光合电子流对光响应的机理模型及其应用

光合电子流对光响应的机理可以揭示植物光合电子流与光强、植物捕光色素分子物理特性之间的关系。该文讨论了光合电子流对光响应的机理模型的特性以及捕光色素分子的物理性质, 并利用此模型拟合了山莴苣(Lagedium sibiricum)、一年蓬(Erigeron annuus)和紫菀(Aster tataricus)的光合电子流对光响应的曲线。由此模型不仅可以得到植物的最大光合电子流、饱和光强、初始斜率等参数, 还可以获得捕光色素分子有效光能吸收截面和处于最低激发态的捕光色素分子数对光的响应关系。结果表明: 随光强的增加, 山莴苣的捕光色素分子的有效光能吸收截面下降最快, 紫苑的下降速度最慢; 山莴苣处于最低激发态的捕光色素分子数增长速度最快, 紫苑的增长速度最小。捕光色素分子的有效光能吸收截面随光强增加而下降、处于最低激发态的捕光色素分子数随光强增加而增加的特性将减少其光能的吸收和激子的传递, 因而有利于减少强光对植物产生的光伤害。     

A mechanistic model for the light response of photosynthetic electron transport rate based on light harvesting properties of photosynthetic pigment molecules

Models describing the light response of photosynthetic electron transport rate (ETR) are routinely used to determine how light absorption influences energy, reducing power and yields of primary productivity; however, no single model is currently able to provide insight into the fundamental processes that implicitly govern the variability of light absorption. Here we present development and application of a new mechanistic model of ETR for photosystem II based on the light harvesting (absorption and transfer to the core ‘reaction centres’) characteristics of photosynthetic pigment molecules. Within this model a series of equations are used to describe novel biophysical and biochemical characteristics of photosynthetic pigment molecules and in turn light harvesting; specifically, the eigen-absorption cross-section and the minimum average lifetime of photosynthetic pigment molecules in the excited state, which describe the ability of light absorption of photosynthetic pigment molecules and retention time of excitons in the excited state but are difficult to be measured directly. We applied this model to a series of previously collected fluorescence data and demonstrated that our model described well the light response curves of ETR, regardless of whether dynamic down-regulation of PSII occurs, for a range of photosynthetic organisms (Abies alba, Picea abies, Pinus mugo and Emiliania huxleyi). Inherent estimated parameters (e.g. maximum ETR and the saturation irradiance) by our model are in very close agreement with the measured data. Overall, our mechanistic model potentially provides novel insights into the regulation of ETR by light harvesting properties as well as dynamical down-regulation of PSII.     

不同CO2浓度下番茄幼苗叶片的光能利用效率

光能利用效率(LUE)是评价植物叶片光能利用能力的重要参数,更是影响生态系统生产力大小和质量的主要因素.本文基于植物光合作用对光响应的机理模型推导出植物叶片的光能利用效率模型以及叶片的最大光能利用效率(LUE_max)和对应的饱和光强(I_L-sat)的数学表达式,并用光能利用效率模型研究了番茄幼苗叶片在CO₂浓度分别为350、450、550和650 μmol·mol^-1下的光能利用效率.拟合结果表明: 所推导的叶片光能利用效率模型可以很好地描述4种CO₂浓度条件下番茄幼苗叶片的光能利用效率;在这4种CO₂浓度条件下,番茄的LUE_max在光合有效辐射(I)为70～90 μmol·m^-2·s^-1时就可以达到;番茄在CO₂浓度为550和650 μmol·mol^-1时叶片的LUE_max和I_L-sat没有差异.产生这种现象的原因可能是由于番茄幼苗长时间处于较低光强下,番茄幼苗叶片的光合功能已经适应了低光强环境,以致于高CO₂浓度难以改变它的捕光色素分子的内秉特性,如有效光能吸收截面以及处于激发态和基态色素的比例等.     

