Carbon balance during CAM: an assessment of respiratory CO2 recycling in the epiphytic bromeliads Aechmea nudicaulis and Aechmea fendleri

Abstract The regulation of crassulacean acid metabolism (CAM) under controlled environmental conditions has been investigated for two tropical epiphytes, relating plant water and carbon balance to growth form and habitat preference under natural conditions. Aechmea fendleri is restricted to wet, upper montane regions of Trinidad, while A. nudicaulis has a wider distribution extending into more arid regions of the island. Morphological characteristics of these plants are related to habitat preference in terms of leaf succulence (0.44 and 0.94 kg m^?2 for the two species respectively) and a distinct layer of water storage parenchyma in A. nudicaulis In contrast, the thinner leaves of A. fendleri contain little water-storage parenchyma and less chlorenchyma per unit area, but the plants have a more open leaf rosette. The two species differ in expression of CAM, since the proportion of respiratory CO₂ recycled as part of CAM had been found to be much lower in A. fendleri This study compared the efficiency of water use and role of respiratory CO₂ recycling under two PAR regimes (300 and 120 μnol m^?2 s^?1) and three night temperatures (12, 18 and 25 °C). Dark CO₂ uptake rates for both species were comparable to plants in the field (maximum of 2.3 ± 0.2 μmol m^?2s^?1± SD, n= 3). Total net CO₂ uptake at night increased on leaf area basis with temperature for both species under higher PAR, although under the low PAR regime CO₂ uptake was maximal at 18 °C. Water-use efficiency (WUE) increased at 18 °C and 25 °C during dark CO₂ uptake (Phase I) and also during late afternoon photosynthesis (Phase IV) in both species. For A. fendleri, dawn to dusk changes in titrable acidity (ΔH ⁺) were similar under high and low PAR, although ΔH⁺ was correlated to night temperature and PAR in A. nudicaulis. The proportion of ΔH⁺ derived from respiratory CO₂ also varied with experimental conditions. Thus percentage recycling was lower in A. fendleri under high PAR (0–10%), but was only reduced at 18 °C under low PAR. Recycling by A. nudicaulis ranged from 32–42% under high PAR, but was also reduced to 6% under low PAR at 18 °C; at 12 °C and 25 °C, recycling was 37% and 52% respectively. Previous studies have suggested a relationship between the proportion of recycling and degree of water stress. This study indicated that CAM as a CO₂ concentrating mechanism regulates both water-use efficiency and plant carbon balance in these epiphytes, in response to PAR and night temperature. However, the precise relationship between respiratory processes and the balance between external and internal sources of CO₂ is as yet unresolved.     

Interaction between light and chilling temperature on the inhibition of photosynthesis in chilling-sensitive plants*

Abstract. Fully expanded leaves of 25°C grown Phaseolus vulgaris and six other species were exposed for 3 h to chilling temperatures at photon flux densities equivalent to full sunlight. In four of the species this treatment resulted in substantial inhibition of the subsequent quantum yield of CO₂ uptake, indicating reduction of the photochemical efficiency of photosynthesis. The extent of inhibition was dependent on the photon flux density during chilling and no inhibition occurred when chilling occurred at a low photon flux density. No inhibition occurred at temperatures above 11.5°C, even in the presence of the equivalent of full sunlight. This interaction between chilling and light to cause inhibition of photosynthesis was promoted by the presence of oxygen at normal air partial pressures and was unaffected by the CO₂ partial pressure present when chilling occurred in air. When chilling occurred at low O₂ partial pressures, CO₂ was effective in reducing the degree of inhibition. Apparently, when leaves of chilling-sensitive plants are exposed to chilling temperatures in air of normal composition then light is instrumental in inducing rapid damage to the photochemical efficiency of photosynthesis.     

