Molecular basis of photoprotection and control of photosynthetic light-harvesting

In order to maximize their use of light energy in photosynthesis, plants have molecules that act as light-harvesting antennae, which collect light quanta and deliver them to the reaction centres, where energy conversion into a chemical form takes place. The functioning of the antenna responds to the extreme changes in the intensity of sunlight encountered in nature. In shade, light is efficiently harvested in photosynthesis. However, in full sunlight, much of the energy absorbed is not needed and there are vitally important switches to specific antenna states, which safely dissipate the excess energy as heat. This is essential for plant survival, because it provides protection against the potential photo-damage of the photosynthetic membrane. But whereas the features that establish high photosynthetic efficiency have been highlighted, almost nothing is known about the molecular nature of the dissipative states. Recently, the atomic structure of the major plant light-harvesting antenna protein, LHCII, has been determined by X-ray crystallography. Here we demonstrate that this is the structure of a dissipative state of LHCII. We present a spectroscopic analysis of this crystal form, and identify the specific changes in configuration of its pigment population that give LHCII the intrinsic capability to regulate energy flow. This provides a molecular basis for understanding the control of photosynthetic light-harvesting.     

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.     

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.     

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.     

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.     

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.     

Low-light-adapted Prochlorococcus species possess specific antennae for each photosystem

Prochlorococcus, the most abundant genus of photosynthetic organisms, owes its remarkably large depth distribution in the oceans to the occurrence of distinct genotypes adapted to either low- or high-light niches. The pcb genes, encoding the major chlorophyll-binding, light-harvesting antenna proteins in this genus, are present in multiple copies in low-light strains but as a single copy in high-light strains. The basis of this differentiation, however, has remained obscure. Here we show that the moderate low-light-adapted strain Prochlorococcus sp. MIT 9313 has one iron-stress-induced pcb gene encoding an antenna protein serving photosystem I (PSI)--comparable to isiA genes from cyanobacteria--and a constitutively expressed pcb gene encoding a photosystem II (PSII) antenna protein. By comparison, the very low-light-adapted strain SS120 has seven pcb genes encoding constitutive PSI and PSII antennae, plus one PSI iron-regulated pcb gene, whereas the high-light-adapted strain MED4 has only a constitutive PSII antenna. Thus, it seems that the adaptation of Prochlorococcus to low light environments has triggered a multiplication and specialization of Pcb proteins comparable to that found for Cab proteins in plants and green algae.     

弱光和Tris处理条件下PSⅡ放氧核心复合物的损伤

弱光条件下(120μmol·m-2·s-1)用Tris(0.8mol/L,pH6.5～10.0)处理具有放氧活性的PSⅡ核心复合物,可引起33kD锰稳定蛋白的释放和锰复合物的破坏,并导致核心复合物的结构发生明显的改变.温和电泳、SDS-PAGE和双向电泳分析表明,主要是PSⅡ核心复合物的二聚体和单体减少,并且复合物部分解体;除反应中心D1和D2蛋白外,核心天线CP43和CP7的量也减少,33kD锰稳定蛋白要发生降解.避光时,PSⅡ核心复合物不受影响.     

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.     

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.     

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.     

The application of lights-conversed polyethylene film for agriculture

Lanthanum-rhodamine (6G and B) complexes were synthesized by Rheological Phase Reaction Method. Lanthanum-rhodamine (6G and B) complexes doped polyethylene films which have a function of lights-conversion were prepared. The emission and excitation spectra were measured. The experiments of growing seedling and culture were carried out in the shed built with doped and undoped polyethylene films. Lanthanum-rhodamine doped polyethylene films which have a function of lights-conversion can efficiently convert the green light in the sunlight to the red light for photosynthesis of crops, to promote the maturing of crops and raise the yield of crops.     

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...     

Structure and function of the AAA+ protein CbbX, a red-type Rubisco activase

Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyses the fixation of atmospheric CO(2) in photosynthesis, but tends to form inactive complexes with its substrate ribulose 1,5-bisphosphate (RuBP). In plants, Rubisco is reactivated by the AAA(+) (ATPases associated with various cellular activities) protein Rubisco activase (Rca), but no such protein is known for the Rubisco of red algae. Here we identify the protein CbbX as an activase of red-type Rubisco. The 3.0-? crystal structure of unassembled CbbX from Rhodobacter sphaeroides revealed an AAA(+) protein architecture. Electron microscopy and biochemical analysis showed that ATP and RuBP must bind to convert CbbX into functionally active, hexameric rings. The CbbX ATPase is strongly stimulated by RuBP and Rubisco. Mutational analysis suggests that CbbX functions by transiently pulling the carboxy-terminal peptide of the Rubisco large subunit into the hexamer pore, resulting in the release of the inhibitory RuBP. Understanding Rubisco activation may facilitate efforts to improve CO(2) uptake and biomass production by photosynthetic organisms.     

