Force field development on pigments of photosystem 2 reaction centre

We developed new parameters for molecular dynamics (MD) simulations, namely partial atomic charges, equilibrium bond-lengths, angles, dihedrals, atom types, and force constants of chlorophyll a (Chl) and plastoquinone (PQ), and both reduced and neutral form of primary acceptor (PHO) molecule. These parameters are essential for MD simulations that can interpret various structure functional relationships during primary processes of charge separation and stabilization in photosystem 2 reaction centres.     

Thermostability of photosynthesis in two new chlorophyllb-less rice mutants

Leaves of the two new chlorophyllb-less rice mutants VG28-1, VG30-5 and the wild type rice cv. Zhonghua 11 were subjected to temperatures 28, 36, 40, 44 and 48°C in the dark for 30 min or gradually elevated temperature from 30°C to 80°C at 0.5°C/min. The thermostability of photosynthetic apparatus was estimated by the changes in chlorophyll fluorescence parameters, photosynthetic rate and pigment content, chloroplast ultrastructure and tissue location of H₂O₂ accumulation. There were different patterns of F_o-temperature curves between the Chlb-less mutants and the wild type plant, and the temperature of F_o rising threshold was shifted 3°C lower in the Chlb-less mutants (48°C) than in the wild type (51°C). At temperature up to about 45°C, chloroplasts were swollen and thylakoid grana became misty accompanied with the complete loss of photosynthetic oxygen evolution in the two Chlb-less mutants, but chloroplast ultrastructure in the wild type showed no obvious alteration. After 55°C exposure, the disordered thylakoid and significant H₂O₂ accumulation in leaves were found in the two Chlb-less mutants, whereas in the wild type plant, less H₂O₂ was accumulated and the swollen thylakoid still maintained a certain extent of stacking. A large extent of the changes in qP, NPQ and F_v/F_m was consistent with the Pn decreasing rate in the Chlb-less mutants during high temperature treatment as compared with the wild type. The results indicated that the Chlb-less mutants showed a tendency for higher thermosensitivity, and loss of Chlb in LHC II could lead to less thermostability of PSII structure and function. Heat damage to photosynthetic apparatus might be partially attributed to the internal oxidative stress produced at severely high temperature.     

Spectrofluorimetric method for the determination of large chlorophyll a/b ratios

A practical spectrofluorimetric calibration method at room temperature is described for determining large chlorophyll a/b ratios on direct extracts from plant material of pigments in 80% aqueous acetone, dimethyl sulfoxide, and N,N-dimethylformamide. The method is based on the work of Boardman and Thorne [Boardman NK and Thorne SW (1971) Biochim Biophys Acta 253: 222–231] who used diethyl ether as solvent. We repeated the calibration in diethyl ether and find significantly different parameters for the calibration curve. The range of standards in this work included solutions with chlorophyll a/b ratios of 10–125 in dimethyl sulfoxide, and of 10–220 in the other solvents. Fluorescence emission spectra were found to be a highly sensitive method for assessing chlorophyll purity. We determined the limits of sensitivity for each solvent from the calibration data. The empirically determined slope of the calibration curve was shown to be related to intrinsic properties of the chlorophylls in solution; this allows predictions of the performance of the method in other solvents.     

Photosynthesis in leaves of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L. infected with tobacco mosaic virus

