Plants lacking the main light-harvesting complex retain photosystem II macro-organization

Photosystem II (PSII) is a key component of photosynthesis, the process of converting sunlight into the chemical energy of life. In plant cells, it forms a unique oligomeric macrostructure in membranes of the chloroplasts. Several light-harvesting antenna complexes are organized precisely in the PSII macrostructure-the major trimeric complexes (LHCII) that bind 70% of PSII chlorophyll and three minor monomeric complexes-which together form PSII supercomplexes. The antenna complexes are essential for collecting sunlight and regulating photosynthesis, but the relationship between these functions and their molecular architecture is unresolved. Here we report that antisense Arabidopsis plants lacking the proteins that form LHCII trimers have PSII supercomplexes with almost identical abundance and structure to those found in wild-type plants. The place of LHCII is taken by a normally minor and monomeric complex, CP26, which is synthesized in large amounts and organized into trimers. Trimerization is clearly not a specific attribute of LHCII. Our results highlight the importance of the PSII macrostructure: in the absence of one of its main components, another protein is recruited to allow it to assemble and function.     

Identification of a mechanism of photoprotective energy dissipation in higher plants

Under conditions of excess sunlight the efficient light-harvesting antenna found in the chloroplast membranes of plants is rapidly and reversibly switched into a photoprotected quenched state in which potentially harmful absorbed energy is dissipated as heat, a process measured as the non-photochemical quenching of chlorophyll fluorescence or qE. Although the biological significance of qE is established, the molecular mechanisms involved are not. LHCII, the main light-harvesting complex, has an inbuilt capability to undergo transformation into a dissipative state by conformational change and it was suggested that this provides a molecular basis for qE, but it is not known if such events occur in vivo or how energy is dissipated in this state. The transition into the dissipative state is associated with a twist in the configuration of the LHCII-bound carotenoid neoxanthin, identified using resonance Raman spectroscopy. Applying this technique to study isolated chloroplasts and whole leaves, we show here that the same change in neoxanthin configuration occurs in vivo, to an extent consistent with the magnitude of energy dissipation. Femtosecond transient absorption spectroscopy, performed on purified LHCII in the dissipative state, shows that energy is transferred from chlorophyll a to a low-lying carotenoid excited state, identified as one of the two luteins (lutein 1) in LHCII. Hence, it is experimentally demonstrated that a change in conformation of LHCII occurs in vivo, which opens a channel for energy dissipation by transfer to a bound carotenoid. We suggest that this is the principal mechanism of photoprotection.     

The PSI-H subunit of photosystem I is essential for state transitions in plant photosynthesis

Photosynthesis in plants involves two photosystems responsible for converting light energy into redox processes. The photosystems, PSI and PSII, operate largely in series, and therefore their excitation must be balanced in order to optimize photosynthetic performance. When plants are exposed to illumination favouring either PSII or PSI they can redistribute excitation towards the light-limited photosystem. Long-term changes in illumination lead to changes in photosystem stoichiometry. In contrast, state transition is a dynamic mechanism that enables plants to respond rapidly to changes in illumination. When PSII is favoured (state 2), the redox conditions in the thylakoids change and result in activation of a protein kinase. The kinase phosphorylates the main light-harvesting complex (LHCII) and the mobile antenna complex is detached from PSII. It has not been clear if attachment of LHCII to PSI in state 2 is important in state transitions. Here we show that in the absence of a specific PSI subunit, PSI-H, LHCII cannot transfer energy to PSI, and state transitions are impaired.     

Photosystem II core phosphorylation and photosynthetic acclimation require two different protein kinases

Illumination changes elicit modifications of thylakoid proteins and reorganization of the photosynthetic machinery. This involves, in the short term, phosphorylation of photosystem II (PSII) and light-harvesting (LHCII) proteins. PSII phosphorylation is thought to be relevant for PSII turnover, whereas LHCII phosphorylation is associated with the relocation of LHCII and the redistribution of excitation energy (state transitions) between photosystems. In the long term, imbalances in energy distribution between photosystems are counteracted by adjusting photosystem stoichiometry. In the green alga Chlamydomonas and the plant Arabidopsis, state transitions require the orthologous protein kinases STT7 and STN7, respectively. Here we show that in Arabidopsis a second protein kinase, STN8, is required for the quantitative phosphorylation of PSII core proteins. However, PSII activity under high-intensity light is affected only slightly in stn8 mutants, and D1 turnover is indistinguishable from the wild type, implying that reversible protein phosphorylation is not essential for PSII repair. Acclimation to changes in light quality is defective in stn7 but not in stn8 mutants, indicating that short-term and long-term photosynthetic adaptations are coupled. Therefore the phosphorylation of LHCII, or of an unknown substrate of STN7, is also crucial for the control of photosynthetic gene expression.     