不同CO2浓度下番茄幼苗叶片的光能利用效率

光能利用效率(LUE)是评价植物叶片光能利用能力的重要参数,更是影响生态系统生产力大小和质量的主要因素.本文基于植物光合作用对光响应的机理模型推导出植物叶片的光能利用效率模型以及叶片的最大光能利用效率(LUE_max)和对应的饱和光强(I_L-sat)的数学表达式,并用光能利用效率模型研究了番茄幼苗叶片在CO₂浓度分别为350、450、550和650 μmol·mol^-1下的光能利用效率.拟合结果表明: 所推导的叶片光能利用效率模型可以很好地描述4种CO₂浓度条件下番茄幼苗叶片的光能利用效率;在这4种CO₂浓度条件下,番茄的LUE_max在光合有效辐射(I)为70～90 μmol·m^-2·s^-1时就可以达到;番茄在CO₂浓度为550和650 μmol·mol^-1时叶片的LUE_max和I_L-sat没有差异.产生这种现象的原因可能是由于番茄幼苗长时间处于较低光强下,番茄幼苗叶片的光合功能已经适应了低光强环境,以致于高CO₂浓度难以改变它的捕光色素分子的内秉特性,如有效光能吸收截面以及处于激发态和基态色素的比例等.     

光合作用对光和CO2响应模型的研究进展

光合作用对光和CO₂响应模型是研究植物生理和植物生态学的重要工具, 可为植物光合特性对主要环境因子的响应提供科学依据。该文综述了当前光合作用对光和CO₂响应模型的研究进展和存在的问题, 并在此基础上探讨了这些模型的可能发展趋势。光合作用涉及光能的吸收、能量转换、电子传递、ATP合成、CO₂固定等一系列复杂的物理和化学反应过程。光合作用由原初反应、同化力形成和碳同化3个基本过程构成, 任一个过程均可对光合作用速率产生直接的影响。光合作用对光响应模型只涉及光能的转换, 而光合作用的生化模型包含了同化力形成和碳同化这两个基本过程。把光合作用的原初反应, 即把参与光能吸收、传递和转换的捕光色素分子的物理参数(如捕光色素分子数、捕光色素分子光能吸收截面、捕光色素分子处于激发态的平均寿命等)结合到生化模型中, 可能是今后光合作用对光响应机理模型的发展方向。     

Fluorescence decay kinetics of chlorophyll in photosynthetic membranes

The absorption of light by the pigments of photosynthetic organisms results in electronic excitation that provides the energy to drive the energy-storing light reactions. A small fraction of this excitation gives rise to fluorescence emission, which serves as a sensitive probe of the energetics and kinetics of the excited states. The wavelength dependence of the excitation and emission spectra can be used to characterize the nature of the absorbing and fluorescing molecules and to monitor the process of sensitization of the excitation transfer from one pigment to another. This excitation transfer process can also be followed by the progressive depolarization of the emitted radiation. Using time-resolved fluorescence rise and decay kinetics, measurements of these processes can now be characterized to as short as a few picoseconds. Typically, excitation transfer among the antenna or light harvesting pigments occurs within 100 psec, whereupon the excitation has reached a photosynthetic reaction center capable of initiating electron transport. When this trap is functional and capable of charge separation, the fluorescence intensity is quenched and only rapidly decaying kinetic components resulting from the loss of excitation in transit in the antenna pigment bed are observed. When the reaction centers are blocked or saturated by high light intensities, the photochemical quenching is relieved, the fluorescence intensity rises severalfold, and an additional slower decay component appears and eventually dominates the decay kinetics. This slower (1-2 nsec) decay results from initial charge separation followed by recombination in the blocked reaction centers and repopulation of the excited electronic state, leading to a rapid delayed fluorescence component that is the origin of variable fluorescence. Recent growth in the literature in this area is reviewed here, with an emphasis on new information obtained on excitation transfer, trapping, and communication between different portions of the photosynthetic membranes.     

