Regulation levels of photosynthesis processes

The basic mechanisms of kinetic regulation of photosynthetic processes are considered, which provide a strict light regulation of electron transfer in photosynthetic reaction centers and a more flexible regulation at the level of interaction of photosystems, transmembrane ion fluxes and coupling with dark reactions of the Calvin cycle. A generalized model was developed, which integrates the modern knowledge about photosynthetic processes of higher plants. The general principles of multilevel regulation in photosynthetic systems are discussed.     

EFFECTS OF LONG TERM EXPOSURE TO LOW TEMPERATURE ON THE PHOTOSYNTHETIC APPARATUS OF DUNALIELLA TERTIOLECTA (CHLOROPHYCEAE)1

The effects of long term exposure to suboptimal growth temperature on the photosynthetic apparatus of Dunaliella tertiolecta Butcher were investigated using carbon fixation rate versus irradiance curves and the variable fluorescence induction method. Carbon fixation rates per unite chlorophyll a at saturating (p^B_m) and subsaturating (α^B) irradiances were 55% and 39% lower, respectively, at 12° C than at 20° C. Chlorophyll a quotas and the spectrally averaged in vivo absorption cross section normalized to chlorophyll a (a^*) were not significantly different at these two temperatures. Analysis of the fluorescence kinetics revealed 1) no significant variations of the amount of PSII photoactive reaction centers per unit chlorophyll a, 2) a 14% decrease of the PSII quantum yield(+) and 3) a 29% decrease of the energy transfer efficiency between the light harvesting chlorophyll a pigment bed and the PSII reaction centers. The decrease in energy transfer efficiency between the antennae and the PSII reaction centers at 12° C was interpreted as a mechanism to avoid photoinhibition.     

Photosynthetic Light Utilization Efficiency, Photosystem II Heterogeneity, and Fluorescence Quenching in Chlamydomonas reinhardtii during the Induction of the CO(2)-Concentrating Mechanism

The photosynthetic light-response curve, the relative amounts of the different photosystem II (PSII) units, and fluorescence quenching were altered in an adaptive manner when CO₂-enriched wild-type Chlamydomonas reinhardtii cells were transferred to low levels of CO₂. This treatment is known to result in the induction of an energy-dependent CO₂-concentrating mechanism (CCM) that increases the internal inorganic carbon concentration and thus the photosynthetic CO₂ utilization efficiency. After 3 to 6 h of low inorganic carbon treatment, several changes in the photosynthetic energy-transducing reactions appeared and proceeded for about 12 h. After this time, the fluorescence parameter variable/maximal fluorescence yield and the amounts of both PSIIα and PSIIβ (secondary quinone electron acceptor of PSII-reducing) centers had decreased, whereas the amount of PSIIβ (secondary quinone electron acceptor of PSII-nonreducing) centers had increased. The yield of noncyclic electron transport also decreased during the induction of the CCM, whereas both photochemical and nonphotochemical quenching of PSII fluorescence increased. Concurrent with these changes, the photosynthetic light-utilization efficiency also decreased significantly, largely attributed to a decline in the curvature parameter θ, the convexity of the photosynthetic light-response curve. Thus, it is concluded that the increased CO₂ utilization efficiency in algal cells possessing the CCM is maintained at the cost of a reduced light utilization efficiency, most probably due to the reduced energy flow through PSII.     

Modeling of primary photosynthetic processes using the kinetic Monte Carlo method

Processes that occur in the ensemble of photosynthetic electron transport systems have been modeled using a kinetic Monte Carlo method. The size of a simulated ensemble (3–5 million elementary photosynthetic chains) corresponds to the number of photosynthetic reaction centers in a plant cell. The method enables one to modify the structure of a model system according to different concepts of the organization of processes in a photosynthetic membrane. Using this model, the experimental kinetics of the chlorophyll fluorescence induction associated with the Photosystem II and the redox transformations of a photoactive pigment of the Photosystem I have been successfully reproduced. The model was verified by comparing the calculated fluorescence induction curves to experimental curves, obtained in the presence of various photosynthesis inhibitors and under temperature inactivation of the Photosystem II donor side.     