Crystal structure of plant photosystem I

Oxygenic photosynthesis is the principal producer of both oxygen and organic matter on Earth. The conversion of sunlight into chemical energy is driven by two multisubunit membrane protein complexes named photosystem I and II. We determined the crystal structure of the complete photosystem I (PSI) from a higher plant (Pisum sativum var. alaska) to 4.4 A resolution. Its intricate structure shows 12 core subunits, 4 different light-harvesting membrane proteins (LHCI) assembled in a half-moon shape on one side of the core, 45 transmembrane helices, 167 chlorophylls, 3 Fe-S clusters and 2 phylloquinones. About 20 chlorophylls are positioned in strategic locations in the cleft between LHCI and the core. This structure provides a framework for exploration not only of energy and electron transfer but also of the evolutionary forces that shaped the photosynthetic apparatus of terrestrial plants after the divergence of chloroplasts from marine cyanobacteria one billion years ago.     

Two-dimensional spectroscopy of electronic couplings in photosynthesis

Time-resolved optical spectroscopy is widely used to study vibrational and electronic dynamics by monitoring transient changes in excited state populations on a femtosecond timescale. Yet the fundamental cause of electronic and vibrational dynamics--the coupling between the different energy levels involved--is usually inferred only indirectly. Two-dimensional femtosecond infrared spectroscopy based on the heterodyne detection of three-pulse photon echoes has recently allowed the direct mapping of vibrational couplings, yielding transient structural information. Here we extend the approach to the visible range and directly measure electronic couplings in a molecular complex, the Fenna-Matthews-Olson photosynthetic light-harvesting protein. As in all photosynthetic systems, the conversion of light into chemical energy is driven by electronic couplings that ensure the efficient transport of energy from light-capturing antenna pigments to the reaction centre. We monitor this process as a function of time and frequency and show that excitation energy does not simply cascade stepwise down the energy ladder. We find instead distinct energy transport pathways that depend sensitively on the detailed spatial properties of the delocalized excited-state wavefunctions of the whole pigment-protein complex.     

A giant chlorophyll-protein complex induced by iron deficiency in cyanobacteria

Cyanobacteria are abundant throughout most of the world's water bodies and contribute significantly to global primary productivity through oxygenic photosynthesis. This reaction is catalysed by two membrane-bound protein complexes, photosystem I (PSI) and photosystem II (PSII), which both contain chlorophyll-binding subunits functioning as an internal antenna. In addition, phycobilisomes act as peripheral antenna systems, but no additional light-harvesting systems have been found under normal growth conditions. Iron deficiency, which is often the limiting factor for cyanobacterial growth in aquatic ecosystems, leads to the induction of additional proteins such as IsiA (ref. 3). Although IsiA has been implicated in chlorophyll storage, energy absorption and protection against excessive light, its precise molecular function and association to other proteins is unknown. Here we report the purification of a specific PSI-IsiA supercomplex, which is abundant under conditions of iron limitation. Electron microscopy shows that this supercomplex consists of trimeric PSI surrounded by a closed ring of 18 IsiA proteins binding around 180 chlorophyll molecules. We provide a structural characterization of an additional chlorophyll-containing, membrane-integral antenna in a cyanobacterial photosystem.     

内蒙古高原荒漠区几种锦鸡儿属优势植物叶绿素荧光参数的比较研究

比较了内蒙古高原荒漠区4种锦鸡儿属优势植物——柠条锦鸡儿、狭叶锦鸡儿、垫状锦鸡儿和荒漠锦鸡儿叶绿素荧光参数的日进程.对光合有效辐射（PAR）、最小荧光（Fo）、最大荧光（Fm）、光系统Ⅱ（PSⅡ）最大光能转换效率（Fv/Fm）、PSⅡ光能捕获效率（Fv′/Fm′）、PSⅡ实际光化学量子效率（ΦPSⅡ）、电子传递速率（ETR）和叶片光合功能的相对限制L（PFD）的分析表明：高的PAR会导致4种锦鸡儿光合作用的光抑制,但并不造成PSⅡ反应中心的不可逆破坏.光抑制程度荒漠锦鸡儿大于垫状锦鸡儿;狭叶锦鸡儿和柠条锦鸡儿光抑制程度差异不大,光抑制程度最小.随着PAR的升高,4种锦鸡儿对光能捕获、转化和利用能力降低,种间表现为柠条锦鸡儿〉狭叶锦鸡儿〉荒漠锦鸡儿〉垫状锦鸡儿.淬灭分析表明：随着PAR的升高,4种锦鸡儿用于光化学反应的光能减少,耗散的热能增多;其中,柠条锦鸡儿用于光化学反应的光能最多,荒漠锦鸡儿用于耗散的热能最多.对PSⅡ天线色素吸收光能中既没有被光合电子传递所利用,也没有作为热能耗散的部分（Excess）比较研究表明：4种锦鸡儿的Excess都较高,种间差异不明显.     

