Rapid chlorophyll a fluorescence transient of Lemna gibba leaf as an indication of light and hydroxylamine effect on photosystem II activity.

Rapid chlorophyll fluorescence transient induced by saturating flash (3000 micromol of photons m-2 s-1) was investigated when Lemna gibba had been exposed to light (100 micromol of photons m-2 s-1) causing the Kautsky effect or in low light intensity unable to trigger PSII photochemistry. Measurements were made by using, simultaneously, a pulse amplitude modulated fluorometer and plant efficiency analyzer system, either on non-treated L. gibba leaf or those treated with different concentrations of hydroxylamine (1-50 mM) causing gradual inhibition of the water splitting system. When any leaf was exposed to continuous light during the Kautsky effect, a rapid fluorescence transient may reflect current activity of photosystem II within the photosystem II complex. Under those conditions, a variation of transition steps appearing over time was related to a drastic change to the photosystem II functional properties. This value indicated that the energy dissipation through non-photochemical pathways was undergoing extreme change. The change of rapid fluorescence transient, induced under continuous light, when compared to those obtained under very low light intensity, confirmed the ability of photosystem II to be capable to undergo rapid adaptation lasting about two minutes. When the water splitting system was inhibited and electron donation partially substituted by hydroxylamine, the adaptation ability of photosystem II to different light conditions was lost. In this study, the change of rapid fluorescence kinetic and transient appearing over time was shown to be a good indication for the change of the functional properties of photosystem II induced either by light or by hydroxylamine.     

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

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

Light-induced fluorescence quenching in the light-harvesting chlorophyll a/b protein complex

Summary Irradiation of the principal photosystem II light-harvesting chlorophyll-protein antenna complex, LHC II, with high light intensities brings about a pronounced quenching of the chlorophyll fluorescence. Illumination of isolated thylakoids with high light intensities generates the formation of quenching centres within LHC II in vivo, as demonstrated by fluorescence excitation spectroscopy. In the isolated complex it is demonstrated that the light-induced fluorescence quenching: a) shows a partial, biphasic reversibility in the dark; b) is approximately proportional to the light intensity; c) is almost independent of temperature in the range 0–30°C; d) is substantially insensitive to protein modifying reagents and treatments; e) occurs in the absence of oxygen. A possible physiological importance of the phenomenon is discussed in terms of a mechanism capable of dissipating excess excitation energy within the photosystem II antenna.Abbreviations chla chlorophyll a - chlb chlorophyll b - F₀ fluorescence yield with reaction centers open - F_m fluorescence yield with reaction centres closed - F_i fluorescence at the plateau level of the fast induction phase - LHC II light-harvesting chlorophyll a/b protein complex II - PS II photosystem II - PSI photosystem I - Tricine N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine     

Mechanisms of photoinhibition induced by high light in Hosta grown outdoors

Aims It has long been recognized that photoinhibition of photosynthesis is induced by high light. However, our recent studies are not consistent with this traditional view. Therefore, the objective of this study is to explore the induction of photoinhibition and its mechanisms under full sunlight outdoors.
Methods Changes of leaf morphology, gas exchange, and chlorophyll a fluorescence were measured to investigate the induction and mechanisms of photoinhibition under high light in Hosta, which is a typical shade-tolerant plant.
Important findings Hosta plants grown under full sunlight (HT) and low light (LT) developed sun- and shade-type leaf morphological characteristics, respectively. Under a full sunlight, Hosta plants had lower photosynthetic rate and chlorophyll content than under the LT; whereas, there were only slight difference in the maximum quantum yield of photosystem II (F_v/F_m) between the two treatments, suggesting that Hosta plants could grow normally under full sunlight without severe photoinhibition. After transition from the low to a high light (LHT), the photosynthetic rate and maximum quantum yield of photosystem II decreased sharply, reflecting that the LHT treatment led to irreversibly inactivation of photosystem II. Additionally, the shape of chlorophyll a fluorescence transients also changed significantly; the relative fluorescence yield of the K and J steps were reduced by 24.3% and 34.2%, respectively, indicating that the acceptor side of photosystem II was damaged more severely than the donor side. Consequently, we postulate that photoinhibition in Hosta leaves is mainly induced by the sudden enhancement of light intensity outdoors. Hosta can acclimate to high irradiance through leaf development outdoors. Our finding is of great significance in understanding the acclimation of plants to high light and cultivation of shade-tolerant plants in field.     