In tobacco leaves inoculated with tobacco mosaic virus (TMV), changes in chlorophyll (Chl) and carotenoid contents, parameters of slow Chl fluorescence kinetics, i.e. the maximum quantum yield of photosystem (PS2) photochemistry F_v/F_m, the effective quantum yield of photochemical energy conversion in PS2 Φ₂, ratio of quantum yields of photochemical and concurrent non-photochemical processes in PS2 F_v/F₀, non-photochemical quenching (NPQ), and photochemical activities of isolated chloroplasts from systemically infected tobacco leaves were investigated. We compared two successive stages of infection, the first in the stage of vein clearing at 9^th day post inoculation (dpi) and the second at 22^nd dpi when two different regions, i.e. light- (LGI) or dark-green (DGI) islands in the infected leaf were apparent and symptoms were fully developed. These two different regions were measured separately. The Chl and carotenoid contents in infected leaves decreased with a progression of infection and were lowest in LGI in the second stage. Also the ratio of Chl a/b declined in similar manner. The maximum quantum yield of PS2 photochemistry F_v/F_m, was decreased in the following order: first stage, DGI, and LGI. The same is true for the ratio F_v/F₀. The decrease of Φ₂ in infected leaves declined as compared to their controls. On the contrary, NPQ increased in infected leaves, the highest value was found in the first infection stage. Photochemical activities of the whole electron transport chain in isolated chloroplasts dramatically declined with the progression of symptoms, the lowest value was in LGI. Similarly, but to a lesser extent, the activity of PS2 in isolated chloroplasts decreased in infected leaves. Generally, the most marked impairment of the photosynthetic apparatus was manifested in the LGI of infected leaves.     

水稻叶片叶绿素和过氧化氢含量的QTL检测及上位性分析

研究水稻叶片叶绿素和过氧化氢含量的遗传规律,对探讨光合代谢产物遗传规律和开展高产育种具有重要指导意义。利用由日本晴／Kasalath∥日本晴的杂交组合衍生的98个回交重组自交家系(BC1F9)所组成的BIL(backcross inbred lines)群体,在第1、2、3和10染色体上分别检测出5个与叶绿素含量相关的QTL和2个影响剑叶过氧化氢含量的QTL,其中位于第1染色体的RFLP标记C86和C813之间的q-Chll对叶绿素含量的影响最为显著,对表型变异的贡献率达22％,其增效基因来自粳稻品种日本晴;同时在该区间检测到1个与剑叶过氧化氢含量相关的QTL:q-H2O2I,对过氧化氢含量的减效基因来自日本晴品种。上位性分析结果显示影响叶绿素含量及过氧化氢含量的非等位QTL之间存在显著的上位性效应。具有上位性效应的QTL分布于第2、6、11和12染色体上,未检测到与q-Chll或q-H2O2I互作的位点。暗示日本晴品种的RFLP标记C86和C813之间存在1个能够提高叶绿素含量,同时又能降低过氧化氢含量的主效QTL,其加性效应显著而不存在上位性效应。     

Changes in Pigment Composition and Photosynthetic Activity of Aquatic Fern (Azolla Microphylla Kaulf.) Exposed to Low Doses of UV-C (254 nm) Radiation

Changes in the content of pigments and rate of photosynthesis in Azolla microphylla Kaulf. fronds were measured during growth under solar and ultraviolet-C (UV-C) supplemented solar radiation. Maximum content of total chlorophyll (Chl) was observed on the 13^th day (termination of the experiment) of treatment in both control and treated plants. The treated plants had significantly lower total Chl and carotenoid contents than the control plants during the 1^st day of growth. After the 4^th day of exposure to UV-C supplemented solar radiation, the Chl and carotenoids accumulation increased in treated plants, so that the pigment concentration in the treated fronds was nearer to the control values after the 13^th day of treatment. Significant increase in UV absorbing pigments, anthocyanins, and flavonoids was observed at the 13^th day of treatment. In spite of the roughly similar photosynthetic pigment concentration, the photosynthetic activity measured as the rate of electron transport at photosystem 2 was only 65 % of the control values after 13 d of UV-C exposure. This revised version was published online in June 2006 with corrections to the Cover Date.     

Diurnal Fluctuations in Photochemical Activities of Chloroplasts in Field Grown Cyamopsis Tetragonoloba Seedlings

In field-grown Cyamopsis seedlings, distinct changes were found in the rates of photosystems (PS) 2 and 1 activities at different time of the day. Maximum PS2 activity was at around 11:00 h and decreased thereafter. On the contrary, PS1 activity continued to increase up to 14:00 h and declined in evening hours. Significant energy transfer from PS2 to PS1 was evident during the morning and evening hours of the day whereas a slow excitation of PS2 and energy transfer was favoured during noon hours. This revised version was published online in September 2006 with corrections to the Cover Date.     

