Guard cell photosynthesis is critical for stomatal turgor production,yet does not directly mediate CO2‐ and ABA‐induced stomatal closing

Stomata mediate gas exchange between the inter‐cellular spaces of leaves and the atmosphere. CO₂ levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO₂]. [CO₂] in leaves mediates stomatal movements. The role of guard cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll‐deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard cell specific enhancer trap line. Our data show that more than 90% of guard cells were chlorophyll‐deficient. Interestingly, approximately 45% of stomata had an unusual, previously not‐described, morphology of thin‐shaped chlorophyll‐less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole‐leaf photosynthetic parameters (PSII, qP, qN, F_V′/F_M′) were comparable with wild‐type plants. Time‐resolved intact leaf gas‐exchange analyses showed a reduction in stomatal conductance and CO₂‐assimilation rates of the transgenic plants. Normalization of CO₂ responses showed that stomata of transgenic plants respond to [CO₂] shifts. Detailed stomatal aperture measurements of normal kidney‐shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO₂] elevation and abscisic acid (ABA), while thin‐shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO₂] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard cell CO₂ and ABA signal transduction are not directly modulated by guard cell photosynthesis/electron transport. Moreover, the finding that chlorophyll‐less stomata cause a ‘deflated’ thin‐shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard cell turgor production.     

Effects of addition of external nitric oxide on the allocation of photosynthetic electron flux in Rumex K-1 leaves under osmotic shock

Photosynthetic electron flux allocation, stomatal conductance, and the activities of key enzymes involved in photosynthesis were investigated in Rumex K-1 leaves to better understand the role of nitric oxide (NO) in photoprotection under osmotic stress caused by polyethylene glycol. Gas exchange and chlorophyll fluorescence were measured simultaneously with a portable photosynthesis system integrated with a pulse modulated fluorometer to calculate allocation of photosynthetic electron fluxes. Osmotic stress decreased stomatal conductance, photosynthetic carbon assimilation, and nitrate assimilation, increased Mehler reaction, and resulted in photoinhibition. Addition of external NO enhanced the stomatal conductance, photosynthetic rate, activities of glutamine synthetase and nitrate reductase, and reduced Mehler reaction and photoinhibition. These results demonstrated that osmotic stress reduced CO₂ assimilation, decreasing the use of excited energy via CO₂ assimilation which caused significant photoinhibition. Improving stomatal conductance by the addition of external NO enhanced the use of excited energy via CO₂ assimilation. As a result, less excited energy was allocated to Mehler reaction, which reduced production of reactive oxygen species via this pathway. We suppose that Mehler reaction is not promoted unless photosynthesis and nitrogen metabolism are prominently inhibited.     

Seasonal and diurnal changes in photosynthetic limitation of young sweet orange trees

This study tests the hypothesis that potted sweet orange plants show a significant variation in photosynthesis over seasonal and diurnal cycles, even in well-hydrated conditions. This hypothesis was tested by measuring diurnal variations in leaf gas exchange, chlorophyll fluorescence, leaf water potential, and the responses of CO₂ assimilation to increasing air CO₂ concentrations in 1-year-old ‘Valência’ sweet orange scions grafted onto ‘Cleopatra’ mandarin rootstocks during the winter and summer seasons in a subtropical climate. In addition, diurnal leaf gas exchange was evaluated under controlled conditions, with constant environmental conditions during both winter and summer. In relation to our hypothesis, a greater rate of photosynthesis is found during the summer compared to the winter. Reduced photosynthesis during winter was induced by cool night conditions, as the diurnal fluctuation of environmental conditions was not limiting. Low air and soil temperatures caused decreases in the stomatal conductance and in the rates of the biochemical reactions underlying photosynthesis (ribulose-1,5-bisphosphate (RuBP) carboxylation and RuBP regeneration) during the winter compared to the values obtained for those markers in the summer. Citrus photosynthesis during the summer was not impaired by biochemical or photochemical reactions, as CO₂ assimilation was only limited by stomatal conductance due to high leaf-to-air vapor pressure difference (VPD) during the afternoon. During the winter, the reduction in photosynthesis during the afternoon was caused by decreases in RuBP regeneration and stomatal conductance, which are both precipitated by low night temperature.     

