclear. We investigated the influence of the variation potential on photosynthesis in geranium (Pelargonium zonale) under different conditions (control, low external CO₂ concentration, and actinic light absence). The variation potential caused by lamina burning induced a reduction in photosynthesis (decreases in effective quantum yields of photosystem I and II, CO₂ assimilation rate, and stomatal conductance) in unstimulated leaves under control conditions. Changes in the majority of light-stage parameters (photosystem I and II quantum yields, coefficients of photochemical and non-photochemical quenching, quantum yield of non-photochemical energy dissipation in photosystem I due to donor-side limitation) were correlated with a decrease in CO₂ assimilation rate. The changes were similar to those caused by lowering [CO₂]; their magnitudes decreased both under low external CO₂ concentration and without actinic light. These results support the hypothesis that Calvin cycle inactivation plays a key role in photosynthetic response induced by electrical signals. However, a decrease in electron transport through the PSI acceptor side also induced by variation potential was not correlated with a decrease in the CO₂ assimilation rate and did not depend on the external CO₂ concentration or actinic light intensity. Thus, we suggest that there are two different mechanisms of light-stage inactivation induced by the variation potential in geranium: one strongly dependent on dark-stage inactivation and one weakly dependent on dark-stage inactivation.     

Preservation of photosynthetic electron transport from senescence-induced inactivation in primary leaves after decapitation and defoliation of bean plants

The comparative effects of decapitation and defoliation on the senescence-induced inactivation of photosynthetic activity in primary leaves of bean plants were investigated. Decapitation was performed during different phases of bean plant ontogenesis, immediately after the appearance of the 1st, 2nd, 3rd and 4th composite leaf. In addition, we examined a variant with primary leaves and stem with an apical bud, but without composite leaves, i.e. defoliated plants. Analyses of chlorophyll fluorescence, millisecond delayed fluorescence and absorption at 830nm in primary leaves were undertaken to investigate the alterations in photosystems II and I electron transport during the decapitation-induced delayed senescence in the non-detached leaves. Analysis of the OKJIP transients using the JIP-test (see [Strasser R, Srivastava A, Tsimilli-Michael M. Analysis of the chlorophyll a fluorescence transient. In: Papageorgiou G, Govindjee, editors. Chlorophyll a fluorescence: a signature of photosynthesis. The Netherlands: Kluwer Academic Publishers, 2004; pp. 321-362]) showed an increase in several biophysical parameters of photosystem II in decapitated plants, specifically, the density of active reaction centers on a chlorophyll basis, the yields of trapping and electron transport, and the performance index. We also observed a decrease in the absorbed light energy per reaction center. Such a decrease in light absorption could be a result of the photosystem II down regulation that appeared as an increase in Q(B)-non-reducing photosystem II centers. The effect was identical when all leaves except the primary leaves were removed. The variant with a preserved apical bud, the defoliated plant, showed values similar to those of decapitated plants with primary leaves only. The changes in the induction curves of the delayed fluorescence also indicated an acceleration of electron transport beyond photosystem II in the decapitated and in defoliated plants. In these plants, the photosystem I-driven electron transport was accelerated, and the size of the plastoquinone pool was enhanced. It was established that decapitation can retard the senescence of primary leaves, can expand leaf life span and can cause activation of both photosystems I and II electron transport. The decapitation procedure shows similarities to the process of defoliation. The overcompensation effect that is developed after defoliation could initially be manifested as an acceleration of the linear photosynthetic electron flow in the rest of the leaves.     

Processes contributing to photoprotection of grapevine leaves illuminated at low temperature

Photoinactivation of photosystem II (PSII) and energy dissipation at low leaf temperatures were investigated in leaves of glasshouse-grown grapevine ( Vitis vinifera L. cv. Riesling). At low temperatures (< 15°C), photosynthetic rates of CO₂ assimilation were reduced. However, despite a significant increase in the amount of light excessive to that required by photosynthesis, grapevine leaves maintained high intrinsic quantum efficiencies of PSII ( F _v/ F _m) and were highly resistant to photoinactivation compared to other species. Non-photochemical energy dissipation involving xanthophylls and fast D1 repair were the main protective processes reducing the 'gross' rate of photoinactivation and the 'net' rate of photoinactivation, respectively. We developed an improved method of energy dissipation analysis that revealed up to 75% of absorbed light is dissipated thermally via pH- and xanthophyll-mediated non-photochemical quenching at low temperatures (5–15°C) and moderate (800 µmol quanta m⁻² s⁻¹) light. Up to 20% of the energy flux contributing to electron transport was dissipated via photorespiration when taking into account temperature-dependent mesophyll conductance; however, this flux used in photorespiration was only a relatively small amount of the total absorbed light energy. Photoreduction of O₂ at photosystem I (PSI) and subsequent superoxide detoxification (water-water cycle) was more sensitive to inhibition by low temperature than photorespiration. Therefore the water-water cycle represents a negligibly small energy sink below 15°C, irrespective of mesophyll conductance.     