Modulation of gelsolin function by phosphatidylinositol 4,5-bisphosphate

The actin-binding protein gelsolin requires micromolar concentrations of calcium ions to sever actin filaments, to potentiate its binding to the end of the filament and to promote the polymerization of monomeric actin into filaments. Because transient increases in both intracellular [Ca2+] and actin polymerization accompany the cellular response to certain stimuli, it has been suggested that gelsolin regulates the reversible assembly of actin filaments that accompanies such cellular activations. But other evidence suggests that these activities do not need increased cytoplasmic [Ca2+] and that once actin-gelsolin complexes form in the presence of Ca2+ in vitro, removal of free Ca2+ causes dissociation of only one of two bound actin monomers from gelsolin and the resultant binary complexes cannot sever actin filaments. The finding that cellular gelsolin-actin complexes can be dissociated suggests that a Ca2+-independent regulation of gelsolin also occurs. Here we show that, like the dissociation of profilin-actin complexes, phosphatidylinositol 4,5-bisphosphate, which undergoes rapid turnover during cell stimulation, strongly inhibits the actin filament-severing properties of gelsolin, inhibits less strongly the nucleating ability of this protein and restores the potential for filament-severing activity to gelsolin-actin complexes.     

植物抗热性与叶绿体的关系

与其它细胞器相比,叶绿体对热敏感的多,PSH比PSI对热敏感.各种光合反应中,放氧过程对热最敏感。高温下功能性Mn~(2 )的解离,抑制光合放氧。高温胁迫下,叶绿体膜脂的特性是否与抗热性有关,目前看法不一,有些资料表明,叶绿体在热适应过程中,类囊体中产生热保护因子,其性质不甚明了。HSPs可能参与热保护作用。叶绿体中含有丰富的抗氧化物质和自由基清除酶系,目前有很多资料显示,耐热与热敏植物品种或植物在不同的热处理条件下,其各种活性氧含量或活性氧清除能力存在差异。     

浸没式UV-C技术用于自来水厂除藻的研究

设计了一种浸没式UV-C装置,用于自来水厂斜管沉淀池的抑藻、除藻,通过分析其试验数据,认为是因为抑制了藻类的光合作用,从而达到抑藻和除藻目的,还实施了用黑色网格纤维罩在斜管沉淀池上的办法,减弱斜管沉淀池中藻类可吸收的太阳光,同样能有效抑制斜管沉淀池中藻类的生长繁殖。     

海滨锦葵的耐冷性及低温弱光对其光系统的伤害

以海滨锦葵为实验材料,研究不同低温处理对海滨锦葵光合作用的伤害,探求海滨锦葵对低温胁迫的敏感温度,以及低温弱光对海滨锦葵的伤害．结果表明：在低温胁迫下,海滨锦葵的净光合速率（Pn）、光系统II实际光化学效率（ФPSII）、最大光化学效率（Fv／Fm）显著下降,说明随着温度的降低,海滨锦葵光化学活性受到抑制,13℃是其低温胁迫下的临界温度．低温弱光（6℃、200μmol·m^-2s^-1）处理4h后Fv/Fm下降了2．5％,而光系统I活性（△I／I0）下降了18．5％,说明在低温弱光条件下,海滨锦葵光系统I受到的伤害高于光系统II;在恢复过程中,光系统II在8h基本完全恢复,而光系统I和净光合速率在48h后仍没有恢复到正常水平,说明PSI的恢复速率成了光合作用的主要限制因素．     

Effect of Phosphatidylcholine on the Steady State Fluorescence of Chlorophyll in Photosystem Ⅱ Particles

Phosphatidylcholine (PC) accounts for less than 1% of the total lipids in plant photosystem II (PSII) particles.In this experiment, PSII particles were reconstituted with PC to construct PSII-PC vesicles.The effect of PC on the steady state fluorescence of chlorophyll (Chl) in PSII particles was studied.The results show that PC significantly affected the fluorescence intensity, but did not obviously affect the fluorescence emission band peak position.PC also did not obviously affect the absorbance at 436 nm or the amide I band peak position in FT-IR spectroscopy of PSII particles.The results suggest that PC may affect the light energy transfer from the antenna chlorophyll molecules to the reaction center chlorophyll molecule (P680).     

Effect of Phosphatidylcholine on the Steady State Fluorescence of Chlorophyll in Photosystem Ⅱ Particles

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