Analysis of limitations to CO2 assimilation on exposure of leaves of two Brassica napus cultivars to UV-B

Apex and Bristol cultivars of oilseed rape (Brassica napus) were irradiated with 0.63 W m^?2 of UV-B over 5 d. Analyses of the response of net leaf carbon assimilation to intercellular CO₂ concentration were used to examine the potential limitations imposed by stomata, carboxylation velocity and capacity for regeneration of ribulose 1,5-bis-phosphate on leaf photosynthesis. Simultaneous measurements of chlorophyll fluorescence were used to estimate the maximum quantum efficiency of photosystem II (PSII) photochemistry, the quantum efficiency of linear electron transport at steady-state photosynthesis, and the light and CO₂-saturated rate of linear electron transport. Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) content and activities were assayed in vitro. In both cultivars the UV-B treatment resulted in decreases in the light-saturated rate of CO₂ assimilation, which were accompanied by decreases in carboxylation velocity and Rubisco content and activity. No major effects of UV-B were observed on end-product inhibition and stomatal limitation of photosynthesis or the rate of photorespiration relative to CO₂ assimilation. In the Bristol cultivar, photoinhibition of PSII and loss of linear electron transport activity were observed when CO₂ assimilation was severely inhibited. However, the Apex cultivar exhibited no major inhibition of PSII photochemistry or linear electron transport as the rate of CO₂ assimilation decreased. It is concluded that loss of Rubisco is a primary factor in UV-B inhibition of CO₂ assimilation.     

Oxygen sensitivity of C4 photosynthesis: evidence from gas exchange and chlorophyll fluorescence analyses with different C4 subtypes

Because photosynthetic rates in C₄ plants are the same at normal levels of O₂ (c, 20 kPa) and at c, 2 kPa O₂ (a conventional test for evaluating photorespiration in C₃ plants) it has been thought that C₄ photosynthesis is O₂ insensitive. However, we have found a dual effect of O₂ on the net rate of CO₂ assimilation among species representing all three C₄ subtypes from both monocots and dicots. The optimum O₂ partial pressure for C₄ photosynthesis at 30 °C, atmospheric CO₂ level, and half full sunlight (1000 μmol quanta m^?2 s^?1) was about 5–10 kPa. Photosynthesis was inhibited by O₂ below or above the optimum partial pressure. Decreasing CO₂ levels from ambient levels (32.6 Pa) to 9.3 Pa caused a substantial increase in the degree of inhibition of photosynthesis by supra-optimum levels of O₂ and a large decrease in the ratio of quantum yield of CO₂ fixation/quantum yield of photosystem II (PSII) measured by chlorophyll a fluorescence. Photosystem II activity, measured from chlorophyll a fluorescence analysis, was not inhibited at levels of O₂ that were above the optimum for CO₂ assimilation, which is consistent with a compensating, alternative electron How as net CO₂ assimilation is inhibited. At suboptimum levels of O₂, however, the inhibition of photosynthesis was paralleled by an inhibition of PSII quantum yield, increased state of reduction of quinone A, and decreased efficiency of open PSII centres. These results with different C₄ types suggest that inhibition of net CO₂ assimilation with increasing O₂ partial pressure above the optimum is associated with photorespiration, and that inhibition below the optimum O₂ may be caused by a reduced supply of ATP to the C₄ cycle as a result of inhibition of its production photochemically.     

高大气CO2浓度下氮素对小麦叶片光能利用的影响

关于氮素对高大气CO₂浓度下C₃植物光合作用适应现象的调节机理已有较为深入的研究, 但对其光合作用适应现象的光合能量转化和分配机制缺乏系统分析。该文以大气CO₂浓度和施氮量为处理手段, 通过测定小麦(Triticum aestivum)抽穗期叶片的光合作用-胞间CO₂浓度响应曲线以及荧光动力学参数来测算光合电子传递速率和分配去向, 研究了长期高大气CO₂浓度下小麦叶片光合电子传递和分配对施氮量的响应。结果表明, 与正常大气CO₂浓度处理相比, 高大气CO₂浓度下小麦叶片较多的激发能以热量的形式耗散, 增施氮素可使更多的激发能向光化学反应方向的分配, 降低光合能量的热耗散速率; 大气CO₂浓度升高后小麦叶片光化学淬灭系数无明显变化, 高氮叶片的非光化学猝灭降低而低氮叶片明显升高, 施氮促进PSII反应中心的开放比例, 降低光能的热耗散; 高大气CO₂浓度下高氮叶片通过PSII反应中心的光合电子传递速率(J_F)较高, 而且参与光呼吸的非环式电子流速率(J₀)显著降低, 较正常大气CO₂浓度处理的高氮叶片下降了88.40%, 光合速率增加46.47%; 高大气CO₂浓度下小麦叶片J_F-J₀升高而J₀/J_F显著下降, 光呼吸耗能被抑制, 更多的光合电子分配至光合还原过程。因此, 大气CO₂浓度增高条件下, 小麦叶片激发能的热耗散速率增加, 但增施氮素后小麦叶片PSII反应中心开放比例提高, 光化学速率增加, 进入PSII反应中心的电子流速率明显升高, 光呼吸作用被抑制, 光合电子较多地进入光化学过程, 这可能是高氮条件下光合作用适应性下调被缓解的一个原因。     

