A hierarchical Bayesian approach for estimation of photosynthetic parameters of C3 plants

We describe a hierarchical Bayesian (HB) approach to fitting the Farquhar et al. model of photosynthesis to leaf gas exchange data. We illustrate the utility of this approach for estimating photosynthetic parameters using data from desert shrubs. Unique to the HB method is its ability to simultaneously estimate plant- and species-level parameters, adjust for peaked or non-peaked temperature dependence of parameters, explicitly estimate the 'critical' intracellular [CO₂] marking the transition between ribulose 1·5-bisphosphate carboxylase/oxygenase (Rubisco) and ribulose-1,5-bisphosphate (RuBP) limitations, and use both light response and CO₂ response curve data to better inform parameter estimates. The model successfully predicted observed photosynthesis and yielded estimates of photosynthetic parameters and their uncertainty. The model with peaked temperature responses fit the data best, and inclusion of light response data improved estimates for day respiration ( R _d). Species differed in R _d25 ( R _d at 25 °C), maximum rate of electron transport ( J _max25), a Michaelis–Menten constant ( K _c25) and a temperature dependence parameter (Δ S ). Such differences could potentially reflect differential physiological adaptations to environmental variation. Plants differed in R _d25, J _max25, mesophyll conductance ( g _m25) and maximum rate of Rubisco carboxylation ( V _cmax25). These results suggest that plant- and species-level variation should be accounted for when applying the Farquhar et al. model in an inferential or predictive framework.     

Growth in elevated CO2 enhances temperature response of photosynthesis in wheat

The temperature dependence of C₃ photosynthesis may be altered by the growth environment. The effects of long-term growth in elevated CO₂ on photosynthesis temperature response have been investigated in wheat ( Triticum aestivum L.) grown in controlled chambers with 370 or 700 μmol mol⁻¹ CO₂ from sowing through to anthesis. Gas exchange was measured in flag leaves at ear emergence, and the parameters of a biochemical photosynthesis model were determined along with their temperature responses. Elevated CO₂ slightly decreased the CO₂ compensation point and increased the rate of respiration in the light and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) V_cmax, although the latter effect was reversed at 15°C. With elevated CO₂, J_max decreased in the 15–25°C temperature range and increased at 30 and 35°C. The temperature response (activation energy) of V_cmax and J_max increased with growth in elevated CO₂. CO₂ enrichment decreased the ribulose 1,5-bisphosphate (RuBP)-limited photosynthesis rates at lower temperatures and increased Rubisco- and RuBP-limited rates at higher temperatures. The results show that the photosynthesis temperature response is enhanced by growth in elevated CO₂. We conclude that if temperature acclimation and factors such as nutrients or water availability do not modify or negate this enhancement, the effects of future increases in air CO₂ on photosynthetic electron transport and Rubisco kinetics may improve the photosynthetic response of wheat to global warming.     

基于FvCB模型的几种草本和木本植物光合生理生化特性

为研究不同生活型植物的光合能力及其叶片光合机构,采用直角双曲线修正模型和C3植物FvCB模型对7种木本植物和4种草本植物的CO₂响应曲线进行拟合,并对不同木本植物、不同草本植物和2种生活型植物的最大净光合速率(P_{n max})、Rubisco酶最大羧化速率(V_{c max})、最大电子传递速率(J_max)、光合暗呼吸速率(R_d)和叶肉阻力(r_m)等参数进行比较.结果表明: 7种木本植物P_{n max}大小顺序为乌桕、苎麻>润楠、海桐>青冈、苦槠、娜塔栎;乌桕、苎麻、润楠和海桐的V_{c max}显著大于青冈和苦槠;J_max大小顺序为乌桕>苎麻、海桐>娜塔栎、苦槠和青冈;润楠和苦槠的r_m显著大于乌桕、海桐和苎麻.商陆的P_{n max}显著大于藿香蓟和土牛膝;4种草本植物的V_{c max}无显著差异;商陆的J_max显著大于藿香蓟;龙葵和土牛膝的r_m显著大于藿香蓟;商陆的R_d显著大于藿香蓟和土牛膝.木本植物的P_{n max}、V_{c max}、J_max和r_m光合参数均显著大于草本植物,但二者的R_d无显著差异.不同物种之间以及2种生活型植物光合能力的差异主要是由叶片内部Rubisco酶羧化能力、电子传递能力和叶肉阻力等差异引起的.     

