Excitation energy transfer between pigment system II units in blue-green algae

Efficiency in excitation energy transfer from closed to open reaction center II in blue-green and red algae was estimated by the method developed by Joliot and Joliot (C.R. Acad. Sci. (1964) 258, 4622–4625) after slight modification; the number of open reaction centers II was counted from the mean O₂ yield of repetitive short flashes.

The efficiency in energy transfer in Chlorella pyrenoidosa was the same in our measurement as that reported by Joliot and Joliot (0.55 ± 0.02). However, the values obtained with four blue-green algae and one red alga were very small, in a range of 0.00–0.07. The low efficiency was always obtained independently of the size of the apparent photosynthetic unit which was varied by growth conditions. Results indicated that pigment system II forms a unit in which only one reaction center II is operative.     

Excitation transfer between photosynthetic units: the 1964 experiment

We review here the background and the experiments that led to the concept of excitation energy transfer among photosystem (PS) II units. On the basis of a kinetic analysis of oxygen evolution and chlorophyll a fluorescence yield, the authors showed, in 1964, that the PS II photochemical reaction involved in the formation of oxygen is not a first-order process. We concluded that excitation energy localized in a `photosynthetic unit' including a reduced primary acceptor is transferred with a high probability to neighboring PS II units. Here, the beginnings and the original data of this topic are presented. This revised version was published online in August 2006 with corrections to the Cover Date.     

Energy transfer in purple bacterial photosynthetic units from cells grown in various light intensities

Three photosynthetic membranes, called intra-cytoplasmic membranes (ICMs), from wild-type and the ?pucBA_abce mutant of the purple phototrophic bacterium Rps. palustris were investigated using optical spectroscopy. The ICMs contain identical light-harvesting complex 1–reaction centers (LH1–RC) but have various spectral forms of light-harvesting complex 2 (LH2). Spectroscopic studies involving steady-state absorption, fluorescence, and femtosecond time-resolved absorption at room temperature and at 77 K focused on inter-protein excitation energy transfer. The studies investigated how energy transfer is affected by altered spectral features of the LH2 complexes as those develop under growth at different light conditions. The study shows that LH1 → LH2 excitation energy transfer is strongly affected if the LH2 complex alters its spectroscopic signature. The LH1 → LH2 excitation energy transfer rate modeled with the Förster mechanism and kinetic simulations of transient absorption of the ICMs demonstrated that the transfer rate will be 2–3 times larger for ICMs accumulating LH2 complexes with the classical B800–850 spectral signature (grown in high light) compared to the ICMs from the same strain grown in low light. For the ICMs from the ?pucBA_abce mutant, in which the B850 band of the LH2 complex is blue-shifted and almost degenerate with the B800 band, the LH1 → LH2 excitation energy transfer was not observed nor predicted by calculations.     

A theory of excitation transfer in photosynthetic units

A theory of the excitation kinetics in the bacteria photosynthetic unit with regard to its globular structure is presented. It assumes that the excitation transfer between globulae is carried out by means of the mechanism of incoherent excitons, at the same time considering the finite time of the excitation fixation in the reaction center. A method of local perturbations is used with a view to finding a solution to the given problem. The expressions obtained for the fluorescence decay time and its quantum yield are discussed in connection with the multiple experiments considering the cubic as well as the hexagonal probable structure of the photosynthetic unit. The analysis given shows that the period of the excitation transfer between globulae equals 10 to 100 psec and the number of the globulae is less than 35. These conclusions fall in with the initial assumption of the energy transfer between globulae by incoherent excitons. Without considering the globularity, the consistency of the theory with experimental data becomes difficult.     