Changes in leaf photosynthesis of transgenic rice with silenced <Emphasis Type="Italic">OsBP-73</Emphasis> gene

In comparison with its wild type (WT), the transgenic (TG) rice with silenced OsBP-73 gene had significantly lower plant height, grain number per panicle, and leaf net photosynthetic rate (P _N). Also, the TG rice showed significantly lower chlorophyll (Chl), ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO), RuBPCO activase, and RuBP contents, photosystem 2 (PS2) photochemical efficiency (F_v/F_m and ΔF/F_m′), apparent quantum yield of carbon assimilation (Φ_c), carboxylation efficiency (CE), photosynthetic electron transport and photophosphorylation rates as well as sucrose phosphate synthase activity, but higher intercellular CO₂ concentration, sucrose, fructose, and glycerate 3-phosphate contents, and non-photochemical quenching of Chl fluorescence (NPQ). Thus the decreased P _N in the TG rice leaves is related to both RuBP carboxylation and RuBP regeneration limitations, and the latter is a predominant limitation to photosynthesis.     

Water stress induces different levels of photosynthesis and electron transport rate regulation in grapevines

A, net CO₂ assimilation rate
E, leaf transpiration
ETR, electron transport rate
F_s, fluorescence yield at steady state
F_m and F_m', maximal fluorescence levels when all PSII reaction centres are closed in dark- and light-acclimated leaves, respectively
F_o and F_o', initial fluorescence levels when all PSII reaction centres are closed in dark- and light-acclimated leaves, respectively
F_v/F_m, efficiency of excitation capture by open PSII in dark-adapted leaves
ΔF/F_m', actual photochemical efficiency of PSII
g, stomatal conductance
NPQ, non-photochemical quenching of chlorophyll fluorescence
PPFD, photosynthetic photon flux density
Ψ_PD and Ψ_MD, leaf water potential at pre-dawn and midday, respectively
Rl, estimated photorespiration rate
I₁ and I₂, Irrigation treatments
R, Recovery treatment
D₁ and D₂, drought treatments
HD₁ and HD₂, hard drought treatments

Diurnal time courses of chlorophyll fluorescence and gas-exchange rates were measured in young potted grapevines (Vitis vinifera L. cv. Tempranillo) subjected to different conditions of water supply under Mediterranean summer conditions. The irrigated plants exhibited typical diurnal patterns for all measured parameters, showing a correspondence between electron transport rate, net CO₂ assimilation and stomatal conductance. Mild decreases in soil-water availability led to different degrees of down-regulation of photosynthesis and increased nonphotochemical quenching of chlorophyll fluorescence. A good correspondence between electron transport rate and CO₂ assimilation was still maintained, suggesting a coregulation of both photosynthetic processes. In contrast, a severe water deficit induced a drastic down-regulation of photosynthesis and breakage of the above-mentioned link. Both midday net CO₂ assimilation and electron transport rate significantly correlated with pre-dawn water potential (Ψ_PD) (r² = 0·65 and r² = 0·92, P < 0·001, respectively). However, when field data were analysed, the relationship between electron transport rate and Ψ_PD was not maintained, although net CO₂ assimilation was similarly correlated with Ψ_PD. Interestingly, the steady-state chlorophyll fluorescence yield was a good indicator of plant water stress.     

Photosynthesis and chlorophyllafluorescence during flag leaf senescence of field-grown wheat plants

The characteristics of photosynthetic gas exchange, chlorophyll a fluorescence, and xanthophyll cycle pigments during flag leaf senescence of field-grown wheat plants were investigated. With senescence progressing, the light-saturated net CO₂ assimilation rate expressed either on a basis of leaf area or chlorophyll decreased significantly. The apparent quantum yield of net photosynthesis decreased when expressed on a leaf area basis but increased when expressed on a chlorophyll basis. The maximal efficiency of PSII photochemistry decreased very little while actual PSII efficiency, photochemical quenching, and the efficiency of excitation capture by open PSII centers decreased considerably. At the same time, non-photochemical quenching increased significantly. A substantial decrease in the contents of violaxanthin and zeaxanthin, but a slight decrease in the content of antheraxanthin were observed. However, the de-epoxidation status of the xanthophyll cycle was positively correlated with progressive senescence. This increase was due mainly to a smaller decrease in zeaxanthin than in violaxanthin. Our results suggest that PSII apparatus remained functional, but a down-regulation of PSII occurred under the steady state of photosynthesis in senescent flag leaves. Such a down-regulation was associated with the closure of PSII centers and an enhanced xanthophyll cycle-related thermal dissipation in the PSII antennae.     