Light Energy Dissipation under Water Stress Conditions: Contribution of Reassimilation and Evidence for Additional Processes

Using ¹⁴CO₂ gas exchange and metabolite analyses, stomatal as well as total internal CO₂ uptake and evolution were estimated. Pulse modulated fluorescence was measured during induction and steady state of photosynthesis. Leaf water potential of Digitalis lanata EHRH. plants decreased to −2.5 megapascals after withholding irrigation. By osmotic adjustment, leaves remained turgid and fully exposed to irradiance even at severe water stress. Due to the stress-induced reduction of stomatal conductance, the stomatal CO₂ exchange was drastically reduced, whereas the total CO₂ uptake and evolution were less affected. Stomatal closure induced an increase in the reassimilation of internally evolved CO₂. This `CO<sub>2</sub> recycling' consumes a significant amount of light energy in the form of ATP and reducing equivalents. As a consequence, the metabolic demand for light energy is only reduced by about 40%, whereas net photosynthesis is diminished by about 70% under severe stress conditions. By CO<sub>2</sub> recycling, carbon flux, enzymatic substrate turnover and consumption of light energy were maintained at high levels, which enabled the plant to recover rapidly after rewatering. In stressed D. lanata plants a variable fluorescence quenching mechanism, termed `coefficient of actinic light quenching,' was observed. Besides water conservation, light energy dissipation is essential and involves regulated metabolic variations.     

In situ estimation of net CO2 assimilation, photosynthetic electron flow and photorespiration in Turkey oak (Q. cerris L.) leaves: diurnal cycles under different levels of water supply

Diurnal time courses of net CO₂ assimilation rates, stomatal conductance and light-driven electron fluxes were measured in situ on attached leaves of 30-year-old Turkey oak trees (Quercus cerris L.) under natural summer conditions in central Italy. Combined measurements of gas exchange and chlorophyll a fluorescence under low O₂ concentrations allowed the demonstration of a linear relationship between the photochemical efficiency of PSII (fluorescence measurements) and the apparent quantum yield of gross photosynthesis (gas exchange). This relationship was used under normal O₂ to compute total light-driven electron fluxes, and to partition them into fractions used for RuBP carboxylation or RuBP oxygenation. This procedure also yielded an indirect estimate of the rate of photorespiration in vivo. The time courses of light-driven electron flow, net CO₂ assimilation and photorespiration paralleled that of photosynthetic photon flux density, with important afternoon deviations as soon as a severe drought stress occurred, whereas photochemical efficiency and maximal fluorescence underwent large but reversible diurnal decreases. The latter observation indicated the occurrence of a large non-photochemical energy dissipation at PSII. We estimated that less than 60% of the total photosynthetic electron flow was used for carbon assimilation at midday, while about 40% was devoted to photorespiration. The rate of carbon loss by photorespiration (R₁) reached mean levels of 56% of net assimilation rates. The potential application of this technique to analysis of the relative contributions of thermal de-excitation at PSII and photorespiratory carbon recycling in the protection of photosynthesis against stress effects is discussed.     

Photosynthetic and stomatal responses of spinach leaves to salt stress

The gas exchange of spinach plants, salt-stressed by adding NaCl to the nutrient solution in increments of 25 millimolar per day to a final concentration of 200 millimolar, was studied 3 weeks after starting NaCl treatment. Photosynthesis became light saturated at 1100 to 1400 micromoles per square meter per second in salt-treated plants and at approximately 2000 micromoles per square meter per second in control plants. Photosynthetic capacity of the mesophyll measured as a function of intercellular partial pressure of CO₂ at the light intensity prevailing during growth and at light saturation were both decreased in the salttreated plants. The CO₂ compensation points and relative enhancements of photosynthesis at low O₂ were not affected by salinity. The lower photosynthetic rates in salt-treated leaves at 450 micromoles per square meter per second were associated with a 70% reduction in stomatal conductance and low intercellular CO₂ (219 microbars; cf. 285 microbars for controls). Increasing photon flux density to light saturation extended the linear portions of the CO₂ response curves, increased stomatal conductances, increased intercellular CO₂ in the salt-treated plants, but lowered it in controls, and accentuated differences in photosynthetic rate (area basis) between the treatments.