State transitions,photosystem stoichiometry adjustment and non-photochemical quenching in cyanobacterial cells acclimated to light absorbed by photosystem I or photosystem II

Cells of the cyanobacterium Synechococcus 6301 were grown in yellow light absorbed primarily by the phycobilisome (PBS) light-harvesting antenna of photosystem II (PS II), and in red light absorbed primarily by chlorophyll and, therefore, by photosystem I (PS I). Chromatic acclimation of the cells produced a higher phycocyanin/chlorophyll ratio and higher PBS-PS II/PS I ratio in cells grown under PS I-light. State 1-state 2 transitions were demonstrated as changes in the yield of chlorophyll fluorescence in both cell types. The amplitude of state transitions was substantially lower in the PS II-light grown cells, suggesting a specific attenuation of fluorescence yield by a superimposed non-photochemical quenching of excitation. 77 K fluorescence emission spectra of each cell type in state 1 and in state 2 suggested that state transitions regulate excitation energy transfer from the phycobilisome antenna to the reaction centre of PS II and are distinct from photosystem stoichiometry adjustments. The kinetics of photosystem stoichiometry adjustment and the kinetics of the appearance of the non-photochemical quenching process were measured upon switching PS I-light grown cells to PS II-light, and vice versa. Photosystem stoichiometry adjustment was complete within about 48 h, while the non-photochemical quenching occurred within about 25 h. It is proposed that there are at least three distinct phenomena exerting specific effects on the rate of light absorption and light utilization by the two photoreactions: state transitions; photosystem stoichiometry adjustment; and non-photochemical excitation quenching. The relationship between these three distinct processes is discussed.Abbreviations Chl chlorophyll - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F relative fluorescence intensity at emission wavelength nm - F _o fluorescence intensity when all PS II traps are open - light 1 light absorbed preferentially by PS I - light 2 light absorbed preferentially by PS II - PBS phycobilisome - PS photosystem     

Photosystem II Activity, Plastoquinone A Levels, and Fluorescence Characterization of a Virescens Mutant of Barley

Chloroplasts isolated from seedlings of a virescens mutant of barley (Hordeum vulgare L cv Gateway) grown for 6 days under continuous illumination had lower levels of photosystem II activities on a chlorophyll basis than wild-type seedlings. After 8 days, however, the photosystem II rates of the mutant and wild-type were approximately equal. Lower levels of the photosystem II activities in the mutant were correlated with a smaller functional plastoquinone pool size as determined by room temperature fluorescence induction. Higher levels of extractable plastoquinone A on a chlorophyll basis, however, were obtained from mutant chloroplasts. An increased room temperature fluorescence yield in the mutant was shown to be due to a higher level of initial fluorescence. A decreased sigmoidicity in the room temperature fluorescence induction transient in the presence of diuron and an increased 77 K fluorescence emission at 680 nanometers lead us to believe that a certain population of the light harvesting chlorophyll protein complex in the mutant membranes is unconnected to photo-system II reaction centers. Although photochemical activities of the mutant approach wild-type values as the mutant develops, the population of dissociated light harvesting complexes does not appear to change.     

环境强光诱导玉簪叶片光抑制的机制

为进一步阐述光抑制的强光诱导和发生机制, 该文以喜阴植物玉簪(Hosta spp.)为材料研究其光抑制发生规律及其与环境光强的关系。结果表明, 全日照和遮阴条件下玉簪叶片发育分别形成适应强光和弱光的形态特征; 与遮阴处理相比, 强光下生长的玉簪光合速率和叶绿素含量较低, 但两种处理叶片最大光化学效率差异很小, 证明强光下植株可以正常生长且光合机构未发生严重的光抑制。将遮阴处生长的植株转移到全日照下, 光合速率和最大光化学效率急剧下降; 荧光诱导动力学曲线发生明显改变, 而且光系统II供体侧和受体侧荧光产量的变化幅度分别达到24.3%和34.2%, 表明玉簪由弱光转入强光后光系统II发生不可逆失活, 且受体侧受到的伤害较供体侧更严重。因此, 作者认为环境光强骤然提高并超过玉簪生长光强时很容易诱导其光合机构发生严重的光抑制。该研究对于理解植物适应光环境的策略以及喜阴植物的优质栽培有重要意义。     