ULTRASTRUCTURE AND PIGMENTS OF TWO STRAINS OF THE PICOPLANKTONIC ALGA PELAGOCOCCUS SUBVIRIDIS (CHRYSOPHYCEAE)1

The organelle ultrastructure and photosynthetic pigments of a new isolate of the picoplanktonic alga Pelagococcus subviridis Norris from the East Australian Current was compared with the North Pacific Ocean type species. No differences in the ultrastructure of the two isolates were observed. Mitosis was studied in detail in the Australian strain, and showed two unusual features: the de novo appearance of centrioles prior to mitosis, and the formation of a small, extra-nuclear spindle. The major carotenoids in both strains were fucoxanthin and a 19′-butanoyloxyfucoxanthin-like pigment, with diadinoxanthin and diatoxanthin as secondary pigments. Several minor carotenoids have not yet been identified. In addition to chlorophylls a and c₂, a new chlorophyll c derivative (chlorophyll c₃), present in both strains, was separated by high-performance thin-layer chromatography (HPTLC). The Australian isolate, unlike the type material, showed no evidence of chlorophyllase activity during cell harvest and extraction. While the pigment composition suggests affinities with certain newly examined prymnesiophytes, organelle ultrastructure indicates Pelagococcus to be a member of the Chrysophyceae. Mitosis is, however, atypical of both Prymnesiophyceae and Chrysophyceae, and if this picoplanktonic alga is to be retained in the Chrysophyceae it must be seen as a most unusual member.     

Photosystem 2 activity of <Emphasis Type="Italic">Citrus volkameriana</Emphasis> (L.) leaves as affected by Mn nutrition and irradiance

Citrus volkameriana (L.) plants were grown for 43 d in nutrient solutions containing 0, 2, 14, 98, or 686 μM Mn (Mn₀, Mn₂, Mn₁₄, Mn₉₈, and Mn₆₈₆, respectively). To adequately investigate the combined effects of Mn nutrition and irradiance on photosystem 2 (PS2) activity, irradiance response curves for electron transport rate (ETR), nonphotochemical quenching (q_N), photochemical quenching (q_P), and real photochemical efficiency of PS2 (Φ_PS2) were recorded under 10 different irradiances (66, 96, 136, 226, 336, 536, 811, 1 211, 1 911, and 3 111 μmol m⁻² s⁻¹, I₆₆ to I₃₁₁₁, respectively) generated with the PAM-2000 fluorometer. Leaf chlorophyll content was significantly lower under Mn excess (Mn₆₈₆) compared to Mn₀; its highest values were recorded in the treatments Mn₂-Mn₉₈. However, ETR and Φ_PS2 values were significantly lower under Mn₀ compared to the other Mn treatments, when plants were exposed to irradiances ≥96 μmol m⁻² s⁻¹. Furthermore, Mn₀ plants had significantly higher values of q_N and lower values of q_P at irradiances ≤226 and ≥336 μmol m⁻² s⁻¹, respectively, than those grown under Mn₂-Mn₆₈₆. Irrespective of Mn treatment, the values of Φ_PS2 and q_N decreased, while those of q_P increased progressively by increasing irradiance from I₁₃₆ to I₃₁₁₁. Finally, Mn₂-Mn₉₈ plants were less sensitive to photoinhibition of photosynthesis (≥811 μmol m⁻² s⁻¹) than the Mn₆₈₆ (≥536 μmol m⁻² s⁻¹) and Mn₀ (≥336 μmol m⁻² s⁻¹) ones.     