Three-dimensional structure of plant light-harvesting complex determined by electron crystallography

The structure of the light-harvesting chlorophyll a/b-protein complex, a membrane protein serving as the major antenna of solar energy in plant photosynthesis, has been determined at 6 A resolution by electron crystallography. Within the complex, three membrane-spanning alpha helices and 15 chlorophyll molecules are resolved. There is an intramolecular diad relating two of the alpha helices and some of the chlorophylls. The spacing of the chlorophylls suggests energy transfer by delocalized exciton coupling and F?rster mechanisms.     

Towards complete cofactor arrangement in the 3.0 A resolution structure of photosystem II

Oxygenic photosynthesis in plants, algae and cyanobacteria is initiated at photosystem II, a homodimeric multisubunit protein-cofactor complex embedded in the thylakoid membrane. Photosystem II captures sunlight and powers the unique photo-induced oxidation of water to atmospheric oxygen. Crystallographic investigations of cyanobacterial photosystem II have provided several medium-resolution structures (3.8 to 3.2 A) that explain the general arrangement of the protein matrix and cofactors, but do not give a full picture of the complex. Here we describe the most complete cyanobacterial photosystem II structure obtained so far, showing locations of and interactions between 20 protein subunits and 77 cofactors per monomer. Assignment of 11 beta-carotenes yields insights into electron and energy transfer and photo-protection mechanisms in the reaction centre and antenna subunits. The high number of 14 integrally bound lipids reflects the structural and functional importance of these molecules for flexibility within and assembly of photosystem II. A lipophilic pathway is proposed for the diffusion of secondary plastoquinone that transfers redox equivalents from photosystem II to the photosynthetic chain. The structure provides information about the Mn4Ca cluster, where oxidation of water takes place. Our study uncovers near-atomic details necessary to understand the processes that convert light to chemical energy.     

Ecology: a niche for cyanobacteria containing chlorophyll d

The cyanobacterium known as Acaryochloris marina is a unique phototroph that uses chlorophyll d as its principal light-harvesting pigment instead of chlorophyll a, the form commonly found in plants, algae and other cyanobacteria; this means that it depends on far-red light for photosynthesis. Here we demonstrate photosynthetic activity in Acaryochloris-like phototrophs that live underneath minute coral-reef invertebrates (didemnid ascidians) in a shaded niche enriched in near-infrared light. This discovery clarifies how these cyanobacteria are able to thrive as free-living organisms in their natural habitat.     

Quantum control of energy flow in light harvesting

Coherent light sources have been widely used in control schemes that exploit quantum interference effects to direct the outcome of photochemical processes. The adaptive shaping of laser pulses is a particularly powerful tool in this context: experimental output as feedback in an iterative learning loop refines the applied laser field to render it best suited to constraints set by the experimenter. This approach has been experimentally implemented to control a variety of processes, but the extent to which coherent excitation can also be used to direct the dynamics of complex molecular systems in a condensed-phase environment remains unclear. Here we report feedback-optimized coherent control over the energy-flow pathways in the light-harvesting antenna complex LH2 from Rhodopseudomonas acidophila, a photosynthetic purple bacterium. We show that phases imprinted by the light field mediate the branching ratio of energy transfer between intra- and intermolecular channels in the complex's donor acceptor system. This result illustrates that molecular complexity need not prevent coherent control, which can thus be extended to probe and affect biological functions.     

State transitions and light adaptation require chloroplast thylakoid protein kinase STN7

Photosynthetic organisms are able to adjust to changing light conditions through state transitions, a process that involves the redistribution of light excitation energy between photosystem II (PSII) and photosystem I (PSI). Balancing of the light absorption capacity of these two photosystems is achieved through the reversible association of the major antenna complex (LHCII) between PSII and PSI (ref. 3). Excess stimulation of PSII relative to PSI leads to the reduction of the plastoquinone pool and the activation of a kinase; the phosphorylation of LHCII; and the displacement of LHCII from PSII to PSI (state 2). Oxidation of the plastoquinone pool by excess stimulation of PSI reverses this process (state 1). The Chlamydomonas thylakoid-associated Ser-Thr kinase Stt7, which is required for state transitions, has an orthologue named STN7 in Arabidopsis. Here we show that loss of STN7 blocks state transitions and LHCII phosphorylation. In stn7 mutant plants the plastoquinone pool is more reduced and growth is impaired under changing light conditions, indicating that STN7, and probably state transitions, have an important role in response to environmental changes.     