Electron transport is the functional limitation of photosynthesis in field-grown Pima cotton plants at high temperature

Restrictions to photosynthesis can limit plant growth at high temperature in a variety of ways. In addition to increasing photorespiration, moderately high temperatures (35–42 °C) can cause direct injury to the photosynthetic apparatus. Both carbon metabolism and thylakoid reactions have been suggested as the primary site of injury at these temperatures. In the present study this issue was addressed by first characterizing leaf temperature dynamics in Pima cotton (Gossypium barbadense) grown under irrigation in the US desert south‐west. It was found that cotton leaves repeatedly reached temperatures above 40 °C and could fluctuate as much as 8 or 10 °C in a matter of seconds. Laboratory studies revealed a maximum photosynthetic rate at 30–33 °C that declined by 22% at 45 °C. The majority of the inhibition persisted upon return to 30 °C. The mechanism of this limitation was assessed by measuring the response of photosynthesis to CO₂ in the laboratory. The first time a cotton leaf (grown at 30 °C) was exposed to 45 °C, photosynthetic electron transport was stimulated (at high CO₂) because of an increased flux through the photorespiratory pathway. However, upon cooling back to 30 °C, photosynthetic electron transport was inhibited and fell substantially below the level measured before the heat treatment. In the field, the response of assimilation (A) to various internal levels of CO₂ (C_i) revealed that photosynthesis was limited by ribulose‐1,5‐bisphosphate (RuBP) regeneration at normal levels of CO₂ (presumably because of limitations in thylakoid reactions needed to support RuBP regeneration). There was no evidence of a ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) limitation at air levels of CO₂ and at no point on any of 30 A–C_i curves measured on leaves at temperatures from 28 to 39 °C was RuBP regeneration capacity measured to be in substantial excess of the capacity of Rubisco to use RuBP. It is therefore concluded that photosynthesis in field‐grown Pima cotton leaves is functionally limited by photosynthetic electron transport and RuBP regeneration capacity, not Rubisco activity.     

Long-term exposure to elevated [CO2] in a natural Quercus ilex L. community: net photosynthesis and photochemical efficiency of PSII at different levels of water stress

Naturally grown trees of Mediterranean evergreen oak (Quercus ilex L.), representing the climax species of the region, were enclosed in six large open-top chambers and exposed to ambient and elevated CO₂ concentrations during a 3 year period. Maximum daily net photosynthetic rates measured at the two different CO₂ concentrations were from 30 to 100% higher in elevated than in ambient [CO₂] throughout the experimental period. The increase in maximum daily photosynthesis was also accompanied by a 93% rise in the apparent quantum yield of CO₂ assimilation, measured during periods of optimum soil moisture conditions. Hence, no clear evidence of down-regulation of net photosynthetic activity was found. Interactions between atmospheric CO₂ concentration and plant water stress were studied by following the natural evolution of drought in different seasons and years. At each level of water stress, the maximum rate of carbon assimilation was higher in elevated than in ambient [CO₂] by up to 100%. Analysis of in vivo chlorophyll fluorescence parameters in normal (21%) and low (2%) oxygen concentrations provided useful insights into the functioning and stability of the photosynthetic processes. The photochemical efficiency of PSII (F_v/F_m) progressively decreased as drought conditions became more evident; this trend was accentuated under elevated [CO₂]. Thermal de-excitation processes were possibly more significant under elevated than under ambient [CO₂], in a combination of environmental stresses. This research suggests two possible conclusions: (i) a ‘positive’ interaction between elevated [CO₂] and carbon metabolism can be obtained through relief of water stress limitation in the summer months, and (ii) elevated [CO₂], under drought conditions, may also enhance the significance of slow-relaxing quenching.     

Effects of exogenous nitric oxide on the photosynthetic characteristics of bamboo (<Emphasis Type="Italic">Indocalamus barbatus</Emphasis> McClure) seedlings under acid rain stress

