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     

Very high CO2 partially restores photosynthesis in sunflower at low water potentials

We re-examined the question of whether the stomata limit photosynthesis in dehydrated sunflower (Helianthus annuus L.) plants having low leaf water potentials. A gas-exchange apparatus was modified to operate at external CO₂ partial pressures as high as 3000 Pa (3%), which were much higher than previously achieved. This allowed photosynthesis and stomatal behavior to be monitored simultaneously at very high CO₂ in the same leaf. The data were compared with those from leaves treated with abscisic acid (ABA) where effects on photosynthesis are entirely stomatal. Photosynthesis was inhibited at low water potential and was only slightly enhanced by increasing the external CO₂ partial pressure from 34 Pa (normal air) to 300 Pa. Photosynthesis in ABA-treated leaves was similarly inhibited but recovered fully at 300 Pa. In both cases, the stomata closed to the same extent as judged from the average conductance of the leaves. Because the ABA effect resulted from diffusion limitation for CO₂ caused by stomatal closure, the contrasting data show that most of the dehydration effect was nonstomatal at low water potentials. When CO₂ partial pressures were raised further to 3000 Pa, photosynthesis increased somewhat at low water potentials but not in ABA-treated leaves. This indicates that some nonstomatal component of photosynthesis responded differently in leaves at low water potential and leaves treated with ABA. Because this component was only partially restored by very high CO₂, it was likely to be metabolic and was an important source of photosynthetic inhibition.Abbreviations and Symbol ABA abscisic acid - Chl chlorophyll - p_a external partial pressure of CO₂ - P_i intercellular partial pressure of CO₂ - _w water potential This work was supported by grant DE-FG02-87ER13776 from the Department of Energy and a grant from E.I. DuPont de Nemours and Company.     

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.     

Determination of carbon-reduction-cycle intermediates in leaves of Arbutus unedo L. suffering depressions in photosynthesis after application of abscisic acid or exposure to dry air

Gas exchange and contents of photosynthetic intermediates of leaves of Arbutus unedo L. were determined with the aim of recognizing the mechanisms of inhibition that were responsible for the midday depression of photosynthesis following exposure to dry air, and the decline in photosynthetic capacity following application of abscisic acid (ABA). Rapidly killed (<0.1 s) leaf samples were taken when gas analysis showed reduced CO₂ assimilation. Determination of the contents of 3-phosphoglyceric acid (PGA), ribulose 1,5-bisphosphate (RuBP), triose phosphates, fructose 1,6-bisphosphate and hexose phosphates in the samples showed that significant variation occurred only in the level of PGA. As a result, the ratio PGA/RuBP decreased with increasing inhibition of photosynthesis, particularly when application of ABA had been the cause. A comparison of metabolite patterns did not bring out qualitative differences that would have indicated that effects of ABA and of dry air had been caused by separate mechanisms. Depression of photosynthesis occurred in the presence of sufficient RuBP which indicated that the carboxylation reaction of the carbon-reduction-cycle was inhibited after application of ABA or exposure to dry air.Abbreviations and symbols ABA abscisic acid - C _a partial pressure of CO₂ in the ambient air - C _i partial pressure of CO₂ in the intercellular spaces - I quantum flux - PGA 3-phosphoglyceric acid - RuBP ribulose 1,5-bisphosphate - I _L leaf temperature - w water-vapor pressure difference between leaf and air     

Anatomy of non-uniform leaf photosynthesis

Since 1986, non-uniform photosynthesis over the leaf area that may be attributed to patchy stomatal closure, has been an important issue in stress physiology of photosynthesis. In this review, I first outline the gaseous environment within the intercellular spaces, because this is the most fundamental background of this problem. Then, recent studies approaching non-uniform photosynthesis are reviwed. After examining techniques for the detection of non-uniform photosynthesis or non-uniform stomatal aperture, results of the relevant studies are discussed for respective stress factors, seeking causes and consequences of non-uniform photosynthesis. From these, mechanisms responsible for, and consequences of non-uniform photosynthesis, are considered. The problems which should be challenged are also pointed out.Abbreviations A rate of photosynthetic CO₂ fixation (assimilation rate) - ABA abscisic acid - g conductance for CO₂ or H₂O - p_a partial pressure of CO₂ in the ambient air - p_i partial pressure of CO₂ in the intercellular space - p_i p_i estimated by the conventional gas exchange techniques - q_N non-photochemical quenching - q_p photochemical quenching     

Control of photosynthesis by the carbohydrate level in leaves of the C4 plant Amaranthus edulis L.