Uptake of diuron and concomitant loss of photosynthetic activity in leaves as visualized by imaging the red chlorophyll fluorescence

The principles of the chlorophyll (Chl) fluorescence induction kinetics (known as Kautsky effect) and their change by the photosystem II herbicide diuron are presented together with the Chl fluorescence emission spectra of a normal and diuron-inhibited leaf. By imaging the Chl fluorescence emission of green leaves the successive uptake of diuron and the concomitant loss of photosynthetic quantum conversion from the leaf base to the leaf tip are documented.     

The photodynamic action of eosin,a singlet-oxygen generator

Eosin, a known generator of singlet oxygen, applied to leaf discs of Pisum sativum L. sensitized the inhibition of photosynthesis. Analysis of partial photosynthetic electron-transport reactions and of the kinetics of variable chlorophyll fluorescence located the damage at photosystem II. This injury required the presence of oxygen and was also caused by the irradiation of eosin-treated leaf tissue with green light. The role of oxygen and photodynamic reactions in the susceptibility of photosystem II to damage by environmental stresses is discussed.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DCPIP 2,6-dichlorophenolindophenol - DPC 1,5-diphenylcarbazide - PSI photosystem I - PSII photosystem II - ¹O₂ singlet oxygen - Tricine N-[2-hydroxyl-3,1-bis(hydroxymethyl)ethyl]-glycine     

Primary sites of ozone-induced perturbations of photosynthesis in leaves: identification and characterization in Phaseolus vulgaris using high resolution chlorophyll fluorescence imaging

High resolution imaging of chlorophyll a fluorescence was used to identify the sites at which ozone initially induces perturbations of photosynthesis in leaves of Phaseolus vulgaris. Leaves were exposed to 250 and 500 nmol mol(-1) ozone at a photosynthetically active photon flux density of 300 micromol m(-2) s(-1) for 3 h. Images of fluorescence parameters indicated that large decreases in both the maximum and operating quantum efficiencies of photosystem II had occurred in cells adjacent to stomata in the upper, but not lower, leaf surfaces. However, this treatment did not produce any significant changes in the maximum or operating quantum efficiencies of photosystem II in the leaves when estimated from fluorescence parameters measured with a conventional, integrating fluorometer. The localized decreases in photosystem II photochemical efficiencies were accompanied by an increase in the minimal fluorescence level, which is indicative of photoinactivation of photosystem II complexes and a decrease in stomatal conductance. Perturbations of photochemical efficiencies were not observed in cells associated with all of the stomata on the upper leaf surface or within cells distant from the upper leaf surface. It is concluded that ozone penetrates the leaf through stomata and initially damages only cells close to stomatal pores.     

Age dependent alterations in photosystem II acceptor side in Cucumis sativus cotyledonary leaf thylakoids: analysis of binding characteristics of herbicide [14C]-atrazine.

Senescence induced temporal changes in photosystems can be conveniently studied in cotyledonary leaves. We monitored the protein, chlorophyll and electron transport activities in Cucumis sativus cv Poinsette cotyledonary leaves and observed that by 20th day, there was a 50%, 41% and 30-33% decline in the chlorophyll, protein and photosystem II activity respectively when compared to 6th day cotyledonary leaves taken as control. We investigated the changes in photosystem II activity (O2 evolution) as a function of light intensity. The photosystem II functional antenna decreased by 27% and the functional photosystem II units decreased by 30% in 20-day old cotyledonary leaf thylakoids. The herbicide [14C]-atrazine binding assay to monitor specific binding of the herbicide to the acceptor side of photosystem II reaction centre protein, D1, showed an increase in the affinity for atrazine towards D1 protein and decrease in the QB binding sites in 20th day leaf thylakoids when compared to 6th day leaf thylakoids. The western blot analysis also suggested a decrease in steady state levels of D1 protein in 20th day cotyledonary leaf thylakoids as compared to 6th day sample which is in agreement with [14C]-atrazine binding assay and light saturation kinetics.     