Relationship between photosystem II activity and CO2 fixation in leaves

There is now potential to estimate photosystem II (PSII) activity in vivo from chlorophyll fluorescence measurements and thus gauge PSII activity per CO₂ fixed. A measure of the quantum yield of photosystem II, Φ_II (electron/photon absorbed by PSII), can be obtained in leaves under steady-state conditions in the light using a modulated fluorescence system. The rate of electron transport from PSII equals Φ_II times incident light intensity times the fraction of incident light absorbed by PSII. In C₄ plants, there is a linear relationship between PSII activity and CO₂ fixation, since there are no other major sinks for electrons; thus measurements of quantum yield of PSII may be used to estimate rates of photosynthesis in C₄ species. In C₃ plants, both CO₂ fixation and photorespiration are major sinks for electrons from PSII (a minimum of 4 electrons are required per CO₂, or per O₂ reacting with RuBP). The rates of PSII activity associated with photosynthesis in C₃ plants, based on estimates of the rates of carboxylation (v_o) and oxygenation (v_o) at various levels of CO₂ and O₂, largely account for the PSII activity determined from fluorescence measurements. Thus, in C₃ plants, the partitioning of electron flow between photosynthesis and photorespiration can be evaluated from analysis of fluorescence and CO₂ fixation.     

Photomovement of the red alga Porphyridium cruentum (Ag.) Naegeli

Photophobic reactions of the red alga Porphyridium cruentum have been studied by single cell observations and by population experiments with the light trap method. In white light traps photoaccumulation is saturated at about 6000 lx. Experiments with monochromatic light demonstrate the necessity of carefully separating the three basic light reactions, viz. phototaxis, photokinesis and photophobic response by an appropriate experimental set-up: In single-beam experiments trap wavelengths >695 nm cause photodispersal which is not due to photophobic entrance reactions, but is exclusively due to the positive photokinetic effect of the trap light. This photodispersal can be cancelled by a photokinetically active background light. In the short wavelength range not only photokinesis, but also phototaxis interferes with photophobic reactions thus affecting the density of photoaccumulations in the light trap. Phototactic and photokinetic interference can be avoided by a blue background light. The action spectrum measured this way indicates activity of photosystem I and photosystem II pigments in the perception of the step-down photophobic stimulus. Varying the wavelength of the background light at constant trap light absorbed mainly by photosystem I or photosystem II respectively, efficient spill-over of light energy from photosystem II to the light reaction of photosystem I could be demonstrated. From the results it is concluded that phobic reactions are induced by a decrease of the electron flow rate in the linear electron transport chain.     

The PsbU subunit of photosystem II stabilizes energy transfer and primary photochemistry in the phycobilisome-photosystem II assembly of Synechocystis sp. PCC 6803

The PsbU subunit of photosystem II (PSII) is one of three extrinsic polypeptides associated with stabilizing the oxygen evolving machinery of photosynthesis in cyanobacteria. We investigated the influence of PsbU on excitation energy transfer and primary photochemistry by spectroscopic analysis of a PsbU-less (or deltaPsbU) mutant. The absence of PsbU was found to have multiple effects on the excited state dynamics of the phycobilisome and PSII. DeltaPsbU cells exhibited decreased variable fluorescence when excited with light absorbed primarily by allophycocyanin but not when excited with light absorbed primarily by chlorophyll a. Fluorescence emission spectra at 77 K showed evidence for impaired energy transfer from the allophycocyanin terminal phycobilisome emitters to PSII. Picosecond fluorescence decay kinetics revealed changes in both allophycocyanin and PSII associated decay components. These changes were consistent with a decrease in the coupling of phycobilisomes to PSII and an increase in the number of closed PSII reaction centers in the dark-adapted deltaPsbU mutant. Our results are consistent with the assumption that PsbU stabilizes both energy transfer and electron transport in the PBS/PSII assembly.     