Photosynthesis was studied in relation to the carbohydrate status in intact leaves of the C₄ plant Amaranthus edulis. The rate of leaf net CO₂ assimilation, stomatal conductance and intercellular partial pressure of CO₂ remained constant or showed little decline towards the end of an 8-h period of illumination in ambient air (340 bar CO₂, 21% O₂). When sucrose export from the leaf was inhibited by applying a 4-h cold-block treatment (1°C) to the petiole, the rate of photosynthesis rapidly decreased with time. After the removal of the cold block from the petiole, further reduction in photosynthetic rate occurred, and there was no recovery in the subsequent light period. Although stomatal conductance declined with time, intercellular CO₂ partial pressure remained relatively constant, indicating that the inhibition of photosynthesis was not primarily caused by changes in stomatal aperture. Analysis of the leaf carbohydrate status showed a five- to sixfold increase in the soluble sugar fraction (mainly sucrose) in comparison with the untreated controls, whereas the starch content was the same. Leaf osmotic potential increased significantly with the accumulation of soluble sugars upon petiole chilling, and leaf water potential became slightly more negative. After 14 h recovery in the dark, photosynthesis returned to its initial maximum value within 1 h of illumination, and this was associated with a decline in leaf carbohydrate levels overnight. These data show that, in Amaranthus edulis, depression in photosynthesis when translocation is impaired is closely related to the accumulation of soluble sugars (sucrose) in source leaves, indicating feedback control of C₄ photosynthesis. Possible mechanisms by which sucrose accumulation in the leaf may affect the rate of photosynthesis are discussed with regard to the leaf anatomy of C₄ plants.Abbreviations and symbols A net CO₂ assimilation rate - Ci intercellular CO₂ partial pressure - PEP phosphoenolpyruvate - RuBP ribulose-1,5-bisphosphate - water potential - osmotic pressure     

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.     

The midday depression of CO2 assimilation in leaves of Arbutus unedo L.: diurnal changes in photosynthetic capacity related to changes in temperature and humidity

Parts of attached leaves of the sclerophyllous shrub Arbutus unedo were subjected to simulated mediterranean days. Gas exchange was recorded in order to recognize the causes of the midday depression in CO₂ assimilation. Depressions could be induced in part of a leaf: they were local responses. The CO₂-saturation curves of photosynthesis, determined during the morning and afternoon maxima of CO₂ assimilation and during the minimum at midday, established that depressions in CO₂ assimilation were in one-half of the investigated cases totally caused by reversible reductions in the photosynthetic capacity of the leaves, and in the other half almost totally caused by such reductions. An analysis of 37 daily courses showed that morning reductions and afternoon recoveries of stomatal conductance and rate of photosynthesis occurred simultaneously and in proportion to each other, with the result that the partial pressure of CO₂ in the intercellular spaces remained more or less constant. Midday depressions occurred also in detached leaves standing in water. The initiation of a midday depression was not caused by a circadian rhythm, nor was high quantum flux or high temperature a requirement. There was no correlation between the rate of water loss from the leaves, or the amount of water lost, with the degree of reduction of the photosynthetic capacity. However, depressions occurred if an apparent threshold in the water-vapor pressure difference between leaf and air was exceeded. This critical value varied between about 20 and 30 mbar, depending on the leaf investigated. The dominating role of humidity in the induction of the midday depression was further demonstrated when leaf temperature was held constant and the vapor-pressure difference was made to follow the pattern of the mediterranean day: depressions occurred. Depressions however were hardly noticeable when the water-vapor pressure difference was held constant and leaf temperature was allowed to vary. In another set of experiments, leaves were subjected to variations in temperature and humidity independent of the time of the day, under otherwise constant conditions. Photosynthetic capacity and stomatal conductance proved to be almost insensitive to changes in temperature (in a range extending from 20 to 37° C) as long as the water vapor-pressure difference was held constant. If it was not, the rate of photosynthesis began to decline with increasing temperature after a threshold water-vapor pressure difference was exceeded. The position of the resulting apparent temperature optimum of photosynthesis depended on the humidity of the air. We suggest that the ability of A. unedo to respond to a dry atmosphere with a reversible reduction of its photosynthetic capacity (by a still unknown mechanism) is the result of a co-evolution with the development of a strong stomatal sensitivity to changes in humidity.     

