In vivo temperature response functions of parameters required to model RuBP-limited photosynthesis

The leaf model of C₃ photosynthesis of Farquhar, von Caemmerer & Berry (Planta 149, 78–90, 1980) provides the basis for scaling carbon exchange from leaf to canopy and Earth‐System models, and is widely used to project biosphere responses to global change. This scaling requires using the leaf model over a wider temperature range than that for which the model was originally parameterized. The leaf model assumes that photosynthetic CO₂ uptake within a leaf is either limited by the rate of ribulose‐1,5‐bisphosphate (RuBP) regeneration or the activity of RuBP carboxylase‐oxygenase (Rubisco). Previously we reported a re‐parameterization of the temperature responses of Rubisco activity that proved robust when applied to a range of species. Herein this is extended to re‐parameterizing the response of RuBP‐limited photosynthesis to temperature. RuBP‐limited photosynthesis is assumed to depend on the whole chain electron transport rate, which is described as a three‐parameter non‐rectangular hyperbolic function of photon flux. Herein these three parameters are determined from simultaneous measurement of chlorophyll fluorescence and CO₂ exchange of tobacco leaves, at temperatures from 10 to 40 °C. All varied significantly with temperature and were modified further with variation in growth temperature from 15 to 35 °C. These parameters closely predicted the response of RuBP‐limited photosynthesis to temperature measured in both lemon and poplar and showed a significant improvement over predictions based on earlier parameterizations. We provide the necessary equations for use of the model of Farquhar et al. (1980) with our newly derived temperature functions for predicting both Rubisco‐ and RuBP‐limited photosynthesis.     

Low-temperature photosynthetic performance of a C4 grass and a co-occurring C3 grass native to high latitudes

The photosynthetic performance of C₄ plants is generally inferior to that of C₃ species at low temperatures, but the reasons for this are unclear. The present study investigated the hypothesis that the capacity of Rubisco, which largely reflects Rubisco content, limits C₄ photosynthesis at suboptimal temperatures. Photosynthetic gas exchange, chlorophyll a fluorescence, and the in vitro activity of Rubisco between 5 and 35 °C were measured to examine the nature of the low‐temperature photosynthetic performance of the co‐occurring high latitude grasses, Muhlenbergia glomerata (C₄) and Calamogrostis canadensis (C₃). Plants were grown under cool (14/10 °C) and warm (26/22 °C) temperature regimes to examine whether acclimation to cool temperature alters patterns of photosynthetic limitation. Low‐temperature acclimation reduced photosynthetic rates in both species. The catalytic site concentration of Rubisco was approximately 5.0 and 20 µmol m^?2 in M. glomerata and C. canadensis, respectively, regardless of growth temperature. In both species, in vivo electron transport rates below the thermal optimum exceeded what was necessary to support photosynthesis. In warm‐grown C. canadensis, the photosynthesis rate below 15 °C was unaffected by a 90% reduction in O₂ content, indicating photosynthetic capacity was limited by the capacity of P_i‐regeneration. By contrast, the rate of photosynthesis in C. canadensis plants grown at the cooler temperatures was stimulated 20–30% by O₂ reduction, indicating the P_i‐regeneration limitation was removed during low‐temperature acclimation. In M. glomerata, in vitro Rubisco activity and gross CO₂ assimilation rate were equivalent below 25 °C, indicating that the capacity of the enzyme is a major rate limiting step during C₄ photosynthesis at cool temperatures.     

Effects of growth and measurement light intensities on temperature dependence of CO2 assimilation rate in tobacco leaves

Effects of growth light intensity on the temperature dependence of CO₂ assimilation rate were studied in tobacco (Nicotiana tabacum) because growth light intensity alters nitrogen allocation between photosynthetic components. Leaf nitrogen, ribulose 1·5‐bisphosphate carboxylase/oxygenase (Rubisco) and cytochrome f (cyt f) contents increased with increasing growth light intensity, but the cyt f/Rubisco ratio was unaltered. Mesophyll conductance to CO₂ diffusion (g_m) measured with carbon isotope discrimination increased with growth light intensity but not with measuring light intensity. The responses of CO₂ assimilation rate to chloroplast CO₂ concentration (C_c) at different light intensities and temperatures were used to estimate the maximum carboxylation rate of Rubisco (V_cmax) and the chloroplast electron transport rate (J). Maximum electron transport rates were linearly related to cyt f content at any given temperature (e.g. 115 and 179 µmol electrons mol^?1 cyt f s^?1 at 25 and 40 °C, respectively). The chloroplast CO₂ concentration (C_trans) at which the transition from RuBP carboxylation to RuBP regeneration limitation occurred increased with leaf temperature and was independent of growth light intensity, consistent with the constant ratio of cyt f/Rubisco. In tobacco, CO₂ assimilation rate at 380 µmol mol^?1 CO₂ concentration and high light was limited by RuBP carboxylation above 32 °C and by RuBP regeneration below 32 °C.     