Photosynthetic units of sun and shade plants

A computer analysis of fluorescence induction curves of leaves treated with 3-(3,4-dichlorophenyl)-1,1 dimethylurea was done for several species. These measurements gave the ratios of the total chlorophyll to photosystem II reaction centers. This communication is a preliminary survey of sun and shade plants and demonstrates a significant variation in this ratio. In the sun plants, the photosynthetic unit sizes (chlorophyll reaction centers) varied between 220 to 480. The shade plants gave numbers mostly in the range between 630 to 940. The computer analysis of the fluorescence data also gave the connectivity parameter of energy transfer between photosynthetic units of photosystem II which varied between 0.2 and 0.5 but did not show any obvious correlation to the photosynthetic unit size.     

Transfer of excitation energy in photosynthesis: some thoughts

The aim of the present paper is to aid biologists understand the complex physical problems of intramolecular energy transfer, in particular, between antenna (bacterio) chlorophyll molecules in vivo.The author has attempted, in the first part of the paper, to explain complicated processes of excitation transfer in a language understandable to readers with knowledge in fundamentals of general physics, but not in molecular optics.The second part of this paper is a critical review relevant to the specifics of physical theories and their applicability to the problem of energy transfer in antenna (bacterio) chlorophylls ((B) Chls) to reaction centers (RCs) in the photosynthetic organisms.Abbreviations PSU photosynthetic unit - RC reaction center - Chl chlorophyll - BChl bacteriochlorophyll - _r intrinsic radiactive lifetime - _fl fluorescence lifetime - _fl fluorescence quantum yield - S^* singlet excited state of a molecule     

不同剂量~(137)Cs-γ辐射对毛竹幼苗叶片叶绿素荧光参数的影响

利用不同剂量的137Cs-γ射线对毛竹(Phyllostachys heterocycla ‘Pubescens’)种子进行辐射,测定实生苗叶片中的光合色素含量和叶绿素荧光参数等指标,探讨辐射对毛竹幼苗生长的影响,为筛选有利的突变单株奠定良好基础。结果表明:30或60Gy的137Cs-γ射线辐射后,毛竹幼苗的光合色素含量以及最大荧光强度(Fm)、可变荧光强度(Fv)、PSII最大光化学效率(Fv/Fm)、PSII的潜在活性(Fv/Fo)、PSII实际光化学效率(Yield)和表观光合电子传递速率(ETR)等荧光参数值均高于90Gy辐射处理,说明较低剂量辐射后PSII反应中心的能量捕获效率高,且具有较强的光合能力;而90Gy的137Cs-γ射线辐射对毛竹的影响则与之相反。不同处理剂量之间叶片光能耗散程度以及表观光合电子传递速率-光合有效辐射(ETR-PAR)响应曲线的分析结果也进一步证实了以上结论。     

Effect of photosystem II reaction center closure on nanosecond fluorescence relaxation kinetics

The fluorescence decay of chlorophyll in spinach thylakoids was measured as a function of the degree of closure of Photosystem II reaction centers, which was set for the flowed sample by varying either the preillumination by actinic light or the exposure of the sample to the exciting pulsed laser light. Three exponential kinetic components originating in Photosystem II were fitted to the decays; a fourth component arising from Photosystem I was determined to be negligible at the emission wavelength of 685 nm at which the fluorescence decays were measured. Both the lifetimes and the amplitudes of the components vary with reaction center closure. A fast (170–330 ps) component reflects the trapping kinetics of open Photosystem II reaction centers capable of reducing the plastoquinone pool; its amplitude decreases gradually with trap closure, which is incompatible with the concept of photosynthetic unit connectivity where excitation energy which encounters a closed trap can find a different, possibly open one. For a connected system, the amplitude of the fast fluorescence component is expected to remain constant. The slow component (1.7–3.0 ns) is virtually absent when the reaction centers are open, and its growth is attributable to the appearance of closed centers. The middle component (0.4–1.7 ns) with approximately constant amplitude may originate from centers that are not functionally linked to the plastoquinone pool. To explain the continuous increase in the lifetimes of all three components upon reaction center closure, we propose that the transmembrane electric field generated by photosynthetic turnover modulates the trapping kinetics in Photosystem II and thereby affects the excited state lifetime in the antenna in the trap-limited case.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - HEPES 4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid - PQ plastoquinone - PSI and PSII Photosystem I and II - Q_A and Q_B primary and secondary quinone acceptor of PSII     