Leaves from salt-treated plants were thicker but contained about 73% of the chlorophyll per unit area of control plants. When photosynthetic rates were expressed on a chlorophyll basis there was no difference in initial slope of assimilation versus intercellular CO₂ between treatments. Photosynthetic rates (chlorophyll basis) at light saturation differed only by 20% which was also observed earlier with isolated, intact chloroplasts (Robinson et al. 1983 Plant Physiol 73: 238-242).

Measurement of carbon isotope ratio revealed less discrimination against ¹³C with salt treatment and confirmed the persistence of low intercellular partial pressures of CO₂ during plant growth. The development of a thicker leaf with less chlorophyll per unit area during salt treatment permitted stomatal conductance and intercellular partial pressure of CO₂ to decline without restricting photosynthesis and had the benefit of greatly increasing water use efficiency.

     

Flavescence dorée phytoplasma deregulates stomatal control of photosynthesis in Vitis vinifera

Flavescence dorée (FD) is among the major grapevine diseases causing high management costs; curative methods against FD are unavailable. In FD‐infected plants, decrease in photosynthesis is usually recorded, but deregulation in stomatal control of leaf gas exchange during FD infection and recovery is unknown. We measured the seasonal time course of gas exchange rates in two cultivars (‘Barbera’ and ‘Nebbiolo’) during the term of 1 year when grapevines experienced a water stress and another with no drought, with difference in gas exchange rates in response to FD infection and recovery as assessed by symptom observation and phytoplasma detection through PCR analysis. Chlorophyll fluorescence was also evaluated at the time of maximum symptom severity in ‘Barbera’, the cultivar showing the most severe stress response to FD infection, causing the highest damage in vineyards of north‐western Italy. In FD‐infected plants, net photosynthesis and transpiration gradually decreased during the season, more during the no drought year than during drought. During recovery, healthy (PCR negative) plants infected 2 years before, but not those infected an year before, regained the gas exchange performances to the level as measured before infection. The relationships between stomatal conductance and the residual leaf intercellular CO₂ concentration (c_i) discriminated healthy versus FD‐infected and recovered plants; at the same c_i, FD‐infected leaves had higher non‐photochemical quenching than healthy ones. We conclude that metabolic, not stomatal, leaf gas exchange limitation in FD‐infected and recovered grapevines is the basis of plant response to FD disease. In addition, we also suggest that such response is dependent upon water stress, by showing that water stress superimposes on FD infection in terms of stomatal and metabolic non‐stomatal limitations to carbon assimilation.     

Photosynthesis is limited at high leaf to air vapor pressure deficit in a mutant of Arabidopsis thaliana that lacks trienoic fatty acids

We have investigated the role of polyunsaturated fatty acids in photosynthesis using a triple mutant of Arabidopsis thaliana that lacks trienoic fatty acids (fad 3-2 fad 7-2 fad 8). Though this mutant is male sterile, vegetative growth and development under normal conditions are largely unaffected (McConn and Browse, 1996 Plant Cell 8: 403–416). At 0.2–1.0 kPa vapor pressure deficit (low VPD), maximum photosynthetic rates of wild-type and mutant plants were similar while stomatal conductance rates were up to 2 times higher in mutant plants. However, light-saturated rates of carbon assimilation and stomatal conductance in the mutant were lower than in wild-type plants when measured at ambient (35 Pa) CO₂ and 2.0–2.8 kPa vapor pressure deficit (high VPD). The limitation to photosynthesis in the mutant plants at high VPD was overcome by saturating partial pressures of CO₂ suggesting a stomatal limitation. Chlorophyll fluorescence measurements indicate that differences observed in maximum assimilation rates were not due to limitations within the photochemical reactions of photosynthesis. Stomatal response to VPD and intrinsic water use efficiency was drastically different in mutant versus wild-type plants. The results of this investigation indicate that for Arabidopsis, polyunsaturated fatty acids may be an important determinant of responses of photosynthesis and stomatal conductance to environmental stresses such as high VPD. This revised version was published online in June 2006 with corrections to the Cover Date.     

