Mechanisms of non-photochemical chlorophyll fluorescence quenching in higher plants

The excitation energy of pigment molecules in photosynthetic antennae systems is utilised by photochemistry, partly it is thermally dissipated, and partly it is emitted as fluorescence. Changes in the quantum yield of chlorophyll (Chl) fluorescence reflect the changes in quantum yield of photochemical reaction and thermal dissipation of the excitation energy. Decrease of the Chl fluorescence quantum yield is called the Chl fluorescence quenching. The decrease of the quantum yield that is accompanied by photochemical reactions has been termed the photochemical quenching, and the decrease accompanied by thermal dissipation of the excitation energy is called the non-photochemical quenching. This review deals with mechanisms of the non-photochemical quenching.     

Modifications in photosystem II photochemistry in senescent leaves of maize plants

Analyses of CO2 exchange and chlorophyll fluorescence were carried out to assess photosynthetic performance during senescence of maize leaves. Senescent leaves displayed a significant decrease in CO2 assimilatory capacity accompanied by a decrease in stomatal conductance and an increase in intercellular CO2 concentration. The analyses of fluorescence quenching under steady-state photosynthesis showed that senescence resulted in an increase in non-photochemical quenching and a decrease in photo-chemical quenching. It also resulted in a decrease in the efficiency of excitation energy capture by open PSII reaction centres and the quantum yield of PSII electron transport, but had very little effect on the maximal efficiency of PSII photochemistry. The results determined from the fast fluorescence induction kinetics indicated an increase in the proportion of QB-non-reducing PSII reaction centres and a decrease in the rate of QA reduction in senescent leaves. Theoretical analyses of fluorescence parameters under steady-state photosynthesis suggest that the increase in the non-photochemical quenching was due to an increase in the rate constant to thermal dissipation of excitation energy by PSII and that the decrease in the quantum yield of PSII electron transport was associated with a decrease in the rate constant of PSII photochemistry. Based on these results, it is suggested that the decrease in the quantum yield of PSII electron transport in senescent leaves was down-regulated by an increase in the proportion of QB-non-reducing PSII reaction centres and in the non-photochemical quenching. The photosynthetic electron transport would thus match the decreased demand for ATP and NADPH in carbon assimilation which was inhibited significantly in senescent leaves.Key words: Chlorophyll fluorescence, gas exchange, maize (Zea mays L.), photochemical and non-photochemical quenching, photosystem II photochemistry.      

Spatial-temporal variations in rose leaves under water stress conditions studied by chlorophyll fluorescence imaging.

Spatial-temporal changes were examined by imaging chlorophyll (Chl) a fluorescence in four leaf areas, two central and two external of rose plants (Rosa x hybrida) cv. Grand Gala for 9 days, under progressive water stress. New fluorescence parameters based on the lake model have recently been used to determine Q(A) redox state and excitation energy fluxes in order to gain a better understanding of the mechanisms that occur under drought stress. Chlorophyll fluorescence images showed a spatial variation in the leaves. The lower values for F(o), F(M), phi(2), q(P) and q(L) were found in the internal leaf area while higher values of non-photochemical quenching calculated from Stern-Volmer quenching (NPQ) and phi(NPQ). phi(Po) were more homogeneous throughout leaf. Temporal changes were also observed during the experiment, a 10% decrease in relative water content (RWC) (between day 1 and 2), led to a decrease in photochemical quenching and an increase in non-photochemical processes. Chlorophyll fluorescence parameters were more or less constant till day 8. At the end of the experiment (day 9), energy dissipation by downregulation, electron transport and Q(A) redox state, decreased and phi(NO) increased to compensate the change. Chlorophyll fluorescence parameters based on the lake model q(L), phi(NPQ) and phi(NO) have been found more appropriate for estimating the fraction of open centres, the quantum yield of regulated energy dissipation in photosystem II (PSII) and the quantum yield of non-regulated energy dissipation in PSII, respectively. The F(s)/F(o) ratio is strongly correlated with NPQ and phi(NPQ) up to a RWC of 20%. This coincides with a greater decrease in photochemical quenching and non-photochemical quenching and an increase in phi(NO).     