Seasonal changes in photosynthetic characteristics of Anemone raddeana,a spring-active geophyte,in the temperate region of Japan

Summary Seasonal changes in the photosynthetic characteristics of intact involucral leaves of Anemone raddeana were investigated under laboratory conditions. Net photosynthesis and constant water vapor pressure deficit showed almost the same seasonal trend. They increased rapidly from mid-April immediately after unfolding of the leaves and reached the maximum in late-April, before the maximum expansion of the leaves. They retained the maximum values until early-May and then decreased toward late-May with a progress of leaf senescence. The calculated values of intercellular CO₂ concentration and relative stomatal limitation of photosynthesis showed no significant change throughout the season. The carboxylation efficiency as assessed by the initial slope of Ci-photosynthesis curve and the net photosynthesis under a high Ci regime varied seasonally in parallel with the change of the light-saturated photosynthesis. The results indicate that the seasonal changes in light-saturated net photosynthesis are not due to a change of stomatal conductance, but to a change in the photosynthetic capacity of mesophyll. Nevertheless, leaf conductance changed concomitantly with photosynthetic capacity, indicating that the seasonal change in stomatal conductance is modulated by the mesophyll photosynthetic capacity such that the intercellular CO₂ concentrations is maintained constant. The shape of light-photosynthesis curve was similar to that of sun-leaf type. The quantum yield also changed simultaneously with the photosynthetic capacity throughout the season.Contribution No. 2965 from the Institute of Low Temperature Science     

The relationship between contents of photosynthetic metabolites and the rate of photosynthetic carbon assimilation in leaves of Amaranthus edulis L.

The relationship between the gas-exchange characteristics of attached leaves of Amaranthus edulis L. and the contents of photosynthetic intermediates was examined in response to changing irradiance and intercellular partial pressure of CO₂. After determination of the rate of CO₂ assimilation at known intercellular CO₂ pressure and irradiance, the leaf was freeze-clamped and the contents of ribulose-1,5-bisphosphate, glycerate-3-phosphate, fructose-1,6-bisphosphate, glucose-6-phosphate, fructose-6-phosphate, triose phosphates, phosphoenolpyruvate, pyruvate, oxaloacetate, aspartate, alanine, malate and glutamate were measured. A comparison between the sizes of metabolite pools and theoretical calculations of metabolite gradients required for transport between the mesophyll and the bundle-sheath cells showed that aspartate, alanine, glycerate-3-phosphate and triose phosphates were present in sufficient quantities to support transport by diffusion, whereas pyruvate and oxaloacetate were not likely to contribute appreciably to the flux of carbon between the two cell types. The amounts of ribulose-1,5-bisphosphate were high at low intercellular partial pressures of CO₂, and fell rapidly as the CO₂-assimilation rate increased with increasing intercellular partial pressures of CO₂, indicating that bundle-sheath CO₂ concentrations fell at low intercellular partial pressures of CO₂. In contrast, the amount of phosphoenolpyruvate and of C₄-cycle intermediates declined at low intercellular partial pressures of CO₂. This behaviour is discussed in relation to the co-ordination of carbon assimilation between the Calvin and C₄ cycles.Abbreviations PEP phosphoenolpyruvate - PGA glycerate-3-phosphate - p _i intercellular CO₂ pressure - RuBP ribulose-1,5-bisphosphate - triose-P triose phosphates     

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.

     

Seasonal changes in photosynthetic characteristics of Pachysandra terminalis (Buxaceae), an evergreen woodland chamaephyte,in the cool temperate regions of Japan

Summary Seasonal changes in photosynthetic capacity, and photosynthetic responses to intercellular CO₂ concentration and irradiance were investigated under laboratory conditions on intact leaves of Pachysandra terminalis. Photosynthetic capacity and stomatal conductance under saturating light intensity and constant water vapor pressure deficit showed almost the same seasonal trend. They increased from early June just after the expansion of leaves, reached the maximum in late-Septemer, and then decreased to winter. In over-wintering leaves they recovered and increased immediately after snow-melting, reached a first maximum in late April, and then decreased to early July in response to the reduction of light intensity on the forest floor. There-after, they increased from mid August, reached a second maximum in late September, and then decreased to winter. The parallel changes of photosynthesis and stomatal conductane indicate a more or less constant intercellular CO₂ concentration throughout the year. The calculated values of relative stomatal limitation of photosynthesis were nearly constant throughout the year, irrespective of leaf age. The results indicate that the seasonal changes in light-saturated photosynthetic capacity are not due to a change of stomatal conductance, but to a change in the photosynthetic capacity of mesophyll. Indeed, carboxylation efficiency assessed by the inital slope of the Ci-photosynthesis curve changed in proportion to seasonal changes of the photosynthetic capacity in both current-year and over-wintered leaves. High photosynthetic capacity in current-year leaves as compared with one-year-old leaves was also due to the high photosynthetic capacity of mesophyll. Nevertheless, stomatal conductance changed in proportion to photosynthetic capacity, indicating that stomatal conductance is regulated by the mesophyll photosynthetic capacity such that the intercellular CO₂ concentrations are maintained constant. The quantum yield also changed seasonally parallel with that in the photosynthetic capacity.Contribution No. 2893 from the Institute of Low Temperature Science     

