Functioning of photosystems I and II in pea leaves exposed to heat stress in the presence or absence of light

Fluorimetric, photoacoustic, polarographic and absorbance techniques were used to measure in situ various functional aspects of the photochemical apparatus of photosynthesis in intact pea leaves (Pisum sativum L.) after short exposures to a high temperature of 40 ° C. The results indicated (i) that the in-vivo responses of the two photosystems to high-temperature pretreatments were markedly different and in some respects opposite, with photosystem (PS) II activity being inhibited (or down-regulated) and PSI function being stimulated; and (ii) that light strongly interacts with the response of the photosystems, acting as an efficient protector of the photochemical activity against its inactivation by heat. When imposed in the dark, heat provoked a drastic inhibition of photosynthetic oxygen evolution and photochemical energy storage, correlated with a marked loss of variable PSII-chlorophyll fluorescence emission. None of the above changes were observed in leaves which were illuminated during heating. This photoprotection was saturated at rather low light fluence rates (around 10 W · m^–2). Heat stress in darkness appeared to increase the capacity for cyclic electron flow around PSI, as indicated by the enhanced photochemical energy storage in far-red light and the faster decay of P ₇₀₀ ⁺ (oxidized reaction center of PSI) monitored upon sudded interruption of the far-red light. The presence of light during heat stress reduced somewhat this PSI-driven cyclic electron transport. It was also observed that heat stress in darkness resulted in the progressive closure of the PSI reaction centers in leaves under steady illumination whereas PSII traps remained largely open, possibly reflecting the adjustment of the photochemical efficiency of undamaged PSI to the reduced rate of photochemistry in PSII.Abbreviations B₁ and B₂ fraction of closed PSI and PSII reaction centers, respectively - ES photoacoustically measured energy storage - F_o, F_m and F_s initial, maximal and steady-state levels of chlorophyll fluorescence - P₇₀₀ reaction center of PSI - PS (I, II) photosystem (I, II) - V = (F_s – F_o)/(F_m – F_o) relative variable chlorophyll fluorescence We wish to thank Professor R. Lannoye (ULB, Brussels) for the use of this photoacoustic spectrometer and Mrs. M. Eyletters for her help.     

Sites of action of sulphite and bisulphite in the photosynthetic apparatus of sugar maple leaves as studied by photo-acoustic and modulated fluorescence methods

The photosynthetic activity of intact sugar maple leaves has been assessed in the presence of exogenous sulphite, bisulphite and sulphate under varying light conditions using photo-acoustic and modulated fluorescence methods. In the light, bisulphite was found to be more toxic than the other two, sulphate being the least toxic of all. Interestingly, the vitality index, R_rd, measured as the ratio of the modulated fluorescence decrease (F_d) to the steady-state fluorescence (F_s), which indicates the efficiency of the whole photosynthetic activity, was more affected than the total photosynthetic energy storage (PES) of PSII and PSI during linear and cyclic electron transport, and F_v/F_m (PSII activity). The severity of the damage appeared to be a function of light intensity. Bisulphite treatment in darkness resulted in a dramatic decrease in R_rd, a moderate decrease in PES and a marginal decrease in F_v/F_m. As for sulphite, the effect was negligible with a tendency for enhanced activity. It is inferred that the Calvin cycle is a good candidate for the primary site of bisulphite and sulphite action. Recovery of activity, especially at the R_rd level, obtained in the presence of ascorbate, glutathione and L-cysteine, indicated a contribution of O₂ free radicals to the observed inhibition of the photosynthetic activity in the light.     

Down regulation of photosynthesis in Artabotrys hexapetatus by high light

Using variable to maximum fluorescence (F_v/F_m) as the criterion, the down regulation of photosynthesis by high light stress was characterized in the detached leaves of Artabotrys hexapetatus. The decrease in F_v/F_m was corelated with the decrease in oxygen evolution by thylakoids isolated from high light exposed leaves. The decrease in F_v/F_m was linear with increasing time of exposure to high light. A comparison of recovery measured as F_v/F_m, in low light versus dark, revealed that the recovery in darkness was as significant as in low light. Since the relaxation of fluorescence was a rapid response after exposure to high light and the fact that the recovery occurs in total darkness, it is concluded that photoinhibition and down regulation of photosynthesis by high light are independent events.Abbreviation Fpl- initial plateau - F_m- maximum fluorescence - F_o- prompt fluorescence - F_v- variable fluorescence - PFD- photon flux density - PS I (II)- Photosystem I (II)     