The relationship between non-photochemical quenching of chlorophyll fluorescence and the rate of photosystem 2 photochemistry in leaves

It has been suggested previously that non-photochemical quenching of chlorophyll fluorescence is associated with a decrease in the rate of photosystem 2 (PS 2) photochemistry. In this study analyses of fluorescence yield changes, induced by flashes in leaves exhibiting different amounts of non-photochemical quenching of fluorescence, are made to determine the effect of non-photochemical excitation energy quenching processes on the rate of PS 2 photochemistry. It is demonstrated that both the high-energy state and the more slowly relaxing components of non-photochemical quenching reduce the rate of PS 2 photochemistry. Flash dosage response curves for fluorescence yield show that non-photochemical quenching processes effectively decrease the relative effective absorption cross-section for PS 2 photochemistry. It is suggested that non-photochemical quenching processes exert an effect on the rate of PS 2 photochemistry by increasing the dissipation of excitation energy by non-radiative processes in the pigment matrices of PS 2, which consequently results in a decrease in the efficiency of delivery of excitation energy for PS 2 photochemistry.     

Photoadaptation in the Green Alga Spongiochloris Sp. A Three-Fluorometer Study

The dark-adapted cells of the green alga Spongiochloris sp. were exposed to "white light" of 1000 μmol(photon) m⁻² s⁻¹ for 2 h and then dark adapted for 1.5 h. Changes of photochemical activities during photoadaptation were followed by measurement of chlorophyll (Chl) fluorescence kinetics, 77 K emission spectra, photosynthetic oxygen evolution, and pigment composition. We observed a build-up of slowly-relaxing non-photochemical quenching which led to a decrease of the F_v/F_m parameter and the connectivity. In contrast to the depression of F_v/F_m (35 %) and the rise of non-photochemical quenching (∼ 1.6), we observed an increase in effective absorption cross-section (20 %), Hill reaction (30 %), photosynthetic oxygen evolution (80 %), and electron transport rate estimated from the Chl fluorescence analysis (80 %). We showed an inconsistency in the presently used interpretation schemes, and ascribe the discrepancy between the increase of effective absorption cross-section and the photosynthetic activities on one side and the effective non-photochemical quenching on the other side to the build-up of a quenching mechanism which dissipates energy in closed reaction centres. Such a type of quenching changes the ratio between thermal dissipation and fluorescence without any effect on photochemical yield. In this case the F_v/F_m ratio cannot be used as a measure of the maximum photochemical yield of PS2. This revised version was published online in August 2006 with corrections to the Cover Date.     

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。     

Chlorophyll Fluorescence Parameters: The Definitions,Photosynthetic Meaning,and Mutual Relationships

Chlorophyll fluorescence parameters (Chl FPs) derived from the slow (long-term) induction kinetics of modulated Chl a fluorescence are reviewed and analysed with respect to their application in photosynthesis research. Only four mutually independent Chl FPs, calculated from values of five essential Chl fluorescence (ChlF) yields, are distinguished as the basic ones. These are: the maximum quantum yield of PS2 photochemistry (_{P O}), the photochemical quenching of variable ChlF (q_P), the non-photochemical quenching of variable ChlF (q_N), and the relative change of minimum ChlF (q_O). _{P O} refers to the dark-adapted state of a thylakoid membrane, q_P, q_N and q_O characterise the light-adapted state. It is demonstrated that all other Chl FPs can be determined using this quartet of parameters. Moreover, three FPs related to the non-radiative energy dissipation within thylakoid membranes are evaluated, namely: the non-photochemical ChlF quenching (NPQ), the complete non-photochemical quenching of ChlF (q_CN), and the effective quantum yield of non-photochemical processes in PS2 (_N). New FPs, the total quenching of variable ChlF (q_TV) and the absolute quenching of ChlF (q_A) which allow to quantify co-action of the photochemical and non-photochemical processes during a light period are defined and analysed. The interpretation of Chl FPs and recommendations for their application in the photosynthesis research are also given. Some alternative FPs used in the laboratory practice have only an approximate character and can lead to incorrect conclusions if applied to stressed plants. They are reviewed and compared with the standard ones. All formulae and conclusions discussed herein are verified using experimental values obtained on young seedlings of the Norway spruce (Picea abies [L.] Karst.).     