Fluorescence induction curves registered from individual microalgae cenobiums in the process of population growth

Registration of chlorophyll fluorescence induction curves (IC) from individual microalgae cenobiums was performed during Scenedesmus quadricauda culture growth. Emphasis was placed on the analysis of patterns of the slow phase of IC, since these slow fluorescence transitions reflect complex interactions between primary and secondary photosynthetic processes. A classification was performed of the ICs obtained according to the patterns of their slow phase. Four different types of such patterns were distinguished. The microalgae population structure with respect to IC patterns was investigated at different stages of culture growth. The distribution of microalgae cenobiums over the patterns of IC was found to change in accordance with the stage of population development. At the stage of the population growth enhancement, nonmonotonous IC dominated with a high steady-state level of fluorescence. The stage of linear growth was characterized by IC with monotonous decay kinetics and low steady-state level of fluorescence. At the third stage including the phases of growth inhibition, stationary state and the beginning of cell death the population structure was the most heterogeneous, with all IC patterns observed.Abbreviations CO₂ carbon dioxide - ETC electron transfer chain - Fl fluorescence - FNR ferredoxin-NADPH reductase - IC induction curve of chlorophyll fluorescence - PQ plastoquinone - PS I Photosystem I - PS II Photosystem II - Q_A primary quinone acceptor of PS II     

Functional and structural organization of chlorophyll in the developing photosynthetic membranes of Euglena gracilis Z. I. Formation of System II photosynthetic units during greening under optimal light intensity

The relationships between light-harvesting chlorophyll and reaction centers in Photosystem II were analyzed during the chloroplast development of dark-grown, non-dividing Euglena gracilis Z. Comparative measurements included light saturation of photosynthesis, oxygen evolution under flashing-light and fluorescence induction. The results obtained can be summarized as follows: (1) Photosystem II photocenters are formed in parallel with chlorophyll synthesis, but after a longer lag phase. (2) As a consequence, the chlorophyll: reaction center ratio (Emerson's type photosynthetic unit) decreases during greening. (3) This decrease is accompanied by considerable changes in the energy transfer and trapping properties of Photosystem II. Most of the initially synthesized chlorophylls are inactive in the transfer of excitations to active photochemical centers and are shared among newly formed Photosystem II photocenters; as a consequence, the number of chlorophylls functionally connected to each Photosystem II photocenter decreases and cooperativity between these centers appears. Results are discussed in terms of chlorophyll organization in developing photosynthetic membranes with reference to the lake or puddle models of photosynthetic unit organization.     

Functional and structural organization of chlorophyll in the developing photosynthetic membranes of Euglena gracilis Z. II. Formation of system II photosynthetic units during greening under optimal light intensity.

The relationships between light-harvesting chlorophyll and reaction centers in Photosystem II were analyzed during the chloroplast development of dark-grown, non-dividing Euglena gracilis Z. Comparative measurements included light saturation of photosynthesis, oxygen evolution under flashing-light and fluorescence induction. The results obtained can be summarized as follows: (1) Photosystem II photocenters are formed in parallel with chlorophyll synthesis, but after a long lag phase. (2) As a consequence, the chlorophyll reaction center ratio (Emerson's type photosynthetic unit) decreases during greening. (3) This decrease is accompanied by considerable changes in the energy transfer and trapping properties of Photosystem II. Most of the initially synthesized chlorophylls are inactive in the transfer of excitations to active photochemical centers and are shared among newly formed Photosystem II photocenters; as a consequence, the number of chlorophylls functionally connected to each Photosystem II photocenter decreases and cooperatively between these centers appears. Results are discussed in terms of chlorophyll organization in developing photosynthetic membranes with reference to the lake or puddle models of photosynthetic unit organization.     