Low temperature effects on photosynthesis and growth of grapevine

Growth and photosynthesis of grapevine (Vitis vinifera L.) planted on two sloping cool climate vineyards were measured during the early growth season. At both vineyards, a small difference in mean minimum air temperature (1–3 °C) between two microsites accumulated over time, producing differences in shoot growth rate. The growth rates of the warmer (upper) microsite were 34–63% higher than the cooler (lower) site. Photosynthesis measurements of both east and west canopy sides revealed that the difference in carbon gain between the warmer and cooler microsites was due to low temperatures restricting the photosynthetic contribution of east‐facing leaves. East‐facing leaves at the warmer microsite experienced less time at suboptimal temperature while being exposed to high irradiance, contributing to an average 10% greater net carbon gain compared to the east‐facing leaves at the cooler microsite. This chilling‐induced reduction in photosynthesis was not due to net photo‐inhibition. Further analysis revealed that CO₂‐ and light‐saturated photosynthesis of grapevines was restricted by stomatal closure from 15 to 25 °C and by a limitation of RuBP regeneration and/or end‐product limitation from 5 to 15 °C. Changes in photosynthetic carboxylation efficiency implied that Rubisco activity may also play a regulatory role at all temperatures. This restriction of total photosynthetic carbon gain is proposed to be a major contributor to the temperature dependence of growth rate at both vineyards during the early season growth period.     

Photosynthetic performance at low temperature of Bouteloua gracilis Lag., a high-altitude C4 grass from the Rocky Mountains, USA

The mechanisms controlling the photosynthetic performance of C₄ plants at low temperature were investigated using ecotypes of Bouteloua gracilis Lag. from high (3000 m) and low (1500 m) elevation sites in the Rocky Mountains of Colorado. Plants were grown in controlled‐environment cabinets at a photon flux density of 700 μ mol m^?2 s^?1 and day/night temperatures of 26/16 °C or 14/7 °C. The thermal response of the net CO₂ assimilation rate (A) was evaluated using leaf gas‐exchange analysis and activity assays of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco), phosphoenolpyruvate carboxylase (PEPCase) and pyruvate,orthophosphate dikinase (PPDK). In both ecotypes, a reduction in measurement temperature caused the CO₂‐saturated rate of photosynthesis to decline to a greater degree than the initial slope of A versus the intercellular CO₂ response, thereby reducing the photosynthetic CO₂ saturation point. As a consequence, A in normal air was CO₂‐saturated at sub‐optimal temperatures. Ecotypic variation was low when grown at 26/16 °C, with the major difference between the ecotypes being that the low‐elevation plants had higher A; however, the ecotypes responded differently when grown at cool temperature. At temperatures below the thermal optimum, A in high‐elevation plants grown at 14/7 °C was enhanced relative to plants grown at 26/16 °C, while A in low‐elevation plants grown at 14/7 °C was reduced compared to 26/16 °C‐grown plants. Photoinhibition at low growth temperature was minor in both ecotypes as indicated by small reductions in dark‐adapted F_v/F_m. In both ecotypes, the activity of Rubisco was equivalent to A below 17 °C but well in excess of A above 25 °C. Activities of PEPCase and PPDK responded to temperature in a similar proportion relative to Rubisco, and showed no evidence for dissociation that would cause them to become principal limitations at low temperature. Because of the similar temperature response of Rubisco and A, we propose that Rubisco is a major limitation on C₄ photosynthesis in B. gracilis below 17 °C. Based on these results and for theoretical reasons associated with how C₄ plants use Rubisco, we further suggest that Rubisco capacity may be a widespread limitation upon C₄ photosynthesis at low temperature.     