CO2 uptake and fluorescence responses for a shade-tolerant cactus Hylocereus undatus under current and doubled CO2 concentrations

Hylocereus undatus (Haworth) Britton and Rose growing in controlled environment chambers at 370 and 740 μmol CO₂ mol^?1 air showed a Crassulacean acid metabolism (CAM) pattern of CO₂ uptake, with 34% more total daily CO₂ uptake under the doubled CO₂ concentration and most of the increase occurring in the late afternoon. For both CO₂ concentrations, 90% of the maximal daily CO₂ uptake occurred at a total daily photosynthetic photon flux density (PPFD) of only 10 mol m^?2 day^?1 and the best day/night air temperatures were 25/15°C. Enhancement of the daily net CO₂ uptake by doubling the CO₂ concentration was greater under the highest PPFD (30 mol m^?2 day^?1) and extreme day/night air temperatures (15/5 and 45/35°C). After 24 days of drought, daily CO₂ uptake under 370 μmol CO₂ mol^?1 was 25% of that under 740 μmol CO₂ mol^?1. The ratio of variable to maximal chlorophyll fluorescence (F_y/F_m) decreased as the PPFD was raised above 5 mol m^?2 day^?1, at extreme day/night temperatures and during drought, suggesting that stress occurred under these conditions. F_v/F_m was higher under the doubled CO₂ concentration, indicating that the current CO₂ concentration was apparently limiting for photosynthesis. Thus net CO₂ uptake by the shade-tolerant H. undatus, the photosynthetic efficiency of which was greatest at low PPFDs. showed a positive response to doubling the CO₂ concentration, especially under stressful environmental conditions.     

Detection of QTLs associated with shoot wilting and root ammonium uptake under chilling temperatures in an interspecific backcross population from Lycopersicon esculentum ×L. hirsutum

The genetic basis for shoot wilting and root ammonium uptake under chilling temperatures was examined in an interspecific backcross (BC₁) population derived from Lycopersicon esculentum Mill. cv T5 and wild Lycopersicon hirsutum f. typicum accession LA1778. The chilling sensitivity of shoot wilting and ammonium uptake was evaluated in four replicated cuttings from each of 196 BC₁ plants. Wilting was evaluated at two different times: 2 hours (wilting 2 h) and 6 hours (wilting 6 h recovery) after root exposure to 4°C. The BC₁ plants were genotyped with 89 polymorphic RFLP markers, and composite interval mapping was used to detect quantitative trait loci (QTLs). Three QTLs, one each on chromosomes 5, 6 and 9, were detected for wilting 2 h. The presence of a L. hirsutum (H) allele at the QTL on chromosomes 5 and 9 decreased wilting, while the H allele at the QTL on chromosome 6 increased wilting. To analyze plant recovery from wilting at 6 h, subsets of the BC₁ population were selected, based on phenotype and genotype, because not all plants wilted at 2 h. The phenotype subset (wilting 6 h-PS) included plants that wilted to a greater degree at 2 h, and the genotype subsets included plants carrying specific allelic compositions at the QTL for wilting 2 h on chromosomes 5 (wilting 6 h-GS-ch5), 6 (wilting 6 h-GS-ch6), and 9 (wilting 6 h-GS-ch9). On chromosome 6, a QTL was located that was associated with three subsets (wilting 6 h-PS, wilting 6 h-GS-ch5 and wilting 6 h-GS-ch9), while on chromosome 7 a QTL was detected with two subsets (wilting 6 h-PS and wilting 6 h-GS-ch5). Three additional QTLs were detected within a single subset: chromosome 1 (wilting 6 h-GS-ch6), chromosome 11 (wilting 6 h-GS-ch5) and chromosome 12 (wilting 6 h-GS-ch9). The presence of the H allele at the QTL on chromosomes 7 and 12 had a positive effect, enhancing recovery from wilting, while the H allele at the other QTL had a negative effect. Three traits were used to evaluate the chilling sensitivity of root ammonium uptake: ammonium uptake before a chilling episode, ammonium uptake after the chilling episode, and the relative inhibition of uptake (difference in uptake rates before and after chilling divided by the rate before chilling). One QTL was detected on chromosome 3 for the rate before chilling and one on chromosome 6 for the relative inhibition of ammonium uptake. Our results demonstrate that shoot wilting and ammonium uptake under chilling are controlled by multiple QTLs. Received: 10 August 1999 / Accepted: 25 March 2000     

The effects of O2 on net photosynthesis at low temperature (5°C)