抑制AtCLH1表达降低叶片衰老进程中叶绿素a与b的比值但不影响叶绿素降解速率

为了分析叶绿素酶在叶绿素降解过程中的作用,构建了抑制AfCLH1和／或AtCLH2的拟南芥RNM株系。在RNAi株系中,叶绿素酶的表达和活性都被显著抑制。然而,无论是在正常生长情况下还是在黑暗诱导的衰老进程中,RN加株系中的叶绿素降解速率与野生型中的相比均无显著差异。进一步的分析结果表明,AtCLH1 RNAi株系中的叶绿素a/b比值在叶片衰老进程中逐渐降低,而野生型中的却逐渐升高。另外,还观察到所有株系中叶绿素酶的活性在黑暗诱导的衰老过程的初期均呈现出上升的趋势。与这一变化趋势正好相反,在所有株系中,叶绿素酶基因的表达在黑暗诱导衰老后就开始大幅下降。有趣的是,与野生型相比,RNAi株系中的目标叶绿素酶基因仍然维持在一个较低的表达水平。这说明拟南芥中可能存在一个反馈补偿机制。基于这些结果,并结合相关的研究报道,认为叶绿素酶很可能参与了衰老进程早期的叶绿素b向a的转换过程。     

Photochemical Properties of Photosystem 2 in Primary Leaves of Barley Seedlings Grown Under Various Blue or Red Irradiances

Photosystem 2 (PS2)-driven electron transfer was studied in primary leaves of barley (Hordeum vulgare L.) seedlings grown under various photon fluxes (0.3–170.0 mol m^–2 s^–1) of blue (BR) or red (RR) radiation using modulated chlorophyll fluorescence. The F_v/F_m ratio was 0.78–0.79 in leaves of all radiation variants, except in seedlings grown under BR or RR of 0.3 mol m^–2 s^–1. The extent of the photochemical phase of the polyphasic F_v rise induced by very strong white light was similar in leaves of all radiation treatments. Neither radiation quality nor photon flux under plant cultivation influenced the amount of non Q_B-transferring centres of PS2 except in leaves of seedlings grown under BR of 0.3 mol m^–2 s^–1, in which the amount of such centres increased threefold. Both BR and RR stimulated the development of photochemically competent PS2 at photon fluxes as low as 3 mol m^–2 s^–1. Three exponential components with highly different half times were distinguished in the kinetics of F_v dark decay. This indicates different pathways of electron transfer from Q_A ^–, the reduced primary acceptor of PS2, to other acceptors. Relative magnitudes of the individual decay components did not depend on the radiation quality or the photon flux during plant cultivation. Significant differences were found, however, between plants grown under BR or RR in the rate of the middle and fast components of F_v dark decay, which showed 1.5-times faster intersystem linear electron transport in BR-grown leaves.     

Photoinhibition of Photosynthesis in Vitis berlandieri and Vitis rupestris Leaves under Field Conditions

Photoinhibition of photosynthesis was investigated in Vitis berlandieri and Vitis rupestris leaves under field conditions at different sampling time in a day. The degree of photoinhibition was determined by means of the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m) and photosynthetic electron transport measurements. When the photochemical efficiency of PS2, F_v/F_m, markedly declined, F₀ increased significantly in leaves of V. berlandieri, while F₀ did not increase in V. rupestris leaves. Isolated thylakoids of leaves of V. berlandieri showed significant inhibition of whole chain and PS2 activities at midday. A smaller inhibition was observed for V. rupestris. Later, the leaves reached maximum PS2 efficiencies similar to those observed early in the morning during sampling at evening. The artificial exogenous electron donor Mn²⁺ failed to restore PS2 activity in both species, while DPC and NH₂OH significantly restored PS2 activity in V. rupestris midday leaf samples. Quantification of the PS2 reaction centre protein D1 and 33 kDa protein of water splitting complex following midday exposure of leaves showed pronounced differences between V. berlandieri and V. rupestris leaves. The marked loss of PS2 activity noticed in midday samples was mainly due to the marked loss of D1 protein in V. berlandieri while in V. rupestris it was the 33 kDa protein.     