Chlorophyll thermoluminescence of leaf discs: simple instruments and progress in signal interpretation open the way to new ecophysiological indicators

Luminescence from photosynthetic material observed in darkness following illumination is a delayed fluorescence produced by a recombination of charge pairs stored in photosystem II, i.e. the back-reaction of photosynthetic charge separation. Thermoluminescence (TL) is a technique consisting of a rapid cooling followed by the progressive warming of a preilluminated sample to reveal the different types of charge pairs as successive emission bands, which are resolved better than the corresponding decay phases recorded at constant temperatures. Progress in thermoelectric Peltier elements and in compact light detectors made the development of simple, affordable and transportable instruments possible. These instruments take advantage of multifurcated light guides for combined TL, fluorescence and absorbance/reflectance measurements. Meanwhile, experiments on unfrozen leaf discs, with excitation by single turn-over flashes or far red light and infiltration by specific inhibitors/uncouplers, have led to a better understanding of in vivo TL signals. Much like chlorophyll fluorescence and in a complementary way, TL in the 0-60 degrees C temperature range not only informs on the state of photosystem II in leaf tissues and its possible alterations, but also gives a broader insight into the energetic state inside the chloroplast by probing (1) the light-induced or dark-stable thylakoid proton gradient through the protonation of the Mn oxygen-evolving complex, (2) the induction of cyclic/chlororespiratory electron flow towards the plastoquinone pool, (3) the [NADPH+ATP] assimilatory potential. By a different mechanism, warming above 60 degrees C without preillumination reveals chemiluminescence high temperature thermoluminescence (HTL) bands due to the radiative thermolysis of peroxides, which are indicators of oxidative stress in leaves.     

Light-dependent inhibitory action of p-nitrothiophenol on Photosystem II in relation to the redox state of electron carriers

The photosystem-II activity of chloroplasts was inhibited by the treatment with p-nitrothiophenol (NphSH) in the light, and the inhibition was accompanied by a change of the fluorescence spectrum. Aromatic mercaptans examined were active in causing this inhibition and fluorescence change. These effects of p-nitrothiophenol were highly accelerated by blocking the electron transport on the oxidation side of photosystem II by carbonyl cyanide-m-chlorophenylhydrazone (CCCP) or Tris · HCl or heat pre-treatment, whereas these were suppressed by blocking the transport on the reduction side by 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). It was deduced that the site of NphSH action in the electron transport chain is closer to the reaction center of photosystem II that the blocking site of CCCP or Tris · HCl or heat, and that such a site in photosystem II is exposed to be modified with NphSH when electron carriers on the oxidation side of photosystem II are oxidized by illumination.     

A New Mechanism for Adaptation to Changes in Light Intensity and Quality in the Red Alga Porphyra perforata: III. Fluorescence Transients in the Presence of 3-(3,4-Dichlorophenyl)-1,1-dimethylurea

In the red alga Porphyra perforata, the level of chlorophyll fluorescence in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) decreased during illumination of the thallus. The results showed that: (a) this decay was related to the photooxidative activity of photosystem I; (b) Q, the primary electron acceptor of photosystem II, became oxidized during the decay of the fluorescence; (c) reagents which inhibit the back reaction of photosystem II inhibited the decay.

From these results, it is suggested that, when conditions in the chloroplasts of this red alga become too oxidative, excess light energy can be converted to heat as a result of an accelerated back reaction of photosystem II. This may be one of the mechanisms by which this alga can cope with the high salt and high light conditions that can occur in its natural habitat.

     

A model of photosystem II for the analysis of a fast fluorescence rise in plant leaves

The polyphasic patterns of fluorescence induction rise in pea leaves in vivo and after the treatment with ionophores have been studied using a plant efficiency analyzer. To analyze in detail photosystem II (PS II) electron transfer processes, an extended PS II model was applied, which included the sums of exponential functions to specify explicitly the light-driven formation of the transmembrane electric potential (delta psi(t)) as well as pH in the lumen (pHL(t)) and stroma (pHs(t)). PS II model parameters and numerical coefficients in delta psi(t), and pHs(t) were evaluated to fit fluorescence induction data for different experimental conditions: leaf in vivo or after ionophore treatment at low or high light intensity. The model imitated changes in the pattern of fluorescence induction rise due to the elimination of transmembrane potential in the presence of ionophores, when delta psi = 0 and pHL(t), pHS(t) altered to small extent relative to control values in vivo, with maximum delta psi(t) approximately 90 MB and delta psi(t) approximately 40 MB, for the stationary state at deltapH aproximately equal to 1.8. As the light intensity was increased from 300 to 1200 micromol x m(-2) x s(-1), the heat dissipation rate constants increased threefold for nonradiative recombination of P680+Phe- and by approximately 30% for P680+Q(A)-. The parameters delta psi, pH(S) and pH(L) were analyzed as factors of PS II redox state populations and fluorescence yield. The kinetic mechanism of qE quenching is discussed, which is related with light induced pH(L) lumen acidification, when Q(A)- and P680+ recombination probability increases to regulate the QA reduction.     