越冬期广玉兰阳生叶和阴生叶PSⅡ功能及捕光色素分子内禀特性的比较研究

捕光色素分子的内禀特性不仅决定了光能的吸收与传递,也将影响到激发能向光化学反应、热耗散和叶绿素荧光的分配。本文采用叶绿素荧光技术和光合电子流对光响应机理模型,研究了越冬期广玉兰（Magnolia grandiflora）阳生叶和阴生叶两种不同光环境下叶片PSⅡ功能及其捕光色素分子内禀特性的差异,以探索广玉兰越冬的光保护策略。结果表明：越冬期低温导致叶片轻微光抑制的发生,全光照加剧了阳生叶光抑制程度,而弱光环境有利于阴生叶光抑制的恢复。阳生叶可通过降低叶绿素含量和捕光色素分子数量以减少对光能的吸收,并且具有较强的光化学和热耗散能力以保护光合机构免受低温强光伤害。而阴生叶虽然其光化学反应能力相对较弱,但具有较强的热耗散能力,可有效地保护其免受短时曝露在强光下的伤害。     

Photosynthetic and physiological responses of foxtail millet (<Emphasis Type="Italic">Setaria italica</Emphasis> L.) to low-light stress during grain-filling stage

Two foxtail millet (Setaria italica L.) varieties were subjected to different shading intensity treatments during a grain-filling stage in a field experiment in order to clarify physiological mechanisms of low-light effects on the yield. Our results showed that the grain fresh mass per panicle, yield, photosynthetic pigment contents, net photosynthetic rate, stomatal conductance, effective quantum yield of PSII photochemistry, and electron transport rate decreased with the increase of shading intensity, whereas the intercellular CO₂ concentration increased in both varieties. In addition, shading changed a double-peak diurnal variation of photosynthesis to a one-peak curve. In conclusion, the lower yield of foxtail millet was caused mainly by a reduction of grain mass assimilated, a decline in chlorophyll content, and the low photosynthetic rate due to low light during the grain-filling stage. Reduced light energy absorption and conversion, restricted electron transfer, and reduced stomatal conductance might cause the decrease in photosynthesis.     

Photophysics of the carotenoids associated with the xanthophyll cycle in photosynthesis

Green plants use the xanthophyll cycle to regulate the flow of energy to chlorophylla within photosynthetic proteins. Under conditions of low light intensity violaxanthin, a carotenoid possessing nine conjugated double bonds, functions as an antenna pigment by transferring energy from its lowest excited singlet state to that of chlorophylla within light-harvesting proteins. When the light intensity increases, violaxanthin is biochemically transformed into zeaxanthin, a carotenoid that possesses eleven conjugated double bonds. The results presented here show that extension of the conjugation of the polyene lowers the energy of the lowest excited singlet state of the carotenoid below that of chlorophylla. As a consequence zeaxanthin can act as a trap for the excess excitation energy on chlorophylla pigments within the protein, thus regulating the flow of energy within photosynthetic light-harvesting proteins.     

Interaction of carbon nanotubes with curcumin: Effect of temperature and pH on simultaneous static and dynamic fluorescence quenching of curcumin using carbon nanotubes

Curcumin (Cur) has medicinal properties, undergoes hydrolysis, and has low water solubility that limits its bioavailability and industrial usage. Different host molecules such as carbon nanotubes (CNT) can be useful in improving solubility and stabilizing Cur, therefore understanding the interaction of Cur with host molecules such as CNT is crucial. In this study, UV–visible light absorption and fluorescence spectroscopic techniques have been applied to reveal the interaction of Cur with CNT. Visible light absorption of Cur increases with CNT concentration, whereas fluorescence intensity of Cur is quenched in the presence of CNT. The obtained results confirm that fluorescence reduction is due to both static and dynamic quenching and is a result of the ground state and excited‐state complex formation. The pH environment influences the quenching rate due to deprotonation of Cur at higher pH; excess OH‐ ion concentration in the solution further discourages electrostatic interaction between the deprotonated form of Cur with CNT. It is found that at lower temperatures (<35°C) dynamic quenching is much more dominant and at higher temperatures (45°C) static quenching is more dominant. The interaction is further supported using X‐ray diffraction patterns and Fourier transform infrared spectra in the solid state, and suggests encapsulation of curcumin within the CNT. It is further evident that fluorescence quenching of Cur using CNT is further enhanced in the presence of several salts, as increase in ionic strength of the solution pushes the hydrophobic Cur molecule towards the CNT core by increasing the proximity between them.     