Variable chlorophyll fluorescence and its use for assessing physiological condition of plant photosynthetic apparatus

Analysis of plant behavior under diverse environmental conditions would be impossible without the methods for adequate assessment of the processes occurring in plants. The photosynthetic apparatus and its reaction to stress factors provide a reliable source of information on plant condition. One of the most informative methods based on monitoring the plant biophysical characteristics consists in detection and analysis of chlorophyll a fluorescence. Fluorescence is mainly emitted by chlorophyll a from the antenna complexes of photosystem II (PSII). However, fluorescence depends not only on the processes in the pigment matrix or PSII reaction centers but also on the redox reactions at the PSII donor and acceptor sides and even in the entire electron transport chain. Presently, a large variety of fluorometers from various manufacturers are available. Although application of such fluorometers does not require specialized training, the correct interpretation of the results would need sufficient knowledge for converting the instrumental data into the information on the condition of analyzed plants. This review is intended for a wide range of specialists employing fluorescence techniques for monitoring the physiological plant condition. It describes in a comprehensible way the theoretical basis of light emission by chlorophyll molecules, the origin of variable fluorescence, as well as relations between the fluorescence parameters, the redox state of electron carriers, and the light reactions of photosynthesis. Approaches to processing and analyzing the fluorescence induction curves are considered in detail on the basis of energy flux theory in the photosynthetic apparatus developed by Prof. Reto J. Strasser and known as a “JIP-test.” The physical meaning and relation of each calculated parameter to certain photosynthetic characteristics are presented, and examples of using these parameters for the assessment of plant physiological condition are outlined.     

State-transitions facilitate robust quantum yields and cause an over-estimation of electron transport in Dunaliella tertiolecta cells held at the CO2 compensation point and re-supplied with DIC

Photosynthetic energy consumption and non-photosynthetic energy quenching processes are inherently linked. Both processes must be controlled by the cell to allow cell maintenance and growth, but also to avoid photodamage. We used the chlorophyte algae Dunaliella tertiolecta to investigate how the interactive regulation of photosynthetic and non-photosynthetic pathways varies along dissolved inorganic carbon (DIC) and photon flux gradients. Specifically, cells were transferred to DIC-deplete media to reach a CO₂ compensation before being re-supplied with DIC at various concentrations and different photon flux levels. Throughout these experiments we monitored and characterized the photophysiological responses using pulse amplitude modulated fluorescence, oxygen evolution, 77 K fluorescence emission spectra, and fast-repetition rate fluorometry. O₂ uptake was not significantly stimulated at DIC depletion, which suggests that O₂ production rates correspond to assimilatory photosynthesis. Fluorescence-based measures of relative electron transport rates (rETRs) over-estimated oxygen-based photosynthetic measures due to a strong state-transitional response that facilitated high effective quantum yields. Adoption of an alternative fluorescence-based rETR calculation that accounts for state-transitions resulted in improved linear oxygen versus rETR correlation. This study shows the extraordinary capacity of D. tertiolecta to maintain stable effective quantum yields by flexible regulation of state-transitions. Uncertainties about the control mechanisms of state-transitions are presented.     

Theory of fluorescence induction in photosystem II: derivation of analytical expressions in a model including exciton-radical-pair equilibrium and restricted energy transfer between photosynthetic units.

The theoretical relationships between the fluorescence and photochemical yields of PS II and the fraction of open reaction centers are examined in a general model endowed with the following features: i) a homogeneous, infinite PS II domain; ii) exciton-radical-pair equilibrium; and iii) different rates of exciton transfer between core and peripheral antenna beds. Simple analytical relations are derived for the yields and their time courses in induction experiments. The introduction of the exciton-radical-pair equilibrium, for both the open and closed states of the trap, is shown to be equivalent to an irreversible trapping scheme with modified parameters. Variation of the interunit transfer rate allows continuous modulation from the case of separated units to the pure lake model. Broadly used relations for estimating the relative amount of reaction centers from the complementary area of the fluorescence kinetics or the photochemical yield from fluorescence levels are examined in this framework. Their dependence on parameters controlling exciton decay is discussed, allowing assessment of their range of applicability. An experimental induction curve is analyzed, with a discussion of its decomposition into alpha and beta contributions. The sigmoidicity of the induction kinetics is characterized by a single parameter J related to Joliot's p, which is shown to depend on both the connectivity of the photosynthetic units and reaction center parameters. On the other hand, the relation between J and the extreme fluorescence levels (or the deviation from the linear Stern-Volmer dependence of 1/phi f on the fraction of open traps) is controlled only by antenna connectivity. Experimental data are consistent with a model of connected units for PS II alpha, intermediate between the pure lake model of unrestricted exciton transfer and the isolated units model.     

