Reduced levels of cytochrome bf complex in transgenic tobacco leads to marked photochemical reduction of the plastoquinone pool,without significant change in acclimation to irradiance

Tobacco transgenics with decreased amounts of the FeS apoprotein were generated using an antisense RNA construct targeted against the nuclear-encoded Rieske FeS protein of cytochrome bf complex [Price et al. (1995) Aust J Plant Physiol 22: 285–297]. FeS phenotypes ranging from intermediate to low were obtained which had 69% and 26% of the Rieske FeS protein of wild type. Similar reductions in the other subunits of cytochrome bf complex, cytochrome f, cytochrome b₅₆₃and the 17 kDa subunit, were demonstrated in the thylakoids of intermediate and low FeS phenotypes. Confirmation that the levels of assembled cytochrome bf in leaves matched the levels of the FeS protein was demonstrated by laser flash-induced redox absorbance changes in leaves, with the extents of cytochrome f oxidation and cytochrome b₅₆₃reduction being equivalent to the decreased amounts of the subunits in isolated thylakoids of the antisense plants. Despite greatly enhanced photochemical reduction of Q_Aand the plastoquinone pool in the antisense plants, light acclimation of the FeS phenotypes to irradiance did not occur. Furthermore, the state 1–state 2 transitions were identical in wild type and antisense plants. Our results suggest that neither Q_Anor the plastoquinone pool acts alone in either the redox control of gene expression or the regulation of light energy distribution between the photosystems. We suggest rather that reduced plastoquinone acting at the inner Q_psite of cytochrome bf complex is involved in molecular redox signalling.     

Pigments, photosynthesis and photoinhibition in two amphibious plants: consequences of varying carbon availability

In the present study, we investigated the effects of CO(2) availability on photosynthesis, photoinhibition and pigmentation in two species of amphibious plants, Lobelia cardinalis and Nesaea crassicaulis. The plants were grown emergent under atmospheric conditions and submerged under low and high CO(2) availability. Compared with Lobelia, Nesaea had thin leaves and few stomata in all CO(2) treatments. While Lobelia expressed no variation in anthocyanin content among treatments, Nesaea produced high concentrations of anthocyanin when submerged. Lobelia photosynthesis increased in response to increasing CO(2) availability, and photoinhibition was negatively related to xanthophyll content. By contrast, Nesaea photosynthesis was highest under submerged conditions, and there was no relationship between photoinhibition and the xanthophyll content. We conclude that the response of Lobelia to varying CO(2) availability is similar to that of terrestrial plants and that this species relies on the xanthophyll cycle for nonphotochemical quenching (NPQ) and protection against photoinhibition. By contrast, the thin leaves, few stomata and low levels of chlorophylls and accessory pigments in Nesaea, relative to Lobelia, suggest adaptation to a submerged habitat. While Nesaea does not seem to rely on the xanthophyll cycle or other xanthophylls for NPQ, some role of anthocyanins in the protection against photoinhibition cannot be ruled out, owing to its effect as a sunscreen and as an efficient quencher of free radicals.     

Stimulation of the diadinoxanthin cycle by UV-B radiation in the diatom Phaeodactylum tricornutum

In this study we show that the diadinoxanthin cycle in the diatom Phaeodactylum tricornutum is stimulated by mild UV-B radiation. High steady state concentrations of diatoxanthin established during a period of strong actinic illumination with white light (300 mol photons m-2 s-1 PAR) are further increased if weak UV-B (3 mol photons m-2 s-1) is additionally applied. Short term increases in the diatoxanthin concentration caused by UV-B strongly correlate with a stoichiometric decrease in diadinoxanthin. The UV-B dependent increase in diatoxanthin is correlated with a concommitant enhancement of non-photochemical quenching of chlorophyll fluorescence and a decrease in the quantum efficiency of oxygen evolution. This indicates that UV-B induced diatoxanthin functions in thermal energy dissipation. Possible scenarios for a stimulation of the diadinoxanthin cycle by UV-B are discussed.     