Non-Radiative Dissipation of Absorbed Excitation Energy Within Photosynthetic Apparatus of Higher Plants

The review deals with thermal dissipation of absorbed excitation energy within pigment-protein complexes of thylakoid membranes in higher plants. We focus on the de-excitation regulatory processes within photosystem 2 (PS2) that can be monitored as non-photochemical quenching of chlorophyll (Chl) a fluorescence consisting of three components known as energy-dependent quenching (q_E), state-transition quenching (q_T), and photoinhibitory quenching (q_I). We summarize the role of thylakoid lumen pH, xanthophylls, and PS2 proteins in q_E mechanism. Further, both the similarity between q_E and q_I and specific features of q_I are described. The other routes of thermal energy dissipation are also mentioned, that is dissipation within photosystem 1 and dissipation through the triplet Chl pathway. The significance of the individual de-excitation processes in protection against photo-oxidative damage to the photosynthetic apparatus under excess photon supply is stretched.     

Photosynthesis and protective mechanisms during ageing in transgenic tobacco leaves with over-expressed cytokinin oxidase/dehydrogenase and thus lowered cytokinin content

The content of cytokinins (CKs), the plant inhibitors of the final phase of plant development, senescence, is effectively controlled by irreversible degradation catalysed by cytokinin oxidase/dehydrogenase (CKX). In transgenic tobacco, denoted as AtCKX, with over-expressed CKX causing lowered CK content, we investigated changes in the time courses of chlorophyll (Chl) and xanthophyll (violaxanthin, antheraxanthin, zeaxanthin, neoxanthin, and lutein) contents. We also determined parameters of slow Chl fluorescence kinetics such as minimum Chl fluorescence yield in the darkadapted state F₀, maximum quantum yield of PS2 photochemistry (F_v/F_m), maximum ratio of quantum yields of photochemical and concurrent non-photochemical processes in photosystem 2 (PS2), F_v/F₀, non-photochemical quenching (NPQ), and effective quantum yield of photochemical energy conversion in PS2 (Φ₂). We used three different developmental leaf stages, old, mature, and young, and compared this with time courses of these characteristics in leaves with natural CK levels. The parameters F_v/F_m, F_v/F₀, and Φ₂ were unchanged during ageing in AtCKX plants in contrast to control ones where a significant decrease in old leaves was found. In control plants F₀ increased during ageing, but in the oldest leaf a considerable decrease was observed. This could indicate progressive damage to PS2 reaction centres and then detachment and rapid degradation of Chl. This is in agreement with time course of Chl content. NPQ decreased with age and was similar in both plant types. We observed a decline of xanthophyll contents in the oldest leaves in both plant types, but the contents were enhanced in AtCKX compared to control plants, especially of neoxanthin. The higher xanthophyll contents in the transgenic plants contribute to a better photoprotection and the fluorescence parameters indicated that photosynthetic apparatus was in better condition compared to control and it consequently postponed the onset of leaf senescence.     

Effects of shading on photosynthetic characteristics and chlorophyll fluorescence parameters in leaves of Hydrangea macrophylla