Stomatal function,density and pattern,and CO2 assimilation in Arabidopsis thaliana tmm1 and sdd1‐1 mutants

• Stomata modulate the exchange of water and CO₂ between plant and atmosphere. Although stomatal density is known to affect CO₂ diffusion into the leaf and thus photosynthetic rate, the effect of stomatal density and patterning on CO₂ assimilation is not fully understood.
• We used wild types Col‐0 and C24 and stomatal mutants sdd1‐1 and tmm1 of Arabidopsis thaliana, differing in stomatal density and pattern, to study the effects of these variations on both stomatal and mesophyll conductance and CO₂ assimilation rate. Anatomical parameters of stomata, leaf temperature and carbon isotope discrimination were also assessed.
• Our results indicate that increased stomatal density enhanced stomatal conductance in sdd1‐1 plants, with no effect on photosynthesis, due to both unchanged photosynthetic capacity and decreased mesophyll conductance. Clustering (abnormal patterning formed by clusters of two or more stomata) and a highly unequal distribution of stomata between the adaxial and abaxial leaf sides in tmm1 mutants also had no effect on photosynthesis.
• Except at very high stomatal densities, stomatal conductance and water loss were proportional to stomatal density. Stomatal formation in clusters reduced stomatal dynamics and their operational range as well as the efficiency of CO₂ transport.

     

A Microscale Model for Combined CO2 Diffusion and Photosynthesis in Leaves

Transport of CO₂ in leaves was investigated by combining a 2-D, microscale CO₂ transport model with photosynthesis kinetics in wheat (Triticum aestivum L.) leaves. The biophysical microscale model for gas exchange featured an accurate geometric representation of the actual 2-D leaf tissue microstructure and accounted for diffusive mass exchange of CO_2. The resulting gas transport equations were coupled to the biochemical Farquhar-von Caemmerer-Berry model for photosynthesis. The combined model was evaluated using gas exchange and chlorophyll fluorescence measurements on wheat leaves. In general a good agreement between model predictions and measurements was obtained, but a discrepancy was observed for the mesophyll conductance at high CO₂ levels and low irradiance levels. This may indicate that some physiological processes related to photosynthesis are not incorporated in the model. The model provided detailed insight into the mechanisms of gas exchange and the effects of changes in ambient CO₂ concentration or photon flux density on stomatal and mesophyll conductance. It represents an important step forward to study CO₂ diffusion coupled to photosynthesis at the leaf tissue level, taking into account the leaf''s actual microstructure.     

Elevated CO2 alleviates the impact of drought on barley improving water status by lowering stomatal conductance and delaying its effects on photosynthesis

We analysed the impact of elevated CO₂ on water relations, water use efficiency and photosynthetic gas exchange in barley (Hordeum vulgare L.) under wet and drying soil conditions. Soil moisture was less depleted under elevated compared to ambient [CO₂]. Elevated CO₂ had no significant effect on the water relations of irrigated plants, except on whole plant hydraulic conductance, which was markedly decreased at elevated compared to ambient CO₂ concentrations. The values of relative water content, water potential and osmotic potential were higher under elevated CO₂ during the entire drought period. The better water status of water-limited plants grown at elevated CO₂ was the result of stomatal control rather than of osmotic adjustment. Despite the low stomatal conductance produced by elevated CO₂, net photosynthesis was higher under elevated than ambient CO₂ concentrations. With water shortage, photosynthesis was maintained for longer at higher rates under elevated CO₂. The reduction of stomatal conductance and therefore transpiration, and the enhancement of carbon assimilation by elevated CO₂, increased instantaneous and whole plant water use efficiency in both irrigated and droughted plants. Thus, the metabolism of barley plants grown under elevated CO₂ and moderate or mild water deficit conditions is benefited by increased photosynthesis and lower transpiration. The reduction in plant water use results in a marked increase in soil water content which delays the onset and severity of water deficit.     