Light adaptation of cyclic electron transport through Photosystem I in the cyanobacterium Synechococcus sp. PCC 7942

Photosystem I-driven cyclic electron transport was measured in intact cells of Synechococcus sp PCC 7942 grown under different light intensities using photoacoustic and spectroscopic methods. The light-saturated capacity for PS I cyclic electron transport increased relative to chlorophyll concentration, PS I concentration, and linear electron transport capacity as growth light intensity was raised. In cells grown under moderate to high light intensity, PS I cyclic electron transport was nearly insensitive to methyl viologen, indicating that the cyclic electron supply to PS I derived almost exclusively from a thylakoid dehydrogenase. In cells grown under low light intensity, PS I cyclic electron transport was partially inhibited by methyl viologen, indicating that part of the cyclic electron supply to PS I derived directly from ferredoxin. It is proposed that the increased PSI cyclic electron transport observed in cells grown under high light intensity is a response to chronic photoinhibition.Abbreviations DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - ES energy storage - MV methyl viologen - PA_m photoacoustic thermal signal with strong non-modulated background light added - PA_s photoacoustic thermal signal without background light added CIW/DPB Publication No. 1205.     

Performance of active Photosystem II centers in photoinhibited pea leaves

Effects of photoinhibition on photosynthesis in pea (Pisum sativum L.) leaves were investigated by studying the relationship between the severity of a photoinhibitory treatment (measured as F_v/F_m) and several photoacoustic and chlorophyll a fluorescence parameters. Because of the observed linear relationship between the decline of F_v/F_m and the potential oxygen evolution rate determined by the photoacoustic method, the parameter F_v/F_m was used as an indicator for the severity of photoinhibition. Our analysis revealed that part of the Photosystem II (PS II) reaction centers is inactive in oxygen evolution and is also less sensitive to photoinhibition. Correcting the parameter q_P (fraction of open PS II reaction centers) for inactive PS II centers unveiled a strong increase of q_P in severely inhibited pea leaves, indicating that the inactivated active centers do no longer contribute to q_P and that photoinhibition has an all or none effect on PS II centers. Analysis of q_E (energy quenching) demonstrated its initial increase possibly associated with dephosphorylation of LHC II. Analysis of q_I (photoinhibition dependent quenching) showed that the half-time of recovery of q_I increases steeply below an F_v/F_m of 0.65. This increase of the relaxation half-time corresponds with a decrease of the electron transport rate J and tentatively indicates that the supply of ATP, needed for the recovery, starts to decrease. The data indicate the necessity of correcting for inactive centers in order to make valuable conclusions about effects of photoinhibition on photosynthetic parameters.     

Fluoride affects distribution of absorbed excitation energy more in favour of photosystem 1

The effects of fluoride on the photosynthetic electron transport chain have been studied in spinach thylakoid membranes. Inhibition in photosystem (PS) 2 electron transport rates and a subsequent increase in PS 1 electron transport rate indicated a possibility of state transitions being a mechanism of fluoride action. This hypothesis was further confirmed by the increase in fluorescence emission F_735/685 at 77 K, a decrease in variable to maximum fluorescence ratio (F_v/F_m) at room temperature and increase in the absorption cross section of PS 1 suggesting that fluoride affects distribution of the excitation energy in favour of PS 1 at the expense of PS 2.     