Abstract. It has been shown that atmospheric O₂ can either depress or stimulate the rate of apparent photosynthesis of white mustard depending on the environmental conditions: CO₂ concentration, light intensity and temperature. Stimulation by O₂ was observed only under high photon fluence rate and at high CO₂ concentrations. The critical CO₂ concentration below which O₂ was inhibiting and above which it was stimulating was dependent on the temperature of the assay: for plants grown at 12°C the critical CO₂ concentration was 13.35 mmol at 5° C and 21.92 mmol at 10° C. Stimulation by O₂ depended also on the growth temperature: for measurements at 26.31 mmol m^?3 CO₂, O₂ was stimulating at temperatures less than 12°C for plants grown at 12°C and less than 19°C for plants grown at 27°C. The efficiency of the O₂-dependent stimulation of net photosynthesis was maximum at 9.21 mol m^?3 O₂ at 26.31 mmol m^?3 CO₂. Oxygen-stimulation of net photosynthesis was detected in Nicotiana tabacum L. var Samsun, Lycopersicum esculentum L. and Chenopodium album L. At 5°C and under high photon fluence rate, O₂ increased the carboxylation capacity of the photosynthetic apparatus of mustard and decreased its affinity for CO₂. The O₂ inhibition of the net CO₂ uptake observed at low CO₂ concentrations was the result of a decrease in the affinity for carbon dioxide. The nature of the mechanism which causes the stimulation of photosynthesis is discussed.     

Overexpression of bacterial catalase in tomato leaf chloroplasts enhances photo-oxidative stress tolerance

The Escherichia coli gene katE, which is driven by the promoter of the Rubisco small subunit gene of tomato, rbcS3C, was introduced into a tomato (Lycopersicon esculentum Mill.) by Agrobacterium tumefaciens‐mediated transformation. Catalase activity in progeny from transgenic plants was approximately three‐fold higher than that in wild‐type plants. Leaf discs from transgenic plants remained green at 24 h after treatment with 1 µm paraquat under moderate light intensity, whereas leaf discs from wild‐type plants showed severe bleaching after the same treatment. Moreover, ion leakage from transgenic leaf discs was significantly less than that from wild‐type leaf discs at 24 h after treatment with 1 µm paraquat and 10 mm H₂O₂, respectively, under moderate light intensity. To evaluate the efficiency of the E. coli catalase to protect the whole transgenic plant from the oxidative stress, transgenic and wild‐type plants were sprayed with 100 µm paraquat and exposed to high light illumination (800 µmol m^?2 s^?1). After 24 h, the leaves of the transgenic plants were less damaged than the leaves of the wild‐type plants. The catalase activity and the photosynthesis activity (indicated by the F_v/F_m ratio) were less affected by paraquat treatment in leaves of transgenic plants, whereas the activities of the chloroplastic ascorbate peroxidase isoenzymes and the ascorbate content decreased in both lines. In addition, the transgenic plants showed increased tolerance to the oxidative damage (decrease of the CO₂ fixation and photosystem II activity and increase of the lipid peroxidation) caused by drought stress or chilling stress (4 °C) under high light intensity (1000 µmol m^?2 s^?1). These results indicate that the expression of the catalase in chloroplasts has a positive effect on the protection of the transgenic plants from the photo‐oxidative stress invoked by paraquat treatment, drought stress and chilling stress.     

Elevated CO2 mitigates chilling-induced water stress and photosynthetic reduction during chilling

Bean, cucumber and corn plants were grown in controlled-environment chambers at 25/18 °C day/night temperature and either ambient (350 μmol mol^?1) or elevated (700 μmol mol^?1) CO₂ concentration, and at 20–30 d after emergence they were exposed to a 24 h chilling treatment (6.5 ± 1.5 °C) at their growth CO₂ concentration. Whole-plant transpiration rates (per unit leaf area basis) during the first 3 h of chilling were about 26,28 and 13% lower at elevated than at ambient CO₂ for bean, cucumber and corn, respectively. The decline in leaf water potential (ψ_L) and visible wilting of bean and cucumber during chilling were significantly less at elevated than at ambient CO₂. Corn ψ_L was not significantly affected by chilling, and corn did not exhibit any other symptoms of chilling-induced water stress. Leaf osmotic potentials (measured before chilling only) of bean and cucumber were more negative at elevated than at ambient CO₂, and the corresponding calculated leaf turgor potentials were significantly higher at elevated than at ambient CO₂. Leaf relative water content (RWC) during chilling at ambient CO₂fell to 62 and 48% for bean and cucumber, respectively. RWC during chilling at elevated CO₂ was never below 79% for bean or 63% for cucumber. Corn RWC was not measured. After 24 h of chilling at ambient CO₂, net photosynthetic rate (P_N) reductions were 83, 89 and 24% for bean, cucumber and corn, respectively. P_N reductions during chilling were less at elevated CO₂: 53, 40 and 4% for bean, cucumber and corn, respectively. At ambient CO₂, none of the species fully recovered to pre-chilling P_N, but at elevated CO₂ both bean and corn recovered fully. The average percentage leaf area with visible leaf damage due to chilling was 20.6 and 9.6% at ambient and elevated CO₂, respectively, for bean, and 32.4 and 23.6% at ambient and elevated CO₂, respectively, for cucumber. Corn showed no significant permanent leaf damage from chilling at either CO₂ concentration. These results indicate that cucumber was most sensitive to chilling as imposed in this study, followed by bean and corn. The results support the hypothesis that, at least in young plants under controlled-environment conditions, elevated CO₂ improves plant water relations during chilling and can mitigate photosynthetic depression and chilling damage. The implications for long-term growth and reproductive success in managed and natural ecosystems will require testing of this hypothesis under field conditions.     