The effects of the exogenous application of nitric oxide (NO, in the form of sodium nitroprusside, SNP) on the diurnal variation in photosynthesis, chlorophyll content, chlorophyll fluorescence, light response curve and the net assimilation of CO₂ against intercellular CO₂ concentration (A-Ci) curve parameters were investigated in the leaves of bamboo (Indocalamus barbatus McClure) exposed to simulated acid rain (SAR, pH 3.0) stress. According to the results of the diurnal variation in photosynthesis, foliar applications of 100–400 mg/L SNP effectively inhibited the decrease in net photosynthetic rate (Pn) as a result of non-stomatal factors, and mitigated midday depression under acid rain stress. The mitigating effect was most pronounced at 400 mg/L SNP. However, at higher concentrations of SNP (700 and 1000 mg/L), the mitigating effect became weak and even counterproductive. The results of the chlorophyll content, light response and A-Ci curve parameters suggested that the regulating role of NO against acid rain in the photosynthetic processes occurs through improving not only the efficiency of the light-harvesting and the activity of photosynthetic apparatus, but also the absorption of CO₂ and the availability of CO₂ for photosynthesis. The results of the chlorophyll fluorescence investigation further indicated that NO protected PSII activity from the damage of acid rain toxicity by enhancing the electron transport activity and photochemical efficiency, especially concerning the increase in the proportion of PSII open reaction centers. Furthermore, NO induced an increase in photorespiration (Rp), rather than an increase in non-photochemical quenching (NPQ), to dissipate the excessive excitation energy, which provided some protection to the photosynthetic apparatus under acid rain stress.     

Regulation of photosynthetic electron-transport in Phaseolus vulgaris L., as determined by room-temperature chlorophyll a fluorescence

The regulation of photosystem II (PSII) by light-, CO₂-, and O₂-dependent changes in the capacity for carbon metabolism was studied. Estimates of the rate of electron transport through PSII were made from gas-exchange data and from measurements of chlorophyll fluorescence. At subsaturating photon-flux density (PFD), the rate of electron transport was independent of O₂ and CO₂. Feedback on electron transport was observed under two conditions. At saturating PFD and low partial pressure of CO₂, p(CO₂), the rate of electron transport increased with p(CO₂). However, at high p(CO₂), switching from normal to low p(O₂) did not affect the net rate of photosynthetic CO₂ assimilation but the rate of electron-transport decreased by an amount related to the change in the rate of photorespiration. We interpret these effects as 1) regulation of ribulose-1,5-bisphosphatecarboxylase (RuBPCase, EC 4.1.1.39) activity to match the rate of electron transport at limiting PFD, 2) regulation of electron-transport rate to match the rate of RuBPCase at low p(CO₂), and 3) regulation of the electron-transport rate to match the capacity for starch and sucrose synthesis at high p(CO₂) and PFD. These studies provide evidence that PSII is regulated so that the capacity for electron transport is matched to the capacity for other processes required by photosynthesis, such as ribulose-bisphosphate carboxylation and starch and sucrose synthesis. We show that at least two mechanisms contribute to the regulation of PSII activity and that the relative engagement of these mechanisms varies with time following a step change in the capacity for ribulose-bisphosphate carboxylation and starch and sucrose synthesis. Finally, we take advantage of the relatively slow activation of deactivated RuBPCase in vivo to show that the activation level of this enzyme can limit the rate of electron transport as evidenced by increased feedback on PSII following a step change in p(CO₂). As RuBPCase as activated, the feedback on PSII declined.Abbreviations and symbols J_C electron-transport rate calculated from CO₂-assimilation measurements - J_F electron-transport rate calculated from fluorescence parameters - PFD photon-flux density - q_E energy-dependent quenching - PSII photosystem II - q_Q Q-dependent quenching - QY quantum yield - RuBPCase ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) C.I.W. publication No. 1015     

Effect of growth temperature on the electron flow for photorespiration in leaves of tobacco grown in the field

Photorespiration has been indicated as an important mechanism for maintaining CO₂ assimilation and alleviating photodamage under conditions of high light and low CO₂. We tested the hypothesis that plants grown under a high temperature had greater electron flow for photorespiration compared with those grown under a relative low temperature. Responses of photosynthetic electron flow and CO₂ assimilation to incident light intensity and intercellular CO₂ concentration were examined in leaves of tobacco cultivar ‘k326’. Plants were cultivated at three sites with different ambient temperatures (Zhengzhou, Zunyi and Jiangchuan). Under high light, plants grown in Zhengzhou (with the highest growth temperature in the three sites) showed higher effective quantum yield of photosystem II and total electron flow through photosystem II than that in Zunyi and Jiangchuan. However, regardless of light intensity and intercellular CO₂ status, there were no significant differences among sites in the photosynthetic CO₂ assimilation rate or electron flow devoted to the carboxylation of ribulose‐1,5‐bisphosphate (RuBP). As a result, plants grown at high temperature showed higher electron flow devoted to oxygenation of RuBP than plants grown at low temperature. These results suggested that enhancement of electron flow for photorespiration is an important strategy in tobacco for acclimating to high growth temperature.     