大气CO2浓度升高对干旱条件下冬小麦叶片光合适应的影响

大气CO₂浓度升高是全球气候变化的主要特征,但大气CO₂浓度长期升高条件下冬小麦叶片发生光合适应的机制尚不十分清楚。本研究以盆栽冬小麦‘郑麦9023’为试验材料,在人工气候控制室内设置2个CO₂浓度(400和600 μmol·mol^-1)、2个水分条件(田间持水量的80%±5%和55%±5%),测定拔节期和抽穗期的光合特征曲线、叶绿素荧光动力学参数、叶氮含量和收获后的籽粒产量等指标,探讨干旱条件下库源关系改变对叶片光合适应的影响。结果表明: 在小麦拔节期,干旱条件下CO₂浓度升高处理的小麦PSⅡ实际光化学效率没有显著增加,但通过提升最大电子传递速率和电子向光化学方向的传递比例,增强了Rubisco的羧化速率,从而提高了最大净光合速率;在抽穗期,功能叶最大电子传递速率和电子向光化学方向的传递比例虽然较高,但PSⅡ实际光化学转换效率降低,Rubisco羧化速率和丙糖磷酸利用效率下降,以致最大净光合速率降低。干旱条件下,CO₂浓度升高增加了小麦单茎生物量、单穗粒数和穗粒重,降低了不孕小穗数,提高了籽粒产量。土壤干旱条件下,CO₂浓度升高对收获期小麦单茎籽粒产量的促进作用可能主要来自于生长前期的光合产物积累。生长后期光合适应发生的主要原因是功能叶PSⅡ实际光化学转换效率和丙糖磷酸利用效率的降低,而不是最大电子传递速率、光化学方向的电子传递比例和新叶库强的变化。     

Development of an activity-directed selection system enabled significant improvement of the carboxylation efficiency of Rubisco

Photosynthetic CO₂ fixation is the ultimate source of organic carbon on earth and thus is essential for crop production and carbon sequestration. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the first step of photosynthetic CO₂ fixation. However, the extreme low carboxylation efficiency of Rubisco makes it the most attractive target for improving photosynthetic efficiency. Extensive studies have focused on re-engineering a more efficient enzyme, but the effort has been impeded by the limited understanding of its structure-function relationships and the lack of an efficient selection system towards its activity. To address the unsuccessful molecular engineering of Rubisco, we developed an Escherichia coli-based activity-directed selection system which links the growth of host cell solely to the Rubisco activity therein. A Synechococcus sp. PCC7002 Rubisco mutant with E49V and D82G substitutions in the small subunit was selected from a total of 15,000 mutants by one round of evolution. This mutant showed an 85% increase in specific carboxylation activity and a 45% improvement in catalytic efficiency towards CO₂. The small-subunit E49V mutation was speculated to influence holoenzyme catalysis through interaction with the large-subunit Q225. This interaction is conserved among various Rubisco from higher plants and Chlamydomonas reinhardtii. Knowledge of these might provide clues for engineering Rubisco from higher plants, with the potential of increasing the crop yield.     

施氮和大气CO2浓度升高对小麦旗叶光合电子传递和分配的影响

采用开顶式气室盆栽培养小麦,设计2个大气CO₂浓度（正常：400 μmol·mol^-1;高：760 μmol·mol^-1）、2个氮素水平（0和200 mg·kg^-1土）的组合处理,通过测定小麦抽穗期旗叶氮素和叶绿素浓度、光合速率（P_n）-胞间CO₂浓度（C_i）响应曲线及荧光动力学参数,来测算小麦叶片光合电子传递速率等,研究了高大气CO₂浓度下施氮对小麦旗叶光合能量分配的影响.结果表明：与正常大气CO₂浓度相比,高大气CO₂浓度下小麦叶片氮浓度和叶绿素浓度降低,高氮处理的小麦叶片叶绿素a/b升高.施氮后小麦叶片PSⅡ最大光化学效率（F_v/F_m）、PSⅡ反应中心最大量子产额（F_v′/F_m′）、PSⅡ反应中心的开放比例（q_p）和PSⅡ反应中心实际光化学效率（Φ_PSⅡ）在大气CO₂浓度升高后无明显变化,虽然叶片非光化学猝灭系数（NPQ）显著降低,但PSⅡ总电子传递速率（J_F）无明显增加;不施氮处理的F_v′/F_m′、Φ_PSⅡ和NPQ在高大气CO₂浓度下显著降低,尽管F_v/F_m和q_P无明显变化,J_F仍显著下降.施氮后小麦叶片J_F增加,参与光化学反应的非环式电子流传递速率(J_C)明显升高.大气CO₂浓度升高使参与光呼吸的非环式电子流传递速率(J₀)、Rubisco氧化速率（V₀）、光合电子的光呼吸/光化学传递速率比（J₀/J_C）和Rubisco氧化/羧化比（V₀/V_C）降低,但使J_C和Rubisco羧化速率（V_C）增加.因此,高大气CO₂浓度下小麦叶片氮浓度和叶绿素浓度降低,而增施氮素使通过PSⅡ反应中心的电子流速率显著增加,促进了光合电子流向光化学方向的传递,使更多的电子进入Rubisco羧化过程,P_n显著升高.     