Quantitative relationship between two reaction centersin the photosynthetic system of blue-green algae

The quantitative relationship between reaction centers I andII was studied with blue-green algae Anabaena cylindrica, Anabaenavariabilis and Anacystis nidulans grown under different lightconditions. The number of reaction centers I was estimated fromthe P700 content and that of reaction centers II, from the O₂yield of repetitive short flashes. Supplementary determinationswere done with three other blue-green algae and one red alga.The maximum number of reaction centers II counted from the O₂yield of repetitive short flashes was markedly smaller thanthe total number of P700 in all algae tested when the algaewere grown under weak light; in the extreme case (Anabaena cylindrica),the ratio was only 0.258?0.015. This ratio became larger andclose to unity when the algae were grown under stronger light.Variation in the number of reaction centers in a single cellsuggested that reaction center I was a variable component. Ourresults indicate that the proportion of the two reaction centersmay markedly vary in blue-green algae depending on the growthconditions (Received November 13, 1978; )     

Capture frequency of excitations and energy transfer between photosynthetic units in the photosystem II

The connections which exist, in the photosynthetic apparatus, between the spatial arrangement of chlorophyll and the movement of excitation energy are discussed.The capture frequency of excitations in the photosystem II is analysed. At the microscopic level of a photosynthetic unit two stages are studied: the propagation of the excitation to the reaction centre and the photochemical utilization of the excitation by the centre. It is shown that the transport process is not a limiting one. It implies that the capture frequency depends on the reaction centre state. Thus it is possible to distinguish eight states for a photosynthetic unit of system II.At the macroscopic level of a set of units, the analysis of the fluorescence yield-fluctuations shows that these units are not isolated. It also indicates that the fluorescence emitted by the photosynthetic apparatus originates almost entirely from the system II, and that the reaction centres are traps for excitations whatever their states.     

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.     

Energy transfer in a single self-aggregated photosynthetic unit

The primary events of bacterial photosynthesis rely on the interplay of various specialized protein complexes organized in a supramolecular structure commonly termed the photosynthetic unit (PSU), which consists of the photochemical reaction center and of an associated antenna network. Employing single-molecule spectroscopic techniques we have been able to observe the excitation-energy transfer within a single PSU. From these findings we conclude that the building blocks of the PSU spontaneously form stable, functional aggregates in a non-membrane environment.     

Energy migration in purple bacteria. The criterion for discrimination between migration- and trapping-limited photosynthetic units

A criterion has been evolved for distinguishing between migration- and trapping-limited photosynthetic units (PSUs). Its application to purple bacteria has proved their PSUs to be of trapping-limited type. It means that any improvements of the molecular structure of their PSUs cannot noticeably increase the overall rate constant of excitation delivery from antenna BChls to reaction centers (RCs).Abbreviations PSUs photosynthetic units - RCs reaction centers - Chl chlorophyll - BChl bacteriochlorophyll - R intermolecular distance, _e - quantum yields of the primary excitation trapping and wasteful losses respectively - _fl excitation and fluorescence lifetimes respectively     

Relation between pigment content and photosynthetic characteristics in a blue-green algae

1. The blue-green alga Anacystis nidulans was cultured under steady state conditions at 25 and 39°C. and under several different light intensities to give five different types of cells. 2. Cells were submitted to pigment analysis based upon acetone extracts and aqueous extracts obtained by sonic disintegration. The different cell types show a threefold range of chlorophyll content and a fourfold range of phycocyanin content with only minor changes in the chlorophyll/phycocyanin ratio. Cells of highest pigment content were estimated to contain 2.8 per cent chlorophyll a and 24 per cent phycocyanin, the latter on a total chromoproteid basis. 3. Light intensity curves of photosynthesis were obtained for each of the cell types at 25 and at 39°C. The slopes of the light-limited regions of the curves are approximately linear functions of chlorophyll and phycocyanin contents. Maximum light-saturated rates of photosynthesis at 25 and 39° show no simple relation to pigment content.     

Exciton transfer between LH1 antenna complex and photosynthetic reaction center dimer

The exciton transfer between light-harvesting complex 1(LH1) and photosynthetic reaction center dimer is investigated theoretically. We assume a ring shape structure of the LH1 complex with dimer in the ring centre. The kinetic equations which describe the energy transfer between the antenna complex and reaction center dimer were derived. It was shown that the dimer does not act as a photon trap. There is a weak localization of the exciton on the dimer and there is relatively rapid back exciton transfer from dimer to antenna complex which depends on the number of the pigment molecules in the antenna ring. The relation between the rates of the exciton transfer from the antenna complex to dimer and back transfer from dimer to antenna complex has been derived.     