Photosynthetic energy storage of Photosystems I and II in the spectral range of photosynthetically active radiation in intact sugar maple leaves

The relative activity of Photosystems (PS) I and II in the spectral range between 400 and 720 nm was studied by measuring photosynthetic energy storage (ES) of an intact sugar maple leaf using photoacoustic spectroscopy. ES, determined with a modulated (80 Hz) monochromatic light beam in the presence of saturating intensity of background non-modulated white light, indicated the total energy stored by both photosystems (ES_T). Using background far-red light, ES of PS I (ES_{PS I}) was quantified. ES_{PS II} was derived from ES_T-ES_{PS I}. ES_T dependence on intensity and wavelength of modulated light was studied at 470, 560, 640 and 680 nm. ES_T was maximum in red light and minimum in blue light. It decreased with an increase in modulated light intensity. The ratio ES_{PS II}/ES_{PS I}, measured at 640 nm, remained nearly constant with an increase in modulated light intensity. The relative quantum yield of ES_T spectrum showed two peaks around 610 and 660 nm, and declined sharply after 680 nm, revealing a clear red drop. ES_{PS I} spectrum presented peaks around 610 and 670 nm, and a minimum between 440 and 470 nm. ES_{PS I} was observed beyond 700 nm up to 720 nm, indicating the energy stored by cyclic electron transport. ES_{PS II} spectrum showed broad peaks, around 460, 490, 600 and 660 nm, and a shoulder between 530 and 560 nm. ES_{PS II} was always higher than ES_{PS I} between 400 and 690 nm and reached zero around 700 nm.Abbreviations ES energy storage - ES_{PS I} energy storage of PS I - ES_{PS II} energy storage of PS II - ES_T energy storage of PS I and PS II - PA photoacoustic - PS I Photosystem I - PS II Photosystem II - Q_m PA signal in the absence of any background light - Q_ma PA signal in the presence of background white light - Q_mfrl PA signal in the presence of background far-red light - S/N signal to noise     

Fluorescence parameters of White Sea phytoplankton under different nitrogen sources

We investigated dependence of fluorescence parameters and phytoplankton biomass on the nitrogen source and irradiance in enriched flask studies with White Sea plankton from August-September 2007. Phytoplankton was exposed in situ for 18 d with addition of 180 μM/L of nitrogen in the forms of nitrate, urea, ammonia, and glycine under two levels of irradiance. Maximum quantum efficiency of PS2 (F_v/F_m) was determined in the samples adapted to darkness. Rapid light curves were obtained for each sample with the sequential increase of light intensity (8 levels). The maximal relative electron transport rate (rETR_max), the maximum light utilization coefficient (α), and the nonphotochemical quenching (NPQ) were calculated. The phytoplankton abundance increased on nitrogen addition, and the photosynthetic parameters changed. The values F_v/F_m reached 0.64–0.71, which indicated a good physiological state of algae and lack of nitrogen limitation. The dynamics of rETR_max and NPQ depended of the nitrogen source and irradiance, while α did not depend on nitrogen addition.     

A photoacoustic study of water infiltrated leaves

Photoacoustic measurements of photosynthetic energy storage were conducted on water infiltrated pea and sugar maple leaves. The samples were vacuum infiltrated with pure water or with a suitable buffer. The use of such methodology permitted an accurate determination of the energy storage parameter at low modulation frequencies, where in non-infiltrated leaves oxygen evolution dominates the photoacoustic signal and does not allow energy storage measurements. Differences between infiltration media were not essential, however the use of pure water as infiltration medium sometimes caused instability of the measured energy storage, particularly at longer experimental time. Values of energy storage in individual samples ranged mostly between 0.2 to 0.35. Measured as a function of the modulation frequency, energy storage was found to be constant from about 10 to 200 Hz for pea leaves. In sugar maple leaves, the energy storage slightly increased between 100 and 500 Hz. Obtaining an accurate value for energy storage also allowed an accurate estimation of the O₂ evolution contribution to the photoacoustic signal of an unfiltrated leaf. In a maple leaf its frequency dependence showed only the effect of diffusion in the entire frequency range (10–500 Hz). Energy storage transients were observed after long periods (ca. 1/4-2 hrs) of dark adaptation upon the transition to light. In this case the initial energy storage was roughly about 1/2 that of the steady state value indicating strong PS I activity, while PS II was transiently incompetent. Energy-storage increased during illumination in a way to correspond to photosynthetic induction events as previously measured by fluorescence and O₂ evolution. Transients in energy storage were also found following high light to low light transitions (i.e., switch off of the saturating background light), that paralleled similar transients in oxygen evolution, showing initial transient inactivation followed by progressive reactivation of PS II.Abbreviations ES energy storage - PA photoacoustic(s) - PTR photothermal radiometry     

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.     