不同CO2浓度下大豆叶片的光合生理生态特性

CO₂是光合作用的原料和底物,影响着光合作用的进程和光合产物的数量.利用Li-6400-40B同时测量大豆叶片在不同CO₂浓度(300、400、500和600 μmol·mol^-1)下的光合电子传递速率和光合作用对光的响应曲线,并用构建的光合作用对光响应机理模型拟合这些光响应曲线,获得大豆叶片一系列的光合参数、生理生态参数和捕光色素分子的物理参数.结果表明: 电子利用效率、最大电子传递速率和最大净光合速率随CO₂浓度的升高而增加;光补偿点和暗呼吸速率随CO₂浓度的升高而下降;光能利用效率和内禀(瞬时)水分利用效率随CO₂浓度的升高而增加,不同CO₂浓度下的最大光能利用效率和最大内禀(瞬时)水分利用效率之间存在显著差异,但不同CO₂浓度下的最大羧化效率的差异不显著.CO₂浓度的大小对光合作用中原初光反应存在一定程度的影响,即高CO₂浓度有利于减小捕光色素分子处于最低激发态的最小平均寿命,以提高光能传递的速度及增加大豆光合电子流的利用效率.     

不同CO2浓度下大豆叶片的光合生理生态特性

CO₂是光合作用的原料和底物,影响着光合作用的进程和光合产物的数量.利用Li-6400-40B同时测量大豆叶片在不同CO₂浓度(300、400、500和600 μmol·mol^-1)下的光合电子传递速率和光合作用对光的响应曲线,并用构建的光合作用对光响应机理模型拟合这些光响应曲线,获得大豆叶片一系列的光合参数、生理生态参数和捕光色素分子的物理参数.结果表明: 电子利用效率、最大电子传递速率和最大净光合速率随CO₂浓度的升高而增加;光补偿点和暗呼吸速率随CO₂浓度的升高而下降;光能利用效率和内禀(瞬时)水分利用效率随CO₂浓度的升高而增加,不同CO₂浓度下的最大光能利用效率和最大内禀(瞬时)水分利用效率之间存在显著差异,但不同CO₂浓度下的最大羧化效率的差异不显著.CO₂浓度的大小对光合作用中原初光反应存在一定程度的影响,即高CO₂浓度有利于减小捕光色素分子处于最低激发态的最小平均寿命,以提高光能传递的速度及增加大豆光合电子流的利用效率.     