Protective Action of Abscisic Acid Against the Inhibition of Photosynthesis of Barley Leaves by Bisulphite

The inhibition of photosynthetic activity by bisulphite was studied in intact leaves of abscisic acid (ABA)-treated and non-treated (control) barley plants. ABA inhibited the photosynthetic process as evidenced by lower values of chlorophyll fluorescence kinetic parameters F_v/F_m (photosystem 2 activity) and R_fd (vitality index, related to the whole photosynthetic activity) compared with ABA-non-treated plants. After bisulphite treatment, the extent of inhibition was smaller in ABA-treated plants than in the control ones indicating a protective effect of ABA. The protective action sites of ABA were the Q_A reduction and the Calvin cycle. This revised version was published online in September 2006 with corrections to the Cover Date.     

不同氮素浓度下CO2浓度、温度对蒙古栎(Quercus mongolica)幼苗叶绿素含量的影响

叶片中叶绿素含量在光合作用中的光吸收、传递和转换过程中起到重要作用。为了预测未来大气CO2浓度升高并伴随温度上升的情况下,植物在不同的氮素营养水平下光合能力的变化,做了在3种氮素水平下(15 mmol.L-1N,7.5 mmol.L-1N和不施氮)CO2倍增和温度升高4℃对蒙古栎一年生幼苗叶片中叶绿素含量的影响的实验。结果表明:氮素水平对叶绿素含量影响显著,在CO2倍增(700μmol.mol-1)、高温( 4℃)和正常温度、大气CO2浓度条件下,高氮素水平下的叶绿素含量明显高于正常氮素水平和不施氮;CO2浓度和温度对叶绿素含量的影响受到氮素的制约:在高氮的条件下CO2浓度倍增促进叶绿素a、b的合成,而且对叶绿素b合成的促进尤为显著;而温度升高4℃能够促进叶绿素a的合成,但是对叶绿素b含量的影响不显著。在正常氮素条件下叶绿素a、叶绿素b及总量各个处理间的差异均不显著;在不施氮的条件下,CO2倍增和升高适当的温度在一定的程度上可以促进叶绿素a的合成,不能同时保证叶绿素b的合成。CO2浓度升高明显导致蒙古栎幼苗对氮素水平的需求也增加,高温条件下的蒙古栎幼苗也在一定程度上增加了对氮素的需求。     

Conversion of chlorophyll b to chlorophyll a precedes magnesium dechelation for protection against necrosis in Arabidopsis

Chlorophyll is a deleterious molecule that generates reactive oxygen species and must be converted to non‐toxic molecules during plant senescence. The degradation pathway of chlorophyll a has been determined; however, that of chlorophyll b is poorly understood, and multiple pathways of chlorophyll b degradation have been proposed. In this study, we found that chlorophyll b is degraded by a single pathway, and elucidated the importance of this pathway in avoiding cell death. In order to determine the chlorophyll degradation pathway, we first examined the substrate specificity of 7‐hydroxymethyl chlorophyll a reductase. 7‐hydroxymethyl chlorophyll a reductase reduces 7‐hydroxymethyl chlorophyll a but not 7‐hydroxymethyl pheophytin a or 7‐hydroxymethyl pheophorbide a. These results indicate that the first step of chlorophyll b degradation is its conversion to 7‐hydroxymethyl chlorophyll a by chlorophyll b reductase, although chlorophyll b reductase has broad substrate specificity. In vitro experiments showed that chlorophyll b reductase converted all of the chlorophyll b in the light‐harvesting chlorophyll a/b protein complex to 7‐hydroxymethyl chlorophyll a, but did not completely convert chlorophyll b in the core antenna complexes. When plants whose core antennae contained chlorophyll b were incubated in the dark, chlorophyll b was not properly degraded, and the accumulation of 7‐hydroxymethyl pheophorbide a and pheophorbide b resulted in cell death. This result indicates that chlorophyll b is not properly degraded when it exists in core antenna complexes. Based on these results, we discuss the importance of the proper degradation of chlorophyll b.     