Specific reduction of chloroplast glyceraldehyde-3-phosphate dehydrogenase activity by antisense RNA reduces CO2 assimilation via a reduction in ribulose bisphosphate regeneration in transgenic tobacco plants

The reduction of 3-phosphoglycerate (PGA) to triose phosphate is a key step in photosynthesis linking the photochemical events of the thylakoid membranes with the carbon metabolism of the photosynthetic carbon-reduction (PCR) cycle in the stroma. Glyceraldehyde-3-phosphate dehydrogenase: NADP oxidoreductase (GAPDH) is one of the two chloroplast enzymes which catalyse this reversible conversion. We report on the engineering of an antisense RNA construct directed against the tobacco (Nicotiana tabacum L.) chloroplastlocated GAPDH (A subunit). The construct was integrated into the tobacco genome by Agrobacterium-mediated transformation of leaf discs. Of the resulting transformants, five plants were recovered with reduced GAPDH activities ranging from 11 to 24% of wild-type (WT) activities. Segregation analysis of the kanamycin-resistance character in self-pollinated T₁ seed from each of the five transformants revealed that one plant (GAP-R) had two active DNA inserts and the others had one insert. T₁ progeny from GAP-R was used to generate plants with GAPDH activities ranging from WT levels to around 7% of WT levels. These were used to study the effect of variable GAPDH activities on metabolite pools for ribulose1,5-bisphosphate (RuBP) and PGA, and the accompanying effects on the rate of CO₂ assimilation and other gasexchange parameters. The RuBP pool size was linearly related to GAPDH activity once GAPDH activity dropped below the range for WT plants, but the rate of CO₂ assimilation was not affected until RuBP levels dropped to 30–40% of WT levels. That is, the CO₂ assimilation rate fell when RuBP per ribulose-1,5-biphosphate carboxylase-oxygenase (Rubisco) site fell below 2 mol·(mol site)^–1 while the ratio for WT plants was 4–5 mol·m(mol site)^–1. Leaf conductance was not reduced in leaves with reduced GAPDH activities, resulting in an increase in the ratio of intercellular to ambient CO₂ partial pressure. Conductance in plants with reduced GAPDH activities was still sensitive to CO₂ and showed a normal decline with increases in CO₂ partial pressure. Although PGA levels did not fluctuate greatly, the effect of reduced GAPDH activity on RuBP-pool size and assimilation rate can be interpreted as being due to a blockage in the regeneration of RuBP. Concomitant gas-ex change and chlorophyll a fluorescence measurements indicated that photosynthesis changed from being Rubisco-limited to being RuBP-regeneration-limited at a lower CO₂ partial pressure in the antisense plants than in WT plants. Photosynthetic electron transport was down-regulated by the build-up of a large proton gradient and the electron-transport chain did not become over-reduced due to a shortage of NADP. Plants with severely reduced GAPDH activity were not photoinhibited despite the continuous presence of a large thylakoid proton gradient in the light. Along with plant size, Rubisco activity, leaf soluble protein and chlorophyll content were reduced in plants with the lowest GAPDH activities. We conclude that chloroplastic GAPDH activity does not appear to limit steady-state photosynthetic CO₂ assimilation at ambient CO₂. This is because WT leaves maintain the ratio of RuBP per Rubisco site about twofold higher than the level required to achieve a maximal rate of CO₂ assimilation.Abbreviations and Symbols bp base pairs - DHAP dihydroxy-acetone phosphate - GAPDH glyceraldehyde-3-phosphate dehy-drogenase - PCR photosynthetic carbon reduction - PGA 3-phosphoglycerate - p_i intercellular CO₂ partial pressure - q_NP non-photochemical fluorescence quenching - q_Q photochemicalfluorescence quenching - PSII quantum efficiency of electronflow through PSII - Rubisco ribulose-1,5-bisphosphate carboxy-lase-oxygenase - RuBP ribulose-1,5-bisphosphate - WT wild type We thank Karin Harrison, Prue Kell, Anne Gallagher and Barbara Setchell for excellent technical assistance. G.D.P. and S.V.C. acknowledge support from QE II Research Fellowships (Australian Research Council).     

Localization of photosynthetic metabolism in the parasitic angiosperm Cuscuta reflexa

Cells capable of photosynthesis in the parasitic angiosperm Cuscuta reflexa Roxb. (dodder) are highly localized. Immunolocalization of ribulose-1,5 bisphosphate carboxylase-oxygenase (Rubisco) and autofluorescence of chlorophyll in transverse sections of stems showed that they were largely restricted to a band of cells adjacent to the vascular bundles, consequently, the concentrations of Rubisco and chlorophyll were low per unit area or fresh weight. When ¹⁴CO₂ was supplied to stem segments of C. reflexa it preferentially accumulated in these cells adjacent to the vasculature. Although the conductance for CO₂ movement to the cells containing chlorophyll and Rubisco was very low, both the light reactions and dark reactions of photosynthesis appeared to be functional. De-epoxidation of the xanthophyll-cycle pigments after exposure to high light, and the chlorophyll fluorescence parameters, photochemical quenching (qP), non-photochemical quenching (NPQ) and the quantum efficiency of photosystem II (φPSII) responded normally to changes in photon flux density, indicating functional light-driven electron transport. The response of CO₂ exchange to photon flux density followed a typical hyperbolic curve, and positive rates of CO₂ fixation occurred when external CO₂ was increased to 5%. We propose that CO₂ for carbon assimilation is derived from internally respired CO₂ and that this layer of photosynthetic cells makes a positive contribution to the carbon budget of C. reflexa. Received: 23 October 1997 / Accepted: 16 December 1997     