植物叶片最大羧化速率与叶氮含量关系的变异性

叶片最大羧化速率是表征植物光合能力的关键参数, 受到光照、温度、水分、CO₂浓度、叶片氮含量等多个要素的控制。准确地模拟植物叶片最大羧化速率对环境因子的响应是预测未来植被生产力和碳循环过程的前提。目前大多数陆地碳循环过程模型以Farqhuar光合作用模型为基础模拟植物的光合作用, 关于植物叶片的最大羧化速率与叶氮含量关系的模拟方法却各不相同。该文汇总了1990-2013年国内外植物叶片光合速率观测研究文献中叶片最大羧化速率与叶氮含量的关系式及相关数据, 分析了叶片最大羧化速率与叶氮含量关系随不同植被功能型和时间的变化特征, 以及环境因子变化条件下最大羧化速率与叶氮含量关系的变化特征, 探讨了二者关系变异性的可能原因以及影响因子。结果表明: 1)不同功能型植物叶片的最大羧化速率和叶氮含量的关系存在较大差异, 二者线性关系式的斜率平均值变化范围为16.29-50.25 μmol CO₂·g N^-1·s^-1。落叶植被叶片的最大羧化速率随叶氮含量的变化率和光合氮利用效率一般都高于常绿植被, 其变异主要源于植物的比叶重和叶片内部氮素分配的差异。2)叶片最大羧化速率随叶氮含量的变化存在季节和年际变异。在没有受到水分胁迫的年份中, 叶片最大羧化速率随叶氮含量变化的速率一般在春季或夏季最高, 其季节变异与比叶重和叶氮在Rubisco的分配比例的季节变化有关。受到干旱的影响, 叶片最大羧化速率随叶氮含量的变化率会升高。3)当大气CO₂浓度增加时, 由于叶片中Rubisco含量的降低, 多年生针叶叶片最大羧化速率和叶氮关系斜率值会出现降低; 当供氮水平增加时, 叶片最大羧化速率和叶片氮含量均表现出增加趋势, 二者线性关系的斜率也相应增加。在此基础上, 该文指出在模拟叶片最大羧化速率与叶氮含量的关系时, 应考虑叶片比叶重和叶氮在Rubisco中的分配比例的季节变异、水分胁迫、大气CO₂浓度和供氮水平变化对二者关系的影响。囿于数据的有限性, 今后应进一步加强多因子控制实验研究, 深入探讨叶片最大羧化速率与叶氮含量关系的变异性机理, 并获得更系统的观测数据, 以助生态系统过程模型的改进, 提高模型的模拟精度。     

Variations in the relationship between maximum leaf carboxylation rate and leaf nitrogen concentration

叶片最大羧化速率是表征植物光合能力的关键参数, 受到光照、温度、水分、CO₂浓度、叶片氮含量等多个要素的控制。准确地模拟植物叶片最大羧化速率对环境因子的响应是预测未来植被生产力和碳循环过程的前提。目前大多数陆地碳循环过程模型以Farqhuar光合作用模型为基础模拟植物的光合作用, 关于植物叶片的最大羧化速率与叶氮含量关系的模拟方法却各不相同。该文汇总了1990-2013年国内外植物叶片光合速率观测研究文献中叶片最大羧化速率与叶氮含量的关系式及相关数据, 分析了叶片最大羧化速率与叶氮含量关系随不同植被功能型和时间的变化特征, 以及环境因子变化条件下最大羧化速率与叶氮含量关系的变化特征, 探讨了二者关系变异性的可能原因以及影响因子。结果表明: 1)不同功能型植物叶片的最大羧化速率和叶氮含量的关系存在较大差异, 二者线性关系式的斜率平均值变化范围为16.29-50.25 μmol CO₂·g N^-1·s^-1。落叶植被叶片的最大羧化速率随叶氮含量的变化率和光合氮利用效率一般都高于常绿植被, 其变异主要源于植物的比叶重和叶片内部氮素分配的差异。2)叶片最大羧化速率随叶氮含量的变化存在季节和年际变异。在没有受到水分胁迫的年份中, 叶片最大羧化速率随叶氮含量变化的速率一般在春季或夏季最高, 其季节变异与比叶重和叶氮在Rubisco的分配比例的季节变化有关。受到干旱的影响, 叶片最大羧化速率随叶氮含量的变化率会升高。3)当大气CO₂浓度增加时, 由于叶片中Rubisco含量的降低, 多年生针叶叶片最大羧化速率和叶氮关系斜率值会出现降低; 当供氮水平增加时, 叶片最大羧化速率和叶片氮含量均表现出增加趋势, 二者线性关系的斜率也相应增加。在此基础上, 该文指出在模拟叶片最大羧化速率与叶氮含量的关系时, 应考虑叶片比叶重和叶氮在Rubisco中的分配比例的季节变异、水分胁迫、大气CO₂浓度和供氮水平变化对二者关系的影响。囿于数据的有限性, 今后应进一步加强多因子控制实验研究, 深入探讨叶片最大羧化速率与叶氮含量关系的变异性机理, 并获得更系统的观测数据, 以助生态系统过程模型的改进, 提高模型的模拟精度。     

Photosynthetic acclimation in rice leaves to free-air CO2 enrichment related to both ribulose-1,5-bisphosphate carboxylation limitation and ribulose-1,5-bisphosphate regeneration limitation