Long- and short-term effects of decreased sink demand on carbohydrate levels and photosynthesis in wheat leaves

Wheat (Triticum aestivum L.) ears were removed to investigate long-term regulation of photosynthesis by sink demand at ambient CO₂ and 22 °C. The CO₂ level was also increased to 660 μmol mol^?1 and temperature was lowered to 5 °C to examine short-term responses of photosynthesis to low sink demand. Sink removal inhibited photosynthesis and increased leaf levels of glucose, fructose and ribulose-1, 5-bisphosphate (RuBP), and the glucose-6-phosphate (G6P)/fructose-6-phosphate (F6P) and RuBP/3-phosphoglycerate (PGA) ratios under growth conditions, but had no effect on the activity and activation state of ribulose-1, 5-bisphosphate carboxylase oxygenase (Rubisco) either under growth or short-term conditions, suggesting an inhibition of photosynthesis by decreased in vivo catalysis of Rubisco. Photosynthesis increased similarly in eared and earless shoots after a rise in CO₂ concentration, and the ratio of triose-phosphates (glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, TP) to PGA was similar or higher for removed than intact ears, suggesting that feedback inhibition of photosynthesis was not caused by a limitation of ATP synthesis in chloroplasts. Under short-term conditions (660 μmol mol^?1 CO₂, 5 °C), TP and RuBP levels and the TP/PGA and TP/RuBP ratios were increased by sink removal, indicating an additional limitation of photosynthesis by the rate of RuBP regeneration.     

Effect of heat stress on the inhibition and recovery of the ribulose-1,5-bisphosphate carboxylase/oxygenase activation state

Experiments were conducted to determine the relative contributions of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) activation state vis-à-vis Rubisco activase and metabolite levels to the inhibition of cotton (Gossypium hirsutum L.) photosynthesis by heat stress. Exposure of leaf tissue in the light to temperatures of 40 or 45 °C decreased the activation state of Rubisco to levels that were 65 or 10%, respectively, of the 28 °C control. Ribulose-1,5-bisphosphate (RuBP) levels increased in heat-stressed leaves, whereas the 3-phosphoglyceric acid pool was depleted. Heat stress did not affect Rubisco per se, as full activity could be restored by incubation with CO₂ and Mg²⁺. Inhibition and recovery of Rubisco activation state and carbon dioxide exchange rate (CER) were closely related under moderate heat stress (up to 42.5 °C). Moderate heat stress had negligible effect on Fv/Fm, the maximal quantum yield of photosystem II. In contrast, severe heat stress (45 °C) caused significant and irreversible damage to Rubisco activation, CER, and Fv/Fm. The rate of Rubisco activation after alleviating moderate heat stress was comparable to that of controls, indicating rapid reversibility of the process. However, moderate heat stress decreased both the rate and final extent of CER activation during dark-to-light transition. Treatment of cotton leaves with methyl viologen or an oxygen-enriched atmosphere reduced the effect of heat stress on Rubisco inactivation. Both treatments also reduced tissue RuBP levels, indicating that the amount of RuBP present during heat stress may influence the degree of Rubisco inactivation. Under both photorespiratory and non-photorespiratory conditions, the inhibition of the CER during heat stress could be completely reversed by increasing the internal partial pressure of CO₂ (Ci). However, the inhibition of the CER by nigericin, a K⁺ ionophore, was not reversible when the Ci was increased at ambient or high temperature. Our results indicate that inhibition of photosynthesis by moderate heat stress is not caused by inhibition of the capacity for RuBP regeneration. We conclude that heat stress inhibits Rubisco activation via a rapid and direct effect on Rubisco activase, possibly by perturbing Rubisco activase subunit interactions with each other or with Rubisco. Received: 25 February 2000 / Accepted: 13 May 2000     

In situ temperature relationships of biochemical and stomatal controls of photosynthesis in four lowland tropical tree species