Photosynthesis in the water-stressed C4 grass Setaria sphacelata is mainly limited by stomata with both rapidly and slowly imposed water deficits

A comparison of the effects of a rapid and a slowly imposed water deficit on photosynthesis was performed in Setaria sphacelata var. splendida (Stapf) Clayton, a C₄ NADP‐ME grass. Gas exchange was measured in rapidly and slowly dehydrated adult leaves either under atmospheric CO₂ partial pressure with an infrared gas analyser or under saturating CO₂ partial pressure with a leaf disc oxygen electrode. These measurements were used to calculate stomatal and non‐stomatal limitations to photosynthesis. These were further investigated using modulated chlorophyll a fluorescence measurements and photosynthetic pigment quantification. The decrease of net photosynthesis, leaf conductance and water use efficiency was more pronounced under rapid stress than in slow stress. However, photosynthesis is always mainly limited by stomata in both types of stress, albeit the contribution of non‐stomatal limitations increases at severe water deficits in slow stress experiments. The substomatal CO₂ partial pressure significantly increased in both types of stress, suggesting an increased resistance due to an internal barrier to CO₂ diffusion. Physical alterations in the structure of the intercellular spaces due to leaf shrinkage may account for these results. The maximal photochemical efficiency of photosystem II (PSII) was remarkably resistant to stress, as the F_v/F_m ratio decreased only at severe water deficit. On the contrary, the effective photochemical efficiency of PSII (ΔF/F′_m) measured under high actinic light decreased linearly in both types of stress, although in a more pronounced way under rapid stress. A similar variation in photochemical quenching suggests that the decrease of ΔF/F′_m is mainly due to the closure of PSII reaction centres. The non‐photochemical quenching did not change significantly except under severe dehydration indicating that the energization state of thylakoids remained stable under stress. The decrease observed in photosynthetic pigments may be an adaptation to stress rather than a limiting factor to photosynthesis. Results suggests that, although intrinsic mesophyll metabolic inhibitions occur, stomatal limitation to CO₂ diffusion is the main reason for the decrease in photosynthesis.     

Photosynthetic activity of homoiochlorophyllous desiccation tolerant plant <Emphasis Type="Italic">Haberlea rhodopensis</Emphasis> during dehydration and rehydration

The functional state of the photosynthetic apparatus of flowering homoiochlorophyllous desiccation tolerant plant Haberlea rhodopensis during dehydration and subsequent rehydration was investigated in order to characterize some of the mechanisms by which resurrection plants survive drought stress. The changes in the CO₂ assimilation rate, chlorophyll fluorescence parameters, thermoluminescence, fluorescence imaging and electrophoretic characteristics of the chloroplast proteins were measured in control, moderately dehydrated (50% water content), desiccated (5% water content) and rehydrated plants. During the first phase of desiccation the net CO₂ assimilation decline was influenced by stomatal closure. Further lowering of net CO₂ assimilation was caused by both the decrease in stomatal conductance and in the photochemical activity of photosystem II. Severe dehydration caused inhibition of quantum yield of PSII electron transport, disappearance of thermoluminescence B band and mainly charge recombination related to S₂Q_A⁻ takes place. The blue and green fluorescence emission in desiccated leaves strongly increased. It could be suggested that unchanged chlorophyll content and amounts of chlorophyll–proteins, reversible modifications in PSII electron transport and enhanced probability for non-radiative energy dissipation as well as increased polyphenolic synthesis during desiccation of Haberlea contribute to drought resistance and fast recovery after rehydration.     

