Low carbon dioxide concentrations can reverse stomatal closure during water stress

Leaf water potentials below threshold values result in reduced stomatal conductance (g_s). Stomatal closure at low leaf water potentials may serve to protect against cavitation of xylem. Possible control of g_s by leaf water potential or hydraulic conductance was tested by drying the rooting medium in four herbaceous annual species until g_s was reduced and then lowering the [CO₂] to determine whether g_s and transpiration rate could be increased and leaf water potential decreased and whether hydraulic conductance was reduced at the resulting lower leaf water potential. In all species, low [CO₂] could reverse the stomatal closure because of drying despite further reductions in leaf water potential, and the resulting lower leaf water potentials did not result in reductions in hydraulic conductance. The relative sensitivity of g_s to internal [CO₂] in the leaves of dry plants of each species averaged three to four times higher than in leaves of wet plants. Two species in which g_s was reputed to be insensitive to [CO₂] were examined to determine whether high leaf to air water vapor pressure differences (D) resulted in increased stomatal sensitivity to [CO₂]. In both species, stomatal sensitivity to [CO₂] was indeed negligible at low D, but increased with D, and low [CO₂] partly or fully reversed closure caused by high D. In no case did low leaf water potential or low hydraulic conductance during drying of the air or the rooting medium prevent low [CO₂] from increasing g_s and transpiration rate.     

Stomatal responses of pearl millet (Pennisetum americanum [L.] Leeke) to leaf water status and environmental factors in the field

Abstract. Factors affecting stomatal conductance (g₁) of pearl millet ( Pennisetum americanum [L.] Leeke), cultivar BJ 104, were examined in the field in India during the dry season.
Diurnal changes in g₁ were evaluated for upper expanded leaves at flowering on two occasions using plants subjected to varying degrees of water stress. Except for the most severely stressed treatment, diurnal changes in g₁ closely matched changes in irradiance ( I ), the promotive effect of which largely overcame opposing influences on g₁ of increasing atmospheric vapour pressure deficit, and decreasing leaf water and turgor potentials (Ψ, Ψ_p).
Two main effects of water stress on g₁ were evident: (i) a decrease in the amplitude of the mid-day peak in g₁, and (ii) a decrease in the time over which high g₁ was maintained, resulting in early (mid-day) closure and hysteresis in the relationship between g₁ and I .
Leaf conductance was greatest for upper leaves and decreased down the canopy. At equivalent depths in the canopy g₁ was higher in flowering than in photoperiodically-retarded plants of the same age. The magnitude of water stress-induced stomatal closure increased down the plant, and was more marked in retarded than in flowering plants.
Within individual stress treatments Ψ of upper leaves decreased linearly as transpiration flux increased. It is concluded that stomatal behaviour of upper leaves of pearl millet at flowering largely operates to maximize assimilation rather than to minimize water loss.     

Physiological function of water channels as affected by salinity in roots of paprika pepper

The sap flow (Jv) and the osmotic pressure-dependent hydraulic conductance (L₀) of detached exuding root systems from paprika pepper plants (cv. Albar) were measured. Plants stressed with NaCl (30 m M ) and with six times the macronutrients of the Hoagland nutrient solution (6×HNS) were compared with controls grown in complete Hoagland nutrient solution. Jv of +NaCl and +6×HNS plants decreased markedly, but recovered to values similar to those of controls after removal of the treatments. Hydraulic conductance L₀ was always less in NaCl plants than in controls and 6×HNS. A total increase in the ion concentration of the xylem (except Na⁺ and Cl⁻) was observed with both treatments. In control and 6×HNS plants, HgCl₂ treatment (50 μ M ) caused a sharp decline in L₀ to values similar to those of NaCl-stressed roots, but were restored by treating with 5 m M dithiothreitol (DTT). However, in NaCl roots only a slight effect of Hg²⁺ and DTT was observed. In each treatment, there was no difference in the flux of K⁺ into the xylem after HgCl₂ and DTT application. The results suggest that NaCl decreased L₀ of the roots by reducing either the activity or abundance of Hg-sensitive water channels. The putative reduction in water-channel function of NaCl-treated plants did not seem to be due to the osmotic effect.     

Changes in leaf hydraulics and stomatal conductance following drought stress and irrigation in Ceratonia siliqua (Carob tree)

Changes in leaf hydraulic conductance (K) were measured using the vacuum chamber technique during dehydration and rehydration of potted plants of Ceratonia siliqua . K of whole, compound leaves as well as that of rachides and leaflets decreased by 20–30% at leaf water potentials (Ψ_L) of −1.5 and −2.0 MPa, i.e. at Ψ_L values commonly recorded in field-growing plants of the species. Higher K losses (up to 50%) were measured for leaves at Ψ_L of −2.5 and −3.0 MPa, i.e. near or beyond the leaf turgor loss point. Leaves of plants rehydrated while in the dark for 30 min, 90 min and 12 h recovered from K loss with characteristic times and to extents inversely proportional to the initial water stress applied. Leaf conductance to water vapour of plants dehydrated to decreasing Ψ_L and rehydrated at low transpiration was inversely related to loss of K, thus suggesting that leaf vein embolism and refilling (and related changes in leaf hydraulics) may play a significant role in the stomatal response.     