Net photosynthetic carbon uptake of Panamanian lowland tropical forest species is typically optimal at 30–32 °C. The processes responsible for the decrease in photosynthesis at higher temperatures are not fully understood for tropical trees. We determined temperature responses of maximum rates of RuBP‐carboxylation (V_CMax) and RuBP‐regeneration (J_Max), stomatal conductance (G_s), and respiration in the light (R_Light) in situ for 4 lowland tropical tree species in Panama. G_s had the lowest temperature optimum (T_Opt), similar to that of net photosynthesis, and photosynthesis became increasingly limited by stomatal conductance as temperature increased. J_Max peaked at 34–37 °C and V_CMax ~2 °C above that, except in the late‐successional species Calophyllum longifolium, in which both peaked at ~33 °C. R_Light significantly increased with increasing temperature, but simulations with a photosynthesis model indicated that this had only a small effect on net photosynthesis. We found no evidence for Rubisco‐activase limitation of photosynthesis. T_Opt of V_CMax and J_Max fell within the observed in situ leaf temperature range, but our study nonetheless suggests that net photosynthesis of tropical trees is more strongly influenced by the indirect effects of high temperature—for example, through elevated vapour pressure deficit and resulting decreases in stomatal conductance—than by direct temperature effects on photosynthetic biochemistry and respiration.     

The temporal and species dynamics of photosynthetic acclimation in flag leaves of rice (Oryza sativa) and wheat (Triticum aestivum) under elevated carbon dioxide

In this study, we tested for the temporal occurrence of photosynthetic acclimation to elevated [CO₂] in the flag leaf of two important cereal crops, rice and wheat. In order to characterize the temporal onset of acclimation and the basis for any observed decline in photosynthetic rate, we characterized net photosynthesis, g_s, g_m, C_i/C_a, C_i/C_c, V_cmax, J_max, cell wall thickness, content of Rubisco, cytochrome (Cyt) f, N, chlorophyll and carbohydrate, mRNA expression for rbcL and petA, activity for Rubisco, sucrose phosphate synthase (SPS) and sucrose synthase (SS) at full flag expansion, mid‐anthesis and the late grain‐filling stage. No acclimation was observed for either crop at full flag leaf expansion. However, at the mid‐anthesis stage, photosynthetic acclimation in rice was associated with RuBP carboxylation and regeneration limitations, while wheat only had the carboxylation limitation. By grain maturation, the decline of Rubisco content and activity had contributed to RuBP carboxylation limitation of photosynthesis in both crops at elevated [CO₂]; however, the sharp decrease of Rubisco enzyme activity played a more important role in wheat. Although an increase in non‐structural carbohydrates did occur during these later stages, it was not consistently associated with changes in SPS and SS or photosynthetic acclimation. Rather, over time elevated [CO₂] appeared to enhance the rate of N degradation and senescence so that by late‐grain fill, photosynthetic acclimation to elevated [CO₂] in the flag leaf of either species was complete. These data suggest that the basis for photosynthetic acclimation with elevated [CO₂] may be more closely associated with enhanced rates of senescence, and, as a consequence, may be temporally dynamic, with significant species variation.     

Temperature acclimation of photosynthesis in spinach leaves: analyses of photosynthetic components and temperature dependencies of photosynthetic partial reactions

Spinach (Spinacia oleracea) plants were grown under the day/night temperature regime of 15/10 °C (LT) or 30/25 °C (HT). The plants were also transferred from HT to LT when the sample leaves were at particular developmental stages (HL-transfer). With fully mature leaves, the light-saturated photosynthetic rate (A) at the ambient CO₂ concentration (C_a) of 1500 µL L⁻¹ (A₁₅₀₀) and the initial slope of A versus intercellular CO₂ concentration (C_i) at low C_i region (IS) were obtained to assess capacities of RuBP regeneration and carboxylation. Photosynthetic components including Rubisco and cytochrome f (Cyt f) were also determined. The optimum temperatures for A at C_a of 360 µL L⁻¹ (A₃₆₀), A₁₅₀₀ and IS in HT leaves were 27, 36 and 24 °C, whereas those in LT leaves were 18, 30 and 18 °C. The optimum temperatures in HL-transfer leaves approached those of LT leaves with the increase in the duration at LT. The shift in the optimum temperature was greater and quicker for IS than A₁₅₀₀. By the HL-transfer, the maximum values of A₁₅₀₀ and IS also increased. The maximum A₁₅₀₀ and Cyt f content increased more promptly than IS and Rubisco content. Changes in the Cyt f/Rubisco ratio were reflected to those in the A₁₅₀₀/IS ratio. Taken together, photosynthetic acclimation to low temperature in spinach leaves was due not only to the change in the balance of the absolute rates of RuBP regeneration and carboxylation but also to the large change in the optimum temperature of RuBP carboxylation.     