ACCLIMATION OF PHOTOSYNTHESIS AND PIGMENTS DURING AND AFTER SIX MONTHS OF DARKNESS IN PALMARIA DECIPIENS (RHODOPHYTA): A STUDY TO SIMULATE ANTARCTIC WINTER SEA ICE COVER1

The influence of seasonally fluctuating photoperiods on the photosynthetic apparatus of Palmaria decipiens (Reinsch) Ricker was studied in a year‐round culture experiment. The optimal quantum yield (F_v/F_m) and the maximal relative electron transport rate (ETR_max), measured by in vivo chl fluorescence and pigment content, were determined monthly. During darkness, an initial increase in pigment content was observed. After 3 months in darkness, ETR_max and F_v/F_m started to decrease considerably. After 4 months in darkness, degradation of the light‐harvesting antennae, the phycobilisomes, began, and 1 month later the light harvesting complex I and/or the reaction centers of PSII and/or PSI degraded. Pigment content and photosynthetic performance were at their minimum at the end of the 6‐month dark period. Within 24 h after re‐illumination, P. decipiens started to accumulate chl a and to photosynthesize. The phycobiliprotein accumulation began after a time lag of about 7 days. Palmaria decipiens reached ETR_max values comparable with the values before darkness 7 days after re‐illumination and maximal values after 30 days of re‐illumination. Over the summer, P. decipiens reduced its photosynthetic performance and pigment content, probably to avoid photodamage caused by excess light energy. The data show that P. decipiens is able to adapt to the short period of favorable light conditions and to the darkness experienced in the field.     

低温弱光胁迫及恢复对切花菊光合作用和叶绿素荧光参数的影响

以切花菊品种‘神马’为试材,在偏低温弱光(16℃/12℃,PFD100μmol.m-2.s-1)和临界低温弱光(12℃/8℃,PFD60μmol.m-2.s-1)下分别胁迫11d,然后转入正常条件(22℃/18℃,PFD450μmol.m-2.s-1)恢复11d,研究不同低温弱光强度及恢复对菊花光合作用和叶绿素荧光参数的影响.结果表明:低温弱光导致菊花叶片的净光合速率(Pn)和气孔限制值(Ls)下降,而胞间CO2浓度(Ci)上升.偏低温弱光胁迫下菊花叶片暗适应下最大光化学效率(Fv/Fm)和初始荧光(Fo)无明显变化,但光适应下最大光化学效率(Fv′/Fm′)在处理前期略有下降,后期则有所回升;而临界低温弱光处理的Fo明显升高,Fv/Fm和Fv′/Fm′显著降低.PSⅡ光合电子传递量子效率(ΦPSⅡ)、光化学猝灭系数(qP)和表观光合电子传递速率(ETR)均随着低温弱光胁迫程度的增加和时间的延长而降低;偏低温弱光处理植株在解除胁迫后能迅速恢复到对照水平,而临界低温弱光处理植株回升速度较慢;同时,低温弱光胁迫下吸收光强用于分配光化学反应部分(Prate)的比例减少,而天线热耗散(Drate)和反应中心的能量耗散(Ex)比例上升,但天线热耗散为过剩光能的主要分配途径.     

Changes in Chlorophyll Fluorescence During the Course of Photoperiod and in Response to Drought in Casuarina equisetifolia Forst. and Forst.

The effects of drought and the diurnal changes in photosynthetic electron transport were studied in non-nodulated plants of Casuarina equisetifolia. The induction of fluorescence showed a slightly higher I step in water-stressed than control plants, and the time from the start of irradiation to the P step of induction was significantly shortened by drought. The quantum efficiency of photosystem 2 (PS2) in the dark-adapted state (F_v/F_m) was generally not affected by drought, whereas it decreased during the central hours of the day. The decrease in quantum yield of PS2 electron transport (₂) in water-stressed plants was associated with decreases in the photochemical efficiency of open (oxidised) PS2 centres (F_v'/F_m') and increases in non-photochemical quenching (q_N) rather than with increased closure of PS2 centres (lowered photochemical quenching, q_P). In contrast, the changes in quantum yield of electron transport during the day were related to changes in q_P rather than in F_v'/F_m'. When chlorophyll fluorescence was measured at the same irradiance during the day, a greater q_N was observed at the end of the drying cycle than after watering, and early and late in the photoperiod than in the central hours of the day. The greater q_N at the beginning and end of the day did not prevent an increase in energy not used photochemically nor dissipated non-photochemically. Drought did not affect this excess of photon energy.     

