Sulfite inhibition of photochemical activity of intact pea leaves

Sulfite treatment of pea leaf disks in light caused a significant decrease in the relative quantum yield of photosynthetic oxygen evolution and energy storage (ES) as measured by photoacoustic (PA) spectroscopy. The inhibition was concentration dependent and was less in darkness than in light, indicating light-dependent inhibitory site(s) on the photosynthetic electron transport chain. Further, in darksulfite-treated leaves, the energy storage was more affected than the relative quantum yield of oxygen evolution, suggesting that photophosphorylation and/or cyclic electron transport around PS I are sites of sulfite action in darkness. The R_fd values, the ratio of fluorescence decrease (fd) to the steady-state fluorescence (fs), decreased significantly in leaves treated with sulfite in light but were not affected in dark-treated ones, confirming the photoacoustic observations. Similarly, the ratio of variable fluorescence (F_v) to maximum fluorescence (F_m), a measure of PS II photochemical efficiency, was affected by sulfite treatment in light and not changed by treatment in darkness. An attempt was made to explain the mechanism of sulfite action on photosynthetic electron transport in light and in darkness.Abbreviations A_PT amplitude of photothermal signal - A_ox amplitude of oxygen signal - ES energy storage - fd fluorescence decrease - fs steady-state fluorescence - F_m maximum fluorescence - F_v variable fluorescence - PA photoacoustic(s)     

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     

Inhibition of photosynthetic activities under slow water stress measured in vivo by the photoacoustic method

A slow water stress over several days was imposed on tobacco plants (Nicotiana tabacum L. var. Xanthi) by withholding water from the soil. Photosynthesis was measured in leaves from those water-stressed plants by the photoacoustic method. Slow drought induced marked changes in the photoacoustic signals, which were largely similar to those observed previously in leaves subjected to rapid desiccation in air (over 3–4 h), reflecting two simultaneous changes: 1) Modification of the heat and oxygen diffusion characteristics of the leaves due to changes in their anatomical structure [shown by the change in the slope of the plot of the oxygen (A_OX) to photothermal signal (A_PT) ratio vs the square root of the modulation frequency]; 2) Inhibition of gross photosynthesis measured by the extrapolation of the A_OX/A_PT ratio to zero frequency. However, in contrast to rapid water stress in detached leaves, where it was shown that mainly the oxidizing side of photosystem II (PS II) was damaged, we found a slower and more complex phenomenology having largely biphasic kinetics. During the first 6 days, there was a strong reduction in the photochemical energy storage, but the inhibition of oxygen evolution was relatively mild. The Emerson enhancement in state 1 dropped considerably, indicating lowering of the apparent absorption cross-section of PS II. Fluorescence measurements suggest that PS II reaction center itseIf may be the primary site of the damage. PS I activity, judged by cytochrome f photooxidation, remained largely intact. The subsequent days were associated with a further spectacular decrease in the oxygen evolution quantum yield with both photosystems damaged. The photochemical energy storage continued to decrease further. The Emerson enhancement ratio of the remaining activities in both State 1 and 2 showed a marked increase, indicating the reestablishment of a strong imbalance in the distribution of excitation energy within the photochemical apparatus in favor of PS II. All the photoacoustic changes observed in response to drought were completely reversible within 2–3 days upon rewatering of the soil.     

SO(2) Effect on Photosynthetic Activities of Intact Sugar Maple Leaves as Detected by Photoacoustic Spectroscopy

Short-term (4 hours) effect of different concentrations of SO₂ fumigation on in vivo photochemical activities of sugar maple (Acer saccharum Marsh.) leaves was investigated using photoacoustic spectroscopy. The relative quantum yield of O₂ evolution (ratio of O₂ signal to the photothermal signal) and photochemical energy storage are increased by 0.05 microliter per liter of SO₂. This increase is more pronounced in 5 to 7 year old saplings than in 3 month old seedlings. Both oxygen-relative quantum yield and energy storage of seedlings are inhibited by increased concentrations of SO₂ and the inhibition is concentration dependent. The inhibition is greater in seedlings than in saplings at 2 microliters per liter of SO₂, indicating the more susceptible nature of seedlings. The present study indicates a concentration dependent differential effect of SO₂ on photochemical activities of sugar maple leaves.     