Strong thermal acclimation of photosynthesis in tropical and temperate wet‐forest tree species: the importance of altered Rubisco content

Understanding of the extent of acclimation of light‐saturated net photosynthesis (A_n) to temperature (T), and associated underlying mechanisms, remains limited. This is a key knowledge gap given the importance of thermal acclimation for plant functioning, both under current and future higher temperatures, limiting the accuracy and realism of Earth system model (ESM) predictions. Given this, we analysed and modelled T‐dependent changes in photosynthetic capacity in 10 wet‐forest tree species: six from temperate forests and four from tropical forests. Temperate and tropical species were each acclimated to three daytime growth temperatures (T_growth): temperate – 15, 20 and 25 °C; tropical – 25, 30 and 35 °C. CO₂ response curves of A_n were used to model maximal rates of RuBP (ribulose‐1,5‐bisphosphate) carboxylation (V_cmax) and electron transport (J_max) at each treatment's respective T_growth and at a common measurement T (25 °C). SDS‐PAGE gels were used to determine abundance of the CO₂‐fixing enzyme, Rubisco. Leaf chlorophyll, nitrogen (N) and mass per unit leaf area (LMA) were also determined. For all species and T_growth, A_n at current atmospheric CO₂ partial pressure was Rubisco‐limited. Across all species, LMA decreased with increasing T_growth. Similarly, area‐based rates of V_cmax at a measurement T of 25 °C (V_cmax²⁵) linearly declined with increasing T_growth, linked to a concomitant decline in total leaf protein per unit leaf area and Rubisco as a percentage of leaf N. The decline in Rubisco constrained V_cmax and A_n for leaves developed at higher T_growth and resulted in poor predictions of photosynthesis by currently widely used models that do not account for T_growth‐mediated changes in Rubisco abundance that underpin the thermal acclimation response of photosynthesis in wet‐forest tree species. A new model is proposed that accounts for the effect of T_growth‐mediated declines in V_cmax²⁵ on A_n, complementing current photosynthetic thermal acclimation models that do not account for T sensitivity of V_cmax²⁵.     

The temperature response of CO2 assimilation, photochemical activities and Rubisco activation in Camelina sativa, a potential bioenergy crop with limited capacity for acclimation to heat stress

The temperature optimum of photosynthesis coincides with the average daytime temperature in a species’ native environment. Moderate heat stress occurs when temperatures exceed the optimum, inhibiting photosynthesis and decreasing productivity. In the present study, the temperature response of photosynthesis and the potential for heat acclimation was evaluated for Camelina sativa, a bioenergy crop. The temperature optimum of net CO₂ assimilation rate (A) under atmospheric conditions was 30–32?°C and was only slightly higher under non-photorespiratory conditions. The activation state of Rubisco was closely correlated with A at supra-optimal temperatures, exhibiting a parallel decrease with increasing leaf temperature. At both control and elevated temperatures, the modeled response of A to intercellular CO₂ concentration was consistent with Rubisco limiting A at ambient CO₂. Rubisco activation and photochemical activities were affected by moderate heat stress at lower temperatures in camelina than in the warm-adapted species cotton and tobacco. Growth under conditions that imposed a daily interval of moderate heat stress caused a 63?% reduction in camelina seed yield. Levels of cpn60 protein were elevated under the higher growth temperature, but acclimation of photosynthesis was minimal. Inactivation of Rubisco in camelina at temperatures above 35?°C was consistent with the temperature response of Rubisco activase activity and indicated that Rubisco activase was a prime target of inhibition by moderate heat stress in camelina. That photosynthesis exhibited no acclimation to moderate heat stress will likely impact the development of camelina and other cool season Brassicaceae as sources of bioenergy in a warmer world.     

Photosynthetic downregulation in leaves of the Japanese white birch grown under elevated CO2 concentration does not change their temperature‐dependent susceptibility to photoinhibition