两种生境条件下6种牧草叶绿素含量及荧光参数的比较

昆仑山前山牧场海拔较高, 策勒绿洲海拔相对较低, 两者生境差异较大。以昆仑山前山牧场和策勒绿洲边缘两种不同生境条件下生长的6种牧草: 冰草(Agropyron cristatum)、无芒雀麦(Bromus inermis)、矮生高羊茅(Festuca elata)、披碱草(Elymus dahuricus )、红豆草(Onobrychis pulchella)及和田大叶(Medicago sativa var. luxurians)为试验材料, 研究了不同生境条件下牧草叶片叶绿素含量及叶绿素荧光动力学参数的变化情况。结果显示: (1)在两种生境条件下, 昆仑山前山牧场生境生长的牧草叶绿素a、叶绿素b、总叶绿素的含量明显较高, 生长在策勒绿洲生境的牧草品种叶绿素a/b值较高; (2)昆仑山前山牧场生境牧草最大荧光、光系统II (PSII)最大光化学效率、PSII潜在活性和单位面积反应中心的数量的值明显高于策勒绿洲生境品种, 而初始荧光、单位反应中心吸收的光能、单位反应中心捕获的能量、单位反应中心耗散的能量、荧光诱导曲线初始斜率值则低于策勒绿洲生境品种。因此, 两种生境下环境因子发生了改变, 对牧草产生综合的胁迫作用; 策勒绿洲生境明显对牧草生长产生了抑制, 策勒绿洲生境牧草的色素含量降低以及PSII的机构遭到损坏, 导致反应中心一部分失活或裂解, 剩余有活性的反应中心的效率增加, 昆仑山生境则相对比较适宜牧草生长; 两种生境不同牧草叶绿素含量和叶绿素荧光参数的变化幅度不同。     

Photosynthetic characteristics of five rice cultivars grown under increased solar UV-B radiation

The impact of increased solar UV-B radiation on photosynthetic characteristics of rice (Oryza sativa L.) cultivars ADT36, IR20, IR50, J13 and MDU4 has been studied. In all the cultivars concentrations of photosynthetic pigments decreased under increased UV-B radiation. Even low enhancement of UV-B reduced the photochemical activities in all the cultivars except MDU4 and changed chlorophyll a fluorescence. Enhanced UV-B radiation caused a dose-dependent changes in chloroplast proteins in most of the cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Differential Responses of Growth and Photosynthesis in Cyamopsis Tetragonoloba L. Grown under Ultraviolet-B and supplemental long-wavelength radiations

Cyamopsis tetragonoloba L. seedlings were subjected to continuous ultraviolet (UV)-B radiation for 18 h and post-irradiated with "white light" (WL) and UV-A enhanced fluorescent radiations. UV-B treatment alone reduced plant growth, pigment content, and photosynthetic activities. Supplementation of UV-A promoted the overall seedling growth and enhanced the synthesis of chlorophyll and carotenoids with a relatively high photosystem 1 activity. Post UV-B irradiation under WL failed to photoreactive the UV-B damage whereas a positive photoregulatory effect of UV-A was noticed in electron transport rates and low temperature fluorescence emission spectra.     

强光和短期高浓度CO2对玉米和大豆光能转化效率的影响

盆栽和人工光源条件下,玉米叶片在普通空气中对强光照射不敏感,在高浓度CO2中强光照射0～5h后,光合速率(Pn)逐渐降低,无机磷(Pi)限制是其主要原因之一;大豆叶片在普通空气中受强光照射5h后,Fv／Fm、Pn、羧化效率(CE)和表观量子效率(AOY)明显降低,Fo升高,在高浓度CO2和强光下大豆Fo上升、Fv／Fm和气体交换参数下降的幅度减小。研究表明,高浓度CO2可减轻强光对植物尤其是C3植物光合功能的损伤,有限地缓解光抑制,但不能完全消除强光导致的大豆Pn和气孔导度(Gs)的降低。