Acclimation of leaf carotenoid composition and ascorbate levels to gradients in the light environment within an Australian rainforest

The influence of the growth photon flux density (PFD) on the size and composition of the carotenoid pool and the size of the reduced ascorbate pool was determined across a light gradient from the forest floor to the canopy and the forest edge of a sub-tropical rainforest in New South Wales, Australia. Nineteen plant species (most collected from multiple sites) representing a broad taxonomic range consistently possessed larger total carotenoid pools when found growing in more exposed sites. There was a significant positive correlation between β-carotene content and growth PFD and a significant negative correlation between α-carotene content and growth PFD. Neoxanthin content exhibited no significant trend while the trend in lutein content varied with mode of expression. The pigments of the xanthophyll cycle (violaxanthin, antheraxanthin and zeaxanthin) exhibited the most pronounced response to growth PFD; they comprised a much greater portion of the total carotenoid pool in high light-acclimated plants. The pool of reduced ascorbate was also several-fold greater in high light-acclimated plants. These acclimatory changes in carotenoid and ascorbate content are consistent with a need for a greater capacity to dissipate excessive absorbed light energy in high light-acclimated plants.     

The role of calcium in the pH-dependent control of Photosystem II

pH-dependent inactivation of Photosystem (PS) II and related quenching of chlorophyll-a-fluorescence have been investigated in isolated thylakoids and PS II-particles and related to calcium release at the donor side of PS II. The capacity of oxygen evolution (measured under light saturation) decreases when the pH is high and the pH in the thylakoid lumen decreases below 5.5. Oxygen evolution recovers upon uncoupling. The pH-response of inactivation can be described by a 1 H⁺-transition with an apparent pK-value of about 4.7. The yield of variable fluorescence decreases in parallel to the inactivation of oxygen evolution. pH-dependent quenching requires light and can be inhibited by DCMU. In PS II-particles, inactivation is accompanied by a reversible release of Ca²⁺-ions (one Ca²⁺ released per 200 Chl). In isolated thylakoids, where a pH was created by ATP-hydrolysis, both inactivation of oxygen evolution (and related fluorescence quenching) by internal acidification and the recovery of that inactivation can be suppressed by calcium-channel blockers. In the presence of the Ca²⁺-ionophore A23187, recovery of Chl-fluorescence (after relaxation of the pH) is stimulated by external Ca²⁺ and retarded by EGTA. As shown previously (Krieger and Weis 1993), inactivation of oxygen evolution at low pH is accompanied by an upward shift of the midpoint redox-potential, E_m, of Q_A. Here, we show that in isolated PS II particles the pH-dependent redox-shift (about 160 mV, as measured from redox titration of Chl-fluorescence) is suppressed by Ca²⁺-channel blockers and DCMU. When a redox potential of –80 to –120mV was established in a suspension of isolated thylakoids, the primary quinone acceptor, Q_A, was largely reduced in presence of a pH (created by ATP-hydrolysis) but oxidized in presence of an uncoupler. Ca²⁺-binding at the lumen side seems to control redox processes at the lumen- and stroma-side of PS II. We discuss Ca²⁺-release to be involved in the physiological process of high energy quenching.     

Electron transport to oxygen mitigates against the photoinactivation of Photosystem II in vivo

The role of electron transport to O₂ in mitigating against photoinactivation of Photosystem (PS) II was investigated in leaves of pea (Pisum sativum L.) grown in moderate light (250 mol m^–2 s^–1). During short-term illumination, the electron flux at PS II and non-radiative dissipation of absorbed quanta, calculated from chlorophyll fluorescence quenching, increased with increasing O₂ concentration at each light regime tested. The photoinactivation of PS II in pea leaves was monitored by the oxygen yield per repetitive flash as a function of photon exposure (mol photons m^–2). The number of functional PS II complexes decreased nonlinearly with increasing photon exposure, with greater photoinactivation of PS II at a lower O₂ concentration. The results suggest that electron transport to O₂, via the twin processes of oxygenase photorespiration and the Mehler reaction, mitigates against the photoinactivation of PS II in vivo, through both utilization of photons in electron transport and increased nonradiative dissipation of excitation. Photoprotection via electron transport to O₂ in vivo is a useful addition to the large extent of photoprotection mediated by carbon-assimilatory electron transport in 1.1% CO₂ alone.Abbreviations Fm, Fo, Fv- maximal, initial (corresponding to open PS II traps) and variable chlorophyll fluorescence yield, respectively - NPQ- non-photochemical quenching - PS- photosystem - Q_A- primary quinone acceptor - qP- photochemical quenching coefficient     

Changes in Photosystem II fluorescence in Chlamydomonas reinhardtii exposed to increasing levels of irradiance in relationship to the photosynthetic response to light