The effects of development at sub-optimal growth temperatures on photosynthetic capacity and susceptibility to chilling-dependent photoinhibition in Zea mays

When plants of Zea mays L. cv. LG11 that have been grown at optimal temperatures are transferred to chilling temperatures (0–12°C) photoinhibition of photosynthetic CO₂ assimilation can occur. This study examines how growth at sub-optimal temperatures alters both photosynthetic capacity and resistance to chilling-dependent photoinhibition. Plants of Z. mays cv. LG11 were grown in controlled environments at 14, 17, 20 and 25°C. As a measure of the capacity for photosynthesis under light limiting conditions, the maximum quantum yields of CO₂ assimilation (φ^a.c) and O₂ evolution (φ^a.o) were determined for the laminae of the second leaves at photon fluxes of 50–150 μmol m^-2s^-1. To determine photosynthetic capacity at photon fluxes approaching light saturation, rates of CO₂ uptake (A₁₅₀₀) and O₂ evolution (A₁₅₀₀) were determined in a photon flux of 1500 μmol m^-2s^-1. In leaves developed at 14°C, φ and φ were 26 and 43%, respectively, of the values for leaves grown at 25°C. Leaves grown at 17°C showed intermediate reductions in φ and φ, whilst leaves developed at 20°C showed no significant differences from those grown at 25°C. Similar patterns of decrease were observed for A₁₅₀₀ and A_1500.0 with decreasing growth temperature. Leaves developed at 25°C showed higher rates of CO₂ assimilation at all light levels and measurement temperatures in comparison to leaves developed at 17 and 14°C. A greater reduction in A₁₅₀₀ relative to A_1500.0 with decreasing growth temperature was attributed to increased stomatal limitation. Exposure of leaves to 800–1000 μmol m^-2 s^-1 when plant temperature was depressed to ca 6.5°C produced a photoinhibition of photosynthetic CO₂ assimilation in all leaves. However, in leaves developed at 17°C the decrease in A₁₅₀₀ following this chilling treatment was only 25% compared to 90% in leaves developed at 25°C. Recovery following chilling was completed earlier in leaves developed at 17°C. The results suggest that growth at sub-optimal temperatures induces increased tolerance to exposure to high light at chilling temperatures. This is offset by the large loss in photosynthetic capacity imposed by leaf development at sub-optimal temperatures.     

Prior temperature exposure affects subsequent chilling sensitivity

The chilling sensitivity of small discs or segments of tissue excised from chillingsensitive species was significantly altered by prior temperature exposure subsequent to holding the tissue at chilling temperatures as measured by a number of physiological processes sensitive to chilling. This temperature conditioning was reversible by an additional temperature exposure before chilling, and mature-green and red-ripe tomato tissue exhibit similar chilling sensitivities. Exposing pericarp discs excised from tomato fruit (Lycopersicon esculentum Mill. cv. Castelmart), a chilling-sensitive species, to temperatures from 0 to 37°C for 6 h before chilling the discs at 2.5°C for 4 days significantly altered the rate of ion leakage from the discs, but had no effect on the rate of ion leakage before chilling and only a minimal effect on discs held at a non-chilling temperature of 12°C. Exposing chillingsensitive tissue to temperatures below that required to induce heat-shock proteins but above 20°C significantly increased chilling sensitivity as compared to tissue exposed to temperatures between 10 and 20°C. Rates of ion leakage after 4 days of chilling at 2.5°C were higher from fruit and vegetative tissue of chilling-sensitive species (Cucumis sativus L. cv. Poinsett 76, and Cucurbita pepo L. cv. Young Beauty) that were previously exposed for 6 h to 32°C than from similar tissue exposed to 12°C. Exposure to 32 and 12°C had no effect on the rate of ion leakage from fruit tissue of chilling tolerant species (Malus domestica Borkh. cv. Golden Delicious, Pyrus communis L. cv. Bartlett). Ethylene and CO₂ production were higher and lycopene synthesis was lower in chilled tomato pericarp discs that were previously exposed for 6 h to 32°C than the values from tissue exposed to 12°C for 6 h before chilling. Increased chilling sensitivity induced by a 6 h exposure to 32°C could be reversed by subsequent exposure to 12°C for 6 h.     