Analysis of chlorophyll a fluorescence and proteomic differences of rice leaves in response to photooxidation

This study investigated the effects of increased sunlight on photooxidation of rice leaf mutant 812HS and its wild-type 812S under field conditions. Light is important for plant growth and development. However, when the absorbed energy exceeds the capacity of utilization of photosynthesis, it leads to the accumulation of singlet oxygen molecules and other reactive oxygen species, which causes oxidative damage. Chlorophyll a fluorescence was applied to examine photosystem II photochemistry. The results demonstrated that intensive light had a negative influence on plant photosynthetic processes. However, the electron transport chain was inhibited and energy dissipation was increased, which can minimize photooxidative damage to the optical system. Meanwhile, proteomic analysis showed that the differential expression of proteins in response to photooxidation participated in multiple pathways, including ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) large subunit, RuBisCO large chain precursor, RuBisCO activase, flavodoxin-like quinone reductase 1, l-ascorbate peroxidase S, oxygen-evolving complex protein 1, and glycolate oxidase. The results indicated that photooxidation induced a response in the rice via the stress-related pathway. The aforementioned proteins, identified by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), may be very useful in comprehending how plants respond to photooxidation and can be used as characteristics of stress-induced signals. The results of chlorophyll fluorescence parameter analysis demonstrated the negative influence of intense light on plant photosynthetic processes. This was evidenced by the dissipation of excessive energy and the suppression of the electron transport chain to minimize photooxidative damage to the proteins. Future studies should compare the proteomic difference with parallel gene expression and metabolite profiles.     

缺铁对大豆叶片光合作用和光系统Ⅱ功能的影响

通过气体交换和叶绿素荧光测定研究了缺铁对大豆叶片碳同化和光系统Ⅱ的影响。缺铁条件下大豆光合速率(Pn)大幅下降;最大光化学效率(po)下降幅度较小;荧光诱导动力学曲线发生明显的变化,其中电子传递活性明显下降,K相(VK)相对荧光产量提高。缺铁大豆的天线转化效率(Fv'/Fm')、光化学猝灭系数(qP)和光系统Ⅱ实际光化学效率(ΦPSⅡ)降低,而非光化学猝灭(NPQ)则明显增加。此外,缺铁大豆的光后荧光上升增强。据此,认为铁缺乏伤害了光系统Ⅱ复合物供体侧和受体侧的电子传递;缺铁条件下光系统I环式电子传递的增强可能在维持激发能耗散和ATP供给方面起一定作用。     

Regulation of photosynthetic electron transport and photophosphorylation in intact chloroplasts and leaves of Spinacia oleracea L.

Oxygen ist reduced by the electron transport chain of chloroplasts during CO₂ reduction. The rate of electron flow to oxygen is low. Since antimycin A inhibited CO₂-dependent oxygen evolution, it is concluded that cyclic photophosphorylation contributes ATP to photosynthesis in chloroplasts which cannot satisfy the ATP requirement of CO₂ reduction by electron flow to NADP and to oxygen. Inhibition of photosynthesis by antimycin A was more significant at high than at low light intensities suggesting that cyclic photophosphorylation contributes to photosynthesis particularly at high intensities. Cyclic electron flow in intact chloroplasts is under the control of electron acceptors. At low light intensities or under far-red illumination it is decreased by substrates which accept electrons from photosystem I such as oxaloacetate, nitrite or oxygen. Obviously, the cyclic electron transport pathway is sensitive to electron drainage. In the absence of electron acceptors, cyclic electron flow is supported by far-red illumination and inhibited by red light. The inhibition by light exciting photosystem II demonstrated that the cyclic electron transport pathway is accessible to electrons from photosystem II. Inhibition can be relieved by oxygen which appears to prevent over-reduction of electron carriers of the cyclic pathway and thus has an important regulatory function. The data show that cyclic electron transport is under delicate redox control. Inhibition is caused both by excessive oxidation and by over-reduction of electron carriers of the pathway.     

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.     

Allocation of absorbed light energy in PSII to thermal dissipations in the presence or absence of PsbS subunits of rice

The thermal dissipation (TD) of absorbed light energy in PSII is considered to be an important photoprotection process in photosynthesis. A major portion of TD has been visualized through the analysis of Chl fluorescence as energy quenching (qE) which depends on the presence of the PsbS subunit. Although the physiological importance of qE-associated TD (qE-TD) has been widely accepted, it is not yet clear how much of the absorbed light energy is dissipated through a qE-associated mechanism. In this study, the fates of absorbed light energy in PSII with regard to different TD processes, including qE-TD, were quantitatively estimated by the typical energy allocation models using transgenic rice in which psbS genes were silenced by RNA interference (RNAi). The silencing of psbS genes resulted in a decrease in the light-inducible portion of TD, whereas the allocation of energy to electron transport did not change over a wide range of light intensities. The allocation models indicate that the energy allocated to qE-TD under saturating light is 30-50%. We also showed that a large portion of absorbed light energy is thermally dissipated in manners that are independent of qE. The nature of such dissipations is discussed.     