Heat Emission as a Protective Mechanism against High-irradiance Stress in Sugar Maple Leaves

High-irradiance (HI) induced changes in heat emission, fluorescence, and photosynthetic energy storage (EST) of shade grown sugar maple (Acer saccharum Marsh.) saplings were followed using modulated photoacoustic and fluorescence spectroscopic techniques. HI-treatment at 900-4400 µmol m-2 s-1 for 15 min caused an increase in heat emission and a decrease in EST. In some leaves, HI-treatment of 900 µmol m-2 s-1 for 1 min induced a rapid increase in heat emission with a marginal decrease in EST. Parallel to the increase in heat emission, there was a decrease in fluorescence, and this phenomenon was reversible in darkness. Quenching of thermal energy dissipation and a recovery in EST were observed during the first 15 min after the HI-treatment. This down-regulation of photochemical activity and its recovery may be one of the photoprotective mechanisms in shade grown sugar maple plants. The increase in thermal energy dissipation was greater in the red absorbing long wavelength (640-700 nm) region than in the blue absorbing short wavelength region of photosynthetically active excitation radiation. The photochemical activity was affected more in short wavelengths (400-520 nm) than in the long wavelength region of the spectrum. This can be due to the migration of light-harvesting chlorophyll (Chl) a/b protein complex from photosystem (PS) 2 to PS1 and/or to the disconnection of carotenoid pool from Chls in the pigment bed of photosynthetic apparatus.     

Increased thermal deactivation of excited pigments in pea leaves subjected to photoinhibitory treatments

The photoacoustic technique was used to monitor thermal deexcitation of the photosynthetic pigments in intact pea leaves (Pisum sativum L.) submitted to photoinhibitory treatments. When the leaves were exposed to photon flux densities above 1000 micromoles per square meter per second, the amplitude of the photothermal component of the in vivo photoacoustic signal strongly increased. This high-light-induced stimulation of nonradiative energy dissipation (heat emission) was accompanied by an inverse change in the O₂ evolution activity and in the steady state emission of 685 nanometer chlorophyll fluorescence. The time course of these effects was shown to be very rapid, with a t_1/2 of around 15 minutes. When high-light-treated leaves were readapted to the dark, the heat emission changes were reversed, following somewhat slower kinetics. A reversible increase in the rate of light energy dissipation via radiationless transitions could be a photoprotective mechanism eliminating excess excitation energy from the photosynthetic reaction centers. Interestingly, this process does not operate at temperatures below about 12°C.     

Photoacoustic measurements of the energy-conversion efficiency of photosynthesis in thalli of the green alga Bryopsis maxima

The efficiency of photosynthetic energy conversion in thalli of the green alga Bryopsis maxima was studied with the photoacoustic technique. Photosynthetic O₂ evolution did not interfere with the photoacoustic measurements in this material, most probably owing to its coenocytic cellular organization. The energy yield (defined as the fraction of absorbed photon energy that is stored in photosynthetic products or intermediates relative to the total absorbed photon energy) was estimated from the photoacoustic signals by applying the background-illumination method to obtain a reference without the photochemical capacity (Lasser-Ross, N., Malkin, S. and Cahen, D. (1980) Biochim. Biophys. Acta 593, 330–341). With the monitoring light modulated at 60 Hz, photon energy is mainly stored by redox changes in electron-transport chains because the energy yield was strongly reduced by 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea and heat treatment of the thalli, whereas KCN, an inhibitor of CO₂ reduction, had no effect, and because a significant lowering of the energy yield occurred in the presence of methyl viologen but the effect of the Photosystem I acceptor was largely reversed on the addition of an uncoupler, methylamine. The maximum energy yield of 0.4 that was obtained with a saturating background light and with a sufficiently weak monitoring light modulated at 100 Hz is explained in terms of electron transfer from electron-donor pools to acceptor pools of the two photosystems with the quantum yield close to unity. A lowering of the modulation frequency decreased the energy yield, indicating that less energy is stored in more stable intermediates.     

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.     

Simultaneous detection of photosynthetic energy storage and oxygen evolution in leaves by photothermal radiometry and photoacoustics

We have measured simultaneously the photothermal radiometry and the photoacoustic signals from intact leaves. We have confirmed that while the former senses that part of the modulated absorbed radiation not used in photosynthesis, but converted into heat, the latter, at low modulation frequencies, senses not only this heat but also the modulated oxygen evolution resulting from photosynthesis in the leaf. When photosynthetic activity is saturated upon additional excitation with strong non-modulated light, the photothermal radiometry signal increases (virtually all absorbed modulated light being converted into heat), while at the same time the photoacoustic signal decreases, because virtually no modulated oxygen evolution occurs any more. At higher modulation frequencies the behaviour of the photoacoustic signal closely follows that of the photothermal radiometry signal. We have used combined photothermal radiometry / photoacoustic measurements to estimate directly the yield of chemical energy storage in various plant species which applies for different times after excitation. Measurement of light saturation curves for wheat and Siberian pea bush leaves and of action spectra for the latter confirm the similarity between photothermal radiometry and high-frequency photoacoustic signals, and their difference from the low-frequency photoacoustic signal. Combined use of photothermal radiometry (or high-frequency photoacoustics) and low-frequency photoacoustics can thus provide more information than any one method alone. Experiments on intact chloroplasts and on a blue-green alga demonstrate that photothermal radiometry and photoacoustic methodologies can also be used for these tissues.     

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.     

Induction kinetics of heat emission before and after photoinhibition in cotyledons of Raphanus sativus

Heat emitted during non-radiative de-excitation was determined in vivo by the photoacoustic method. The dependence of the photoacoustic signal on the length of the pulses (modulation frequency) of the excitation light and the effect of continuous light, which saturates photosynthesis but does not directly contribute to the signal, are described. The induction kinetic of heat emission measured with intact leaves differed only slightly from the induction kinetic of fluorescence (Kautsky effect) detected in parallel. The photoacoustic signal at high modulation frequencies (279 Hz), which represents the signal of heat emission, and the photoacoustic signal at low modulation frequencies (17 Hz), interpreted as a signal of pulsed oxygen evolution superimposed on the heat emission, were measured with leaves before and after photoinhibition. It was demonstrated that after photoinhibition the decrease in fluorescence yield and in photosynthetic activity (here detected as photoacoustic signal at 17 Hz) are paralleled by an increase in the yield of non-radiative deexcitation (photoacoustic signal at 279 Hz). The increase of heat emission, which has been hypothized for photoinhibited leaves, could now be proved by measuring the induction kinetics of the photoacoustic signal.     

Detection of photosynthetic energy storage in a photosystem I reaction center preparation by photoacoustic spectroscopy

Thermal emission and photochemical energy storage were examined in photosystem I reaction center/core antenna complexes (about 40 Chl a/P700) using photoacoustic spectroscopy. Satisfactory signals could only be obtained from samples bound to hydroxyapatite and all samples had a low signal-to-noise ratio compared to either PS I or PS II in thylakoid membranes. The energy storage signal was saturated at low intensity (half saturation at 1.5 W m^-2) and predicted a photochemical quantum yield of >90%. Exogenous donors and acceptors had no effect on the signal amplitudes indicating that energy storage is the result of charge separation between endogenous components. Fe(CN)₆ ^-3 oxidation of P700 and dithionite-induced reduction of acceptors F_A-F_B inhibited energy storage. These data are compatible with the hypothesis that energy storage in PS I arises from charge separation between P700 and Fe-S centers F_A-F_B that is stable on the time scale of the photoacoustic modulation. High intensity background light (160 W m^-2) caused an irreversible loss of energy storage and correlated with a decrease in oxidizable P700; both are probably the result of high light-induced photoinhibition. By analogy to the low fluorescence yield of PS I, the low signal-to-noise ratio in these preparations is attributed to the short lifetime of Chl singlet excited states in PS I-40 and its indirect effect on the yield of thermal emission.Abbreviations FFT fast Föurier transform - HA hydroxyapatite - I₅₀ half saturation intensity for energy storage - PA photoacoustic - PS photosystem - PS I-40 photosystem I reaction center/core antenna complex containing about 40 Chl a/P700 - 201-1 photoacoustic energy storage signal - S/N signal-to-noise     

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     

Computer-controlled pulse modulation system for analysis of photoacoustic signals in the time domain

A newly developed photoacoustic system for measurement of photosynthetic reactions in intact leaves is described. The system is based on pulsed light-emitting diodes, the pulse program and pulse response analysis being computer controlled. Separation of various components in the overall photoacoustic signal is achieved by curve fitting analysis of the responses following individual measuring light pulses in the millisecond time domain. This procedure is in distinction to the conventionally used analysis in the frequency domain, with the advantage that various signal components are obtained by on-line deconvolution, yielding simultaneous recordings of photothermal (complement of energy storage) and photobaric (evolution and uptake) signals. The basic components of the new system are described by block diagrams and the principal steps for deconvolution of the overall photoacoustic response are outlined. An example of application with simultaneous recording of chlorophyll fluorescence is given. It is apparent that the photobaric uptake component represents a significant part of the overall signal, particularly during induction of photosynthesis after dark-adaptation. This component probably contains not only O₂-uptake but uptake of CO₂ as well.Abbreviations PA photoacoustic - LED light-emitting-diode - RAM random access memory     

Low frequency photoacoustics for monitoring the photobaric component in vivo of green leaves

The photoacoustic frequency spectrum under steady-state conditions from Philodendron green leaves attached to the plant was measured in the 0.2–200 Hz frequency range. The PA amplitude spectrum showed a maximum at low frequency (around 1 Hz) which was attributed to an optimum frequency for oxygen evolution. The signal decreased at a lower frequency, where the oxygen or carbondioxide uptake starts to become important. Efficiency of the oxygen evolution as a function of excitation light intensity was determined for different levels of background light.     