The regulation of component processes of photosynthesis in transgenic tobacco with decreased phosphoribulokinase activity

Tobacco plants (Nicotiana tabacum L.) transformed with an inverted cDNA encoding ribulose 5-phosphate kinase (phosphoribulokinase,PRK; EC 2.7.1.19) were employed to study the in vivo relationship between photosynthetic electron transport and the partitioning of electron transport products to major carbon metabolism sinks under conditions of elevated ATP concentrations and limited ribulose 1,5-bisphosphate (RuBP) regeneration. Simultaneous measurements of room temperature chlorophyll fluorescence and CO₂ gas exchange were conducted on intact leaves. Under ambient CO₂ concentrations and light intensities above those at which the plants were grown, transformants with only 5% of PRK activity showed down-regulation of PS II activity and electron transport in response to a decrease in net carbon assimilation when compared to wild-type. This was manifested as a decline in the efficiency of PS II electron transport (_{PS II}), an increase in dissipation of excess absorbed light in the antennae of PS II and a decline in: total linear electron transport (J₁), electron transport dedicated to carbon assimilation (J_A) and electron transport allocated to photorespiration (J_L). The transformants showed no alteration in the Rubisco specificity factor measured in vitro and calculated in vivo but had a relatively smaller ratio of RuBP oxygenation to carboxylation rates (v_o/v_c), due to a higher CO₂ concentration at the carboxylation site (C_c). The relationship between _{PS II} and _{CO 2}was similar in transformants and wild-type under photorespiratory conditions demonstrating no change in the intrinsic relationship between PS II function and carbon assimilation, however, a novel result of this study is that this similar relationship occurred at different values of quantum flux, J₁, J_A, J_L and v_o/v_c in the transformant. For both wild-type and transformants, an assessment was made of the possible presence of a third major sink for electron transport products, beside RuBP oxygenation and carboxylation, the data provided no evidence for such a sink.Abbreviations C_c CO₂ concentration at the site of carboxylation - C_i intercellular CO₂ concentration - g_m mesophyll conductance to CO₂ - J₁ total linear electron flow - J_A linear electron flow allocated to CO₂ assimilation - J_c linear electron flow supporting carbon reduction and oxidation cycles - J_L linear electron flow allocated to photorespiration (RuBP oxygenation and fixation of released photorespiratory CO₂) - PRK phosphoribulokinase - q_P, q_N coefficients for photochemical and non-photochemical quenching of fluorescence respectively - Rubisco ribulose 1,5-bisphosphate carboxylase-oxygenase - S Rubisco specificity to CO₂/O₂ - v_c, v_o rates of RuBP carboxylation and RuBP oxygenation, respectively - _{CO 2} relative quantum yield of CO₂ assimilation - _C maximum _{CO 2} under non-photorespiratory conditions - _exc the efficiency of excitation capture by open PS II centres - _{PS II} relative quantum yield of PS II electron transport     

Studies on the limitations to photosynthesis in leaves of the atrazine-resistant mutant ofSenecio vulgaris L.

In leaves of an atrazine-resistant mutant ofSenecio vulgaris the quantum efficiency of CO₂ assimilation was reduced by 21% compared to the atrazine-susceptible wild type, and at a light level twice that required to saturate photosynthesis in the wild type the CO₂ fixation rate in the mutant was decreased by 15%. In leaves at steady-state photosynthesis there was a measurable increase in the reduction state of the photosystem II (PSII) primary quinone acceptor,Q _A. Although this would lead to a decreased rate of PSII electron transport and may thus explain the decrease in quantum efficiency, this cannot account for the fall in the maximum rate of CO₂ fixation. The atrazine-resistant mutant showed an appreciably longer photosynthetic induction time which indicates an effect on carbon metabolism; however, the response of CO₂-fixation rate to intercellular CO₂ concentration revealed no differences in carboxylation efficiency. There were also no differences in the ability to perform a State 1–State 2 transition between the atrazine-resistant and susceptible biotypes and no difference in the profiles of phosphorylated thylakoid polypeptides. It is concluded that the alteration of the redox equilibrium between PSII quinone electron acceptors in the atrazine-resistant biotype limits appreciably the photosynthetic efficiency in non-saturating light. Additionally, there is a further, as yet unidentified, limitation which decreases photosynthesis in the resistant mutant under light-saturating conditions.Abbreviations and symbols DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F _max maximum fluorescence emission - F _o2 minimal fluorescence emission upon exposure to saturating light flash - F _v variable fluorescence emission - F _v2 variable fluorescence emission upon exposure to saturating light flash - kDa kilodalton - PSI, II photosystems I, II - Q _A primary quinone acceptor of PSH - Q _B secondary quinone acceptor of PSII - RuBP ribulose-1,5-bisphosphate     

In vivo photosynthetic electron transport does not limit photosynthetic capacity in phosphate-deficient sunflower and maize leaves