Net photosynthetic rates (Pns) in leaves were compared between rice plants grown in ambient air control and free-air CO2 enrichment (FACE, about 200 micromol mol(-1) above ambient) treatment rings. When measured at the same CO2 concentration, the Pn of FACE leaves decreased significantly, indicating that photosynthetic acclimation to high CO2 occurs. Although stomatal conductance (Gs) in FACE leaves was markedly decreased, intercellular CO2 concentrations (Ci) were almost the same in FACE and ambient leaves, indicating that the photosynthetic acclimation is not caused by the decreased Gs. Furthermore, carboxylation efficiency and maximal Pn, both light and CO2-saturated Pn, were decreased in FACE leaves, as shown by the Pn-Ci curves. In addition, the soluble protein, Rubisco (ribulose-1,5-bisphosphate caboxylase/oxygenase), and its activase contents as well as the sucrose-phosphate synthase activity decreased significantly, while some soluble sugar, inorganic phosphate, chlorophyll and light-harvesting complex II (LHC II) contents increased in FACE leaves. It appears that the photosynthetic acclimation in rice leaves is related to both ribulose-1,5-bisphosphate (RuBP) carboxylation limitation and RuBP regeneration limitation.     

Rate-limiting factors in leaf photosynthesis. I. Carbon fluxes in the calvin cycle

Rates of photosynthesis of spinach leaves were varied by varying light intensity and CO₂ concentration. Metabolism of the leaves was then arrested by freezing them in liquid nitrogen. Chloroplasts were isolated by a nonaqueous procedure. In the chloroplast fractions, levels of intermediates of the carbon reduction cycle were determined and considered in relation to the photosynthetic flux situation of the leaves at the time before freezing. During induction of photosynthesis, ribulose 1,5-bisphosphate levels increased in parallel with CO₂ fixation. In the steady state, a similar relation between ribulose 1,5-bisphosphate levels and CO₂ uptake was observed at light intensities between 0 and 50 W·m⁻². A further increase in light intensity increased CO₂ fixation rates but not ribulose 1,5-bisphosphate levels. Increasing the CO₂ concentration resulted in increased CO₂ uptake, whereas ribulose 1,5-bisphosphate levels decreased. Even under CO₂ saturation, ribulose 1,5-bisphosphate levels were about 100 nmol/mg chlorophyll corresponding to about 3.5 mM ribulose 1,5-bisphosphate in the chloroplast stroma. This suggests that even under CO₂ saturation, ribulose-1,5-bisphosphate carboxylase limits photosynhetic CO₂ uptake. Mass action ratios calculated from measured metabolite levels demonstrated that the thermodynamic gradient required for the regeneration of ribulose 1,5-bisphosphate from hexosephosphate and triosephosphate increased considerably as photosynthetic flux increased. Similar calculations revealed that the enzymatic apparatus responsible for the reduction of 3-phosphoglycerate to dihydroxyacetone phosphate is not displaced much from equilibrium even under maximum rates of photosynthesis at saturating CO₂. The same is true for aldolase. Fructose-1,6-bisphosphatase also did not limit Calvin cycle turnover. Only at very low light intensities and during the first minutes of the induction period was the ratio of fructose 1,6-bisphosphate to fructose 6-phosphate high. This observation was more readily explained in terms of fructose 1,6-bisphosphate binding to ribulose-1,5-bisphosphate carboxylase than by a rate limitation imposed by insufficient activation of fructose-1,6-bisphosphatase.     