Entropy consumption in primary photosynthesis

Knox and Parson have objected to our previous conclusion on possible negative entropy production during primary photochemistry, i.e., from photon absorption to primary charge separation, by considering a pigment system in which primary photochemistry is not specifically considered. This approach does not address our proposal. They suggest that when a pigment absorbs light and passes to an excited state, its entropy increases by hν/T. This point is discussed in two ways: (i) from considerations based on the energy gap law for excited state relaxation; (ii) using classical thermodynamics, in which free energy is introduced into the pigment (antenna) system by photon absorption. Both approaches lead us to conclude that the excited state and the ground state are isoentropic, in disagreement with Knox and Parson. A discussion on total entropy changes specifically during the charge separation process itself indicates that this process may be almost isoentropic and thus our conclusions on possible negentropy production associated with the sequence of reactions which go from light absorption to the first primary charge separation event, due to its very high thermodynamic efficiency, remain unchanged.     

Entropy consumption in primary photosynthesis

Knox and Parson have objected to our previous conclusion on possible negative entropy production during primary photochemistry, i.e., from photon absorption to primary charge separation, by considering a pigment system in which primary photochemistry is not specifically considered. This approach does not address our proposal. They suggest that when a pigment absorbs light and passes to an excited state, its entropy increases by hnu/T. This point is discussed in two ways: (i) from considerations based on the energy gap law for excited state relaxation; (ii) using classical thermodynamics, in which free energy is introduced into the pigment (antenna) system by photon absorption. Both approaches lead us to conclude that the excited state and the ground state are isoentropic, in disagreement with Knox and Parson. A discussion on total entropy changes specifically during the charge separation process itself indicates that this process may be almost isoentropic and thus our conclusions on possible negentropy production associated with the sequence of reactions which go from light absorption to the first primary charge separation event, due to its very high thermodynamic efficiency, remain unchanged.     

Molecular modeling of two-photon absorption and third-order nonlinearities of polymethine dyes for all-optical switching

Stimulated by a recent experimental report [Hales JM et al. (2010) Science 327:1485-1488], two-photon absorption and third-order optical nonlinearities of selenopyrylium- and bis(dioxaborine)-terminated polymethine dyes (called SE-7C and DOB-9C) used for all-optical switching were investigated theoretically with time-dependent DFT (TD-DFT) and response theory as well as visualized real-space analysis. The calculated results for the first hyperpolarizability and second hyperpolarizability demonstrated that the two molecules both have large third-order optical nonlinearities. Using real-space analysis, we were able to visually determine that in the one-photon absorption (OPA) process, the first singlet excited state of SE-7C and DOB-9C is an intramolecular charge transfer (ICT) excited state with strong absorption, while the second excited state of these dyes (also termed the "ICT state") shows weak absorption. However, in the two-photon absorption (TPA) process, a larger TPA absorption cross-section was predicted for the second excited state. In this paper, we describe the properties of the S2 excited state, incorporating charge transfer and the transition moment, via real-space analysis, which was very important for understanding the TPA characteristics of the S(2) state.     