Regulation of cyanobacterial photosynthesis determined from variable fluorescence yields of photosystem II

Abstract The cyanobacteria Fremyella diplosiphon 7601 and Synechocystis 6701 were grown in continuous cultures with monochromatic red light (680 nm). The distribution of light energy over photosystem I and II was determined from changes in PS II fluorescence at 685 nm. In both organisms, wavelengths absorbed primarily by chlorophyll a caused the high fluorescent state of PS II (State 1), while wavelengths absorbed by the phycobilisome led to low PS II fluorescence (State 2). Superimposing continuous light 2 on the excitation light yielded State 2 fluorescence patterns for Synechocystis 6701, while F. diplosiphon 7601 showed fluorescence patterns similar to state 1 → 2 transitions and changes in fluorescence yield were related to the intensity of the background light. Some ecological implications of energy (re)distribution in cyanobacterial photosynthesis are discussed.     

Effect of cadmium on photosystem II activity in <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>: alteration of O–J–I–P fluorescence transients indicating the change of apparent activation energies within photosystem II

In this study, we evaluated how cadmium inhibitory effect on photosystem II and I electron transport may affect light energy conversion into electron transport by photosystem II. To induce cadmium effect on the photosynthetic apparatus, we exposed Chlamydomonas reinhardtii 24 h to 0–4.62 μM Cd²⁺. By evaluating the half time of fluorescence transients O–J–I–P at different temperatures (20–30°C), we were able to determine the photosystem II apparent activation energies for different reduction steps of photosystem II, indicated by the O–J–I–P fluorescence transients. The decrease of the apparent activation energies for PSII electron transport was found to be strongly related to the cadmium-induced inhibition of photosynthetic electron transport. We found a strong correlation between the photosystem II apparent activation energies and photosystem II oxygen evolution rate and photosystem I activity. Different levels of cadmium inhibition at photosystem II water-splitting system and photosystem I activity showed that photosystem II apparent activation energies are strongly dependent to photosystem II donor and acceptor sides. Therefore, the oxido-reduction state of whole photosystem II and I electron transport chain affects the conversion of light energy from antenna complex to photosystem II electron transport.     

Photorespiration is More Effective than the Mehler Reaction in Protecting the Photosynthetic Apparatus against Photoinhibition

I. Isolated intact chloroplasts: Photosystem II, but not photosystem I, of the electron transport chain is rapidly photoinactivated even by very low intensities of red light when no large proton gradient can be formed and the electron transport chain becomes over-reduced in the absence of oxygen and other reducable substrates. Electron acceptors including oxygen provide protection against photoinactivation. Nevertheless, photosystem II is rapidly, and photosystem I more slowly, photoinactivated by high intensities of red light when oxygen is the only electron acceptor available. Increased damage is observed at increased oxygen concentrations although catalase is added to destroy H₂O₂ formed during oxygen reduction in the Mehler reaction. Photoinactivation can be decreased, but not prevented by ascorbate which reduces hydrogen peroxide inside the chloroplasts and increases coupled electron flow. II. Leaves: Simple measurements of chlorophyll fluorescence permit assessment of damage to photosystem II after exposure of leaves to high intensity illumination. In contrast to isolated chloroplasts, chloroplasts suffer more damage in situ at reduced than at elevated oxygen concentrations. The difference in the responses is due to photorespiration which is active in leaves, but not in isolated chloroplasts. After photosynthesis and photorespiration are inhibited by feeding glyceraldehyde to leaves, photoinactivation is markedly increased, although oxygen reduction in the Mehler reaction is not affected by glyceraldehyde. In the presence of reduced CO₂ levels, photorespiratory reactions, but not the Mehler reaction, can prevent the overreduction of the electron transport chain. Over-reduction indicates ineffective control of photosystem II activity. Effective control is needed for protection of the electron transport chain against photoinactivation. It is suggested to be made possible by coupled cyclic electron flow around photosystem I which is facilitated by the redox poising resulting from the interplay between photorespiratory carbohydrate oxidation and the refixation of evolved CO₂.     