Ecophysiological studies of Sonoran Desert plants

Summary The gas exchange of two Sonoran Desert plants was measured near optimum soil water conditions occurring in the summer and winter. Photosynthesis and stomatal conductance rates of a drought-deciduous shrub, Ambrosia deltoidea, and an evergreen non-riparian tree, Olneya tesota, are mainly affected by plant water potential. During such periods the diurnal gas exchange patterns are characterized by maximum rates of photosynthesis and stomatal conductance occurring early in the day, which decrease progressively thereafter. The effect of plant water potential on gas exchange is both direct and indirect. Decreasing plant water potential indirectly affects ¹⁴CO₂ assimilation by closure of stomata, and the effect is similar in both species. However, the direct effects of decreasing plant water potential are dissimilar in the two species. Plants of the shrub species have a higher potential maximum photosynthesis, but are more sensitive to plant water stress than are plants of the tree species. Both species respond to favorable growth conditions in the summer and winter, and have the potential for rapid carbon input into the Sonoran Desert ecosystem.This research was supported by National Science Foundation Grant BMS 74-02671-A04, through the U.S./I.B.P. Desert Biome     

Reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase content by antisense RNA reduces photosynthesis in transgenic tobacco plants

A complementary DNA for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) was cloned from tobacco (Nicotiana tabacum) and fused in the antisense orientation to the cauliflower mosaic virus 35S promoter. This antisense gene was introduced into the tobacco genome, and the resulting transgenic plants were analyzed to assess the effect of the antisense RNA on Rubisco activity and photosynthesis. The mean content of extractable Rubisco activity from the leaves of 10 antisense plants was 18% of the mean level of activity of control plants. The soluble protein content of the leaves of anti-small subunit plants was reduced by the amount equivalent to the reduction in Rubisco. There was little change in phosphoribulokinase activity, electron transport, and chlorophyll content, indicating that the loss of Rubisco did not affect these other components of photosynthesis. However, there was a significant reduction in carbonic anhydrase activity. The rate of CO₂ assimilation measured at 1000 micromoles quanta per square meter per second, 350 microbars CO₂, and 25°C was reduced by 63% (mean value) in the antisense plants and was limited by Rubisco activity over a wide range of intercellular CO₂ partial pressures (p_i). In control leaves, Rubisco activity only limited the rate of CO₂ assimilation below a p_i of 400 microbars. Despite the decrease in photosynthesis, there was no reduction in stomatal conductance in the antisense plants, and the stomata still responded to changes in p_i. The unchanged conductance and lower CO₂ assimilation resulted in a higher p_i, which was reflected in greater carbon isotope discrimination in the leaves of the antisense plants. These results suggest that stomatal function is independent of total leaf Rubisco activity.     

Relationships between leaf conductance to CO2 diffusion and photosynthesis in micropropagated grapevine plants, before and after ex vitro acclimatization

In vitro-cultured plants typically show a low photosynthetic activity, which is considered detrimental to subsequent ex vitro acclimatization. Studies conducted so far have approached this problem by analysing the biochemical and photochemical aspects of photosynthesis, while very little attention has been paid to the role of leaf conductance to CO(2) diffusion, which often represents an important constraint to CO(2) assimilation in naturally grown plants. Mesophyll conductance, in particular, has never been determined in in vitro plants, and no information exists as to whether it represents a limitation to carbon assimilation during in vitro growth and subsequent ex vitro acclimatization. In this study, by means of simultaneous gas exchange and chlorophyll fluorescence measurements, the stomatal and mesophyll conductance to CO(2) diffusion were assessed in in vitro-cultured plants of the grapevine rootstock '41B' (Vitis vinifera 'Chasselas'xVitis berlandieri), prior to and after ex vitro acclimatization. Their impact on electron transport rate partitioning and on limitation of potential net assimilation rate was analysed. In vitro plants had a high stomatal conductance, 155 versus 50 mmol m(-2) s(-1) in acclimatized plants, which ensured a higher CO(2) concentration in the chloroplasts, and a 7% higher electron flow to the carbon reduction pathway. The high stomatal conductance was counterbalanced by a low mesophyll conductance, 43 versus 285 mmol m(-2) s(-1), which accounted for a 14.5% estimated relative limitation to photosynthesis against 2.1% estimated in acclimatized plants. It was concluded that mesophyll conductance represents an important limitation for in vitro plant photosynthesis, and that in acclimatization studies the correct comparison of photosynthetic activity between in vitro and acclimatized plants must take into account the contribution of both stomatal and mesophyll conductance.     