Aims The objectives were to investigate the effects of different light intensities on photosynthetic characteristics and chlorophyll fluorescence parameters, to clarify the physiological responses and photo-protective mechanisms of Hydrangea macrophylla to changes in light regimes in view of the distribution of energy absorbed and photosynthetic characteristics.Methods Three light regimes including natural and shade (shading rate 50% and 75% of natural light) were applied to plants for 60 days. After the treatment, the gas-exchange, chlorophyll a fluorescence and photosynthesis-light curves were measured by a portable leaf gas exchange system (LI-6400).Important findings The results showed that the weak light intensity treatment reduced dark respiration rate, light compensation point and light saturation point of plant, but increased apparent quantum yield, suggesting that plants had the physiological strategy to utilize the weakening light by reducing respiration. The net photosynthetic rate, intercellular CO₂ concentration, transpiration rate and water use efficiency of plants grown below 50% of natural light showed significant difference compared with natural and shading rate 75% of natural light. There were significant difference between natural and shade treatments in the maximal quantum efficiency of PSII (F_v/F_m), as indicated that it was significantly less at full light than that at 50% of natural light. Initial fluorescence intensity (F_o) of plants was higher at full light than that at 50% of natural light, suggesting that photoinhibition occurred in natural light. The non-photochemical quenching (NQP) decreased with the aggravation of shade stress, indicating that shading decreased the efficiency of photochemical reaction by reducing the fraction of incident light in photochemical energy utilization and decreased thermal dissipation through regulating energy distribution in photosystem II (PSII) in the leaves of Hydrangea macrophylla. In general, the 70% of incident light in photochemical energy utilization was distributed to thermal dissipation, 20% was distributed to non-regulated energy dissipation and 4% was distributed to effective photochemical reaction. In conclusion, responses of plants to increased irradiance are governed by strategy: to utilize a high fraction of incident light in photochemistry and regulate energy dissipation in PSII and weaken the accumulation of excess excitation energy in PSII to protect the photosynthetic apparatus in the leaves of H. macrophylla under saturated radiation.     

Carotenoid-triggered energy dissipation in phycobilisomes of Synechocystis sp. PCC 6803 diverts excitation away from reaction centers of both photosystems

Cyanobacteria are capable of using dissipation of phycobilisome-absorbed energy into heat as part of their photoprotective strategy. Non-photochemical quenching in cyanobacteria cells is triggered by absorption of blue-green light by the carotenoid-binding protein, and involves quenching of phycobilisome fluorescence. In this study, we find direct evidence that the quenching is accompanied by a considerable reduction of energy flow to the photosystems. We present light saturation curves of photosystems’ activity in quenched and non-quenched states in the cyanobacterium Synechocystis sp. PCC 6803. In the quenched state, the quantum efficiency of light absorbed by phycobilisomes drops by about 30-40% for both photoreactions—P700 photooxidation in the photosystem II-less strain and photosystem II fluorescence induction in the photosystem I-less strain of Synechocystis. A similar decrease of the excitation pressure on both photosystems leads us to believe that the core-membrane linker allophycocyanin APC-L_CM is at or beyond the point of non-photochemical quenching. We analyze 77 K fluorescence spectra and suggest that the quenching center is formed at the level of the short-wavelength allophycocyanin trimers. It seems that both chlorophyll and APC-L_CM may dissipate excess energy via uphill energy transfer at physiological temperatures, but neither of the two is at the heart of the carotenoid-binding protein-dependent non-photochemical quenching mechanism.     

State of the phycobilisome determines effective absorption cross-section of Photosystem II in Synechocystis sp. PCC 6803

Quenching of excess excitation energy is necessary for the photoprotection of light-harvesting complexes. In cyanobacteria, quenching of phycobilisome (PBS) excitation energy is induced by the Orange Carotenoid Protein (OCP), which becomes photoactivated under high light conditions. A decrease in energy transfer efficiency from the PBSs to the reaction centers decreases photosystem II (PS II) activity. However, quantitative analysis of OCP-induced photoprotection in vivo is complicated by similar effects of both photochemical and non-photochemical quenching on the quantum yield of the PBS fluorescence overlapping with the emission of chlorophyll. In the present study, we have analyzed chlorophyll a fluorescence induction to estimate the effective cross-section of PS II and compared the effects of reversible OCP-dependent quenching of PBS fluorescence with reduction of PBS content upon nitrogen starvation or mutations of key PBS components. This approach allowed us to estimate the dependency of the rate constant of PS II primary electron acceptor reduction on the amount of PBSs in the cell. We found that OCP-dependent quenching triggered by blue light affects approximately half of PBSs coupled to PS II, indicating that under normal conditions, the concentration of OCP is not sufficient for quenching of all PBSs coupled to PS II.     