The effects of extreme phosphate (Pi) deficiency during growth on the contents of adenylates and pyridine nucleotides and the in vivo photochemical activity of photosystem II (PSII) were determined in leaves of Helianthus annuus and Zea mays grown under controlled environmental conditions. Phosphate deficiency decreased the amounts of ATP and ADP per unit leaf area and the adenylate energy charge of leaves. The amounts of oxidized pyridine nucleotides per unit leaf area decreased with Pi deficiency, but not those of reduced pyridine nucleotides. This resulted in an increase in the ratio of reduced to oxidized pyridine nucleotides in Pi-deficient leaves. Analysis of chlorophyll a fluorescence at room temperature showed that Pi deficiency decreased the efficiency of excitation capture by open PSII reaction centres (φ_e), the in vivo quantum yield of PSII photochemistry (φ_PSII) and the photochemical quenching co-efficient (q_P), and increased the non-photochemical quenching co-efficient (q_N) indicating possible photoinhibitory damage to PSII. Supplying Pi to Pi-deficient sunflower leaves reversed the long-term effects of Pi-deficiency on PSII photochemistry. Feeding Pi-sufficient sunflower leaves with mannose or FCCP rapidly produced effects on chlorophyll a fluorescence similar to long-term Pi-deficiency. Our results suggest a direct role of Pi and photophosphorylation on PSII photochemistry in both long-and short-term responses of photosynthetic machinery to Pi deficiency. The relationship between φ_PSII and the apparent quantum yield of CO₂ assimilation determined at varying light intensity and 21 kPa O₂ and 35 Pa CO₂ partial pressures in the ambient air was linear in Pi-sufficient and Pi-deficient leaves of sunflower and maize. Calculations show that there was relatively more PSII activity per mole of CO₂ assimilated by the Pi-deficient leaves. This indicates that in these leaves a greater proportion of photosynthetic electrons transported across PSII was used for processes other than CO₂ reduction. Therefore, we conclude that in vivo photosynthetic electron transport through PSII did not limit photosynthesis in Pi-deficient leaves of sunflower and maize and that the decreased CO₂ assimilation was a consequence of a smaller ATP content and lower energy charge which restricted production of ribulose, 1-5, bisphosphate, the acceptor for CO₂.     

Photorespiration and photoprotection of grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L. cv. Cabernet Sauvignon) under water stress

In order to investigate the photoprotective function of photorespiration in grapevine under water stress, potted grapevines (Vitis vinifera L. cv. Cabernet Sauvignon) were randomly divided into three uniform groups for well-watered [watered every morning to keep the relative water content (RWC) of soil over 70 %], water-stress adapted (drought-adapted at 30 % relative soil water content for 30 days), and water stress without adaptation treatment (water-stressed to 30 % relative soil water content for 3 days). Net assimilation rate (A _N), stomatal conductance (g _s), substomatal CO₂ concentration (C _i), transpiration rate (E), actual photochemical efficiency of PSII (Φ_PSII), and maximum photochemical efficiency of PSII (F_v/F_m) were recorded by combining measurements of gas exchange and chlorophyll fluorescence. Gross photorespiration (P_r), photosynthetic electron partitioning (J_C/J_T), photochemical quenching coefficient (qP), and non-photochemical quenching (NPQ) were also calculated. The ratio of net assimilation rate to transpiration rate (A _N/E) was used as an indicator of water use efficiency (WUE). A _N, apparent P_r, Φ_PSII, F_v/F_m, qp, and g _s decreased, NPQ increased, and gross P_r sustained at a high level under water stress. This suggests that both photorespiration and energy dissipation play important roles in protecting photosynthetic apparatus against photoinhibition. C _i in water-stressed plants without adaptation treatment increased, which indicates the leaves suffered a non-stomatal limitation, while the water-stress adaped plants only suffered a stomatal limitation indicated by low C _i.     

Sinks for photosynthetic electron flow in green petioles and pedicels of Zantedeschia aethiopica: evidence for innately high photorespiration and cyclic electron flow rates

A combination of gas exchange and various chlorophyll fluorescence measurements under varying O₂ and CO₂ partial pressures were used to characterize photosynthesis in green, stomata-bearing petioles of Zantedeschia aethiopica (calla lily) while corresponding leaves served as controls. Compared to leaves, petioles displayed considerably lower CO₂ assimilation rates, limited by both stomatal and mesophyll components. Further analysis of mesophyll limitations indicated lower carboxylating efficiencies and insufficient RuBP regeneration but almost similar rates of linear electron transport. Accordingly, higher oxygenation/carboxylation ratios were assumed for petioles and confirmed by experiments under non-photorespiratory conditions. Higher photorespiration rates in petioles were accompanied by higher cyclic electron flow around PSI, the latter being possibly linked to limitations in electron transport from intermediate electron carriers to end acceptors and low contents of PSI. Based on chlorophyll fluorescence methods, similar conclusions can be drawn for green pedicels, although gas exchange in these organs could not be applied due to their bulky size. Since our test plants were not subjected to stress we argue that higher photorespiration and cyclic electron flow rates are innate attributes of photosynthesis in stalks of calla lily. Active nitrogen metabolism may be inferred, while increased cyclic electron flow may provide the additional ATP required for the enhanced photorespiratory activity in petiole and pedicel chloroplasts and/or the decarboxylation of malate ascending from roots.