Prolonged chilling under moderate light: effect on photosynthetic activity measured with the photoacoustic method

Abstract Exposure of tomato plants to a mild chilling temperature and relatively low ambient photon flux density for an extended period (10°C and 400 μmol photons m^?2 s^?1 d and 5°C night for 6 d) resulted in a significant decrease in the variable chlorophyll fluorescence, the quantum yield of oxygen evolution and the amount of total absorbed energy stored in photochemical intermediates, but not in the chlorophyll concentration or in the activity of ribulose biphosphate carboxylase. These results indicate that photochemical processes involving PSII were affected, and might reflect photoinhibitory effects on the photosynthetic apparatus. Chilling treatment had relatively small influence on the maximal extent of the Emerson effect. This observation, together with the sharp decrease found in the quantum yield of oxygen evolution, could be reconciled with the above results only if some dependency between the two photosystems was assumed. On the basis of this interpretation, it was concluded that the strong Emerson effect after chilling still reflects the typical imbalance between PSI and PSII centres, even though populations of such unaffected pairs are smaller than in the untreated plants. The relatively new photoacoustic technique employed in this study is shown to be useful both as a diagnostic tool and as a means of investigating changes in photochemical activity in the study of environmental stress effects on photosynthesis. The results support the view that photoinhibition can play an important role in limiting photosynthetic activity, and therefore productivity, in chilling-sensitive plants such as the tomato under the natural conditions that prevail during the winter in mediterranean climates.     

Differential response of domestic and wild Lycopersicon species to chilling under low light: growth, carbohydrate content, photosynthesis and the xanthophyll cycle

The response of five Lycopersicon species to 14 days moderate chilling at 10°C under low light (75 μmol m^?2 s^?1) and subsequent recovery was examined by measurements on relative shoot growth rate, leaf dry matter and carbohydrate content, CO₂-exchange and pigment composition. In addition, the susceptibility to dark chilling and temperature dependence of chloroplast electron transport were analyzed by Chl a fluorescence measurements. During 7 days of recovery at 25/20°C subsequent to chilling, the domestic tomato Lycopersiconesculentum (L.) Mill. cv. Abunda exhibited a small capacity for shoot regrowth (39%) compared to the low-altitude wild species L. pimpinellifolium (Jusl.) Mill. PI187002 (82%) and three wild species originating from high altitude: L. peruvianum Mill. LA 385 (92%), L. hirsutum Humb. & Bonpl. LA 1777 (67%) and L. chilense Dunn. LA 1970 (71%). The inter-specific differences in chilling sensitivity at the chloroplast level, analyzed by the decline of the maximum rate of induced Chl fluorescence rise (F_R) after 40 h at 0°C and the temperature at which q_P reached the value 0.5, correlated in general well with the measured differences at whole plant level, measured by the post-chilling regrowth capacity. Chilling resulted in a larger increase in leaf dry matter content in L. esculentum (45%) and L. pimpinellifolium (37%) compared to the high-altitude species (13–16%), which could be attributed to a stronger accumulation of both soluble sugars and starch in mature leaves of the domestic and low-altitude species. Photosynthetic and dark respiration rates during chilling could not account for this difference. The recovery of photosynthesis was better in the high-altitude species. Chl content per unit leaf area decreased more throughout the experiment in the domestic and low-altitude species (63–73%) than in their relatives from high altitude (8–29%). In response to chilling, the domestic and low-altitude species showed an increase in the total xanthophyll cycle pool on Chl basis, whereas the de-epoxidation state of the xanthophyll cycle increased in the high-altitude wild species. Both responses resulted in increased zeaxanthin levels in chilled leaves of all Lycopersicon species.     