Spatio-temporal heterogeneity in Arabidopsis thaliana leaves under drought stress

Using chlorophyll (chl) fluorescence imaging, we studied the effect of mild (MiDS), moderate (MoDS) and severe (SDS) drought stress on photosystem II (PSII) photochemistry of 4-week-old Arabidopsis thaliana. Spatio-temporal heterogeneity in all chl fluorescence parameters was maintained throughout water stress. After exposure to drought stress, maximum quantum yield of PSII photochemistry (F(v)/F(m)) and quantum efficiency of PSII photochemistry (Φ(PSΙΙ)) decreased less in the proximal (base) than in the distal (tip) leaf. The chl fluorescence parameter F(v) /F(m) decreased less after MoDS than MiDS. Under MoDS, the antioxidant mechanism of A. thaliana leaves seemed to be sufficient in scavenging reactive oxygen species, as evident by the decreased lipid peroxidation, the more excitation energy dissipated by non-photochemical quenching (NPQ) and decreased excitation pressure (1-q(p)). Arabidopsis leaves appear to function normally under MoDS, but do not seem to have particular metabolic tolerance mechanisms under MiDS and SDS, as revealed by the level of lipid peroxidation and decreased quantum yield for dissipation after down-regulation in PSII (Φ(NPQ)), indicating that energy dissipation by down-regulation did not function and electron transport (ETR) was depressed. The simultaneous increased quantum yield of non-regulated energy dissipation (Φ(NO)) indicated that both the photochemical energy conversion and protective regulatory mechanism were insufficient. The non-uniform photosynthetic pattern under drought stress may reflect different zones of leaf anatomy and mesophyll development. The data demonstrate that the effect of different degrees of drought stress on A. thaliana leaves show spatio-temporal heterogeneity, implying that common single time point or single point leaf analyses are inadequate.     

膜荚黄芪与蒙古黄芪幼苗的光能利用及其耐光性研究

以盆栽膜荚黄芪和蒙古黄芪幼苗为试验材料,测量2种黄芪叶片的光合速率、叶绿素荧光参数、光能利用率及耐光性等指标的日变化,以探讨它们的光能利用、热耗散以及耐光性差异.结果表明:(1)随着太阳光合有效辐射的日变化进程推进,2种黄芪的初始荧光(F0)、非光化学淬灭(NPQ)、PSⅡ调节性能量耗散量子产最(YNPQ)先升高后降低...     

模拟酸雨对白筋叶片抗氧化酶活性及叶绿素荧光参数的影响

采用棚内盆栽方法, 设置pH值5.6 （对照）、4.0、3.0和2.0的模拟酸雨胁迫试验, 探讨其对白簕幼苗叶片MDA含量、保护酶活性、叶绿素含量、气体交换参数和叶绿素荧光参数的影响。结果表明, 随着模拟酸雨pH值的降低, MDA含量呈先降低后升高的趋势; SOD活性逐渐降低, POD活性逐渐升高, APX活性呈先升高后降低的变化。叶绿素a、叶绿素b、总叶绿素含量均比对照高, 在pH 4.0时达最大值。气孔限制值（Ls）、PSII实际光化学量子产量（ΦPSII）、光化学淬灭系数（qP）均随pH值的降低而下降, 净光速率（Pn）、气孔导度（Gs）、蒸腾速率（Tr）、水分利用效率（WUE）、PSII最大光化学效率（Fv/Fm）、PSII的潜在活性（Fv/Fo）、非光化学淬灭系数（qN）呈先升高后降低趋势, 且也都在pH 4.0时达最大值。由此推测, pH 4.0的酸雨处理有利于白簕幼苗的生长, 表明白簕幼苗可能喜欢生活在微酸环境中, 但是随着酸度加强, 反而起到抑制作用。     