Growth, photosynthesis and podophyllotoxin accumulation of Dysosma versipellis in response to a light gradient and conservation implications

Dysosma versipellis (Hance) M. Cheng ex Ying (Berberidaceae) is a rare and vulnerable, perennial herb endemic to China with pharmaceutical significance. Increasing habitat loss and over-exploitation of the plant has severely affected the plant’s in situ conservation, necessitating ex situ conservation and commercial cultivation. The light regime is a critical environmental factor contributing to successful ex situ conservation via efficient production of biomass and secondary metabolites. We investigated th...     

A pigment-binding protein essential for regulation of photosynthetic light harvesting

Photosynthetic light harvesting in plants is regulated in response to changes in incident light intensity. Absorption of light that exceeds a plant's capacity for fixation of CO2 results in thermal dissipation of excitation energy in the pigment antenna of photosystem II by a poorly understood mechanism. This regulatory process, termed nonphotochemical quenching, maintains the balance between dissipation and utilization of light energy to minimize generation of oxidizing molecules, thereby protecting the plant against photo-oxidative damage. To identify specific proteins that are involved in nonphotochemical quenching, we have isolated mutants of Arabidopsis thaliana that cannot dissipate excess absorbed light energy. Here we show that the gene encoding PsbS, an intrinsic chlorophyll-binding protein of photosystem II, is necessary for nonphotochemical quenching but not for efficient light harvesting and photosynthesis. These results indicate that PsbS may be the site for nonphotochemical quenching, a finding that has implications for the functional evolution of pigment-binding proteins.     

Genetically modified photosynthetic antenna complexes with blueshifted absorbance bands.

Light energy for photosynthesis is collected by the antenna system, creating an excited state which migrates energetically 'downhill'. To achieve efficient migration of energy the antenna is populated with a series of pigments absorbing at progressively redshifted wavelengths. This variety in absorbing species in vivo has been created in a biosynthetically economical fashion by modulating the absorbance behaviour of one kind of (bacterio)chlorophyll molecule. This modulation is poorly understood but has been ascribed to pigment-pigment and pigment-protein interactions. We have examined the relationship between aromatic residues in antenna polypeptides and pigment absorption, by studying the effects of site-directed mutagenesis on a bacterial antenna complex. A clear correlation was observed between the absorbance of bacteriochlorophyll a and the presence of two tyrosine residues, alpha Tyr44 and alpha Tyr45, in the alpha subunit of the peripheral light-harvesting complex of Rhodobacter sphaeroides, a purple photosynthetic bacterium that provides a well characterized system for site-specific mutagenesis. By constructing single (alpha Tyr44, alpha Tyr45----PheTyr) and then double (alpha Tyr44, alpha Tyr45----PheLeu) site-specific mutants, the absorbance of bacteriochlorophyll was blueshifted by 11 and 24 nm at 77 K, respectively. The results suggest that there is a close approach of tyrosine residues to bacteriochlorophyll, and that this proximity may promote redshifts in vivo.     

不同遮阴强度对猕猴桃“桂海4号”光合特性及果实品质的影响

以中华猕猴桃“桂海4号”（Actinidia chinensis Planch cv．Guihaia 4）为研究对象,采用LI-6400便携式光合测定系统测定不同程度（0,40％和60％）遮阴对其光合、荧光特性和果实品质的影响。结果表明,与全光照相比,40％和60％的遮阴强度显著降低了猕猴桃的光饱和速率（Amax）、表观量子效率（AQY）、光饱和点（LSP）、光补偿点（LCP）和暗呼吸速率（Rd）（P〈0．05）,对其水分利用效率（WUE）和叶绿素含量（Chlcontent）均无显著影响（P〉0．05）,但是潜在水分利用效率（WUEi）随着遮阴强度的增加而增加。猕猴桃的初始荧光（Fo）、可变荧光（Fv）、最大荧光（Fm）和PSⅡ最大光能转换效率（Fv/Fm）在全光照和不同遮阴处理间无显著差异（P〉0．05）,表明一定程度的遮刚对其Fo、Fc、Fm、Fv／Fm,无明显影响,但是与全光照相比,40％的遮阴强度显著降低了PSⅡ光能捕获效率（Fc／Fm）、PSⅡ电子传递量子效率（PhiPS2）和光化学猝灭系数（gP）,而60％遮阴强度其F＇v／Fm、PhiPS2和qP与全光照下的无显著差异（P〉0．05）。全光照和遮阴条件下猕猴桃光合和荧光参数相关关系存在明显差异,表明猕猴桃“桂海4号”随着环境条件中光照的改变其光合器官进行了一定的调整,使其适应变化了的环境条件。遮阴对猕猴桃“桂海4号”的果实外观无显著影响,但是却显著影响了其果实品质,40％遮阴强度显著降低其维C、含量、总糖、可溶性固体物和酸含量。     