Dependence of fluorescence spectra of chloroplasts from the activity of photosystem 2]

By means of high sensitive spectrofluorometer the fluorescence spectra have been measured of normal chloroplasts and those with blocked photosystem 2 activity due to photoinhibition or treatment with 0.6 M tris-buffer. At room temperature fluorescence spectra of inactivated chloroplasts are similar to the spectrum of normal chloroplasts measured at low light intensity. Under excitation by intense light a decrease of intensity at 685 nm is appeared (about 3-4 times) in the fluorescence spectra of inactivated chloroplasts as compared to the spectrum of normal chloroplasts. The sharp intensity decrease of maxima at 685 and 695 nm (3-4 times) and small decrease at 680 and 730 nm (by 30-50%) are observed in low temperature fluorescence spectra of inactivated chloroplasts. Thus, the damage of photosystem 2 reaction centres is not accompanied by the preferential decrease of the only fluorescence band. The similarity of fluorescence difference spectra of chloroplasts distinguished by the state of photosystem 2 reaction centre, and the complex structure of difference spectra indicate that the variable fluorescence of chloroplasts during the induction is due to the emission of bulk chlorophyll alpha of the photosystem 2.     

Nuclear pea mutants deficient in chlorophyll b and the major polypeptide of the light-harvesting chlorophyll a/b protein of photosystem II

Eight chlorophyll b deficient nuclear mutants of pea (Pisum sativum L.) have been characterized by low temperature fluorescence emission spectra of their leaves and by the ultrastructure, photochemical activities and polypeptide compositions of the thylakoid membranes. The room temperature fluorescence induction kinetics of leaves and isolated thylakoids have also been recorded. In addition, the effects of Mg²⁺ on the fluorescence kinetics of the membranes have been investigated. The mutants are all deficient in the major polypeptide of the light-harvesting chlorophyll a/b protein of photosystem II. The low temperature fluorescence emission spectra of aurea-5106, xantha-5371 and –5820 show little or no fluorescence around 730 nm (photosystem I fluorescence), but possess maxima at 685 and 695 nm (photosystem II fluorescence). These three mutants have low photosystem II activities, but significant photosystem I activities. The long-wavelength fluorescence maximum is reduced for three other mutants. The Mg²⁺ effect on the variable component of the room temperature fluorescence (685 nm) induction kinetics is reduced in all mutants, and completely absent in aurea-5106 and xantha-5820. The thylakoid membranes of these 2 mutants are appressed pairwise in 2-disc grana of large diameter. Chlorotica-1-206A and–130A have significant long-wavelength maxima in the fluorescence spectra and show the largest Mg²⁺ enhancement of the variable part of the fluorescence kinetics. These two mutants have rather normally structured chloroplast membranes, though the stroma regions are reduced. The four remaining mutants are in several respects of an intermediate type.Abbreviations Chl chlorophyll - CPI Chi-protein complex I, F_o, F_v - F_m parameters of room temperature chlorophyll fluorescence induction kinetics - F685, F695 and F-1 components of low temperature chlorophyll emission with maximum at 685, 695 and ca 735 nm, respectively - PSI photosystem I - PSII photosystem II - LHCI and LHCII light-harvesting chlorophyll a/b complexes associated with PSI and PSII, respectively - SDS sodium dodecyl sulfate     