During photosynthetic induction,biochemical and stomatal limitations differ between Brassica crops

Interventions to increase crop radiation use efficiency rely on understanding of how biochemical and stomatal limitations affect photosynthesis. When leaves transition from shade to high light, slow increases in maximum Rubisco carboxylation rate and stomatal conductance limit net CO₂ assimilation for several minutes. However, as stomata open intercellular [CO₂] increases, so electron transport rate could also become limiting. Photosynthetic limitations were evaluated in three important Brassica crops: Brassica rapa, Brassica oleracea and Brassica napus. Measurements of induction after a period of shade showed that net CO₂ assimilation by B. rapa and B. napus saturated by 10 min. A new method of analyzing limitations to induction by varying intercellular [CO₂] showed this was due to co-limitation by Rubisco and electron transport. By contrast, in B. oleracea persistent Rubisco limitation meant that CO₂ assimilation was still recovering 15 min after induction. Correspondingly, B. oleracea had the lowest Rubisco total activity. The methodology developed, and its application here, shows a means to identify the basis of variation in photosynthetic efficiency in fluctuating light, which could be exploited in breeding and bioengineering to improve crop productivity.     

Stomatal patchiness in Mediterranean evergreen sclerophylls

Midday depression of net photosynthesis and transpiration in the Mediterranean sclerophylls Arbutus unedo L. and Quercus suber L. occurs with a depression of mesophyll photosynthetic activity as indicated by calculated carboxylation efficiency (CE) and constant diurnal calculated leaf intercellular partial pressure of CO₂ (C_i). This work examines the hypothesis that this midday depression can be explained by the distribution of patches of either wide-open or closed stomata on the leaf surface, independent of a coupling mechanism between stomata and mesophyll that results in a midday depression of photosynthetic activity of the mesophyll. Pressure infiltration of four liquids differing in their surface tension was used as a method to show the occurrence of stomatal patchiness and to determine the status of stomatal aperture within the patches. Liquids were selected such that the threshold leaf conductance necessary for infiltration through the stomatal pores covered the expected diurnal range of calculated leaf conductance (g) for these species. Infiltration experiments were carried out with leaves of potted plants under simulated Mediterranean summer conditions in a growth chamber. For all four liquids, leaves of both species were found to be fully infiltratable in the morning and in the late afternoon while during the periods leading up to and away from midday the leaves showed a pronounced patchy distribution of infiltratable and non-infiltratable areas. Similar linear relationships between the amount of liquid infiltrated and g (measured by porometry prior to detachment and infiltration) for all liquids clearly revealed the existence of pneumatically isolated patches containing only wide-open or closed stomata. The good correspondence between the midday depression of CE, calculated under the assumption of no stomatal patchiness, and the diurnal changes in non-infiltratable leaf area strongly indicates that the apparent reduction in mesophyll activity results from assuming no stomatal patchiness. It is suggested that simultaneous responses of stomata and mesophyll activity reported for other species may also be attributed to the occurrence of stomatal patchiness. In Quercus coccifera L., where the lack of constant diurnal calculated C_i and major depression of measured CE at noontime indicates different stomatal behavior, non-linear and dissimilar relationships between g and the infiltratable quantities of the four liquids were found. This indicates a wide distribution of stomatal aperture on the leaf surface rather than only wide-open or closed stomata.Dedicated to Professor Otto L. Lange on the occasion of his 65th birthday     