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.     

A gas-permeable photoacoustic cell

A photoacoustic cell assembly is described that is permeable to CO₂ and other gases but not water vapor. As a replacement for the usually employed solid cover, this cell uses a cover containing a small fritted glass disk that holds a small piece of 6.4 m Teflon film against the sample.With the above arrangement it was possible to increase the rate of O₂ evolution measured photoacoustically about 3 times in Zea mays leaves and about 1.7 times in Phaseolus vulgaris leaves upon adding CO₂ to the gas stream. The extent of energy storage was also enhanced with supplemental CO₂ in Zea and Ulva but less so in Phaseolus. The maximum improvements of photosynthetic activities were obtained when the gas stream contained 2.5–5% CO₂. These high concentrations were presumably necessary as the result of a high resistance to diffusion through the gas-permeable cover.Abbreviations ES energy storage - PA photoacoustic - RuBP ribulose-1,5-bisphosphate carboxylase     

Photoacoustic Measurements of Cyclic Electron Flow around Photosystem I in Leaves Adapted to Light-States 1 and 2

The photoacoustic technique was used to study in vivo cyclicelectron flow through PS I in intact plant leaves irradiatedwith far-red light. Appreciable PS I-cyclic ATP formation wasshown to occur in both C-3 and C-4 plants. It was also observedthat various environmental/experimental conditions leading toa significant inhibition of the linear photosynthetic electrontransport were associated with a stimulation of PS I-cyclicenergy storage. In contrast, in vivo adaptation of leaves tolight-states 1 and 2 did not induce any photoacoustically measurablechanges in the capacity of PS I for cyclic electron transfer.Consequently, the presented data do not support the recent hypothesisthat the main function of light-induced state transitions isto regulate the balance between linear (PS I+PS II) and cyclic(PS I) electron transport in the chloroplasts. (Received February 24, 1992; Accepted May 15, 1992)     

High-irradiance Stress and Photochemical Activities of Photosystems 1 and 2 in vivo

High-irradiance (HI) stress induced changes in the photosynthetic energy storage (ES) of photosystems 1 (ESPS1) and 2 (ESPS2) were studied with 650 nm modulated radiation in intact sugar maple (Acer saccharum Marsh.) leaves. HI-treatment (420 W m-2, 1 h) caused an inhibition of about 40 % in ESPS2 and an enhancement of about 60 % in ESPS1. The rate of PS1 cyclic electron transport, measured with 705 nm modulated radiation, also increased in HI-treated leaves. There was a clear state 1- state 2 transition in HI-treated leaves. ESPS1 increased significantly and ESPS2 decreased drastically in leaves preadapted to state 1 after HI (600 W m-2, 30 min) treatment. Thus, the increase in PS1 activity observed immediately after HI-treatment in leaves preadapted to state 1 can be due to the coupling of LHC2 to PS1 during the HI-treatment. Further, the dissociation of LHC2 from PS2 during the HI-treatment resulted in apparently (about 15 %) greater inhibition than the "true" inhibition of PS2 activity. The presence of LHC2 with PS2 (state 1) at the time of HI-treatment caused no additional damage to PS2 or its coupling to PS1 offered no apparent HI-treatment. Further, the dissociation of LHC2 from PS2 during the HI-treatment resulted in apparently (about 15 %) greater inhibition than the "true" inhibition of PS2 activity. The presence of LHC2 with PS2 (state 1) at the time of HI-treatment caused no additional damage to PS2 or its coupling to PS1 offered no apparent protection to the photosynthetic apparatus.     

On the nature of the photosynthetic energy storage monitored by photoacoustic spectroscopy

The photosynthetic energy storage yield of uncoupled thylakoid membranes was monitored by photoacoustic spectroscopy at various measuring beam intensities. The energy storage rate as evaluated by the half-saturation measuring beam intensity (i₅₀) was inhibited by 3-(3,4-dichlorophenyl)-1,1 dimethylurea, by heat inactivation or by artificial electron acceptors specific for photosystem I or photosystem II; and was activated by electron donors to photosystem I. The reactions involving both photosystems were all characterized by a similar maximal energy storage yield of 16±2 percent. The data could be interpreted if we assumed that the energy storage elicited by the photosystems at 35 Hz is detected at the level of the plastoquinone pool.Abbreviations PS photosystem - Tes N-Tris [hydroxymethl] methyl-2-aminoethanesulfonic acid - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DCIP 2,6-dichlorophenolindophenol - FeCN potassium ferricyanide - DCBQ 2,5-dichlorobenzoquinone - TMPD N,N,N-tetramethyl-p-phenilenediamine