Improving photosynthesis of microalgae by changing the ratio of light-harvesting pigments

Changing the ratio of light-harvesting pigments was regarded as an efficient way to improve the photosynthesis rate in microalgae, but the underlying mechanism is still unclear. In the present study, a mutant of Anabeana simensis （called SP） was selected from retrieved satellite cultures. Several parameters related with photosynthesis, such as the growth, photosynthesis rate, the content of photosynthetic pigment, low temperature fluorescence spectrum （77K） and electron transport rate, were compared with those of the wild type. It was found that the change in the ratio of light-harvesting pigments in the mutant led to more efficient light energy transfer and usage in mutant than in the wild type. This may be the reason why the mutant had higher photosynthesis and growth rates.     

Bacteriophytochrome controls photosystem synthesis in anoxygenic bacteria

Plants use a set of light sensors to control their growth and development in response to changes in ambient light. In particular, phytochromes exert their regulatory activity by switching between a biologically inactive red-light-absorbing form (Pr) and an active far-red-light absorbing form (Pfr). Recently, biochemical and genetic studies have demonstrated the occurrence of phytochrome-like proteins in photosynthetic and non-photosynthetic bacteria--but little is known about their functions. Here we report the discovery of a bacteriophytochrome located downstream from the photosynthesis gene cluster in a Bradyrhizobium strain symbiont of Aeschynomene. The synthesis of the complete photosynthetic apparatus is totally under the control of this bacteriophytochrome. A similar behaviour is observed for the closely related species Rhodopseudomonas palustris, but not for the more distant anoxygenic photosynthetic bacteria of the genus Rhodobacter, Rubrivivax or Rhodospirillum. Unlike other (bacterio)phytochromes, the carboxy-terminal domain of this bacteriophytochrome contains no histidine kinase features. This suggests a light signalling pathway involving direct protein-protein interaction with no phosphorelay cascade. This specific mechanism of regulation may represent an important ecological adaptation to optimize the plant-bacteria interaction.     

水曲柳幼苗水力结构和光合生理对光强梯度变化的耦合响应

【目的】 研究水曲柳(Fraxinus mandschurica)幼苗对不同光照强度的响应及适应机理,为育苗造林和林下天然更新研究提供参考。【方法】 以2年生盆栽水曲柳幼苗为材料进行不同光照处理,1年后测定幼苗在4种强度日光处理(相对光强100%、60%、30%和15%)下的生长、光合生理和水力性状等指标。【结果】 遮阴处理显著影响了水曲柳幼苗的生长、光合和水分生理特性。与60%和30%相对光强处理相比,100%相对光强下水曲柳具有更高的气孔导度(G_s)、净光合速率(P_n)、根系水力导度(K_root)、地上部分水力导度(K_shoot)和整株水力导度(K_plant)。随光照强度的减弱,水曲柳的生长速率显著降低,根生物量占比(R_MR)减少,最大净光合速率(P_n,max)、光补偿点(L_CP)和光饱和点(L_SP)降低,而茎、叶生物量占比(S_MR、L_MR)以及表观量子效率(A_QY)增加;枝条木质部解剖结构在不同光照处理下存在显著差异,相对光强100%处理时的导管密度显著高于相对光强30%和60%处理,而导管直径显著低于30%和60%处理。【结论】 水曲柳幼苗通过调节形态、光合和水分生理特性来适应一定程度的弱光环境,但总体上对光强有较高的需求。水曲柳的光合和水力性状随光照强度的改变都具有较大的可塑性,二者对光照强度梯度变化的响应存在显著耦合关系。这些性状的可塑性响应有利于提高水曲柳幼苗在异质性光环境下的生存适合度,对于其更新阶段在林下不同光照条件下的生存有重要意义。