To determine the effects of elevated CO₂ concentration ([CO₂]) on the temperature‐dependent photosynthetic properties, we measured gas exchange and chlorophyll fluorescence at various leaf temperatures (15, 20, 25, 30, 35 and 40°C) in 1‐year‐old seedlings of the Japanese white birch (Betula platyphylla var. japonica), grown in a phytotron under natural daylight at two [CO₂] levels (ambient: 400 µmol mol^?1 and elevated: 800 µmol mol^?1) and limited N availability (90 mg N plant^?1). Plants grown under elevated [CO₂] exhibited photosynthetic downregulation, indicated by a decrease in the carboxylation capacity of Rubisco. At temperatures above 30°C, the net photosynthetic rates of elevated‐CO₂‐grown plants exceeded those grown under ambient [CO₂] when compared at their growth [CO₂]. Electron transport rates were significantly lower in elevated‐CO₂‐grown plants than ambient‐CO₂‐grown ones at temperatures below 25°C. However, no significant difference was observed in the fraction of excess light energy [(1 ? q_P)× F_v′/F_m′] between CO₂ treatments across the temperature range. The quantum yield of regulated non‐photochemical energy loss was significantly higher in elevated‐CO₂‐grown plants than ambient, when compared at their respective growth [CO₂] below 25°C. These results suggest that elevated‐CO₂‐induced downregulation might not exacerbate the temperature‐dependent susceptibility to photoinhibition, because reduced energy consumption by electron transport was compensated for by increased thermal energy dissipation at low temperatures.     

The temperature acclimation of photosynthetic responses to CO2 in Zea mays and its relationship to the activities of photosynthetic enzymes and the CO2-concentrating mechanism of C4 photosynthesis

Abstract Associations between photosynthetic responses to CO₂ at rate-saturating light and photosynthetic enzyme activities were compared for leaves of maize grown under constant air temperatures of 19, 25 and 31°C. Key photosynthetic enzymes analysed were ribulose bisphosphatc (RuBP) carboxylase, phosphoenolpyruvate (PEP) carboxylase, NADP-malic enzyme and pyruvate, P_i dikinasc. Rates of CO₂-saturated photosynthesis were similar in leaves developed at 19°C and 25°C but were decreased significantly by growth at 31°C. In contrast, carboxylation efficiency differed significantly between all three temperature regimes. Carboxylation efficiency was greatest in leaves developed at 19°C and decreased with increasing temperature during growth. The changes of carboxylation efficiency were highly correlated with changes in the activity of pyruvate, P_i dikinase (r= 0.95), but not with other photosynthetic enzyme activities. The activities of these latter enzymes, including that of RuBP carboxylase, were relatively insensitive to temperature during growth. The sensitivity of quantum yield to O₂ concentration was lower in leaves grown at 19°C than in leaves grown at 31°C. These observations support the novel hypothesis that variation in the capacity for CO₂ delivery to the bundle sheath by the C₄ cycle, relative to the capacity for net assimilation by the C₂ cycle, can be a principal determinant of C₄ photosynthetic responses to CO₂.     

The relationship between CO2 assimilation, photosynthetic electron transport and water–water cycle in chill-exposed cucumber leaves under low light and subsequent recovery

The effects of chilling under low light (9/7 °C, 100 µmol m^?2 s^?1) on the photosynthetic and antioxidant capacities and subsequent recovery were examined in two (one tolerant and one sensitive) cucumber genotypes. Chilling resulted in an irreversible inhibition of net CO₂ assimilation and growth for the sensitive genotype, which was accompanied by decreases in the maximum velocity of RuBP carboxylation by Rubisco (V_cmax), the capacity for ribulose‐1,5‐bisphosphate regeneration (J_max), Rubisco content and activity, and the quantum efficiency of photosystem II, in the absence of any stomatal limitation of CO₂ supply or inorganic phosphate limitation. In contrast, CO₂ assimilation for the tolerant genotype fully recovered after chill. The chill‐induced decrease in the proportion of electron flux for photosynthetic carbon reduction was mostly compensated by an O₂‐dependent alternative electron flux driven by the water–water cycle, especially in the sensitive genotype. Compared with the tolerant genotype, the sensitive genotype after chill showed reduced capacity for scavenging reactive oxygen species and increased accumulation of reactive oxygen species. The balance between O₂‐dependent alternative electron flux and the capacity for scavenging reactive oxygen species in response to chill plays a major role in determining the tolerance of cucumber leaves to this stress factor. It is concluded that the water–water cycle operates at high rates when CO₂ assimilation is restricted in cucumber leaves subjected to chill and low light conditions.     