一个番茄EMS叶色黄化突变体的叶绿素含量及光合作用

本试验以测序番茄品种‘Heinz 1706’的甲基磺酸乙酯(EMS)诱变所获得的叶色黄化突变体(Y55)为试材,分析了突变体的植株生长、叶片叶绿素含量及光合参数.结果表明: Y55的株高、茎粗、鲜质量均显著降低;叶绿素 a、叶绿素 b 、类胡萝卜素、总叶绿素含量以及叶绿素 a/b均显著降低,Y55叶绿素合成的前体物质含量均显著低于野生型,尤其是粪卟啉原Ⅲ及其后前体物质;且Y55叶片净光合速率、蒸腾速率、胞间CO₂浓度、气孔导度均显著低于野生型,最大光合速率、CO₂饱和点与补偿点、光饱和点与补偿点也显著下降;Y55的PSII最大量子效率显著降低,F_o显著升高,PSII与PSI的光合电子产量和电子传递速率显著降低;Y55处于基态的捕光色素分子、捕光色素分子处于最低激发态的平均寿命均显著降低.表明粪卟啉原Ⅲ的合成受阻可能是黄化突变体Y55叶绿素含量下降的主要原因,黄化突变降低了叶片捕光色素分子数量,影响了叶片的光合作用,进而抑制了植株的生长发育.     

生长调节剂对黄连木光合生理指标和荧光参数的影响

以黄连木幼苗为试材,采用叶片喷施的方法,研究了生长调节剂多效唑和矮壮素对黄连木叶片解剖结构、色素含量、光合生理指标和叶绿素荧光参数的影响,以揭示生长调节剂对黄连木光合特性的影响及其内在机制。结果显示:随着两种生长调节剂处理浓度的增加,黄连木幼苗叶片厚度、栅栏组织厚度、海绵组织厚度、色素含量、净光合速率(Pn)、PSⅡ原初光能转化效率(Fv/Fm)、PSⅡ的潜在活性(Fv/F0)、叶片单位面积吸收的能量(ABS/CS)和捕获的能量(TRo/CS)以及电子传递的能量(ETo/CS)均呈先增大后减小的趋势,并均在800mg/L多效唑和500mg/L矮壮素处理时达到最大值,且显著提高了黄连木叶片的净光合速率。研究发现,黄连木幼苗叶片栅/海比例增大和叶绿素含量的增加使叶片对光能的捕获和吸收能力提高,而PSⅡ反应中心开放比例的增大与其活性的增强保证了叶片对光能的高效传递,同时降低了能量的热耗散,是提高苗木光合速率和保证苗木光合作用高效进行的内在机制。     

Assessment of a microscopic photobleaching technique for measuring the spectral absorption efficiency of individual phytoplankton cells

Measurements of the absorptance of photosynthetic pigments inindividual phytoplankton cells were made using an epifluorescencemicroscope equipped with a spectrograph and CCD array detector.Correction for light loss due to scattering was achieved bybleaching the cells with intense light from a mercury arc lamp,and using the bleached cells as a spectrophotometric blank.Absorption efficiency factors were calculated from knowledgeof the geometrical cross-section of the cells obtained fromcalibratedvideo images acquired at the time of measurement. The single-cellefficiency factors were consistent with the average absorptionefficiencies of cell suspensions measured using a spectrophotometerover most of the visible spectrum, but they were significantlylower below 420 nm. Cells of the diatom Cyclotella crypticaand the chlorophyte Chlorella salina showed clear spectral differencesin spectral shape that could be related to taxonomic differencesin pigment content, but absorption efficiency factors of approximately0.4 at 675 nm were found for both species.     

Photo-bleaching and photon saturation in flow cytometry.

In flow cytometry, small particles travel at a high speed through a bright light spot. The high light intensity at the point of measurement causes measurable photon saturation. This observation indicates that the rate at which individual dye molecules emit photons is close to the maximum emission rate. Despite the short exposure time, individual molecules may go through a few hundred excitation cycles while they are in the light beam. The absorbed light dose causes significant dye destruction. This article presents experimental procedures to determine the extent of photon saturation and photo-bleaching of dyes bound to cell nuclei in a flow cytometer. Measurements of Hoechst and propidium iodide bound to chromatin show that the amount of dye bleached per emitted photon is the same at low and high illumination intensities. This finding indicates that photon emission and dye destruction are both the result of the absorption of single excitation photons. The experimental observations allow rough estimates of the lifetime of the excited state and the lifetime of the molecule. The lifetime of the Hoechst 33258 bound to DNA is estimated to be 100 excitation-relaxation cycles. The average propidium iodide molecule lasts approximately 200 excitation-relaxation cycles. The theoretical considerations show that the optimal illumination conditions are different for bleaching and nonbleaching dyes. An optical arrangement for high precision measurements of bleaching dyes is presented.     