Physiological responses of Opuntia ficus-indica to growth temperature

The influences of various day/night air temperatures on net CO₂ uptake and nocturnal acid accumulation were determined for Opuntia ficus-indica, complementing previous studies on the water relations and responses to photosynthetically active radiation (PAR) for this widely cultivated cactus. As for other Crassulacean acid metabolism (CAM) plants, net nocturnal CO₂ uptake had a relatively low optimal temperature, ranging from 11°C for plants grown at day/night air temperatures of 10°C/0°C to 23°C at 45°C/35°C. Stomatal opening, which occurred essentially only at night and was measured by changes in water vapor conductance, progressively decreased as the measurement temperature was raised. The CO₂ residual conductance, which describes chlorenchyma properties, had a temperature optimum a few degrees higher than the optimum for net CO₂ uptake at all growth temperatures. Nocturnal CO₂ uptake and acid accumulation summed over the whole night were maximal for growth temperatures near 25°C/15°C, CO₂ uptake decreasing more rapidly than acid accumulation as the growth temperature was raised. At day/night air temperatures that led to substantial nocturnal acid accumulation (25°C/15°C.). 90% saturation of acid accumulation required a higher total daily PAR than at non-optimal growth temperatures (10°C/0°C and 35°C/25°C). Also, the optimal temperature of net CO₂ uptake shifted downward when the plants were under drought conditions at all three growth temperatures tested, possibly reflecting an increased fractional importance of respiration at the higher temperatures during drought. Thus, water status, ambient PAR, and growth temperatures must all be considered when predicting the temperature response of gas exchange for O. ficus-indica and presumably for other CAM plants.     

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.     

The influence of chilling on photosynthesis and activities of some enzymes of sucrose metabolism in Lycopersicon esculentum Mill

The effects of chilling stress on leaf photosynthesis and sucrose metabolism were investigated in tomato plants (Lycopersicon esculentum Mill. cultivar Marmande). Twenty-one-day-old seedlings were grown in a growth chamber at 25/23 °C (day/night) (control) and at 10/8 °C (day/night) (chilled) for 7 days. The most evident effect of chilling was the marked reduction of plant growth and of CO₂ assimilation as measured after 7 days, the latter being associated with a decrease in stomatal closure and an increase in Ci. The inhibition in photosynthetic rate was also related to an impairment of photochemistry of photosystem II (PSII), as seen from the slight, but significant change in the ratio of F_v/F_m. The capacity of chilled leaves to maintain higher q_P values with respect to the controls suggests that some protection mechanism prevented excess reduction of PSII acceptors. The results of the determination of starch and soluble sugar content could show that chilling impaired sucrose translocation. The activity of leaf invertase increased significantly in chilled plants, while that of other sucrose-metabolizing enzymes was not affected by growing temperature. Furthermore, the increase in invertase (neutral and acid) activity, which is typical of senescent tissue characterized by reduced growth, seems to confirm that tomato is a plant which is not a plant genetically adapted to low temperatures.     

Light, temperature and nutrients as factors in photosynthetic adjustment to an elevated concentration of carbon dioxide

The short-term stimulation of the net rate of carbon dioxide exchange of leaves by elevated concentrations of CO₂ usually observed in C₃ plants sometimes does not persist. Experiments were conducted to test whether the patterns of response to the environment during growth were consistent with the hypotheses that photosynthetic adjustment to elevated CO₂ concentration is due to (1) feedback inhibition or (2) nutrient stress. Soybean [Glycine max (L.) Merr. cv. Williams] and sugar beet (Best vulgaris L. cv. Mono Hye-4) were grown from seed at 350 and 700 μl^? CO₂, at 20 and 25°C, at a photon flux density of 0.5 and 1.0 mmol m^?2 S^?1 and with three nutrient regimes until the third trifoliolate leaf of soybean or the sixth leaf of sugar beet had finished expanding. Net rates of CO₂ exchange of the most recently expanded leaves were then measured at both 350 and 700 μl 1^?1 CO₂. Plants grown at the elevated CO₂ concentration had net rates of leaf CO₂ exchange which were reduced by 33% in sugar beet and 23% in soybean when measured at 350 μl 1^?1 CO₂ and when averaged over all treatments. Negative photosynthetic adjustment to elevated CO₂ concentration was not greater at 20 than at 25°C, was not greater at a photon flux density of 1.0 than at 0.5 mmol m^?2 S^?1 and was not greater with limiting nutrients. Furthermore, in soybean, negative photosynthetic adjustment could be induced by a single night at elevated CO₂ concentration, with net rates of CO₂ exchange the next day equal to those of leaves of plants grown from seed at the elevated concentration of CO₂. These patterns do not support either the feedback-inhibition or the nutrient-stress hypothesis of photosynthetic adjustment to elevated concentrations of CO₂.     