Polygonum sachalinense alters the balance between capacities of regeneration and carboxylation of ribulose‐1,5‐bisphosphate in response to growth CO2 increment but not the nitrogen allocation within the photosynthetic apparatus

The limiting step of photosynthesis changes depending on CO₂ concentration and, in theory, photosynthetic nitrogen use efficiency at a respective CO₂ concentration is maximized if nitrogen is redistributed from non‐limiting to limiting processes. It has been shown that some plants increase the capacity of ribulose‐1,5‐bisphoshate (RuBP) regeneration (evaluated as J_max) relative to the RuBP carboxylation capacity (evaluated as V_cmax) at elevated CO₂, which is in accord with the theory. However, there is no study that tests whether this change is accompanied by redistribution of nitrogen in the photosynthetic apparatus. We raised a perennial plant, Polygonum sachalinense, at two nutrient availabilities under two CO₂ concentrations. The J_max to V_cmax ratio significantly changed with CO₂ increment but the nitrogen allocation among the photosynthetic apparatus did not respond to growth CO₂. Enzymes involved in RuBP regeneration might be more activated at elevated CO₂, leading to the higher J_max to V_cmax ratio. Our result suggests that nitrogen partitioning is not responsive to elevated CO₂ even in species that alters the balance between RuBP regeneration and carboxylation. Nitrogen partitioning seems to be conservative against changes in growth CO₂ concentration.     

Effects of elevated CO2 and temperature on photosynthesis and Rubisco in rice and soybean

Rice (Oryza sativa L. cv. IR-72) and soybean (Glycine max L. Merr. cv. Bragg), which have been reported to differ in acclimation to elevated CO₂, were grown for a season in sunlight at ambient and twice-ambient [CO₂], and under daytime temperature regimes ranging from 28 to 40°C. The objectives of the study were to test whether CO₂ enrichment could compensate for adverse effects of high growth temperatures on photosynthesis, and whether these two C₃ species differed in this regard. Leaf photosynthetic assimilation rates (A) of both species, when measured at the growth [CO₂], were increased by CO₂ enrichment, but decreased by supraoptimal temperatures. However, CO₂ enrichment more than compensated for the temperature-induced decline in A. For soybean, this CO₂ enhancement of A increased in a linear manner by 32–95% with increasing growth temperatures from 28 to 40°C, whereas with rice the degree of enhancement was relatively constant at about 60%, from 32 to 38°C. Both elevated CO₂ and temperature exerted coarse control on the Rubisco protein content, but the two species differed in the degree of responsiveness. CO₂ enrichment and high growth temperatures reduced the Rubisco content of rice by 22 and 23%, respectively, but only by 8 and 17% for soybean. The maximum degree of Rubisco down-regulation appeared to be limited, as in rice the substantial individual effects of these two variables, when combined, were less than additive. Fine control of Rubisco activation was also influenced by both elevated [CO₂] and temperature. In rice, total activity and activation were reduced, but in soybean only activation was lowered. The apparent catalytic turnover rate (K_cat) of rice Rubisco was unaffected by these variables, but in soybean elevated [CO₂] and temperature increased the apparent K_cat by 8 and 22%, respectively. Post-sunset declines in Rubisco activities were accelerated by elevated [CO₂] in rice, but by high temperature in soybean, suggesting that [CO₂] and growth temperature influenced the metabolism of 2-carboxyarabinitol-1-phosphate, and that the effects might be species-specific. The greater capacity of soybean for CO₂ enhancement of A at supraoptimal temperatures was probably not due to changes in stomatal conductance, but may be partially attributed to less down-regulation of Rubisco by elevated [CO₂] in soybean than in rice. However, unidentified species differences in the temperature optimum for photosynthesis also appeared to be important. The responses of photosynthesis and Rubisco in rice and soybean suggest that among C₃ plants species-specific differences will be encountered as a result of future increases in global [CO₂] and air temperatures.     

Increased invasive potential of non‐native Phragmites australis: elevated CO2 and temperature alleviate salinity effects on photosynthesis and growth