外源ATP对NaCl胁迫下菜豆叶片叶绿素荧光特性的调节

盐胁迫是影响植物生长的主要逆境因子之一,外源ATP被发现可作为信号分子参与植物对逆境胁迫生理反应的调节。为了探明外源ATP在植物盐胁迫响应中的作用,以增强植物对土壤盐渍化的耐性,更好地应用于土壤盐渍化修复。该研究以菜豆( Phaseolus vulgaris)为材料,通过叶绿素荧光技术探讨了外源ATP 对菜豆叶片在NaCl胁迫下叶绿素荧光特性的变化规律。结果表明：在NaCl胁迫下,叶片光系统Ⅱ( PSⅡ)潜在最大光化学量子效率( Fv/Fm)、光适应下最大光化学效率( Fv′/Fm′)、PSⅡ光适应下实际光化学效率[ Y (Ⅱ)]、光化学荧光猝灭( qP)、电子传递速率( ETR)与对照组相比均有显著性下降,而非光化学猝灭( NPQ)和( qN)较对照组有显著性增加,这表明NaCl胁迫导致菜豆叶片光系统Ⅱ光化学效率的下降和光能耗散的增加。而外源ATP(eATP)的处理能有效缓解NaCl胁迫所造成的Fv/Fm、Fv′/Fm′、Y(Ⅱ)、qP、ETR下降和NPQ、qN的上升。该研究结果表明在NaCl胁迫下外源ATP可以有效地提高菜豆幼苗光系统Ⅱ( PSⅡ)的光化学反应效率。     

The higher resistance to chilling stress in adaxial side of <Emphasis Type="Italic">Rumex</Emphasis> K-1 leaves is accompanied with higher photochemical and non-photochemical quenching

Responses of two sides of Rumex K-1 leaves to chilling stress (5 °C, photon flux density of 100 μmol m⁻² s⁻¹) were studied by using gas exchange, chlorophyll (Chl) fluorescence, and spectrum reflectance techniques. The Chl and carotenoid contents in the two sides were not affected by chilling treatment, and both were higher in the adaxial side. The maximum quantum yield of photosystem (PS) 2 and fraction of functional PS1 in the abaxial side decreased more markedly than those in the adaxial side during the chilling treatment, indicating that the abaxial side was damaged more significantly than the adaxial side. Before chilling, there were no obvious differences in actual photochemical efficiency of PS2, photosynthesis, and photorespiration between two sides of the leaves. Under chilling stress, the actual photochemical efficiency of PS2, photosynthesis, and photorespiration all declined more significantly in the abaxial side, which was partly attributed to lower carboxylation efficiency in the abaxial side than that in the adaxial side. Non-photochemical quenching was higher in the adaxial side, though the de-epoxidation of xanthophyll cycle pigments’ pool on basis of Chl was higher in the abaxial side. Both the slower decrease in the photochemical quenching and the higher non-photochemical quenching may account for the higher resistance to chilling stress in the adaxial side of Rumex K-1 leaves.     

Photosynthesis and chlorophyllafluorescence during flag leaf senescence of field-grown wheat plants

The characteristics of photosynthetic gas exchange, chlorophyll a fluorescence, and xanthophyll cycle pigments during flag leaf senescence of field-grown wheat plants were investigated. With senescence progressing, the light-saturated net CO₂ assimilation rate expressed either on a basis of leaf area or chlorophyll decreased significantly. The apparent quantum yield of net photosynthesis decreased when expressed on a leaf area basis but increased when expressed on a chlorophyll basis. The maximal efficiency of PSII photochemistry decreased very little while actual PSII efficiency, photochemical quenching, and the efficiency of excitation capture by open PSII centers decreased considerably. At the same time, non-photochemical quenching increased significantly. A substantial decrease in the contents of violaxanthin and zeaxanthin, but a slight decrease in the content of antheraxanthin were observed. However, the de-epoxidation status of the xanthophyll cycle was positively correlated with progressive senescence. This increase was due mainly to a smaller decrease in zeaxanthin than in violaxanthin. Our results suggest that PSII apparatus remained functional, but a down-regulation of PSII occurred under the steady state of photosynthesis in senescent flag leaves. Such a down-regulation was associated with the closure of PSII centers and an enhanced xanthophyll cycle-related thermal dissipation in the PSII antennae.     