外源钙对干旱胁迫下烤烟幼苗光系统Ⅱ功能的影响

以叶绿素快相荧光动力学曲线(OJIP)为探针,研究了外源钙对干旱胁迫下烤烟幼苗光系统Ⅱ(PSⅡ)功能的影响.结果表明:干旱胁迫降低了烤烟幼苗PSⅡ原初光能转换效率(Fv/Fm)和电子传递速率(ETR),抑制了光合作用的原初过程,烤烟幼苗叶片发生了明显的光抑制.叶面喷施10.0 mmol·L-1CaCl2溶液后烤烟叶片的光合电子传递能量比例(ФEo)在干旱胁迫下的降低幅度明显小于对照(喷施清水),电子转运效率(ET0/RC)在干旱胁迫下明显高于对照.叶面喷施CaC12溶液增加了PSⅡ捕获光能用于光合电子传递的比例、剩余有活性反应中心的效率和电子传递链中的能量传递,使烤烟叶片的光系统Ⅱ在干旱胁迫下保持相对较高的活性,从而提高了烤烟幼苗的抗旱能力.     

The effect of inhibitors on the electron flow triggering photo-phobic reactions in cyanophyceae

1. Since photo-phobic reactions in the blue green alga Phormidium uncinatum seem to be triggered by changes of electron flow rates into or out of an electron pool situated in the electron transport chain between photosystem II and I, the effect of inhibitors affecting the electron transport chain has been studied.
2. Dose response curves of the phobic reaction have been measured by varying the trap energy in double beam light trap experiments with constant pairs of monochromatic light. From these dose response curves the effects of the inhibitors on both types of phobic reactions, i.e. exit reactions and entrance reactions, have been calculated.
3. Dibromothymoquinone (DBMIB) inhibits the electron transport between the electron pool and photosystem I by preventing the reoxidation of plastoquinone. The phobic entrance reaction, which results in an emptying of the light trap, is triggered by changes in the electron flow out of the pool; thus it is more effected by DBMIB than the exit reaction, which is mediated by the electron transport into the pool.
4. The phobic exit reaction, which results in accumulations in the light trap, is triggered by changes in the electron flow into the electron pool via photosystem II. 3-[3,4-dichlorophenyl]-1,1-dimethylurea (DCMU) inhibits the electron transport near photosystem II; thus it affects the exit reaction more than the entrance reaction.

     

4种木本植物叶片的光合电子传递和吸收光能分配特性对光强的适应

以气体交换和叶绿素荧光测定相结合的方法研究了亚热带自然林乔木荷树、黧蒴和林下灌木九节、罗伞幼苗的光合电子传递及激发能利用的分配对生长光强的适应特性。4种植物生长于100%、36%和16%的自然光下8个月,叶片的光化学速率和热能耗散速率随光强增大而提高,热能耗散占总的光能吸收的比例也因光强不同而改变,16%光下的相对热耗散率约为40%～45%,100%自然光下增大至50%～75%。叶片总的非环式电子流速率及其分配到光呼吸的比例在100%光强下最高。乔木和灌木的电子传递和光能分配特性在16%光下相似,在100%光下差别较明显。除灌木种有较高的热耗散比例之外,其余的参数皆比乔木的低。结果表明乔木与灌木皆可通过提高激发能热耗散比例和提高光合电子传递向光呼吸的比例来适应于高光强条件。     

Protection of the photosynthetic apparatus against damage by excessive illumination in homoiohydric leaves and poikilohydric mosses and lichens

Experimental work on the control of photosystem II in the photosynthetic apparatus of higher plants, mosses and lichens is reviewed on a background of current literature. Transmembrane proton transport during photoassimilatory and photorespiratory electron flows is considered insufficient for producing the intrathylakoid acidification necessary for control of photosystem II activity under excessive illumination. Oxygen reduction during the Mehler reaction is slow. Together with associated reactions (the water-water cycle), it poises the electron transport chain for coupled cyclic electron transport rather than acting as an efficient electron sink. Coupled electron transport not accompanied by ATP consumption in associated reactions provides the additional thylakoid acidification needed for the binding of zeaxanthin to a chlorophyll-containing thylakoid protein. This results in the formation of energy-dissipating traps in the antennae of photosystem II. Competition for energy capture decreases the activity of photosystem II. In hydrated mosses and lichens, but not in leaves of higher plants, protein protonation and zeaxanthin availability are fully sufficient for effective energy dissipation even when photosystem II reaction centres are open. In leaves, an additional light reaction is required, and energy dissipation occurs not only in the antennae but also in reaction centres. Loss of chlorophyll fluorescence during the drying of predarkened poikilohydric mosses and lichens indicates energy dissipation in the dry state which is unrelated to protonation and zeaxanthin availability. Excitation of photosystem II by sunlight is not destructive in these dry organisms, whereas photosystem II activity of dried leaves is rapidly lost under strong illumination.