The effects of a 60 min exposure to photosynthetic photon flux densities ranging from 300 to 2200 mol m^–2s^–1 on the photosynthetic light response curve and on PS II heterogeneity as reflected in chlorophyll a fluorescence were investigated using the unicellular green alga Chlamydomonas reinhardtii. It was established that exposure to high light acts at three different regulatory or inhibitory levels; 1) regulation occurs from 300 to 780 mol m^–2s^–1 where total amount of PS II centers and the shape of the light response curve is not significantly changed, 2) a first photoinhibitory range above 780 up to 1600 mol m^–2s^–1 where a progressive inhibition of the quantum yield and the rate of bending (convexity) of the light response curve can be related to the loss of Q_B-reducing centers and 3) a second photoinhibitory range above 1600 mol m^–2s^–1 where the rate of light saturated photosynthesis also decreases and convexity reaches zero. This was related to a particularly large decrease in PS II centers and a large increase in spill-over in energy to PS I.Abbreviations Chl chlorophyll - DCMU 3,(3,4-dichlorophenyl)-1,1-dimethylurea - F_M maximal fluorescence yield - F_pl intermediate fluorescence yield plateau level - F₀ non-variable fluorescence yield - F_v total variable fluorescence yield (F_M-F₀) - initial slope to the light response curve, used as an estimate of initial quantum yield - convexity (rate of bending) of the light response curve of photosynthesis - LHC light-harvesting complex - P_max maximum rate of photosynthesis - PQ plastoquinone - Q photosynthetically active photon flux density (400–700 nm, mol m^–2s^–1) - PS photosystem - Q_A and Q_B primary and secondary quinone electron acceptor of PS II     

The inhibited xanthophyll cycle is responsible for the increase in sensitivity to low temperature photoinhibition in rice leaves fed with glutathione

Exposure of intact rice leaves to an irradiance of 1000 μmol m⁻² s⁻¹ at 6 °C for 2 h caused severe photoinhibition of Photosystem II. The rate and extent of photoinhbition were greatly exacerbated in leaves fed with 10 mM reduced glutathione (GSH) or 10 mM cysteine. Analyses of antioxidant enzyme activities as well as the application of protein synthesis inhibitors revealed that the increased sensitivity to photoinhibition following GSH feeding was not related to its effect on cellular antioxidant systems. On the other hand, feeding with GSH markedly suppressed the formation of zeaxanthin and antheraxanthin via the xanthophyll cycle and its associated nonradiative energy dissipation in leaves chilled in high light, suggesting that the stimulating effect of exogenous GSH on photoinhibition may be attributable to its action on the xanthophyll cycle. In vitro experiments using isolated thylakoids indicated that GSH is a weak inhibitor of violaxanthin deepoxidation. The possible implications of these results are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Using chlorophyll fluorescence to assess the fraction of absorbed light allocated to thermal dissipation of excess excitation

In the present study we explored the possibility of assessing the allocation of photons absorbed by photosystem II (PSII) antennae to thermal energy dissipation and photosynthetic electron transport in leaves of several plant species under field conditions. Changes in chlorophyll fluorescence parameters were determined in situ over the course of an entire day in the field in sun-exposed leaves of two species with different maximal rates of photosynthesis, Helianthus annuus (sunflower) and Vinca major. Leaves of Vinca minor (periwinkle) growing in a deeply shaded location were also monitored. We propose using diurnal changes in the efficiency of open PSII centers (F′_v/F′_m) in these sun and shade leaves to (a) assess diurnal changes in the allocation of absorbed light to photochemistry and thermal energy dissipation and, furthermore, (b) make an estimate of changes in the rate of thermal energy dissipation, an analogous expression to the rate of photochemistry. The fraction of light absorbed in PSII antennae that is dissipated thermally (D) is proposed to be estimated from D = 1-F′_v/F′_m, in analogy to the widely used estimation of the fraction of light absorbed in PSII antennae (P) that is utilized in PSII photochemistry from P = F′_v/F′_m× q_P (where q_P is the coefficient for photochemical quenching; Genty, B., Briantais, J.-M. & Baker, N. R. 1989. Biochim. Biophys. Acta 990: 87-92). The rate of thermal dissipation is consequently given by D × PFD (photon flux density), again in analogy to the rate of photochemistry P × PFD, both assuming a matching behavior of photosystems I and II. Characterization of energy dissipation from the efficiency of open PSII centers allows an assessment from a single set of measurements at any time of day; this is particularly useful under field conditions where the fully relaxed reference values of variable or maximal fluorescence needed for the computation of nonphotochemical quenching may not be available. The usefulness of the assessment described above is compared with other currently used parameters to quantify nonphotochemical and photochemical chlorophyll fluorescence quenching.     