Circadian Rhythms in Photosynthesis : Oscillations in Carbon Assimilation and Stomatal Conductance under Constant Conditions

Net carbon assimilation and stomatal conductance to water vapor oscillated repeatedly in red kidney bean, Phaseolus vulgaris L., plants transferred from a natural photoperiod to constant light. In a gas exchange system with automatic regulation of selected environmental and physiological variables, assimilation and conductance oscillated with a free-running period of approximately 24.5 hours. The rhythms in carbon assimilation and stomatal conductance were closely coupled and persisted for more than a week under constant conditions. A rhythm in assimilation occurred when either ambient or intercellular CO₂ partial pressure was held constant, demonstrating that the rhythm in assimilation was not entirely the result of stomatal effects on CO₂ diffusion. Rhythms in assimilation and conductance were not expressed in plants grown under constant light at a constant temperature, demonstrating that the rhythms did not occur spontaneously but were induced by an external stimulus. In plants grown under constant light with a temperature cycle, a rhythm was entrained in stomatal conductance but not in carbon assimilation, indicating that the oscillators driving the rhythms differed in their sensitivity to environmental stimuli.     

Response of photosynthesis and respiration of resurrection plants to desiccation and rehydration

Using non-invasive techniques (CO₂ gas exchange, light scattering, light absorption, chlorophyll fluorescence, chlorophyll luminescence), we have analysed the response of respiration and photosynthesis to dehydration and rehydration of leaves of the resurrection plants Craterostigma plantagineum Hochst., Ramonda mykoni Reichb. and Ceterach officinarum Lam. et DC. and of the drought-sensitive mesophyte spinach (Spinacia oleracea L.). The following observations were made: (i) The rate of water loss during wilting of detached leaves of drought-tolerant resurrection plants was similar to that for leaves of the sensitive mesophyte, spinach. Leaves of Mediterranean xerophytes lost water much more slowly. (ii) Below a residual water content of about 20%, leaves of spinach did not recover turgor on rewatering, whereas leaves of the resurrection plants did. (iii) Respiration was less sensitive to the loss of water during wilting in the resurrection plants than in spinach. (iv) The sensitivity of photosynthesis to dehydration was similar in spinach and the resurrection plants. Up to a water loss of 50% from the leaves, photosynthesis was limited by stomatal closure, not by inhibition of reactions of the photosynthetic apparatus. Photosynthesis was inhibited and stomates reopened when loss of water became excessive. (v) After the leaves had lost 80% of their water or more, the light-dependent reactions of photosynthetic membranes were further inhibited by rewatering in spinach; they recovered in the resurrection plants. (vi) In desiccated leaves of the resurrection plants, slow rehydration reactivated mitochondrial gas exchange faster than photosynthetic membrane reactions. Photosynthetic carbon assimilation recovered only slowly.     

Mechanism of Photosynthesis Decrease by Verticillium dahliae in Potato

Young, visually symptomless leaves from potato (Solanum tuberosum) plants infected with Verticillium dahliae exhibited reduced carbon assimilation rate, stomatal conductance, and intercellular CO₂, but no increase in dark respiration, no change in the relationship between carbon assimilation rate versus intercellular CO₂, and no change in light use efficiency when intercellular CO₂ was held constant. Therefore, the initial decrease in photosynthesis caused by V. dahliae was caused by stomatal closure. Errors in the intercellular CO₂ calculation caused by uneven distribution of carbon assimilation rate across the leaf were tested by ¹⁴CO₂ autoradiography. Patchiness was found at a low frequency. Low stomatal conductance was correlated with low leaf water potentials. Infection did not affect leaf osmotic potentials.     