胡杨异形叶光合作用对光强与CO2浓度的响应

胡杨(Populus euphratica)叶形多变, 随个体生长发育, 植株出现条形、卵形和锯齿阔卵形叶。在新疆塔里木河上游人工胡杨林内选择具有此3种叶形的成年标准株, 将枝条拉至同一高度, 通过活体测定, 比较其光合作用-光与CO₂响应及叶绿素荧光响应特征。结果表明: 胡杨异形叶光合速率对光强/CO₂浓度与电子传递速率对光强的响应曲线均可用直角双曲线修正模型来拟合, 得出的主要光合参数与实测值较吻合。胡杨卵形叶、锯齿阔卵形叶光合速率-光响应参数与生化参数及快速光响应参数与条形叶差异显著, 而光合速率-CO₂响应参数则无显著差异。胡杨异形叶CO₂饱和浓度下的最大净光合速率(P_nmax)较饱和光强下的P_nmax高, 表明胡杨强光下光合速率在很大程度上受CO₂供应和1,5-二磷酸核酮糖(RuBP)再生能力的限制。卵形叶、锯齿阔卵形叶的初始量子效率(α)、初始羧化效率(CE)、P_nmax、光合能力(A_max)与最大羧化速率(V_cmax)均显著高于条形叶; 锯齿叶光饱和点(LSP)、最大电子传递速率(ETR_max)与光呼吸速率(R_p)高于卵形叶, 条形叶光补偿点(LCP)与LSP、α、CE最低。表明荒漠干旱环境下胡杨锯齿叶最耐强光, 高R_p可能是其耗散过剩光能、保护光合机构免于强光破坏的重要途径; 卵形叶高的α、CE、磷酸丙糖利用效率(TPU)、PSII实际光化学效率(Φ_PSII)与低LCP及叶氮分配策略是其保持高光合速率的原因; 条形叶Φ_PSII、ETR、P_n低, 因其制造光合产物不足而难以满足树体生长逐渐减少并处于树冠下部。可见, 胡杨条形叶光合效率低、抗逆性差, 主要以维持生长为主; 随着树体长大, 条形叶难以适应荒漠环境来维系其生长, 出现了卵形叶; 卵形叶光合效率高, 易于快速积累光合产物而加快树体生长, 但其LSP低和耐光抑制能力弱, 逐渐被更耐强光、高温与大气干旱的锯齿叶所取代, 从而使胡杨在极端逆境下得以生存, 这是胡杨从幼苗到成年叶形变化及异形叶着生在树冠不同高度的原因。     

基于FvCB模型分析盐分胁迫对棉花叶片光合作用的影响

为深入理解叶片光合特性对盐胁迫的响应机理,以棉花为试验材料,设置5个盐分(NaCl)浓度处理:0(CK)、50、100、150和200 mmol·L^-1,利用FvCB模型分析盐胁迫对棉花幼苗叶片光合特性的影响。结果表明:与CK相比,50和100 mmol·L^-1盐分处理增加了棉花叶片的最大羧化速率(V_{c max})和最大电子传递速率(J_max),但150和200 mmol·L^-1盐分处理显著降低了V_{c max}和J_max。叶片净光合速率(P_n)、叶肉导度(g_m)和暗呼吸速率(R_d)随盐分浓度升高而下降;与CK相比,50和100 mmol·L^-1盐分处理对g_m无显著影响,但P_n和R_d显著降低。150和200 mmol·L^-1盐分处理明显降低了P_n、g_m和R_d,且与0、50和100 mmol·L^-1盐分处理间存在显著差异;利用FvCB模型模拟了不同盐分胁迫下叶片净光合速率。与不考虑g_m的模拟结果相比,考虑g_m提高模拟值和实测值间的决定系数,并降低了平均绝对误差。棉花幼苗耐盐阈值为100～150 mmol·L^-1,随盐分浓度的增加,光合限制因素由叶肉因素转变为光合机构受损;引入g_m可以提高FvCB模型的模拟精度。     

Use of Transgenic Plants with Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Antisense DNA to Evaluate the Rate Limitation of Photosynthesis under Water Stress

The biochemical lesion that causes impaired chloroplast metabolism (and, hence, photosynthetic capacity) in plants exposed to water deficits is still a subject of controversy. In this study we used tobacco (Nicotiana tabacum L.) transformed with "antisense" ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) DNA sequences to evaluate whether Rubisco or some other enzymic step in the photosynthetic carbon reduction cycle pathway rate limits photosynthesis at low leaf water potential ([psi]w). These transformants, along with the wild-type material, provided a novel model system allowing for an evaluation of photosynthetic response to water stress in near-isogenic plants with widely varying levels of functional Rubisco. It was determined that impaired chloroplast metabolism (rather than decreased leaf conductance to CO2) was the major cause of photosynthetic inhibition as leaf [psi]w declined. Significantly, the extent of photosynthetic inhibition at low [psi]w was identical in wild-type and transformed plants. Decreasing Rubisco activity by 68% did not sensitize photosynthetic capacity to water stress. It was hypothesized that, if water stress effects on Rubisco caused photosynthetic inhibition under stress, an increase in the steady-state level of the substrate for this enzyme, ribulose 1,5-bisphosphate (RuBP), would be associated with stress-induced photosynthetic inhibition. Steady-state levels of RuBP were reduced as leaf [psi]w declined, even in transformed plants with low levels of Rubisco. Based on the similarity in photosynthetic response to water stress in wild-type and transformed plants, the reduction in RuBP as stress developed, and studies that demonstrated that ATP supply did not rate limit photosynthesis under stress, we concluded that stress effects on an enzymic step involved in RuBP regeneration caused impaired chloroplast metabolism and photosynthetic inhibition in plants exposed to water deficits.     

