Competition between annihilation and trapping leads to strongly reduced yields of photochemistry under ps-flash excitation

Excitation of photosynthetic systems with short intense flashes is known to lead to exciton-exciton annihilation processes. Here we quantify the effect of competition between annihilation and trapping for Photosystem II, Photosystem I (thylakoids from peas and membranes from the cyanobacterium Synechocystis sp.), as well as for the purple bacterium Rhodospirillum rubrum. In none of the cases it was possible to reach complete product saturation (i.e. closure of reaction centers) even with an excitation energy exceeding 10 hits per photosynthetic unit. The parameter introduced by Deprez et al. ((1990) Biochim. Biophys. Acta 1015: 295–303) describing the competition between exciton-exciton annihilation and trapping was calculated to range between 4.5 (PS II) and 6 (Rs. rubrum). The rate constants for bimolecular exciton-exciton annihilation ranged between (42 ps)^-1 and (2.5 ps)^-1 for PS II and PS I-membranes of Synechocystis, respectively. The data are interpreted in terms of hopping times (i.e. mean residence time of the excited state on a chromophore) according to random walk in isoenergetic antenna.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - LHC II light harvesting complex II - P primary donor - PS I Photosystem I - PS II Photosystem II - PSU photosynthetic unit - RC reaction center     

Light Intensity-Induced Changes in cab mRNA and Light Harvesting Complex II Apoprotein Levels in the Unicellular Chlorophyte Dunaliella tertiolecta

During a transition from high growth irradiance (700 micromoles quanta per square meter per second) to low growth irradiance (70 micromoles quanta per square meter per second), the unicellular marine chlorophyte Dunaliella tertiolecta Butcher increases the cellular pool size of the light-harvesting complex of photosystem II (LHC II). We showed that the increase in LHC II apoproteins and in chlorophyll content per cell is preceded by an approximately fourfold increase in cab mRNA. The increase in cab mRNA is detectable within 1.5 hours following a shift from high to low light intensity. An increase in the relative abundance of cab mRNA was also found following a shift from high light to darkness and from high light to low light in the presence of gabaculine, a chlorophyll synthesis inhibitor. However, the LHC II apoproteins did not accumulate in the latter experiments, suggesting that LHC II apoprotein synthesis is coupled to chlorophyll synthesis at or beyond translation. We propose that changes in energy balance brought about by a change in light intensity may control a regulatory factor acting to repress cab mRNA expression in high light.     

Stepwise two-photon excited fluorescence from higher excited states of chlorophylls in photosynthetic antenna complexes

Stepwise two-photon excited fluorescence (TPEF) spectra of the photosynthetic antenna complexes PCP, CP47, CP29, and light-harvesting complex II (LHC II) were measured. TPEF emitted from higher excited states of chlorophyll (Chl) a and b was elicited via consecutive absorption of two photons in the Chl a/b Qy range induced by tunable 100-fs laser pulses. Global analyses of the TPEF line shapes with a model function for monomeric Chl a in a proteinaceous environment allow distinction between contributions from monomeric Chls a and b, strongly excitonically coupled Chls a, and Chl a/b heterodimers/-oligomers. The analyses indicate that the longest wavelength-absorbing Chl species in the Qy region of LHC II is a Chl a homodimer with additional contributions from adjacent Chl b. Likewise, in CP47 a spectral form at approximately 680 nm (that is, however, not the red-most species) is also due to strongly coupled Chls a. In contrast to LHC II, the red-most Chl subband of CP29 is due to a monomeric Chl a. The two Chls b in CP29 exhibit marked differences: a Chl b absorbing at approximately 650 nm is not excitonically coupled to other Chls. Based on this finding, the refractive index of its microenvironment can be determined to be 1.48. The second Chl b in CP29 (absorbing at approximately 640 nm) is strongly coupled to Chl a. Implications of the findings with respect to excitation energy transfer pathways and rates are discussed. Moreover, the results will be related to most recent structural analyses.     

Two-photon excited fluorescence from higher electronic states of chlorophylls in photosynthetic antenna complexes: a new approach to detect strong excitonic chlorophyll a/b coupling

Stepwise two-photon excitation of chlorophyll a and b in the higher plant main light-harvesting complex (LHC II) and the minor complex CP29 (as well as in organic solution) with 100-fs pulses in the Q(y) region results in a weak blue fluorescence. The dependence of the spectral shape of the blue fluorescence on excitation wavelength offers a new approach to elucidate the long-standing problem of the origin of spectral "chlorophyll forms" in pigment-protein complexes, in particular the characterization of chlorophyll a/b-heterodimers. As a first result we present evidence for the existence of strong chlorophyll a/b-interactions (excitonically coupled transitions at 650 and 680 nm) in LHC II at ambient temperature. In comparison with LHC II, the experiments with CP29 provide further evidence that the lowest energy chlorophyll a transition (at approximately 680 nm) is not excitonically coupled to chlorophyll b.     

Release and aggregation of the light-harvesting complex in intact leaves subjected to strong CO2 deficit

Tobacco plants were subjected to long-term CO₂ deficit. The stress caused photoinhibition of Photosystem (PS) II photochemistry and the aggregation of the light-harvesting complex of PS II (LHC II). The aggregation was shown by the appearance of the characteristic band at 698–700 nm (F₆₉₉) in 77 K fluorescence emission spectra. LHC II aggregates are considered to quench fluorescence and, therefore, the fluorescence yield was determined to verify their quenching capability. PS II photochemistry, measured as F_V/F_M, was largely depressed during first 4 days of the stress. Unexpectedly, the total fluorescence yield increased in this period. Fitting of emission spectra by Gaussian components approximating emission bands of LHC II, PS II core, PS I and F₆₉₉ revealed that mainly the bands at 680 and 699 nm, representing emission of LHC II aggregates, were responsible for the increase of the fluorescence yield. This shows an interruption of the excitation energy transfer between LHC II and both photosystems and, thus, a physical disconnection of LHC II from photosystems. PS II and PS I emissions were not quenched in this period. Therefore, it was concluded that these LHC II aggregates were accumulated out of PS II antenna, and, thus they cannot be involved in dumping of excess excitation. The total fluorescence yield turned to decrease only after the large depression of PS II photochemistry, when LHC II aggregation was considerably speeded up and the fluorescence yields of PS I and II turned to decline.     

Direct evidence for excitonically coupled chlorophylls a and b in LHC II of higher plants by nonlinear polarization spectroscopy in the frequency domain

Occurrence of excitonic interactions in light-harvesting complex II (LHC II) was investigated by nonlinear polarization spectroscopy in the frequency domain (NLPF) at room temperature. NLPF spectra were obtained upon probing in the chlorophyll (Chl) a/b Soret region and pumping in the Q_y region. The lowest energy Chl a absorbing at 678 nm is strongly excitonically coupled to Chl b.     

Evidence for the role of surface-exposed segments of the light-harvesting complex in cation-mediated control of chloroplast structure and function.

Chloroplast membranes contain a light-harvesting pigment-protein complex (LHC) which binds chlorophylls a and b. A mild trypsin digestion of intact thylakoid membranes has been utilized to specifically alter the apparent molecular weights of polypeptides of this complex. The modified membrane preparations were analyzed for altered functional and structural properties. Cation-induced changes in room temperature fluorescence intensity and low temperature chlorophyll fluorescence emission spectra, and cation regulation of the quantum yield of photosystem I and II partial reactions at limiting light were lost following the trypsin-induced alteration of the LHC. Electron microscopy revealed that cations can neither maintain nor promote grana stacking in membranes which have been subjected to mild trypsin treatment. Freeze-fracture analysis of these membranes showed no significant differences in particle density or average particle size of membrane subunits on the EF fracture face; structural features of the modified lamellae were comparable to membranes which had been unstacked in a “low salt” buffer. Digitonin digestion of trypsin-treated membranes in the presence of cations followed by differential centrifugation resulted in a subchloroplast fractionation pattern similar to that observed when control chloroplasts were detergent treated in cation-free medium. We conclude that: (a) the initial action of trypsin at the thylakoid membrane surface of pea chloroplasts was the specific alteration of the LHC polypeptides, (b) the segment of the LHC polypeptides which was altered by trypsin is necessary for cation-mediated grana stacking and cation regulation of membrane subunit distribution, and (c) cation regulation of excitation energy distribution between photosystem I and II involves the participation of polypeptide segments of the LHC which are exposed at the membrane surface.     

Chlorophyll fluorescence phenomena in chloroplasts on subnanosecond time-scales probed by picosecond pulse pairs

Fluorescence enhancement phenomena and quenching by exciton-exciton annihilation on subnanosecond and nanosecond time-scales were investigated in spinach chloroplasts utilizing picosecond laser pulse pairs (530 nm, 30 ps wide) of equal intensity, spaced apart in time by variable delays of Δt = 0−6 ns. This new method was devised to study the effect of pulse energies (1·10¹⁰–2·10¹⁵ photons per cm²) on the overall fluorescence yield in order to deduce the degree of correlation between the two pulses as a function of Δt. In the case of open reaction centers (F₀ state) in Photosystem II (PS II), it is shown that the quenching effect of excitons generated by the first pulse on the fluorescence yield of the second pulse diminishes with increasing Δt with a characteristic decorrelation time of 140 ± 60 ps. This effect is attributed to either (1) the decay of mobile excitons in the light-harvesting antenna pigment bed as these excitons migrate towards the PS II reaction centers and the associated smaller core antenna pigment pools, or (2) the decay of a quenching state of the reaction center (and/or core antenna) which appears following a rapid (less than 140 ps) trapping of the excitons initially created in the antenna pigment bed. The absence of a significant decay component of exciton quenchers with a lifetime comparable to the 300–600 ps intermediate phase of fluorescence decay kinetics suggests that this phase, although contributing to more than half of the integrated fluorescence emission signal, is not caused by freely mobile exitons migrating in a lake of pigments, but originates instead from smaller pigment pools to which the excitons have migrated. It is proposed that bimolecular exciton-exciton annihilation in these smaller domains dominates annihilation in the larger antenna pigment bed. In the case of closed reaction centers (F_max state), the decorrelation time between the two pulses is increased to 400 ± 100 ps, which is also attributed to either a mobile exciton component or to the decay of a quenching state of the reaction center. At low pulse intensities (below approx. 2 · 10¹² photons per cm²) anomalous fluorescence enhancement effects are noted, which are clearly linked to the existence of initially open PS II reaction centers. These enhancement effects are different from the well-known fluorescence induction phenomena which occur on longer time-scales, and are tentatively attributed to variations in the quenching efficiencies of transitory photochemical states of PS II reaction centers.     

Short-pulse pump-and-probe technique for airborne laser assessment of Photosystem II photochemical characteristics

The development of a technique for laser measurement of fPhotosystem II (PS II) photochemical characteristics of phytoplankton and terrestrial vegetation from an airborne platform is described. Results of theoretical analysis and experimental study of pump-and-probe measurement of the PS II functional absorption cross-section and photochemical quantum yield are presented. The use of 10 ns probe pulses of PS II sub-saturating intensity provides a significant, up to 150-fold, increase in the fluorescence signal compared to conventional `weak-probe' protocol. Little effect on the fluorescence yield from the probe-induced closure of PS II reaction centers is expected over the short pulse duration, and thus a relatively intense probe pulse can be used. On the other hand, a correction must be made for the probe-induced carotenoid triplet quenching and singlet-singlet annihilation. A Stern-Volmer model developed for this correction assumes a linear dependence of the quenching rate on the laser pulse fluence, which was experimentally validated. The PS II saturating pump pulse fluence (532 nm excitation) was found to be 10 and 40 μmol quanta m⁻² for phytoplankton samples and leaves of higher plants, respectively. Thirty μs was determined as the optimal delay in the pump-probe pair. Our results indicate that the short-pulse pump-and-probe measurement of PS II photochemical characteristics can be implemented from an airborne platform using existing laser and LIDAR technologies. This revised version was published online in June 2006 with corrections to the Cover Date.     

Single-molecule electron tunneling spectroscopy of the higher plant light-harvesting complex LHC II

Electronic spectroscopy of a single biological molecule is demonstrated with approximately 4 A spatial resolution. The light-harvesting complex II (LHC II), in the ground and photo-excited states, was studied using scanning tunneling microscopy and spectroscopy of intact Photosystem II complexes. Analysis of the spectra indicates that the main mechanisms of tunneling between the STM tip and the surface involve delocalized electronic states of the LHC II and local vibronic states associated with C=C, C=O, C-H, N-H, and O-H groups near the LHC II surface. Conduction within the bulk LHC II is then due to ohmic and hopping conduction as well as tunneling between amino acid residues. Light activation of LHC II occurs via a photoconductive rather than a photovoltaic mechanism. There is a dramatic light-induced increase in the electronic density of states indicating a light-induced enhancement of energy and electron delocalization which is important for the efficient and rapid transfer of excitation energy from LHC II to the Photosystem II reaction center.     

Complex formation in plant thylakoid membranes. Competition studies on membrane protein interactions using synthetic peptide fragments

Thylakoid membranes of pea were used to study competition between extra-membrane fragments and their parental membrane-bound proteins. Phosphorylated and unphosphorylated fragments of light harvesting complex II (LHC II) from higher plants were used to compete with LHC II for interactions with itself and with other thylakoid protein complexes. Effects of these peptide fragments of LHC II and of control peptides were followed by 80 K chlorophyll fluorescence spectroscopy of isolated thylakoids. The phosphorylated LHC II fragment competes with membrane-bound phosphoproteins in the phosphatase reaction. The same fragment accelerates the process of dark-to-light adaptation and decreases the rate of the light-to-dark adaptation when these are followed by fluorescence spectroscopy. In contrast, the non-phosphorylated LHC II peptide does not affect the rate of adaptation but produces results consistent with inhibition of formation of a quenching complex. In this quenching complex we propose that LHC II remains inaccessible to the LHC II kinase, explaining an observed decrease in LHC II phosphorylation in the later stages of the time-course of phosphorylation. The most conspicuous protein which is steadily phosphorylated during the time-course of phosphorylation is the 9 kDa (psbH) protein. The participation of the phosphorylated form of psbH in the quenching complex, where it is inaccessible to the phosphatase, may explain its anomalously slow dephosphorylation. The significance of the proposed complex of LHC II with phospho-psbH is discussed.Abbreviations LHC II light harvesting complex II - PS II Photosystem II - PS I Photosystem I     

Immunological identification and quantification of light harvesting chlorophyll a/b protein of Chlorella protothecoides

The alteration of photosynthetic membrane proteins in relation to the disappearance of pigments during the heterotrophic growth of Chlorella protothecoides was investigated. Chlorophylls and certain polypeptides associated with the LHC II disappeared after 50 hr of heterotrophic growth but the 24 kDa apoprotein constituting LHC II was not affected. Immunological analysis indicated that the chlorophylls and the light harvesting complex proteins of the thylakoid membranes are not tightly coupled and the latter is retained in its native form irrespective of the presence or absence of the former. The circumstantial evidence that the other photosynthetic membrane polypeptides are degraded along with the pigments due to increased proteolytic activity in the rapidly dividing heterotrophic cells indicate that chlorophyll synthesis is not a pre-requisite for the synthesis of the LHC II apoprotein.     

Regulation of the Size of Light-Harvesting Antennae in Response to Light Intensity in the Green Alga Chlorella pyrenoidosa

Changes in size of the light-harvesting Chl-protein complex(LHC) induced by changes in light intensity were studied withthe green alga Chlorella pyrenoidosa. The Chi a/b ratio, whichis correlated with the size of LHC, varied over a wide range(2.5–5.0) when the light intensity for autotrophic growthwas changed. Comparison of properties of LHC II isolated fromcells grown under light of high and low intensity indicatedthat the large difference in Ch1 a/b ratios in cells grown underlight of different intensities is due mainly to changes in levelsof LHC. Reduction in levels of LHC under light of high intensitydid not occur when proliferation of cells was suppressed. Thisresult indicates that reduction in levels of LHC is not attributableto acceleration of the degradation of LHC under light of highintensity. Stimulation of formation of LHC occurred even underlight of high intensity when formation of photosystems was suppressedby chloram-phenicol (CAP). Analysis of the CAP-induced formationof LHC indicated that (1) such formation of LHC was regulatedby light intensity, being less active under higher intensity,and (2) the suppressive effect of gabaculine, an inhibitor ofthe synthesis of porphyrin, and thus of Ch1, was greater underlight of high intensity while the suppressive effect of cycloheximide,an inhibitor of the synthesis of apoprotein, was slightly greaterunder light of low intensity. The results described in thisreport indicate that (1) intensity-induced changes in the sizeof LHC in Chlorella pyrenoidosa are due to regulation of theassembly of LHC and (2) the regulation occurs primarily at thelevel of the synthesis of Ch1. (Received June 1, 1989; Accepted August 21, 1989)     

Optical effects of sodium dodecyl sulfate treatment of the isolated light harvesting complex of higher plants

The light-harvesting complex (LHC) of higher plants isolated using Triton X-100 has been studied during its transformation into a monomeric form known as CPII. The change was accomplished by gradually increasing the concentration of the detergent, sodium dodecyl sulfate (SDS). Changes in the red spectral region of the absorption, circular dichroism (CD), and linear dichroism spectra occurring during this treatment have been observed at room temperature. According to a current hypothesis the main features of the visible region absorption and CD spectra of CPII can be explained reasonably successfully in terms of an exciton coupling among its chlorophyll (Chl) b molecules. We suggest that the spectral differences between the isolated LHC and the CPII may be understood basically in terms of an exciton coupling between the Chl b core of a given CPII unit and at least one of the Chla's of either the same or the adjacent CPII. We propose that this Chl a-Chl b coupling existing in LHC disappears upon segregation into CPII, probably as a result of a detergent-related overall rotation of the strongly coupled Chl b core which changes the relative orientations of the two types of pigments and thus the nature of their coupling.Abbreviations Chl Chlorophyll - CD Circular dichroism - LD Linear dichroism - LHC Light-harvesting complex - SDS Sodium dodecyl sulfate - CPII A solubilized form of LHC obtained with SDS polyacrylamide gel electrophoresis Dedicated to Prof. L.N.M. Duysens on the occasion of his retirement     

Characterization of a novel Photosystem I-LHCI supercomplex isolated from Chlamydomonas reinhardtii under anaerobic (State II) conditions

A novel supercomplex of Photosystem I (PSI) with light harvesting complex I (LHCI) was isolated from the green alga Chlamydomonas reinhardtii. This novel supercomplex is unique as it is the first stable supercomplex of PSI together with its external antenna. The supercomplex contains 256 chlorophylls per reaction center. The supercomplex was isolated under anaerobic conditions and may represent the State II form of the photosynthetic unit. In contrast to previously reported supercomplexes isolated in State I, which contain only 4 LHC I proteins, this supercomplex contains 10-11 LHC I proteins tightly bound to the PSI core. In contrast to plants, no LHC II is tightly bound to the PSI-LHCI supercomplex in State II. Investigation of the energy transfer from the antenna system to the reaction center core shows that the LHC supercomplexes are tightly coupled to the PSI core, not only structurally but also energetically. The excitation energy transfer kinetics are completely dominated by the fast phase, with a near-complete lack of long-lived fluorescence. This tight coupling is in contrast to all reports of energy transfer in PSI-LHCI supercomplexes (in State I), which have so far been described as weakly coupled supercomplexes with low efficiency for excitation energy transfer. These results indicate that there are large and dynamic changes of the PSI-LHCI supercomplex during the acclimation from aerobic (State I) to anaerobic (State II) conditions in Chlamydomonas.     

The redox state of the plastoquinone pool controls the level of the light-harvesting chlorophyll a/b binding protein complex II (LHC II) during photoacclimation

A cytochrome b ₆ f deficient mutant of Lemna perpusilla maintains a constant and lower level of the light-harvesting chl a/b-binding protein complex II (LHC II) as compared to the wild type plants at low-light intensities. Inhibition of the plastoquinone pool reduction increases the LHC II content of the mutant at both low- and high-light intensities but only at high-light intensity in the wild type plants. Proteolytic activity against LHC II appears during high-light photoacclimation of wild type plants. However, the acclimative protease is present in the mutant at both light intensities. These and additional results suggest that the plastoquinone redox state serves as the major signal-transducing component in the photoacclimation process affecting both, synthesis and degradation of LHC II and appearance of acclimative LHC II proteolysis. The plastoquinol pool cannot be oxidized by linear electron flow in the mutant plants which are locked in a ‘high light’ acclimation state. The cytochrome b ₆ f complex may be involved indirectly in the regulation of photoacclimation via 1) regulation of the plastoquinone redox state; 2) regulation of the redox-controlled thylakoid protein kinase allowing exposure of the dephosphorylated LHC II to acclimative proteolysis. This revised version was published online in June 2006 with corrections to the Cover Date.     

Chlorophyll fluorescence changes at high temperatures induced by linear heating of greening barley leaves

A relative decrease of the high temperature part (above 60°C) of the chlorophyll fluorescence temperature curve during 3 h to 10 h greening period of barley (Hordeum vulgare L.) leaves was found to be concomitant to a decrease of Chl alb ratio and to a gradual increase of LHCP/core ratio found by electrophoresis and the ratio of granal to total length of thylakoid membranes. It is suggested that the high temperature part of the fluorescence temperature curve depends inversely on the relative amount of LHC II in thylakoid membranes.Abbreviations Chl a(b) chlorophyll a(b) - CPa chlorophyll a protein complex of PS II - CP1 P700 chlorophyll a protein complex of PS I - FP free pigments - FTC fluorescence temperature curve - F(T30) fluorescence intensity at 30°C - LHC II light harvesting complex II - LHCP light harvesting chlorophyll protein - LHCP3 (LHCPm) monomeric form of LHC II - LHCPo oligomeric form of LHC II complex - M1 first maximum of FTC - M2 second maximum (region) of FTC - PAA polyacrylamide - PAR photosynthetically active radiation - PS I(II) Photosystem I(II) - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis     

Screening of chlorina mutants of barley (Hordeum vulgare L.) with antibodies against light-harvesting proteins of PS I and PS II: Absence of specific antenna proteins

Twenty-three chlorina (clo) mutants from the barley mutant collection of the Carlsberg Laboratory, Copenhagen, were tested for the presence of the four light-harvesting chlorophyll (Chl) a/b-binding proteins (LHC) of Photosystem I (Lhca1-4) and the PS II antenna proteins Lhcb1-3 (LHC II), Lhcb4-6 (CP29, CP26, CP24) and PsbS (CP22) using monospecific and monoclonal antibodies. Mutants allelic to barley mutant clo-f2, impaired in Chl b synthesis, provided evidence that Lhca4, Lhcb1 and Lhcb6 are unstable in the absence of Chl b, and the accumulation of Lhcb2, Lhcb3 and Lhcb4 is also impaired. Mutants at the locus chlorina-a (clo-a117, clo-a126 and clo-a134) lack or have only trace amounts of Lhca1, Lhca4, Lhcb1 and Lhcb3, whereas a mutant at the locus chlorina-b (clo-b125) had reduced amounts of all Lhca proteins. These two mutations could have an effect in protein import or assembly. Evidence is presented that Lhcb5 is the innermost LHC protein of PS II, and that Lhca1 and Lhca4, which have been supposed to be intimately associated in the LHCI-730 complex, can accumulate independently of each other. 77 K fluorescence emission spectra taken from leaves of clo-f2 101, clo-a126 and clo-b125 indicate that chlorophyll(s) emitting at 742 nm are coupled to the presence of Lhca4 that is bound to the reaction centre, and those emitting around 730 nm are located on Lhca1.     

Kinetics of excited states of pigment clusters in solubilized light-harvesting complex II: photon density-dependent fluorescence yield and transmittance.

Relative fluorescence yield, phi F, and transmittance, T, were measured in solubilized light-harvesting complex II (LHCII) as a function of photon density, Ip, of monochromatic 645-nm laser pulses (duration: approximately 2.5 ns). Special efforts were made in constructing an optical set-up that allows the accurate determination of the fluorescence from an area of constant Ip, phi F(Ip) starts to decline at approximately 10(14) and drops to values below 0.01% at maximum Ip (approximately 10(19) photons cm-2 pulse-1). T(Ip) decreases only slightly at photon densities of approximately 10(15) but increases steeply at values of > 10(17) photons cm-2 pulse-1. The interpretation of the phi F(Ip) data using the saturation limit of Mauzerall's multiple hit model leads to a unit size of about 10-15 chlorophyll molecules. One interpretation is to attribute this result to a very fast exciton-exciton annihilation of multiple excited states generated within this small domain. Alternatively, based on the assumption that delocalized cluster states within the monomeric/trimeric subunit of LHCII exist, the results can be consistently described by a kinetic model comprising ground, monoexcitonic, and biexcitonic states of clusters and a triplet state that is quenched by carotenoids in LHCII. Within the framework of this model the annihilation of multiple excitations is explained as ultrafast radiationless relaxation of higher excited cluster states. Comparative measurements in diluted acetonic Chl a solution are consistently described by the depletion of the ground state, taking the absorption cross section at the used wavelength.     

Assembly of the Light-Harvesting Complexes (LHCs) of Photosystem II (Monomeric LHC IIb Complexes Are Intermediates in the Formation of Oligomeric LHC IIb Complexes)

The light-induced assembly of light-harvesting complex (LHC) II has been followed during the biogenesis of the plastid. Seedlings grown in intermittent light (IML) accumulate only small amounts of chlorophyll b. The minor LHC II apoproteins are present; however, the apoprotein levels of the major LHC II complex, LHC IIb, are severely depressed after exposure to IML. The levels of all LHC II apoproteins increase rapidly upon exposure to continuous illumination. The 25-kD, type 3 LHC IIb subunit appears to be more abundant during the early hours of greening in relation to its level in mature thylakoids. The LHC IIb apoproteins are initially associated with pigments to form monomeric pigment-protein complexes. The abundance of monomeric LHC IIb complexes gradually decreases during exposure to continuous light and a concomitant increase occurs in the amount of the trimeric and higher-order oligomeric forms. Pulse-chase experiments verify that labeled LHC IIb monomeric complexes are intermediates in the formation of trimeric and higher-order oligomeric LHC IIb-pigmented complexes. Therefore, the assembly of LHC II occurs via the initial pigmentation of the apoproteins to form monomeric complexes and proceeds in a sequential manner.