In search of a reversible stage of photoinhibition in a higher plant: No changes in the amount of functional Photosystem II accompany relaxation of variable fluorescence after exposure of lincomycin-treated Cucurbita pepo leaves to high light

Pumpkin (Cucurbita pepo L.) leaves in which chloroplast protein synthesis was inhibited with lincomycin were exposed to strong photoinhibitory light, and changes in F_O, F_M, F_V/F_M and in the amount of functional Photosystem II (O₂ evolution induced by saturating single-turnover flashes) were monitored during the high-light exposure and subsequent dark or low-light incubation. In the course of the photoinhibitory illumination, F_M, F_V/F_M and the amount of functional PS II declined continuously whereas F_O dropped rapidly to some extent and then slowly increased. If the experiments were done at room temperature, termination of the photoinhibitory illumination resulted in partial relaxation of the F_V/F_M ratio and in an increase in F_O and F_M. The relaxation was completed in 10–15 min after short-term (15 min) photoinhibitory treatment but continued 30–40 min if the exposure to high light was longer than 1 h. No changes in the amount of functional PS II accompanied the relaxation of F_V/F_M in darkness or in low light, in the presence of lincomycin. Transferring the leaves to low temperature (+4°C) after the room-temperature illumination (2 h) completely inhibited the relaxation of F_V/F_M. Low temperature did not suppress the relaxation if the photoinhibitory illumination had also been done at low temperature. The results indicate that illumination of lincomycin-poisoned pumpkin leaves at room temperature does not lead to accumulation of a reversibly photoinactivated intermediate.Abbreviations F_O, F_M chlorophyll fluorescence with all reaction centres open or closed, respectively - F_V variable fluorescence (F_V=F_M–F_O) - LHC Light-harvesting complex - PS II Photosystem II - Q_A, Q_B primary and secondary quinone electron acceptors of PS II, respectively - qN_E, qN_T, qN_I non-photochemical quenching due to high-energy state, state transition or photoinhibition, respectively     

Mathematical modelling of photoinhibition and Photosystem II repair cycle. I. Photoinhibition and D1 protein degradation in vitro and in the absence of chloroplast protein synthesis in vivo

The kinetics of photoinhibition of Photosystem II and D1 protein degradation were studied by applying mathematical modelling to new and published data. The word photoinhibition refers here only to such inhibition of PS II activity that requires chloroplast protein synthesis for recovery. It is shown that acceptor-side photoinhibition in vitro as well as in vivo photoinhibition in higher plants and cyanobacteria in the presence of prokaryotic translation inhibitors follow first-order kinetics. Degradation of damaged D1 protein also fits in a first-order reaction equation with respect to the concentration of photoinhibited PS II centres. It is shown that photoprotective lowering of the ratio of variable to maximum fluorescence can be distinguished from the lowering of this ratio associated with photoinhibition.     

Rapid changes in xanthophyll cycle-dependent energy dissipation and photosystem II efficiency in two vines, Stephania japonica and Smilax australis, growing in the understory of an open Eucalyptus forest

Leaves of Stephania japonica and Smilax australis were characterized in situ on the coast of north-eastern New South Wales, Australia, where they were growing naturally in three different light environments: deep shade, in the understory of an open Eucalyptus forest where they received frequent sunflecks of high intensity, and in an exposed site receiving full sunlight. In deep shade the xanthophyll cycle remained epoxidized during the day and the vast majority of absorbed light was utilized for photosynthesis. In the exposed site both deepoxidation and epoxidation of the xanthophyll cycle and changes in the level of xanthophyll-dependent thermal energy dissipation largely tracked the diurnal changes in photon flux density (PFD). In the understory the xanthophyll cycle became largely deepoxidized to zeaxanthin and antheraxanthin upon exposure of the leaves to the first high intensity sunfleck and this high level of deepoxidation was maintained throughout the day both during and between subsequent sunflecks. In contrast, thermal energy dissipation activity, and the efficiency of photosystem II, fluctuated rapidly in response to the changes in incident PFD. These findings suggest a fine level of control over the engagement of zeaxanthin and antheraxanthin in energy dissipation activity, presumably through rapid changes in thylakoid acidification, such that they became rapidly engaged for photoprotection during the sunflecks and rapidly disengaged upon return to low light when continued engagement might limit carbon gain.     