The prospective rise in atmospheric CO₂ and temperature may change the distribution and invasive potential of a species; and intraspecific invasive lineages may respond differently to climate change. In this study, we simulated a future climate scenario with simultaneously elevated atmospheric CO₂ and temperature, and investigated its interaction with soil salinity, to assess the effects of global change on the ecophysiology of two competing haplotypes of the wetland grass Phragmites australis, that are invasive in the coastal marshes of North America. The two haplotypes with the phenotypes ‘EU‐type’ (Eurasian haplotype) and ‘Delta‐type’ (Mediterranean haplotype), were grown at 0‰ and 20‰ soil salinity, and at ambient or elevated climatic conditions (700 ppm CO₂, +5 °C) in a phytotron system. The aboveground growth of both phenotypes was highest at the elevated climatic conditions. Growth at 20‰ salinity resulted in declined aboveground growth, lower transpiration rates (E), stomata conductance (g_s), specific leaf area, photosynthetic pigment concentrations, and a reduced photosynthetic performance. The negative effects of salinity were, however, significantly less severe at elevated CO₂ and temperature than at the ambient climatic conditions. The Delta‐type P. australis had higher shoot elongation rates than the EU‐type P. australis, particularly at high salinity. The Delta‐type also had higher maximum light‐saturated rates of photosynthesis (A_sat), maximum carboxylation rates of Rubisco (V_cmax), maximum electron transport rates (J_max), triose phosphate utilization rates (T_p), stomata conductance (g_s), as well as higher Rubisco carboxylation‐limited, RuBP regeneration‐limited and T_p‐regeneration limited CO₂ assimilation rates than the EU‐type under all growth conditions. Our results suggest that the EU‐type will not become dominant over the Delta‐type, since the Delta‐type has superior ecophysiological traits. However, the projected rise in atmospheric CO₂ and temperature will alleviate the effects of salinity on both phenotypes and facilitate their expansion into more saline areas.     

Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubisco and expression of photosynthetic genes in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Brassinosteroids (BRs) are a new group of plant growth substances that promote plant growth and productivity. We showed in this study that improved growth of cucumber (Cucumis sativus) plants after treatment with 24-epibrassinolide (EBR), an active BR, was associated with increased CO₂ assimilation and quantum yield of PSII (Φ_PSII). Treatment of brassinazole (Brz), a specific inhibitor for BR biosynthesis, reduced plant growth and at the same time decreased CO₂ assimilation and Φ_PSII. Thus, the growth-promoting activity of BRs can be, at least partly, attributed to enhanced plant photosynthesis. To understand how BRs enhance photosynthesis, we have analyzed the effects of EBR and Brz on a number of photosynthetic parameters and their affecting factors, including the contents and activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Northern and Western blotting demonstrated that EBR upregulated, while Brz downregulated, the expressions of rbcL, rbcS and other photosynthetic genes. In addition, EBR had a positive effect on the activation of Rubisco based on increased maximum Rubisco carboxylation rates (V _c,max), total Rubisco activity and, to a greater extent, initial Rubisco activity. The accumulation patterns of Rubisco activase (RCA) based on immunogold-labeling experiments suggested a role of RCA in BR-regulated activation state of Rubisco. Enhanced expression of genes encoding other Calvin cycle genes after EBR treatment may also play a positive role in RuBP regeneration (J _max), thereby increasing maximum carboxylation rate of Rubisco (V _c,max). Thus, BRs promote photosynthesis and growth by positively regulating synthesis and activation of a variety of photosynthetic enzymes including Rubisco in cucumber.     

Thylakoid membrane responses to moderately high leaf temperature in Pima cotton

Photosynthesis is inhibited by high temperatures that plants are likely to experience under natural conditions. Both increased thylakoid membrane ionic conductance and ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) deactivation have been suggested as the primary cause. The moderately heat‐tolerant crop Pima S‐6 cotton (Gossypium barbadense) was used to examine heat stress‐induced inhibition of photosynthesis. Previous field‐work indicated that moderate heat stress (T = 35–45 °C) is associated with very rapid leaf temperature changes. Therefore, a system was devised for rapidly heating intact, attached leaves to mimic natural field heat‐stress conditions and monitored Rubisco activation, carbon‐cycle metabolites, thylakoid ionic conductance, and photosystem I activity. As a proxy for NADPH and stromal redox status the activation state of NADP‐malate dehydrogenase (NADP‐MDH) was measured. In dark‐adapted cotton leaves, heating caused an increase in thylakoid permeability at temperatures as low as 36 °C. The increased permeability did not cause a decline in adenosine 5′‐triphosphate (ATP) levels during steady‐state or transient heating. Rapid heating caused a transient decline in ribulose 1,5‐bisphosphate without a decrease in Rubisco activation. Sustained heating caused a decline in Rubisco activation and also oxidized the stroma as judged by NADP‐MDH activation and this is hypothesized to result from increased cyclic photophosphorylation, explaining the maintenance of ATP content in the face of increased thylakoid membrane ion leakiness.