Biophysical Methods of Ecological Monitoring. Photosynthetic characteristics of tree plants growing in Moscow city

In this work, we studied the influence of ecological factors (distance from thoroughfares) on photosynthetic characteristics of leaves of four tree species growing in Moscow city. Photosynthetic activity of leaves was assayed by instrumental methods of probing the functional state of the photosynthetic apparatus, using electron paramagnetic resonance for measuring the kinetics of photooxidation of P₇₀₀ centers, thermoluminescence, and slow induction of chlorophyll fluorescence. It has been shown that the kinetic parameters of the induction curves, as determined from the kinetics of P₇₀₀ photooxidation and slow fluorescence induction in dark-adapted leaves, are sensitive to variations of plant growth conditions. These parameters can be used as informative characteristics in ecological monitoring.     

A master equation theory of fluorescence induction, photochemical yield, and singlet-triplet exciton quenching in photosynthetic systems.

A master equation theory is formulated to describe the dependence of the fluorescence yield (phi) in photosynthetic systems on the number of photons (Y) absorbed per photosynthetic unit (or domain). This theory is applied to the calculation of the dependence of the fluorescence yield on Y in (a) fluorescence induction, and (b) singlet exciton-triplet excited-state quenching experiments. In both cases, the fluorescence yield depends on the number of previously absorbed photons per domain, and thus evolves in a nonlinear manner with increasing Y. In case a, excitons transform the photosynthetic reaction centers from a quenching state to a nonquenching state, or a lower efficiency of quenching state; subsequently, absorbed photons have a higher probability of decaying by radiative pathways and phi increases as Y increases. In case b, ground-state carotenoid molecules are converted to long-lived triplet excited-state quenchers, and phi decreases as Y increases. It is shown that both types of processes are formally described by the same theoretical equations that relate phi to Y. The calculated phi (Y) curves depend on two parameters m and R, where m is the number of reaction centers (or ground-state carotenoid molecules that can be converted to triplets), and R is the ratio phi (Y leads to infinity)/(Y leads to 0). The finiteness of the photosynthetic units is thus taken into account. The m = 1 case corresponds to the "puddle" model, and m leads to infinity to the "lake," or matrix, model. It is shown that the experimental phi (Y) curves for both fluorescence induction and singlet-triplet exciton quenching experiments are better described by the m leads to infinity cases than the m = 1 case.     

Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes,fluorescence and photosynthesis in leaves of Hedera canariensis

The role of the xanthophyll cycle in regulating the energy flow to the PS II reaction centers and therefore in photoprotection was studied by measurements of light-induced absorbance changes, Chl fluorescence, and photosynthetic O₂ evolution in sun and shade leaves of Hedera canariensis. The light-induced absorbance change at 510 nm (A₅₁₀) was used for continuous monitoring of zeaxanthin formation by de-epoxidation of violaxanthin. Non-radiative energy dissipation (NRD) was estimated from non-photochemical fluorescence quenching (NPQ).High capacity for zeaxanthin formation in sun leaves was accompanied by large NRD in the pigment bed at high PFDs as indicated by a very strong NPQ both when all PS II centers are closed (F'_m) and when all centers are open (F'_o). Such F_o quenching, although present, was less pronounced in shade leaves which have a much smaller xanthophyll cycle pool.Dithiothreitol (DTT) provided through the cut petiole completely blocked zeaxanthin formation. DTT had no detectable effect on photosynthetic O₂ evolution or the photochemical yield of PS II in the short term but fully inhibited the quenching of F_o and 75% of the quenching of F_m, indicating that NRD in the antenna was largely blocked. This inhibition of quenching was accompanied by an increased closure of the PS II reaction centers.In the presence of DTT a photoinhibitory treatment at a PFD of 200 mol m^-2 s^-1, followed by a 45 min recovery period at a low PFD, caused a 35% decrease in the photon yield of O₂ evolution, compared to a decrease of less than 5% in the absence of DTT. The F_v/F_m ratio, measured in darkness showed a much greater decrease in the presence than in the absence of DTT. In the presence of DTT F_o rose by 15–20% whereas no change was detected in control leaves.The results support the conclusion that the xanthophyll cycle has a central role in regulating the energy flow to the PS II reaction centers and also provide direct evidence that zeaxanthin protects against photoinhibitory injury to the photosynthetic system.Abbreviations F, F_m, F_o, F_v Fluorescence yield at actual degree of PS II center closure, when all centers are closed, when all centers are open, variable fluorescence - NPQ non-photochemical fluorescence quenching - NRD non-radiative energy dissipation - PFD photon flux density - Q_A primary acceptor PS II     