Energy transfer in a model of the photosynthetic unit of green plants

A model array is set up to represent a photosynthetic unit of 344 chlorophyll molecules of seven different spectral varieties and in definite orientations. The array is provided with two traps, representing the reaction centers of photosystems I and II. The number of jumps required to obtain a high probability of trapping is lower than on a similar array of undifferentiated chlorophylls by a factor of 15. Most of the molecules fall into two groups which transfer their energy predominantly into one or the other trap, and which may be regarded as functional photosystems I and II. The rate of transfer between these two functional photosystems can be controlled by redirecting the orientation of only six of the molecules, which occupy a key position in the array. The effect on trapping rates of reorientation of these molecules is especially pronounced when one of the traps is closed. This constitutes a model for the control of energy distribution between the two photosystems, as indicated in recent years through fluorescence studies.     

Energy transfer in a bioluminescent system

Many (but not all) of the bioluminescent systems in coelenter-ates involve energy transfer from an excited product molecule of the calcium activated photoprotein to a second species, the green fluorescent protein, with emission at 508 nm from its excited state. Although all the luminescent coelen-terates studied possess photoproteins, not all of them have the green fluorescent protein. This green fluorescent molecule is localized in the luminescent cells; they can thus be easily distinguished by fluorescence microscopy. The active components occur in subcellular particles; these have been isolated in an active form by homogenization in isotonic (to sea water) salt solutions.     

Intergeneric transfer of streptomycin-resistance marker between two blue-green algae

Intergeneric transfer of streptomycin-resistance marker from a unicellular blue-green algaAnacystis nidulans to a filamentous blue-green algaAnabaena doliolum was demonstrated. Mutants ofA. nidulans resistant to streptomycin, occurring spontaneously or mutagenically-induced could be isolated easily. Naturally occurring streptomycin-resistant mutants ofA. doliolum could not be detected. Attempts at isolating such mutants either in nitrogen-free medium or in nitrate containing medium were unsuccessful. However, a streptomycin-resistant strain (recombinant) ofA. doliolum could be isolated in a mixed culture of streptomycin-sensitiveA. doliolum and streptomycin-resistantA. nidulans.     

Energy transfer in the photosynthetic unit. I. The concept of independent units for photosystem II analyzed by flash yields for dichlorophenolindophenol reduction

The problem of the interconnection of photosynthetic units is dealt with. Flash yield results together with quantum yield measurements of DCPIP reduction in isolated chloroplasis indicate that photosynthetic units of PS-II are essentially independent and probably morphological entities. This is shown by the exponential dependence of the flash yield on the flash intensity and a high quantum yield of excitation trapping. Deviation from exponentiality observed in some samples is explained in terms of energy transfer between the units when they are densely packed.     

温度变化对藻类光合电子传递与光合放氧关系的影响

由于直接测定藻类的光合速率耗时且不方便,研究者们常通过测定藻类光合电子传递速率的方式来间接反映其光合速率,理论上,以氧气产生来度量的总光合速率（PGross）与电子传递速率(ETR)之间应该存在很好的线性关系。然而,由于温度的变化会影响藻类的光呼吸等耗氧的生理过程从而影响光合作用中的氧气释放,因此温度可能会对PGross与ETR之间的线性关系产生影响。研究了温度变化对蛋白核小球藻（Cholorella pyrenoidosa）、菱形藻（Nitzschia sp.）和水生集胞藻（Synechocystis aquetilis Sauv.）的总光合放氧速率（PGross）与电子传递速率(ETR)之间比率的影响,结果表明PGross/ETR随温度的升高而降低,低温条件下PGross/ETR比值较高,说明在相同的电子传递速率的情况下水的光裂解产生的氧有更多的可以释放出来;在高温条件下PGross/ETR比值相对较低,说明高温条件下可能有相对更多的水光裂解产生的氧被用于耗氧的生理过程而没有释放出来。研究表明当温度发生变化时,光合放氧与电子传递之间并不呈线性关系,这说明将ETR作为实际光合生产的评价指标时要谨慎,不能不加分析地直接应用。