Regularities of formation of chlorophyll-human serum albumin functionally active complexes in the aqueous medium]

In the system with constant content of the chlorophyll a and increasing amounts of human serum albumin, dependence of pigment incorporation into the complex upon interaction of its aqueous associates with protein solutions was studied by applying the gel filtration on Sephadex G-75 and by measuring light scattering and rate of sensitized photoreduction of the methyl red by ascorbic-acid. The curves were obtained after extraction of the chlorophyll by acetone from dry pigment-protein films formed after desiccation of the aqueous systems. Sigmoid character of the above dependences, their linearization in Hill's coordinates and the value of cooperativity coefficient close to 2 testifies in favour of the cooperative character of the complex formation, two pigment molecules reacting with a single protein molecule. Measurement of adsorption isotherms and their treatment with use of the Brunauer-Emmett-Teller theory of polymolecular adsorption make it possible to evaluate the maximum molar ratio of the pigment to the protein in the complex (close to 2). The pigment-pigment interaction suggests that the chlorophyll molecules adsorbed on the protein are in the state of loosely packed dimers. Deaggregation of aqueus pigment associates by the protein in the course of complex formation results in a considerable increase of the protosensitizing chlorophyll activity.     

UV-B辐射增强对芦苇光合生理及叶绿体超微结构的影响

在UV-B辐照增强条件下,研究不同辐照梯度对芦苇光合特性、光合色素含量及叶绿体超微结构的影响. 结果表明: 与自然光照相比,UV-B辐照增强显著降低芦苇叶片净光合速率、气孔导度、蒸腾速率,且随辐照强度增大,降低程度加剧,胞间CO₂浓度升高,光合效率显著降低;与自然光照相比,UV-B辐照增强显著降低芦苇叶片光合色素含量(包括叶绿素a、叶绿素b、总叶绿素和类胡萝卜素),且随辐照强度增大,降低程度加剧;UV-B辐射增强条件下,叶绿体超微结构遭到破坏,表现为叶绿体结构变形,类囊体片层排列稀疏紊乱、膨胀甚至模糊不清,并且UV-B辐射强度越大,损伤越大,高强度UV-B辐射对叶绿体超微结构的影响大于低强度辐射.     

光系统II实际光化学量子效率对光的响应模型的比较

为了比较光系统II实际光化学量子效率(Φ_PSII)对光的响应机理模型(简称机理模型)、负指数模型和指数模型的优缺点, 用LI-6400-40B光合作用测定仪控制CO₂浓度和温度, 测量了剑叶金鸡菊(Coreopsis lanceolata)、黄荆(Vitex negundo)和大狼杷草(Bidens frondosa)的电子传递速率(ETR)对光的响应曲线(ETR-I)和Φ_PSII对光的响应曲线(Φ_PSII-I), 然后用这3个模型分别拟合了这些数据。拟合结果表明: 3个模型都可以较好地拟合这3种植物的ETR-I的响应数据和Φ_PSII-I的响应数据, 但由指数模型拟合ETR-I和Φ_PSII-I的响应数据得到相应的饱和光强(PAR_sat)和光系统II最大光能利用效率(F_v/F_m)之间存在显著差异, 且估算的饱和光强远低于实测值。由机理模型可知, Φ_PSII不仅与光强的函数有关, 还与植物的内禀特性有关, 即与天线色素分子的本征光能吸收截面、激子的传递效率、能级的简并度、光化学反应常数、热耗散常数和处于最低激发态的平均寿命等参数有关。此外, 由机理模型还可知, Φ_PSII随光强的增加而下降的原因是捕光色素分子的有效光能吸收截面随光强增加而降低。