Energy transfer and fluorescence mechanisms in photosynthetic membranes

Energy transfer in photosynthetic membranes involves the migration of excitons from light‐harvesting antenna chlorophyll‐protein complexes to the reaction center complexes. Recent efforts have focused on determining the time of arrival of excitons (trapping times) at the reaction centers following excitation with a single picosecond laser pulse. Three different approaches have been utilized: (1) determination of appearance of separated charges within the reaction centers by differential absorbtion spectroscopy, (2) determination of appearance of separated charges by fast photoemf measurements, and (3) kinetics of decay of fluorescence. The first two methods provide more direct information on exciton trapping by reaction centers than fluorescence methods, but are experimentally difficult to realize. Therefore, much activity has centered around the accurate measurement and analysis of fluorescence‐decay profiles by single‐photon counting methods. In green plants, about three different components with lifetimes of about 100 psec, 200 to 500 psec, and >1 nsec, have been reported. The first two components are believed to be related to trapping rates by reaction centers, while the third component is attributed to a charge recombination (Klimov) mechanism. Results from photoemf and exciton‐exciton annihilation experiments are consistent with the interpretation that the first decay component reflects exciton‐trapping rates. A critical analysis and discussion of these fast energy‐transfer phenomena in photosynthetic membranes of green plants are offered in this review.     

Theoretical fluorescence induction curves derived from coupled differential equations describing the primary photochemistry of photosystem II by an exciton-radical pair equilibrium

Fluorescence induction curves were calculated from a molecular model for the primary photophysical and photochemical processes of photosystem II that includes reversible exciton trapping by open (PHQ_A) and closed (PHQ^-_A) reaction centers (RCs), charge stabilization as well as quenching by oxidized (P⁺HQ^(-)_A) RCs. For the limiting case of perfectly connected photosynthetic units (“lake model”) and thermal equilibrium between the primary radical pair (P⁺H^-) and the excited singlet state, the primary reactions can be mathematically formulated by a set of coupled ordinary differential equations (ODE). These were numerically solved for weak flashes in a recursive way to simulate experiments with continuous illumination. Using recently published values for the molecular rate constants, this procedure yielded the time dependence of closed RCs as well as of the fluorescence yield (= fluorescence induction curves). The theoretical curves displayed the same sigmoidal shapes as experimental fluorescence induction curves. From the time development of closed RCs and the fluorescence yield, it was possible to check currently assumed proportionalities between the fraction of closed RCs and either (a) the variable fluorescence, (b) the complementary area above the fluorescence induction curve, or (c) the complementary area normalized to the variable fluorescence. By changing selected molecular rate constants, it is shown that, in contrast to current beliefs, none of these correlations obeys simple laws. The time dependence of these quantities is strongly nonexponential. In the presence of substances that quench the excited state, the model predicts straight lines in Stern-Volmer plots. We further conclude that it is impossible to estimate the degree of physical interunit energy transfer from the sigmoidicity of the fluorescence induction curve or from the curvature of the variable fluorescence plotted versus the fraction of closed RCs.     

Effects of freezing out protein vibrational modes on electron transfer kinetics in bacterial reaction centers

We propose a model that some vibrational modes of the protein in bacterial photosynthetic reaction centers may be frozen at low temperatures. The freezing of the protein-environmental motion can affect the electron transfer rate through changes in the reorganization energy and the free energy gap. We offer a qualitative explanation of the different kinetics of the ET processes in reaction centers which are cooled in the dark and cooled under illumination.