Balancing carboxylation and regeneration of ribulose-1,5- bisphosphate in leaf photosynthesis: temperature acclimation of an evergreen tree, Quercus myrsinaefolia

Changes in the temperature dependence of the photosynthetic rate depending on growth temperature were investigated for a temperate evergreen tree, Quercus myrsinaefolia . Plants were grown at 250 μ mol quanta m^–2 s^–1 under two temperature conditions, 15 and 30 °C. The optimal temperature that maximizes the light-saturated rate of photosynthesis at 350 μ L L^–1 CO₂ was found to be 20–25 and 30–35 °C for leaves grown at 15 and 30 °C, respectively. We focused on two processes, carboxylation and regeneration of ribulose-1,5-bisphosphate (RuBP), which potentially limit photosynthetic rates. Because the former process is known to limit photosynthesis at lower CO₂ concentrations while the latter limits it at higher CO₂ concentrations, we determined the temperature dependence of the photosynthetic rate at 200 and 1000 μ L L^–1 CO₂ under saturated light. It was revealed that the temperature dependence of both processes varied depending on the growth temperature. Using a biochemical model, we estimated the capacity of the two processes at various temperatures under ambient CO₂ concentration. It was suggested that, in leaves grown at low temperature (15 °C), the photosynthetic rate was limited solely by RuBP carboxylation under any temperature. On the other hand, it was suggested that, in leaves grown at high temperature (30 °C), the photosynthetic rate was limited by RuBP regeneration below 22 °C, but limited by RuBP carboxylation above 22 °C. We concluded that: (1) the changes in the temperature dependence of carboxylation and regeneration of RuBP and (2) the changes in the balance of these two processes altered the temperature dependence of the photosynthetic rate.     

褪黑素对盐碱复合胁迫下黄瓜幼苗光合特性和渗透调节物质含量的影响

为明确外源褪黑素(MT)对黄瓜盐碱胁迫的缓解效应,以‘新春四号’黄瓜为试材,采用复合盐碱(NaCl∶Na₂SO₄∶Na₂CO₃∶NaHCO₃=1∶9∶1∶9)模拟胁迫,测定外源根施MT和盐碱胁迫下黄瓜叶片光合特性和渗透调节物质含量。结果表明: 与正常生长黄瓜幼苗相比,在40 mmol·L^-1盐碱胁迫下,外源施加10 μmol·L^-1 MT能够显著增加黄瓜幼苗叶片的叶绿素、可溶性糖和可溶性蛋白质含量,提高净光合速率、气孔导度、蒸腾速率、光系统Ⅱ最大光化学效率、实际光化学效率、表观光合电子传递速率和光化学淬灭系数,而胞间CO₂浓度、非光化学淬灭系数、蔗糖、果糖、淀粉和脯氨酸含量减小了11.1%、13.8%、12.7%、27.5%、1.3%和32.8%,同时,碳同化关键酶核酮糖-1,5-二磷酸羧化/加氧酶、果糖-1,6-二磷酸酯酶活性显著升高,且Rubisco亚基(CsrbcS和CsrbcL)、CsFBA、CsRCA、CsFBPase、CsTK的mRNA水平均下调表达。综上,外源MT能够提高盐碱胁迫下黄瓜幼苗的叶绿素、渗透调节物质含量、光合化学效率和碳同化关键酶活性,增强幼苗光合能力,减轻复合盐碱胁迫对植株的伤害。研究结果可为抗盐碱栽培提供理论依据。     

Climatic warming changes plant photosynthesis and its temperature dependence in a temperate steppe of northern China