Correlation between chlorophyll fluorescence and photoacoustic signal transients in spinach leaves

Chlorophyll fluorescence and photoacoustic transients from dark adapted spinach leaves were measured and analyzed using the saturating pulse technique. Except for the first 30 s of photosynthetic induction, a good correlation was found between photoacoustically detected oxygen evolution at 35 Hz modulation frequency and electron flow calculated from the fluorescence quenching coefficients q_P and q_N. The induction kinetics of the photothermal signal, i.e., the photoacoustic signal at 370 Hz, reveal a fast (t _r <10 ms) and a slow (t _r 1 s) rise component. The fast component is suggested to be composed of the minimal thermal losses in photosynthesis and thermal losses from non-photosynthetic processes. The slow phase is attributed to variable thermal losses in photosynthesis. The variable thermal losses were normalized by measuring the minimal photothermal signal (H₀) in the dark-adapted state and the maximal photothermal signal (H_m) during a saturating light pulse. The kinetics of the normalized photochemical loss (H-H₀)/(H_m-H₀) obtained from high-frequency PA measurements were found to correlate with the kinetics of oxygen evolution measured at low frequency.Abbreviations F_m maximum fluorescence - F₀ initial fluorescence - F_v variable fluorescence - H photothermal signal - I in-phase - LED light emitting diode - PA photoacoustic - PL photochemical loss - Q quadrature - q_N non-photochemical quenching - q_P photochemical quenching - VCLS voltage controlled light source     

Short-term responses of Photosystem I to heat stress

When 23°C-grown potato leaves (Solanum tuberosum L.) were exposed for 15 min to elevated temperatures in weak light, a dramatic and preferential inactivation of Photosystem (PS) II was observed at temperatures higher than about 38°C. In vivo photoacoustic measurements indicated that, concomitantly with the loss of PS II activity, heat stress induced a marked gas-uptake activity both in far-red light (>715 nm) exciting only PS I and in broadband light (350–600 nm) exciting PS I and PS II. In view of its suppression by nitrogen gas and oxygen and its stimulation by high carbon-dioxide concentrations, the bulk of the photoacoustically measured gas uptake by heat-stressed leaves was ascribed to rapid carbon-dioxide solubilization in response to light-modulated stroma alkalization coupled to PS I-driven electron transport. Heat-induced gas uptake was observed to be insensitive to the PS II inhibitor diuron, sensitive to the plastocyanin inhibitor HgCl₂ and saturated at a rather high photon flux density of around 1200 E m^–2 s^–1. Upon transition from far-red light to darkness, the oxidized reaction center P700⁺ of PS I was re-reduced very slowly in control leaves (with a half time t_1/2 higher than 500 ms), as measured by leaf absorbance changes at around 820 nm. Heat stress caused a spectacular acceleration of the postillumination P700⁺ reduction, with t_1/2 falling to a value lower than 50 ms (after leaf exposure to 48°C). The decreased t_1/2 was sensitive to HgCl₂ and insensitive to diuron, methyl viologen (an electron acceptor of PS I competing with the endogenous acceptor ferredoxin) and anaerobiosis. This acceleration of the P700⁺ reduction was very rapidly induced by heat treatment (within less than 5 min) and persisted even after prolonged irradiation of the leaves with far-red light. After heat stress, the plastoquinone pool exhibited reduction in darkness as indicated by the increase in the apparent Fo level of chlorophyll fluorescence which could be quenched by far-red light. Application (for 1 min) of far-red light to heat-pretreated leaves also induced a reversible quenching of the maximal fluorescence level Fm, suggesting formation of a pH gradient in far-red light. Taken together, the presented data indicate that PS I responded to the heat-induced loss of PS II photochemical activity by catalyzing an electron flow from stromal reductants. Heat-stress-induced PS I electron transport independent of PS II seems to constitute a protective mechanism since block of this electron pathway in anaerobiosis was observed to result in a dramatic photoinactivation of PS I.Abbreviations PFD photon flux density - PS Photosystem - Apt and Aox amplitude of the photothermal and photobaric components of the photoacoustic signal, respectively - P700 reaction center pigment of PS I - Fo and Fm initial and maximal levels of chlorophyll fluorescence, respectively - Fv=Fm Fo-variable chlorophyll fluorescence - Q_A primary (stable) electron acceptor of PS II - DCMU (diuron) 3-(3,4-dichlorophenyl)-1,1-dimethylurea - Cyt cytochrome     