The influence of elevated CO2 and temperature on biomass production of continuously defoliated white clover

Clonal plants of white clover (Trifolium repens L.), grown singly in pots of Perlite and solely dependent for nitrogen on root nodule N₂ fixation, were maintained in controlled environments which provided four environments: 18/13 °C day/night temperature at 340 and 680 μmol mol^?1 CO₂ and 20·5/15·5°C day/night temperature at 340 and 680 μmol mol^?1 CO₂. The daylength was 12 h and the photon flux density 500±25 μmol m^?2 s^?1 (PFD). All plants were defoliated for about 80d, nominally every alternate day, to leave the youngest expanded leaf intact on 50% of stolons, plus expanding leaves (simulated grazing). Elevated CO₂ increased the yield of biomass removed at defoliation by a constant 45% during the second 40d of the experiment and by a varying amount in the first half of the experiment. Elevated temperature had little effect on biomass yield. Nitrogen, as a proportion of the harvested biomass, was only fractionally affected by elevated CO₂ or temperature. In contrast, N₂ fixation increased in concert with the promoting effect of elevated CO₂ on biomass production. The increased yield of biomass harvested in 680 μmol mol^?1 CO₂ was primarily due to the early development and continued maintenance of more stolons. However, the stolons of plants grown in elevated CO₂ also developed leaves which were heavier and slightly larger in area than their counterparts in ambient CO₂. The conclusion is that, when white clover plants are maintained at constant mass by simulated grazing, they continue to respond to elevated CO₂ in terms of a sustained increase in biomass production.     

Photosynthetic characteristics of Amaranthus tricolor,a C4 tropical leafy vegetable

The gas exchange characteristics are reported for Amaranthus tricolor, a C₄ vegetable amaranth of southeastern Asia. Maximum photosynthetic capacity was 48.3±1.0μmol CO₂ m^?2s^?1 and the temperature optimum was 35°C. The calculated intercellular CO₂ concentration at this leaf temperature and an incident photon flux (400–700 mm) of 2 mmol m^?2s^?1 averaged 208±14 μl l^?1, abnormally high for a C₄ species. The photosynthetic rate, intercellular CO₂ concentration, and leaf conductance all decreased with an increase in water vapor pressure deficit. However, the decrease in leaf conductance which resulted in a decrease in intercellular CO₂ concentration accounted for only one fourth of the observed decrease in photosynthetic rate as water vapor pressure deficit was increased. Subsequent measurements indicated that the depence of net photosynthesis on intercellular CO₂ concetration changed with water vapor pressure deficit.     

Genotypic variation within Zea mays for susceptibility to and rate of recovery from chill-induced photoinhibition of photosynthesis

Six genotypes of Zea mays L. were grown in pots inside a glasshouse at a mean temperature of 22±2°C and a minimum photosynthetic photon flux density (Q) during the daylight period of 400 μmol m^?2 s^?1. Chilling-dependent photoinhibition was induced by exposing plants to a temperature of 7°C and a Q of 1 000 μmol m^?2 s^?1 for 6 h. Recovery from photoinhibition was then followed at a temperature of 25°C and a Q of 200 μmol m^?2 s^?1. Leaf gas exchange and chlorophyll fluorescence were measured on attached leaves at room temperature prior to the photoinhibitory treatments and at 6 sampling intervals from 0 to 24 h during the recovery period. The relative water content (RWC) was also measured during the recovery period. The results showed a significant genotypic variation in the susceptibility to and rate of recovery from chilling-dependent photoinhibition of photosynthesis in Zea mays seedlings. The Highland Pool 1a from highland sites in Mexico was the least susceptible to chill-induced photoinhibition, but had the slowest rate of recovery. The hybrid variety LG11 showed the highest rate of recovery, whilst the inbred line ZPF307 was the most susceptible to chill-induced photoinhibition. Susceptibility to photoinhibition and subsequent recovery were at least partially independent, suggesting that selection for improved genotypes will require independent selection for both tolerance and capacity for recovery. Although chlorophyll fluorescence provided a more rapid method of assessing the occurrence of photoinhibition, it was not as effective as direct gas-exchange measurements of the maximum quantum yield of photosynthesis (φ) in separating genotypes with respect to their susceptibility to photoinhibition, especially in the most vulnerable genotypes such as ZPF307. Water stress induced by chilling and high Q treatments appeared to impair the recovery processes. Decreases in stomatal conductance (g_s) produce a significant decrease in intercellular CO₂ concentration (C_i), although this decrease was never so extreme that it limited photosynthetic rates at the light intensities used to determine φ. Nevertheless, closure of stomata in patches, producing local restriction of CO₂ supply, would explain the poor correlation between chlorophyll fluorescence and quantum yield measurements in some genotypes immediately after photoinhibitory treatments.     