黑暗诱导衰老对不同年代冬小麦品种旗叶光系统Ⅱ功能的影响

小麦品种的更新换代是小麦产量不断提高的重要因素,阐明小麦品种演替过程中不同生理特性的变化对新品种选育具有重要参考价值.旗叶衰老速率快慢是影响小麦产量水平的关键因素,目前对于不同小麦品种衰老过程中旗叶光系统Ⅱ功能的变化规律尚不清楚.本试验选用1941-2014年间河南地区不同时期种植的31个品种,通过黑暗诱导离体叶片衰老,测定旗叶叶绿素荧光诱导动力学参数、叶绿素相对含量的变化,分析了光系统Ⅱ功能的变化规律.结果表明:品种演替过程中旗叶的叶绿素含量逐渐提高,衰老过程中近代品种叶绿素的降解速率低于较早年代品种;旗叶衰老过程中,近代品种荧光诱导动力学曲线的J点上升幅度小于I点;品种更替过程中光系统Ⅱ最大光化学效率和单位面积有活性反应中心数目逐渐增加,但是近代品种降低速率低于较早年代品种.叶绿素含量的变化与未衰老叶片中F_v/F_m没有显著相关性,但是随着衰老程度增加,相关性逐渐增大,且趋势线斜率逐渐提高;光系统Ⅱ单位面积有活性反应中心数目与品种育成时间呈显著正相关,且随着衰老程度增加,相关程度和趋势线斜率均显著提高.综上,小麦品种演替过程中,旗叶叶绿素含量逐渐升高,降解速率逐渐减缓,光合电子传递过程中Q_A到Q_B电子传递的抗衰老能力得到改善,从而减缓了衰老过程中光系统Ⅱ最大光化学效率和有活性反应中心的衰减速率,同时,叶绿素含量的提高和旗叶光系统Ⅱ抗衰老能力的增强也是品种更替过程中产量逐渐提高的重要因素.     

Effect of High Cation Concentrations on Photosystem II Activities

The effects of wide concentration ranges of NaCl, KCl, and MgCl₂ on ferricyanide reduction and the fluorescence induction curve of isolated spinach (Spinacia oleracea) chloroplasts were investigated. Concentrations of the monovalent salts above 100 mm and MgCl₂ above 25 mm produced a decrease in the rate of ferricyanide reduction by thylakoids uncoupled with 2.5 mm NH₄Cl which cannot be attributed to changes in the primary photochemical capacity of photosystem II. Salt-induced decreases in the effective concentration of the secondary electron acceptor of photosystem II, plastoquinone, reduce the capacity for secondary photochemistry of photosystem II and this could contribute to the reduction in ferricyanide reduction by uncoupled thylakoids at high salinities. The rate of ferricyanide reduction by coupled thylakoids is little affected by salinity changes, indicating that the rate-limiting phosphorylation mechanism in electron flow from water to ferricyanide in coupled thylakoids is salt-tolerant, whereas the rate-limiting reaction in uncoupled ferricyanide reduction is considerably affected by salinity changes. Salt-induced changes in the fluorescence induction curve are interpreted in terms of changes in the rate constants for excitation decay by radiationless transitions, exciton transfer from photosystem II chlorophylls to other associated chlorophyll species, and photochemistry.     

Detection of rapid induction kinetics with a new type of high-frequency modulated chlorophyll fluorometer

A newly developed modulation fluorometer is described which operates with 1 sec light pulses from a light-emitting diode (LED) at 100 KHz. Special amplification circuits assure a highly selective recording of pulse fluorescence signals against a vast background of non-modulated light. The system tolerates ratios of up to 1:10⁷ between measuring light and actinic light. Thus it is possible to measure the dark fluorescence yield and record the kinetics of light-induced changes. A high time resolution allows the recording of the rapid relaxation kinetic following a saturating single turnover flash. Examples of system performance are given. It is shown that following a flash the reoxidation kinetics of photosystem II acceptors are slowed down not only by the inhibitor DCMU, but by a number of other treatments as well. From a light intensity dependency of the induction kinetics the existence of two saturated intermediate levels (I₁ and I₂) is apparent, which indicates the removal of three distinct types of fluorescence quenching in the overall fluorescence rise from F₀ to F_max.Abbreviations Q_A and Q_B consecutive electron acceptors of photosystem II - PS II photosystem II - P 680 reaction center chlorophyll of photosystem II - F₀ minimum fluorescence yield following dark adaptation - F_max maximum fluorescence yield - DCMU 3-(3, 4-dichlorophenyl)-1, 1-dimethyl-urea - DCCD N,N-dicyclohexylcarbodiimide - PQ plastoquinone - DAD diaminodurene Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement.