Photosynthesis Research (v.99, #3)
The discovery of state transitions in photosynthesis 40 years ago by Norio Murata (155-160).
In the late 1960s, I identified an aspect of the kinetics of chlorophyll fluorescence in algal cells that I was unable to explain in terms of photochemical quenching. I proposed a novel regulatory mechanism for the distribution of light energy to photosystems I and II, which is now known by the term of “state transitions.” I also examined the “cation-dependent redistribution of light energy” to photosystems I and II and the “energy-dependent quenching” of chlorophyll fluorescence. At that time, financial constraints prevented me from measuring the emission and action spectra of chlorophyll fluorescence at liquid-nitrogen temperature and the light quality-dependent changes in the yield of chlorophyll fluorescence at room temperature. The financial problems were solved by constructing several pieces of electronic equipment using skills obtained by repairing radios when I was a high-school and college student.
Keywords: Cation-dependent distribution of light energy; Chlorophyll fluorescence at liquid-nitrogen temperature; Energy-dependent quenching; Induction kinetics; State transitions
Effect of phosphorylation on the thermal and light stability of the thylakoid membranes by Zsuzsanna Várkonyi; Gergely Nagy; Petar Lambrev; Anett Z. Kiss; Noémi Székely; László Rosta; Gyözö Garab (161-171).
Higher plant thylakoid membranes contain a protein kinase that phosphorylates certain threonine residues of light-harvesting complex II (LHCII), the main light-harvesting antenna complexes of photosystem II (PSII) and some other phosphoproteins (Allen, Biochim Biophys Acta 1098:275, 1992). While it has been established that phosphorylation induces a conformational change of LHCII and also brings about changes in the lateral organization of the thylakoid membrane, it is not clear how phosphorylation affects the dynamic architecture of the thylakoid membranes. In order to contribute to the elucidation of this complex question, we have investigated the effect of duroquinol-induced phosphorylation on the membrane ultrastructure and the thermal and light stability of the chiral macrodomains and of the trimeric organization of LHCII. As shown by small angle neutron scattering on thylakoid membranes, duroquinol treatment induced a moderate (~10%) increase in the repeat distance of stroma membranes, and phosphorylation caused an additional loss of the scattering intensity, which is probably associated with the partial unstacking of the granum membranes. Circular dichroism (CD) measurements also revealed only minor changes in the chiral macro-organization of the complexes and in the oligomerization state of LHCII. However, temperature dependences of characteristic CD bands showed that phosphorylation significantly decreased the thermal stability of the chiral macrodomains in phosphorylated compared to the non-phosphorylated samples (in leaves and isolated thylakoid membranes, from 48.3°C to 42.6°C and from 47.5°C to 44.3°C, respectively). As shown by non-denaturing PAGE of thylakoid membranes and CD spectroscopy on EDTA washed membranes, phosphorylation decreased by about 5°C, the trimer-to-monomer transition temperature of LHCII. It also enhanced the light-induced disassembly of the chiral macrodomains and the monomerization of the LHCII trimers at 25°C. These data strongly suggest that phosphorylation of the membranes considerably facilitates the heat- and light-inducible reorganizations in the thylakoid membranes and thus enhances the structural flexibility of the membrane architecture.
Keywords: Circular dichroism; LHCII; Light stability; Phosphorylation; Thermal stability; Thermo-optic effect
Dynamics of higher plant photosystem cross-section associated with state transitions by Alexander V. Ruban; Matthew P. Johnson (173-183).
Photosynthetic state transitions are a well-known phenomenon of short-term adaptation of the photosynthetic membrane to changes in spectral quality of light in low light environments. The principles of the monitoring and quantification of the process in higher plants are revised here. The use of the low-temperature excitation fluorescence spectroscopy for analysis of the photosystem I antenna cross-section dynamics is described. This cross section was found to increase by 20–25% exclusively due to the migration and attachment of LHCIIb complex in State 2. Analysis of the fine structure of the additional PSI cross-section spectrum revealed the 510 nm band, characteristic of Lutein 2 of LHCIIb and present only when the complex is in a trimeric state. The excitation fluorescence spectrum of the phospho-LHCII resembles the spectrum of aggregated and hence quenched LHCII. This novel observation could explain the fact that at no point in the course of the state transition high fluorescence and long lifetime components of detached trimeric LHCII have ever been observed. In the plants lacking Lhcb1 and 2 proteins and unable to perform state transitions, compensatory sustained adjustments of the photosystem I and II antennae have been revealed. Whilst the major part of the photosystem II antenna is built largely of CP26 trimers, possessing less chlorophyll b and more of the red-shifted chlorophyll a, photosystem I in these plants contains more than 20% of extra LHCI antenna enriched in chlorophyll b. Hence, both photosystems in the plants lacking state transitions have less spectrally distinct antennae, which enable to avoid energy imbalance due to the changes in the light quality. These alterations reveal remarkable plasticity of the higher plant photosynthetic antenna design providing the basis for a flexible adaptation to the light environment.
Keywords: The state transitions; Cross-section; Phospho-LHCII; Excitation fluorescence spectrum
Effect of monogalactosyldiacylglycerol on the interaction between photosystem II core complex and its antenna complexes in liposomes of thylakoid lipids by Feng Zhou; Shuang Liu; Zhaohui Hu; Tingyun Kuang; Harald Paulsen; Chunhong Yang (185-193).
The non-bilayer lipid monogalactosyldiacylglycerol (MGDG) is the most abundant type of lipid in the thylakoid membrane and plays an important role in regulating the structure and function of photosynthetic membrane proteins. In this study, we have reconstituted the isolated major light-harvesting complexes of photosystem II (PSII) (LHCIIb) and a preparation consisting of PSII core complexes and minor LHCII of PSII (PSIICC) into liposomes that consisted of digalactosyldiacylglycerol (DGDG), sulfoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol (PG), with or without MGDG. Transmission electron microscopy and freeze-fracture studies showed unilamellar proteoliposomes, and demonstrated that most of the MGDG is incorporated into bilayer structures. The impact of MGDG on the functional interaction between LHCIIb and PSIICC was investigated by low temperature (77 K) fluorescence emission spectra and the photochemical activity of PSII. The additional incorporation of LHCIIb into liposomes containing PSIICC markedly increased oxygen evolution of PSIICC. Excitation at 480 nm of chlorophyll (Chl) b in LHCIIb stimulated a characteristic fluorescence emission of the Chl a in PSII (684.2 nm), rather than that of the Chl a in LHCIIb (680 nm) in the LHCIIb–PSIICC proteoliposomes, which indicated that the energy was transferred from LHCIIb to PSIICC in liposome membranes. Increasing the percentage of MGDG in the PSIICC–LHCIIb proteoliposomes enhanced the photochemical activity of PSII, due to a more efficient energy transfer from LHCIIb to PSIICC and, thus, an enlarged antenna cross section of PSII.
Keywords: Photosystem II core complex; Light-harvesting complex II; Monogalactosyldiacylglycerol; Liposome; Photochemical activity
Salinity affects the photoacclimation of Chlamydomonas raudensis Ettl UWO241 by Kenji Takizawa; Shinichiro Takahashi; Norman P. A. Hüner; Jun Minagawa (195-203).
Chlamydomonas raudensis Ettl UWO241, a natural variant of C. raudensis, is deficient in state transitions. Its habitat, the deepest layer of Lake Bonney in Antarctica, features low irradiance, low temperature, and high salinity. Although psychrophily and low-light acclimation of this green alga has been described, very little information is available on the effect of salinity. Here, we demonstrate that this psychrophile is halotolerant, not halophilic, and it shows energy redistribution between photosystem I and II based on energy spillover under low-salt conditions. Furthermore, we revealed that C. raudensis exhibits higher non-photochemical quenching in comparison with the mesophile Chlamydomonas reinhardtii, when grown with low-salt, which is due to the lower proton conductivity across the thylakoid membrane. Significance of the C. raudensis UWO241 traits found in the low salinity culture are implicated with their natural habitats, including the high salinity and extremely stable light environments.
Keywords: Light-harvesting complex; NPQ; Photoacclimation; Psychrophile; State transition
Towards efficient hydrogen production: the impact of antenna size and external factors on electron transport dynamics in Synechocystis PCC 6803 by Gábor Bernát; Nadine Waschewski; Matthias Rögner (205-216).
Three Synechocystis PCC 6803 strains with different levels of phycobilisome antenna-deficiency have been investigated for their impact on photosynthetic electron transport and response to environmental factors (i.e. light-quality, -quantity and composition of growth media). Oxygen yield and P700 reduction kinetic measurements showed enhanced linear electron transport rates—especially under photoautotrophic conditions—with impaired antenna-size, starting from wild type (WT) (full antenna) over ΔapcE- (phycobilisomes functionally dissociated) and Olive (lacking phycocyanin) up to the PAL mutant (lacking the whole phycobilisome). In contrast to mixotrophic conditions (up to 80% contribution), cyclic electron transport plays only a minor role (below 10%) under photoautotrophic conditions for all the strains, while linear electron transport increased up to 5.5-fold from WT to PAL mutant. The minor contribution of the cyclic electron transport was proportionally increased with the linear one in the ΔapcE and Olive mutant, but was not altered in the PAL mutant, indicating that upregulation of the linear route does not have to be correlated with downregulation of the cyclic electron transport. Antenna-deficiency involves higher linear electron transport rates by tuning the PS2/PS1 ratio from 1:5 in WT up to 1:1 in the PAL mutant. While state transitions were observed only in the WT and Olive mutant, a further ~30% increase in the PS2/PS1 ratio was achieved in all the strains by long-term adaptation to far red light (720 nm). These results are discussed in the context of using these cells for future H2 production in direct combination with the photosynthetic electron transport and suggest both Olive and PAL as potential candidates for future manipulations toward this goal. In conclusion, the highest rates can be expected if mutants deficient in phycobilisome antennas are grown under photoautotrophic conditions in combination with uncoupling of electron transport and an illumination which excites preferably PS1.
Keywords: Light adaptation; Linear and cyclic electron flow; Photo-biological hydrogen production; Phycobilisomes; Synechocystis
Distinct roles of CpcG1-phycobilisome and CpcG2-phycobilisome in state transitions in a cyanobacterium Synechocystis sp. PCC 6803 by Kumiko Kondo; Conrad W. Mullineaux; Masahiko Ikeuchi (217-225).
State transitions in cyanobacteria regulate the relative energy transfer from phycobilisome to photosystem I and II. Although it has been shown that phycobilisome mobility is essential for phycobilisome-dependent state transitions, the biochemical mechanism is not known. Previously we reported that two distinct forms of phycobilisome are assembled with different CpcG copies, which have been referred to as “rod-core linker,” in a cyanobacterium Synechocystis sp. PCC 6803. CpcG2-phycobilisome is devoid of a typical central core, while CpcG1-phycobilisome is equivalent to the conventional phycobilisome supercomplex. Here, we demonstrated that the cpcG1 disruptant has a severe specific defect in the phycobilisome-dependent state transition. However, fluorescence recovery after photobleaching measurements showed no obvious difference in phycobilisome mobility between the wild type and the cpcG1 disruptant. This suggests that both CpcG1 and CpcG2 phycobilisomes have an unstable interaction with the reaction centres. However, only CpcG1 phycobilisomes are involved in state transitions. This suggests that state transitions require the phycobilisome core.
Keywords: Phycobilisome; Cyanobacteria; CpcG; Photosynthesis; Rod-core linker
Fluorescence changes accompanying short-term light adaptations in photosystem I and photosystem II of the cyanobacterium Synechocystis sp. PCC 6803 and phycobiliprotein-impaired mutants: State 1/State 2 transitions and carotenoid-induced quenching of phycobilisomes by Igor N. Stadnichuk; Evgeny P. Lukashev; Irina V. Elanskaya (227-241).
The features of the two types of short-term light-adaptations of photosynthetic apparatus, State 1/State 2 transitions, and non-photochemical fluorescence quenching of phycobilisomes (PBS) by orange carotene-protein (OCP) were compared in the cyanobacterium Synechocystis sp. PCC 6803 wild type, CK pigment mutant lacking phycocyanin, and PAL mutant totally devoid of phycobiliproteins. The permanent presence of PBS-specific peaks in the in situ action spectra of photosystem I (PSI) and photosystem II (PSII), as well as in the 77 K fluorescence excitation spectra for chlorophyll emission at 690 nm (PSII) and 725 nm (PSI) showed that PBS are constitutive antenna complexes of both photosystems. The mutant strains compensated the lack of phycobiliproteins by higher PSII content and by intensification of photosynthetic linear electron transfer. The detectable changes of energy migration from PBS to the PSI and PSII in the Synechocystis wild type and the CK mutant in State 1 and State 2 according to the fluorescence excitation spectra measurements were not registered. The constant level of fluorescence emission of PSI during State 1/State 2 transitions and simultaneous increase of chlorophyll fluorescence emission of PSII in State 1 in Synechocystis PAL mutant allowed to propose that spillover is an unlikely mechanism of state transitions. Blue–green light absorbed by OCP diminished the rout of energy from PBS to PSI while energy migration from PBS to PSII was less influenced. Therefore, the main role of OCP-induced quenching of PBS is the limitation of PSI activity and cyclic electron transport under relatively high light conditions.
Keywords: Cyanobacterium; Fluorescence emission; Fluorescence excitation; Photosynthetic action spectra; Photosystem I; Photosystem II; Phycobilisome(s); State transitions
Dark-to-light transition in Synechococcus sp. PCC 7942 cells studied by fluorescence kinetics assesses plastoquinone redox poise in the dark and photosystem II fluorescence component and dynamics during state 2 to state 1 transition by Merope Tsimilli-Michael; Kostas Stamatakis; George C. Papageorgiou (243-255).
We investigated the dark-to-light transition in Synechococcus sp. PCC 7942 cells by a detailed analysis of fluorescence transients induced by strong red light. The transients, recorded with high data-acquisition, revealed all the steps of the fast (OJIP; 10−5–1 s) and slow phase (PSM(T); 1–103 s), kinetically distinguished with precision. Focusing on the OJIP-rise, we show, for the first time, how the variable to initial fluorescence ratio and the relative height of J-level can serve as indexes of the plastoquinone redox poise and the established state in the dark; hence, differences among cyanobacteria can be recognised in a simple way. Applying intermittent illumination (20-s light pulses separated by 10-s dark intervals) to induce dark-to-light transition and analysing the individual transients, we establish a method by which we determine the fluorescence component not originating from photosystem (PS) II and we assess PSII dynamics during state 2 to state 1 transition. The development of photochemical and non-photochemical quenching is also discussed, as well as evidences favouring the mobile antenna model.
Keywords: Chlorophyll a fluorescence; Cyanobacteria; OJIP fluorescence transient; Photosystem II; State transitions; Phycobilisomes; Plastoquinone pool redox poise