Effects of Exogenous Putrescine on Chlorophyll Fluorescence Imaging and Heat Dissipation Capacity in Cucumber (Cucumis sativus L.) Under Salt Stress

The objective of this study was to identify the effects of exogenous putrescine on photosynthetic performance and heat dissipation capacity in cucumber seedlings under salt stress. The stress of 75 mM NaCl for 7 days caused a significant decrease in net photosynthetic rate (P _N ). The experiment employed a chlorophyll fluorescence imaging technique and demonstrated that the maximal quantum yield of photosystem II photochemistry (Fv/Fm) and the actual photochemical efficiency of photosystem II (ΦPSII) were reduced by salt stress. Moreover, salt stress markedly reduced the photochemical quenching coefficient (qP) and non-photochemical quenching coefficient (qN), and significantly increased non-regulated heat dissipation (ΦNO). However, stressed plants supplied with exogenous putrescine exhibited higher P _N and ΦPSII, which indicated that putrescine can alleviate the detrimental effects on photosynthesis induced by salt stress. Putrescine sprayed on stressed plants significantly enhanced the regulated energy dissipation (ΦNPQ) and decreased ΦNO. Application of exogenous putrescine also changed the levels of xanthophyll cycle components and further enhanced the de-epoxidation state of xanthophyll cycle pigments under salt stress. Under control conditions, putrescine exerted little influence on the photosynthetic parameters in cucumber leaves. In conclusion, the application of exogenous putrescine may improve the heat dissipation capacity by promoting the xanthophyll cycle to reduce the damage caused by excess excitation energy, thus enhancing the salt tolerance of cucumber seedlings.     

Analysis of LhcSR3, a protein essential for feedback de-excitation in the green alga Chlamydomonas reinhardtii

In photosynthetic organisms, feedback dissipation of excess absorbed light energy balances harvesting of light with metabolic energy consumption. This mechanism prevents photodamage caused by reactive oxygen species produced by the reaction of chlorophyll (Chl) triplet states with O₂. Plants have been found to perform the heat dissipation in specific proteins, binding Chls and carotenoids (Cars), that belong to the Lhc family, while triggering of the process is performed by the PsbS subunit, needed for lumenal pH detection. PsbS is not found in algae, suggesting important differences in energy-dependent quenching (qE) machinery. Consistent with this suggestion, a different Lhc-like gene product, called LhcSR3 (formerly known as LI818) has been found to be essential for qE in Chlamydomonas reinhardtii. In this work, we report the production of two recombinant LhcSR isoforms from C. reinhardtii and their biochemical and spectroscopic characterization. We found the following: (i) LhcSR isoforms are Chl a/b– and xanthophyll-binding proteins, contrary to higher plant PsbS; (ii) the LhcSR3 isoform, accumulating in high light, is a strong quencher of Chl excited states, exhibiting a very fast fluorescence decay, with lifetimes below 100 ps, capable of dissipating excitation energy from neighbor antenna proteins; (iii) the LhcSR3 isoform is highly active in the transient formation of Car radical cation, a species proposed to act as a quencher in the heat dissipation process. Remarkably, the radical cation signal is detected at wavelengths corresponding to the Car lutein, rather than to zeaxanthin, implying that the latter, predominant in plants, is not essential; (iv) LhcSR3 is responsive to low pH, the trigger of non-photochemical quenching, since it binds the non-photochemical quenching inhibitor dicyclohexylcarbodiimide, and increases its energy dissipation properties upon acidification. This is the first report of an isolated Lhc protein constitutively active in energy dissipation in its purified form, opening the way to detailed molecular analysis. Owing to its protonatable residues and constitutive excitation energy dissipation, this protein appears to merge both pH-sensing and energy-quenching functions, accomplished respectively by PsbS and monomeric Lhcb proteins in plants.