High Temperatures and Net CO2 Uptake, Growth, and Stem Damage for the Hemiepiphytic Cactus Hylocereus undatus1

Hylocereus undatus, which is native to tropical forests experiencing moderate temperatures, would not be expected to tolerate the extremely high temperatures that can be tolerated by cacti native to deserts. Nevertheless, total daily net CO₂ uptake by this hemiepiphytic cactus, which is widely cultivated for its fruits, was optimal at day/night air temperatures of 30/20°C, temperatures that are higher than those optimal for daily net CO₂ uptake by cacti native to arid and semiarid areas. Exposure to 35/25°C for 30 weeks led to lower net CO₂ uptake than at 10 weeks; exposure to 40/30°C led to considerable necrosis visible on the stems at 6 weeks and nearly complete browning of the stems by 19 weeks. Dry mass gain over 31 weeks was greatest for plants at 30/20°C, with root growth being especially noteworthy and root dry mass gain representing an increasing percentage of plant dry mass gain as day/night air temperatures were increased. Viability of chlorenchyma cells, assayed by the uptake of the vital stain neutral red into the central vacuoles, was decreased 50 percent by a one‐hour treatment at 55°C compared with an average of 64°C for 18 species of cacti native to deserts. The lower high‐temperature tolerance for H. undatus reflected its low high‐temperature acclimation of only 1.4°C as growth temperatures were raised by 10°C compared with an average acclimation of 5.3°C for the other 18 species of cacti. Thus, this tropical hemiepiphytic cactus is not adapted to day/night air temperatures above ca 40/30°C, although its net CO₂ uptake is optimal at the relatively high day/night air temperatures of 30/20°C.     

Enriched rhizosphere CO2 concentrations can ameliorate the influence of salinity on hydroponically grown tomato plants

Our previous work indicated that salinity caused a shift in the predominant site of nitrate reduction and assimilation from the shoot to the root in tomato plants. In the present work we tested whether an enhanced supply of dissolved inorganic carbon (DIC, CO₂+ HCO₃) to the root solution could increase anaplerotic provision of carbon compounds for the increased nitrogen assimilation in the root of salinity-stressed Lycopersicon esculentum (L.) Mill. cv. F144. The seedlings were grown in hydroponic culture with 0 or 100mM NaCl and aeration of the root solution with either ambient or CO₂-enriched air (5000 μmol mol^?1). The salinity-treated plants accumulated more dry weight and higher total N when the roots were supplied with CO₂-enriched aeration than when aerated with ambient air. Plants grown with salinity and enriched DIC also had higher rates of NO^?₃ uptake and translocated more NO^?₃ and reduced N in the xylem sap than did equivalent plants grown with ambient DIC. Incorporation of DIC was measured by supplying a 1 -h pulse of H¹⁴CO^?₃ to the roots followed by extraction with 80% ethanol. Enriched DIC increased root incorporation of DIC 10-fold in both salinized and non-salinized plants. In salinity-stressed plants, the products of dissolved inorganic ¹⁴C were preferentially diverted into amino acid synthesis to a greater extent than in non-salinized plants in which label was accumulated in organic acids. It was concluded that enriched DIC can increase the supply of N and anaplerotic carbon for amino acid synthesis in roots of salinized plants. Thus enriched DIC could relieve the limitation of carbon supply for ammonium assimilation and thus ameliorate the influence of salinity on NO^?₃ uptake and assimilation as well as on plant growth.