Photoinhibition of photosystem II in vivo is preceded by down-regulation through light-induced acidification of the lumen: Consequences for the mechanism of photoinhibition in vivo

The mechanism of photoinhibition of photosystem II (PSII) was studied in intact leaf discs of Spinacia oleracea L. and detached leaves of Vigna unguiculata L. The leaf material was exposed to different photon flux densities (PFDs) for 100 min, while non-photochemical (q_N) and photochemical quenching (q_p) of chlorophyll fluorescence were monitored. The ‘energy’ and redox state of PSII were manipulated quite independently of the PFD by application of different temperatures (5–20° C), [CO₂] and [O₂] at different PFDs. A linear or curvilinear relationship between q_p and photoinhibition of PSII was observed. When [CO₂] and [O₂] were both low (30 μl · l^?1 and 2%, respectively), PSII was less susceptible at a given q_p than at ambient or higher [CO₂] and photoinhibition became only substantial when q_p decreased below 0.3. When high levels of energy-dependent quenching (q_E) (between 0.6 and 0.8) were reached, a further increase of the PFD or a further decrease of the metabolic demand for ATP and NADPH led to a shift from q_E to photoinhibitory quenching (q_I). This shift indicated that photoinhibition was preceded by down-regulation through light-induced acidification of the lumen. We propose that photoinhibition took place in the centers down-regulated by q_E. The shift from q_E to q_I occurred concomitant with q_P decreasing to zero. The results clearly show that photoinhibition does not primarily depend on the photon density in the antenna, but that photoinhibition depends on the energy state of the membrane in combination with the redox balance of PSII. The results are discussed with regard to the mechanism of photoinhibition of PSII, considering, in particular, effects of light-induced acidification on the donor side of PSII. Interestingly, cold-acclimation of spinach leaves did not significantly affect the relationship between q_P, q_E and photoinhibition of PSII at low temperature.     

Studies on the cytochrome b6/f complex. II. Localization of the complex in the thylakoid membranes from spinach and Chlamydomonas reinhardtii by immunocytochemistry and freeze-fracture analysis of b6/f mutants

The distribution of the b₆/f complex among stacked and unstacked thylakoid membranes was studied by immunocytochemistry and freeze-fracture analysis of mutants of Chlamydomonas reinhardtii lacking the complex. Immunogold labeling demonstrates the presence of b₆/f complex in both regions of the thylakoid membrane in spinach and in C. reinhardtii. Numerous modifications were observed in the ultrastructure of the thylakoid membranes of mutants from C. reinhardtii lacking the complex. These modifications are consistent with the presence of b₆/f complexes in different states of association in the stacked and unstacked regions of the thylakoid membrane. In particular we present evidence for an association of some b₆/f complexes with the reaction centers of Photosystem I and II in large PFu and EFs particles, respectively.     

Photosystem II in a mutant of Chlamydomonas reinhardtii lacking the 23 kDa psbP protein shows increased sensitivity to photoinhibition in the absence of chloride

The psbP gene product, the so called 23 kDa extrinsic protein, is involved in water oxidation carried out by Photosystem II. However, the protein is not absolutely required for water oxidation. Here we have studied Photosystem II mediated electron transfer in a mutant of Chlamydomonas reinhardtii, the FUD 39 mutant, that lacks the psbP protein. When grown in dim light the Photosystem II content in thylakoid membranes of FUD 39 is approximately similar to that in the wild-type. The oxygen evolution is dependent on the presence of chloride as a cofactor, which activates the water oxidation with a dissociation constant of about 4 mM. In the mutant, the oxygen evolution is very sensitive to photoinhibition when assayed at low chloride concentrations while chloride protects against photoinhibition with a dissociation constant of about 5 mM. The photoinhibition is irreversible as oxygen evolution cannot be restored by the addition of chloride to inhibited samples. In addition the inhibition seems to be targeted primarily to the Mn-cluster in Photosystem II as the electron transfer through the remaining part of Photosystem II is photoinhibited with slower kinetics. Thus, this mutant provides an experimental system in which effects of photoinhibition induced by lesions at the donor side of Photosystem II can be studied in vivo.Abbreviations Chl chlorophyll - DCIP 2,6-dichlorophenolindophenol - DPC 2,2-diphenylcarbonic dihydrazide - HEPES 4-(2-hydroxyethyl)-1-piperazinethanesulfonic acid - P₆₈₀ the primary electron donor to PS II - PpBQ phenyl-p-benzoquinone - PS II Photosystem II - Q_A the first quinone acceptor of PS II - Q_B the second quinone acceptor of PS II - SDS sodium dodecyl sulfate - Tris tris(hydroxymethyl)aminomethane - Tyr_D accessory electron donor on the D₂-protein - Tyr_Z tyrosine residue, acting as electron carrier between P₆₈₀ and the water oxidizing system     