Stomatal and Mesophyll Limitations of Photosynthesis in Phosphate Deficient Sunflower, Maize and Wheat Plants

The effects of phosphate deficiency on the composition and photosyntheticCO₂ assimilation rates of fully expanded leaves of sunflower,maize and wheat plants are described. The regulation of photosynthesisby stomatal and mesophyll characteristics of leaves of differentphosphate status is analysed and related to structure. Phosphatedeficient leaves had small concentrations of inorganic phosphate,Pi, in the tissue water. Rate of photosynthesis in leaves andstomatal conductance were smaller in plants grown with inadequatephosphate when measured under any given light intensity or CO₂partial pressure. Despite the decrease in stomatal conductance(and without evidence of patchy stomatal closure), the relativestomatal limitation of photosynthesis was similar in the plantsgrown with deficient or abundant phosphate. However, the mesophyllcapacity for photosynthesis was greatly limited by phosphatedeficiency. Leaves deficient in phosphate had larger numbersof small size cells per unit leaf area than leaves with adequatephosphate. The total soluble protein content of leaves decreasedwith phosphate deficiency in all three species; however, theleaf chlorophyll content was decreased only in sunflower andmaize and not in wheat. These results suggest that stomatalconductance did not restrict the CO₂ diffusion rate, ratherthe metabolism of the mesophyll was the limiting factor. Thisis shown by poor carboxylation efficiency and decreased apparentquantum yield for CO₂ assimilation, both of which contributedto the increase in relative mesophyll limitation of photosynthesisin phosphate deficient plants. Key words: Apparent quantum yield, carboxylation efficiency, phosphate nutrition, photosynthesis, stomatal and mesophyll limitation     

The efficiency of water use in water stressed plants is increased due to ABA induced stomatal closure

Gas exchange and abscisic acid content of Digitalis lanata EHRH. have been examined at different levels of plant water stress. Net photosynthesis, transpiration and conductance of attached leaves declined rapidly at first, then more slowly following the withholding of irrigation. The intercellular partial pressure of CO₂ decreased slightly. The concentration of 2-cis(S)ABA increased about eight-fold in the leaves of non-irrigated plants as compared with well-watered controls. A close linear correlation was found between the ABA content of the leaves and their conductance on a leaf area basis. In contrast, the plot of net assimilation versus ABA concentration was curvilinear, leading to an increased efficiency of water use during stress. After rewatering, photosynthesis reached control values earlier than transpiration, leaf conductance and ABA content. From these data it is concluded that transpiration through the stomata is directly controlled by the ABA content, whereas net photosynthesis is influenced additionally by other factors.Possible reasons for the responses of photosynthesis and water use efficiency to different stress and ABA levels are discussed.Abbreviations A net CO₂ assimilation - ABA abscisic acid - C_i intercellular CO₂ concentration - g stomatal conductance - T transpiration - WUE water use efficiency     

Effects of long-term exposure to elevated UV-B radiation on the photosynthetic performance of five broad-leaved tree species

As part of an ongoing investigation into the effects of long-term UV-B radiation exposure on the growth and morphology of woody perennials, the gas exchange and photosynthesis of five common deciduous tree species were measured. All five tree species had been exposed to UV-B radiation for 5 years, in the field, at an enhancement level equivalent to an 18% ozone depletion. Measurements made during the fifth year of UV-B irradiation recorded reductions in light-saturated photosynthesis, transpiration and water use efficiencies. These changes were accompanied by marked reductions in individual leaf areas, stomatal density, stomatal conductance and carboxylation efficiency. There were no significant changes in the maximum variable fluorescence ratio, the quantum requirement of oxygen evolution, or light-saturated O₂ production. Analysis of the response of net carbon assimilation to changing intercellular CO₂ concentration (A/c_i response) demonstrated no significant change in stomatal limitation. Reductions in photosynthesis were consistent with decreased carboxylation efficiency. Although all five tree species were similarly affected by UV-B radiation treatment, the magnitude of the responses was species-specific. These findings demonstrate the need for more long-term experimentation and also suggest that changes in water use efficiency may be a significant factor in plants' responses to UV-B radiation. This revised version was published online in June 2006 with corrections to the Cover Date.