The photosynthetic characteristics of papyrus in a tropical swamp

Summary Photosynthesis and transpiration was measured in the large emergent C₄ sedge Cyperus papyrus (papyrus) which occupies wide areas of wetland on the African continent. The maximum observed value of net assimilation was 35 mol CO₂ m^-2 s^-1 at full sunlight but light saturation of photosynthesis did not occur. The quantum yield of photosynthesis obtained from the initial slope of the light response curves (0.06 mol mol^-1 incident light) was relatively high and close to previously recorded values for some C₄ grasses. Measurements made over two days showed that stomatal conductance was sensitive to the ambient air vapour pressure deficit (VPD) and was consistently lower on the day when VPD's were higher. There was, however, no marked midday closure of the stomata. Photosynthesis was also reduced on the day when VPD's were higher. The relationship between net photosynthesis and stomatal conductance was close to linear over the range of measurement conditions, with the result that intercellular CO₂ concentrations (C _i ) did not vary markedly. There was some evidence that C _i decreased at high VPD's. The regulation of stomatal movement in papyrus appears to minimise excessive water loss while not severely limiting photosynthesis. The significance of this strategy for a wetland species with plentiful supplies of water is discussed.     

Limitation of photosynthesis by changes in temperature

The aim of this work was to examine the effect of abrupt changes in temperature in the range 5 to 30°C upon the rate of photosynthetic carbon assimilation in leaves of barley (Hordeum vulgare L.). Measurement of the CO₂-assimilation rate in relation to the intercellular partial pressure of CO₂ at different temperatures and O₂ concentrations and at saturating irradiance showed that as the temperature was decreased photosynthesis was saturated at progressively lower CO₂ partial pressures and that the transition between the CO₂-limited and ribulose-1,5-bisphosphate-regeneration-limited rate became more abrupt. Feeding of orthophosphate to leaves resulted in an increased rate of CO₂ assimilation at lower temperatures at around ambient or higher CO₂ partial pressures both in 20% O₂ and in 2% O₂ and it removed the abruptness in the transition between the CO₂-limited and ribulose-1,5-bisphosphate-regeneration-limited rates. Phosphate feeding tended to inhibit carbon assimilation at higher temperatures. The response of carbon assimilation to temperature was altered by feeding orthophosphate, by changing the concentrations of CO₂ or of O₂ or by leaving plants in the dark at 4°C for several hours. Similarly, the response of carbon assimilation to phosphate feeding or to changes in 2% O₂ was altered by leaving the plants in the dark at 4°C. The mechanism of limitation of photosynthesis by an abrupt lowering of temperature is discussed in the light of the results.Abbreviations A rate of CO₂ assimilation - P _i intercellular partial pressure of CO₂ - RuBP ribulose-1,5-bisphosphate     

Transpiration-induced changes in the photosynthetic capacity of leaves

High transpiration rates were found to affect the photosynthetic capacity of Xanthium strumarium L. leaves in a manner analagous to that of low soil water potential. The effect was also looked for and found in Gossypium hirsutum L., Agathis robusta (C. Moore ex Muell.) Bailey, Eucalyptus microcarpa Maiden, Larrea divaricata Cav., the wilty flacca tomato mutant (Lycopersicon esculentum (L.) Mill.) and Scrophularia desertorum (Munz) Shaw. Two methods were used to distinguish between effects on stomatal conductance, which can lower assimilation by reducing CO₂ availability, and effects on the photosynthetic capacity of the mesophyll. First, the response of assimilation to intercellular CO₂ pressure (C _i) was compared under conditions of high and low transpiration. Second, in addition to estimating C _i using the usual Ohm's law analogy, C _i was measured directly using the closed-loop technique of T.D. Sharkey, K. Imai, G.D. Farquhar and I.R. Cowan (1982, Plant Physiol, 60, 657–659). Transpiration stress responses of Xanthium strumarium were compared with soil drought effects. Both stresses reduced photosynthesis at high C _i but not at low C _i; transpiration stress increased the quantum requirement of photosynthesis. Transpiration stress could be induced in small sections of leaves. Total transpiration from the plant did not influence the photosynthetic capacity of a leaf kept under constant conditions, indicating that water deficits develop over small areas within the leaf. The effect of high transpiration on photosynthesis was reversed approximately half-way by returning the plants to low-transpiration conditions. This reversal occurred as fast as measurements could be made (5 min), but little further recovery was observed in subsequent hours.Abbreviations and symbols A photosynthetic CO₂ assimilation rate - C _a ambient CO₂ partial pressure - C _i partial pressure of CO₂ inside the leaf - VPD leaf-to-air water-vapor pressure difference This research was begun while the author was a Postdoctoral Research Fellow at the Australian National University, Canberra     

Benzoxazolin-2-(3<Emphasis Type="Italic">H</Emphasis>)-one reduces photosynthetic activity and chlorophyll fluorescence in soybean