Irradiance prior to and during desiccation improves the tolerance to excess irradiance in the desiccated state of the old forest lichen Lobaria pulmonaria

Hydrated thalli of the lichen Lobaria pulmonaria were either preconditioned to dim irradiance (DI, 5 μmol m⁻² s⁻¹) or medium irradiance (MI, 200 μmol m⁻² s⁻¹) for 6 h. After this 6 h period, the thalli were allowed to desiccate under the two respective irradiances. Thereafter, these dry lichens were exposed to high irradiance (HI, 1 000 μmol m⁻² s⁻¹) for 60 h. After this HI treatment, the maximal photochemical quantum yield (F_V/F_M) and the de-epoxidation state of xanthophyll cycle pigments (DEPS) were highest in thalli preconditioned to MI. Hence irradiance in the last hydrated period before sampling is significant for the physiological state of lichens. A standardized irradiance pre-treatment before start of experiments is recommended.     

Membrane phosphorylation leads to the partial detachment of the chlorophyll a/b protein from Photosystem II

The hypothesis that the chlorophyll fluorescence decline due to membrane phosphorylation is caused principally by the detachment and removal of LHCP from the LHCP-PS II matrix is examined. It is demonstrated that when membranes are phosphorylated in the dark (a) the fluorescence decline is greater when excited by light enriched in wavelengths absorbed mainly by LHCP (475 nm) than when excited by light absorbed to a large extent also by the PS II complex (435 nm), (b) titration with different artificial quenchers of chlorophyll fluorescence is unchanged after the phosphorylation-induced fluorescence decline, and (c) the F_v/F_m ratio does not change after the phosphorylation-induced fluorescence decline. These data indicate that it is indeed principally LHCP that interacts with the quencher (PS I presumably). This interaction involves a small fraction of the total PS II-coupled LHCP, which becomes functionally detached from the LHCP-PS II matrix.     

Studies on the cytochrome b6/f complex. I. Characterization of the complex subunits in Chlamydomonas reinhardtii

We have analyzed the heme-associated peroxidase activity in thylakoid membranes from the green algae Chlamydomonas reinhardtii after electrophoresis in the presence of sodium dodecyl sulfate. Besides cytochrome f and cytochrome b₆, we observed peroxidase activity in two other bands, of 34 and 11 kDa, of unknown origin. Characterization of the b₆/f complex subunits was undertaken by means of a comparison of the polypeptide deficiencies in several b₆/f mutants with the polypeptide content of preparations enriched in b₆/f complexes. We conclude that the b₆/f complex consists of five subunits. Using site-specific translation inhibitors, we show that cytochrome f, cytochrome b₆ and subunit IV are of chloroplast origin, whereas the Rieske protein and probably subunit V are translated on cytoplasmic ribosomes. A model of assembly of the complex is proposed: a cytochrome moiety, comprising the subunits of chloroplast origin, is assembled in the thylakoid membranes prior to the insertion and assembly of the subunits encoded in the nuclear genome.     

Analysis of the slow phases of the in vivo chlorophyll fluorescence induction curve. Changes in the redox state of Photosystem II electron acceptors and fluorescence emission from Photosystems I and II

An analysis of the photo-induced decline in the in vivo chlorophyll a fluorescence emission (Kautsky phenomenon) from the bean leaf is presented. The redox state of PS II electron acceptors and the fluorescence emission from PS I and PS II were monitored during quenching of fluorescence from the maximum level at P to the steady state level at T. Simultaneous measurement of the kinetics of fluorescence emission associated with PS I and PS II indicated that the ratio of PS I/PS II emission changed in an antiparallel fashion to PS II emission throughout the induction curve. Estimation of the redox state of PS II electron acceptors at given points during P to T quenching was made by exposing the leaf to additional excitation irradiation and determining the amount of variable PS II fluorescence generated. An inverse relationship was found between the proportion of PS II electron acceptors in the oxidised state and PS II fluorescence emission. The interrelationships between the redox state of PS II electron acceptors and fluorescence emission from PS I and PS II remained similar when the shape of the induction curve from P to T was modified by increasing the excitation photon flux density. The contributions of photochemical and non-photochemical quenching to the in vivo fluorescence decline from P to T are discussed.