Peroxidation of lipids and growth inhibition induced by UV-B irradiation

Cotyledons excised from dark-grown seedlings of cucumber (Cucumis sativus L.) were cultured in vitro under UV radiation at different wavelengths, obtained by passage of light through cut-off filters with different transmittance properties. Growth and the synthesis of chlorophyll (Chl) in cotyledons were inhibited and malondialdehyde was accumulated upon irradiation at wavelengths below 320 nm. Exogenous application of scavengers of free radicals reversed the growth inhibition induced by UV-B. Measurement of the fluorescence of Chl a suggested that electron transfer in photosystems was affected by UV-B irradiation. On the basis of these results, the involvement is postulated of active species of oxygen in damages to thylakoid membranes and the growth inhibition that are induced by UV-B irradiation.Abbreviations Chl chlorophyll - F_m maximal fluorescence (dark) - F_m maximal fluorescence (light) - F_v variable fluorescence (dark) - F_v variable fluorescence (light) - MDA malondialdehyde - O₂ Superoxide radical - PS photosystem - q_N non-photochemical quenching of fluorescence - q_P photochemical quenching of fluorescence - UV-B_BE biologically effective UV-B radiation - WL(T = 0.5) wavelength at which 50% transmittance occurs     

Effects of dehydration on the electron transport of Chlorella. An in vivo fluorescence study

Chlorella was used to study the effects of dehydration on photosynthetic activities. The use of unicellular green algae assured that the extent of dehydration was uniform throughout the whole cell population during the course of desiccation. Changes in the activities of the cells were monitored by measurements of fluorescence induction kinetics. It was found that inhibition of most of the photosynthetic activities started at a similar level of cellular water content. They included CO₂ fixation, photochemical activity of Photosystem II and electron transport through Photosystem I. The blockage of electron flow through Photosystem I was complete and the whole transition occurred within a relative short time of dehydration. On the other hand, the suppression of Photosystem II activity was incomplete and the transition took a longer time of dehydration. Upon rehydration, the inhibition of Photosystem II activity was fully reversible when samples were in the middle of the transition, but was not thereafter. The electron transport through Photosystem I was also reversible during the transition, but was only partially afterward.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl urea - F_m maximum fluorescence yield - F₀ non-variable fluorescence level emitted when all PS II centers are open - F_v variable part of fluorescence - PS photosystem - Q_A primary quinone acceptor of Photosystem II     

Theoretical assessment of alternative mechanisms for non-photochemical quenching of PS II fluorescence in barley leaves

The components of non-photochemical chlorophyll fluorescence quenching (qN) in barley leaves have been quantified by a combination of relaxation kinetics analysis and 77 K fluorescence measurements (Walters RG and Horton P 1991). Analysis of the behaviour of chlorophyll fluorescence parameters and oxygen evolution at low light (when only state transitions — measured as qN_t — are present) and at high light (when only photoinhibition — measured as qN_i — is increasing) showed that the parameter qN_t represents quenching processes located in the antenna and that qN_i measures quenching processes located in the reaction centre but which operate significantly only when those centres are closed. The theoretical predictions of a variety of models describing possible mechanisms for high-energy-state quenching, measured as the residual quenching, qN_e, were then tested against the experimental data for both fluorescence quenching and quantum yield of oxygen evolution. Only one model was found to agree with these data, one in which antennae exist in two states, efficient in either energy transfer or energy dissipation, and in which those photosynthetic units in a dissipative state are unable to exchange energy with non-dissipative units.Abbreviations: F_o, F_m room-temperature chlorophyll fluorescence yield with all centres open, closed - F_v variable fluorescence yield - LHC II light-harvesting chlorophyll-protein complex of PS II - PS I, PS II Photosystem I, II - P700, P680 primary donor in Photosystem I, II - Q_A primary electron acceptor of PS II - P_max maximum quantum yield of oxygen evolution - qN coefficient of non-photochemical quenching of variable fluorescence - qN_e, qN_t, qN_i coefficient of non-photochemical quenching due to high-energy-state, state transition, photoinhibition - qO coefficient of quenching of dark level fluorescence - qP coefficient of photochemical quenching of variable fluorescence - _P intrinsic quantum yield of open PS II reaction centres = _s/qP - _{PS 2} quantum yield of PS = qP × F_v/F_m - _S quantum yield of oxygen evolution = rate of oxygen evolution/light intensity     

The effects of low temperature acclimation and photoinhibitory treatments on Photosystem 2 studied by thermoluminescence and fluorescence decay kinetics

The effects of low temperature acclimation and photoinhibitory treatment on Photosystem 2 (PS 2) have been studied by thermoluminescence and chlorophyll fluorescence decay kinetics after a single turnover saturating flash. A comparison of unhardened and hardened leaves showed that, in the hardened case, a decrease in overall and B-band thermoluminescence emissions occurred, indicating the presence of fewer active PS 2 reaction centers. A modification in the form of the B-band emission was also observed and is attributed to a decrease in the apparent activation energy of recombination in the hardened leaves. The acclimated leaves also produced slower Q_A ^– reoxidation kinetics as judged from the chlorophyll fluorescence decay kinetics. This change was mainly seen in an increased lifetime of the slow reoxidation component with only a small increase in its amplitude. Similar changes in both thermoluminescence and fluorescence decay kinetics were observed when unhardened leaves were given a high light photoinhibitory treatment at 4°C, whereas the hardened leaves were affected to a much lesser extent by a similar treatment. These results suggest that the acclimated plants undergo photoinhibition at 4°C even at low light intensities and that a subsequent high light treatment produces only a small additive photoinhibitory effect. Furthermore, it can be seen that photoinhibition eventually gives rise to PS 2 reaction centers which are no longer functional and which do not produce thermoluminescence or variable chlorophyll fluorescence.Abbreviations D1 The 32 kDa protein of Photosystem 2 reaction center - F_m maximum chlorophyll fluorescence yield - F₀ minimal chlorophyll fluorescence yield obtained when all PS 2 centers are open - F_i intermediate fluorescence level corresponding to PS 2 centers which are loosely or not connected to plastoquinone (non-B centers) - F_v maximum variable chlorophyll fluorescence yield (F_v=F_m–F₀) - PS 2 Photosystem 2 - Q_A and Q_B respectively, primary and secondary quinonic acceptors of PS 2 - S1, S2 and S3 respectively, the one, two and three positively charged states of the oxygen evolving system - Z secondary donor of PS 2     

Age-related differences in frost sensitivity of the photosynthetic apparatus of two Plagiomnium species

Shoots of two species of moss, Plagiomnium undulatum (Hedw.) Kop. and Plagiomnium affine (Funck) Kop., were subjected to freezing at various temperatures. After thawing, the activities of different photosynthetic reactions were determined in relation to the ages of the leaves. Analysis of the fast kinetics of chlorophyll-a fluorescence of individual leaves showed that young and old tissues were considerably less frost tolerant than mature ones. In principle, the pattern of freeze inactivation of photosynthetic reactions resembles that observed in higher plants. The decreases in the amplitude of F_v (variable fluorescence) and the ratio of F_v to F_m (maximum fluorescence) with increasing freezing stress reflect a progressive inactivation of photosystem II (PSII)-mediated electron transport, i.e. inhibition of photoreaction to photochemistry and-or electron donation to the photochemical reaction, and thus a decline in the potential photochemical efficiency of PSII. The insignificant change in the F₀ (constant fluorescence) level during progressive decline of F_v indicates that the excitation-energy transfer between antenna pigments and from those to reaction centres of PSII was little impaired by lethal freezing stress. Sugar analyses of various stem sections showed that ontogenetic variation in the frost tolerance of leaves cannot be attributed to differences in the cellular levels of sucrose, glucose and fructose.Abbreviations and Symbols DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_m maximum fluorescence - F₀ constant (initial) fluorescence - F_v variable fluorescence