Warming responses of photosynthesis and its temperature dependence in two C₃ grass (Agropyron cristatum, Stipa krylovii), one C₄ grass (Pennisetum centrasiaticum), and two C₃ forb (Artemisia capillaris, Potentilla acaulis) species in a temperate steppe of northern China were investigated in a field experiment. Experimental warming with infrared heater significantly increased daily mean assimilation rate (A) in P. centrasiaticum and A. capillaris by 30 and 43%, respectively, but had no effects on other three species. Seasonal mean A was 13, 15, and 19% higher in the warmed than control plants for P. centrasiaticum, A. capillaries, and S. krylovii, respectively. The mean assimilation rate in A. cristatum and P. acaulis was not impacted by experimental warming. All the five species showed photosynthetic acclimation to temperature. The optimum temperature for photosynthesis (T_opt) and the assimilation rate at T_opt in the five species increased by 0.33–0.78 °C and 4–27%, respectively, under experimental warming. Elevated temperature tended to increase the maximum rate of ribulose-1,5-bisphosphate (RuBP) carboxylation (V_cmax) and the RuBP regeneration capacity (J_max) in the C₃ plants and carboxylation efficiency and the CO₂-saturated photosynthetic rate in the C₄ plant at higher leaf temperature, as well as the optimum temperatures for the four parameters. Our results indicated that photosynthetic responses to warming were species-specific and that most of the species in the temperate steppe of northern China could acclimate to a warmer environment. The changes in the temperature dependence of V_cmax and J_max, as well as the balance of these two processes altered the temperature dependence of photosynthesis under climatic warming.     

Estimation of rate constants of the partial reactions of carboxylation of ribulose-1,5-bisphosphate in vivo

An in vivo method for the estimation of kinetic parameters of partial reactions of carboxylation of ribulose 1,5-bisphosphate (RuBP) catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is described. Rubisco in barley, wheat and bean is different in the ability of its active centers to bind RuBP. The rate constant of the formation of the Rubisco-RuBP complex in these plants at 25 °C is 0.414, 0.245 and 0.660 mM^-1 s^-1, respectively. The rate constant of the reaction of the Rubisco-bound enediol with CO₂ does not differ significantly in barley and wheat, and averages 66 mM^-1 s^-1. Decreased irradiance inhibits Rubisco in two ways: by reducing the concentration of operating catalytic sites and by decreasing the rate constant of binding of RuBP to Rubisco. High concentrations of CO₂ inhibit Rubisco by decreasing the concentration of competent carboxylation centers, without any s ignificant influence upon the rate constants of partial reactions.     

Dependence of photosynthesis of sunflower and maize leaves on phosphate supply, ribulose-1,5-bisphosphate carboxylase/oxygenase activity, and ribulose-1,5-bisphosphate pool size

Sunflower (Helianthus annuus L. cv Asmer) and maize (Zea mays L. cv Eta) plants were grown under controlled environmental conditions with a nutrient solution containing 0, 0.5, or 10 millimolar inorganic phosphate. Phosphate-deficient leaves had lower photosynthetic rates at ambient and saturating CO₂ and much smaller carboxylation efficiencies than those of plants grown with ample phosphate. In addition, phosphate-deficient leaves contained smaller quantities of total soluble proteins and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) per unit area, although the relative proportions of these components remained unchanged. The specific activity of Rubisco (estimated in the crude extracts of leaves) was significantly reduced by phosphate deficiency in sunflower but not in maize. Thus, there was a strong dependence of carboxylation efficiency and CO₂-saturated photosynthetic rate on Rubisco activity only in sunflower. Phosphate deficiency decreased the 3-phosphoglycerate and ribulose-1,5-bisphosphate (RuBP) contents of the leaf in both species. The ratio of 3-phosphoglycerate to RuBP decreased in sunflower but increased in maize with phosphate deficiency. The calculated concentrations of RuBP and RuBP-binding sites in the chloroplast stroma decreased markedly with phosphate deficiency. The ratio of the stromal concentration of RuBP to that of RuBP-binding sites decreased in sunflower but was not affected in maize with phosphate deficiency. We suggest that a decrease in this ratio made the RuBP-binding sites more vulnerable to blockage or inactivation by tight-binding metabolites/inhibitors, causing a decrease in the initial specific activity of Rubisco in the crude extract from phosphate-deficient sunflower leaves. However, the decrease in Rubisco specific activity was much less than the decrease in the RuBP content in the leaf and its concentration in the stroma. A large ratio of RuBP to RuBP-binding sites may have maintained the Rubisco-specific activity in phosphate-deficient maize leaves. We conclude that the effect of phosphate deficiency is more on RuBP regeneration than on Rubisco activity in both sunflower and maize.