Benzoxazolin-2-(3H)-one (BOA) has been tested in many plants species, but not in soybean (Glycine max). Thus, a hydroponic experiment was conducted to assess the effects of BOA on soybean photosynthesis. BOA reduced net photosynthetic rate, stomatal conductance, and effective quantum yield of PSII photochemistry without affecting intercellular CO₂ concentration or maximal quantum yield of PSII photochemistry. Results revealed that the reduced stomatal conductance restricted entry of CO₂ into substomatal spaces, thus limiting CO₂ assimilation. No change found in intercellular CO₂ concentration and reduced effective quantum yield of PSII photochemistry revealed that CO₂ was not efficiently consumed by the plants. Our data indicated that the effects of BOA on soybean photosynthesis occurred due to the reduced stomatal conductance and decreased efficiency of carbon assimilation. The accumulation of BOA in soybean leaves reinforced these findings.     

Rapid field determination of photosynthetic capacity of cut spruce twigs (Picea abies) at saturating ambient CO2

Summary Routine field determination of the parameters characterizing the activity of the photosynthetic apparatus is often difficult when attached branches of tall trees have to be used for gas exchange measurement. If severed twigs could be used, determining these parameters would be greatly facilitated. Because stomatal conductance changes when twigs or leaves are detached, CO₂ assimilation is usually altered. Thus, measurements made at ambient CO₂ concentration fail to accurately assess the activity of the photosynthetic apparatus because photosynthetic rates greatly depend on the supply of carbon dioxide. However, when photosynthetic carboxylation reactions are saturated by increased CO₂ partial pressure in the mesophyll, CO₂ assimilation rates no longer depend on instantaneous stomatal conductance, as shown by gas exchange measurements of spruce (Picea abies) twigs prior to and following detachment. Because net photosynthesis following detachment at saturating CO₂ remains constant for a minimum of 15 min, photosynthetic measurements of severed twigs may be reliable. This length of time is sufficient for detaching and recutting the twig, assembling a portable minicuvette system, re-establishing steady-state conditions with the gas analyser system, and reading the data over a reasonable period of time. The method described measures the maximal photosynthetic CO₂ assimilation of spruce needles of a single age-class from detached spruce twigs under the following conditions: saturating light, saturating external CO₂-partial pressure, standardized temperature and air humidity in the field. The method is applicable as a routine procedure to characterize the status of the photosynthetic apparatus of spruce trees that may be damaged in the process of forest decline.     

Effect of temperature on net CO2 assimilation and photosystem II quantum yield of electron transfer of French bean (Phaseolus vulgaris L.) leaves during drought stress

The effect of leaf temperature on stomatal conductance and net CO₂ uptake was studied on French bean (Phaseolus vulgaris L.) using either dehydrated attached leaves (25–40% water deficit) or cut leaves supplied with 10^–4 M abscisic acid (ABA) solution to the transpiration stream. Decreasing leaf temperature caused stomatal opening and increased net CO₂ uptake (which was close to zero at around 25° C) to a level identical to that of control leaves (without water deficit) at around 15° C. (i) The ABA effect on stomatal closure was modulated by temperature and, presumably, ABA is at least partly responsible for stomatal closure of french bean submitted to a drought stress. (ii) For leaf temperatures lower than 15° C, net CO₂ uptake was no longer limited by water deficit even on very dehydrated leaves. This shows that dehydrated leaves retain a substantial part of their photosynthetic capacity which can be revealed at normal CO₂ concentrations when stomata open at low temperature. In contrast to leaves fed with ABA, decreasing the O₂ concentration from 21% to 1% O₂ did not increase either the rate of net CO₂ uptake or the thermal optimum for photosynthesis of dehydrated leaves. The quantum yield of PSII electron flow (measured by F/F_m) was lower in 1% O₂ than in 21% O₂ for each leaf pretreatment given (non-dehydrated leaves, dehydrated leaves, and leaves fed with ABA) even within a temperature range in which leaf photosynthesis at normal CO₂ concentration was the same in these two O₂ concentrations. It is concluded that this probably indicates an heterogeneity of photosynthesis, since this difference in quantum yield disappears when using high CO₂ concentrations during measurements.Abbreviations and Symbols ABA abscisic acid - F_m maximum chlorophyll fluorescence - F difference between steady-state chlorophyll fluorescence and F_m - PPFD photosynthetic photon flux density We would like to thank Dr. J.-M. Briantais (Laboratoire d'écologie végétale, Orsay, France) for help during fluorescence measurements and Ms. J. Liebert for technical assistance.