Latent manganese deficiency increases transpiration in barley (Hordeum vulgare)

To investigate if latent manganese (Mn) deficiency leads to increased transpiration, barley plants were grown for 10 weeks in hydroponics with daily additions of Mn in the low n M range. The Mn-starved plants did not exhibit visual leaf symptoms of Mn deficiency, but Chl a fluorescence measurements revealed that the quantum yield efficiency of PSII (F_v/F_m) was reduced from 0.83 in Mn-sufficient control plants to below 0.5 in Mn-starved plants. Leaf Mn concentrations declined from 30 to 7 μg Mn g⁻¹ dry weight in control and Mn-starved plants, respectively. Mn-starved plants had up to four-fold higher transpiration than control plants. Stomatal closure and opening upon light/dark transitions took place at the same rate in both Mn treatments, but the nocturnal leaf conductance for water vapour was still twice as high in Mn-starved plants compared with the control. The observed increase in transpiration was substantiated by ¹³C-isotope discrimination analysis and gravimetric measurement of the water consumption, showing significantly lower water use efficiency in Mn-starved plants. The extractable wax content of leaves of Mn-starved plants was approximately 40% lower than that in control plants, and it is concluded that the increased leaf conductance and higher transpirational water loss are correlated with a reduction in the epicuticular wax layer under Mn deficiency.     

Effects of low concentrations of O3, leaf age and water stress on leaf diffusive conductance and water use efficiency in soybean

Ozone, leaf age and water stress each affected leaf conductance in soybean [ Glycine max (L.) Merr. Hodgson], but there were no interactions among these factors. Exposure to increased concentrations of O₃ (0.01, 0.05, 0.09. and 0.13 μl l⁻¹) resulted in linear declines in abaxial and adaxial conductances in leaves of all ages. There were no differences in relative response to O₃ between the two leaf surfaces. For well-watered plants, water use efficiency also decreased with exposure to increased O₃ concentrations (water-stressed plants were not tested). Abaxial conductance increased as leaves aged from 4 to 10 days and then declined with further aging. Adaxial conductance decreased with all increases in leaf age beyond 4 days, and the ratio of abaxial/adaxial conductance increased continuously throughout the leaf lifespan. During water-stress cycles (water withheld for 2–3 days) leaves of water-stressed plants had lower conductances than those from well-watered plants, and there was no difference in relative response between abaxial and adaxial stomata.     

Effects of tree height on branch hydraulics, leaf structure and gas exchange in California redwoods

We examined changes in branch hydraulic, leaf structure and gas exchange properties in coast redwood ( Sequoia sempervirens ) and giant sequoia ( Sequoiadendron giganteum ) trees of different sizes. Leaf-specific hydraulic conductivity ( k _L) increased with height in S. sempervirens but not in S. giganteum , while xylem cavitation resistance increased with height in both species. Despite hydraulic adjustments, leaf mass per unit area (LMA) and leaf carbon isotope ratios ( δ ¹³C) increased, and maximum mass-based stomatal conductance ( g _mass) and photosynthesis ( A _mass) decreased with height in both species. As a result, both A _mass and g _mass were negatively correlated with branch hydraulic properties in S. sempervirens and uncorrelated in S. giganteum . In addition, A _mass and g _mass were negatively correlated with LMA in both species, which we attributed to the effects of decreasing leaf internal CO₂ conductance ( g _i). Species-level differences in wood density, LMA and area-based gas exchange capacity constrained other structural and physiological properties, with S. sempervirens exhibiting increased branch water transport efficiency and S. giganteum exhibiting increased leaf-level water-use efficiency with increasing height. Our results reveal different adaptive strategies for the two redwoods that help them compensate for constraints associated with growing taller, and reflect contrasting environmental conditions each species faces in its native habitat.     

Effects of root restriction on the growth and physiology of cucumber plants

Cucumber plants ( Cucumis sativus L. cv. Athene F1) were grown with four treatments: unrestricted root volume fruiting (UF); unrestricted root volume non-fruiting (UN); restricted root volume fruiting (RF); and restricted root volume non-fruiting (RN). Restricting root volume to 40 ml reduced leaf area, and by day 60 leaf area was only 20% that of unrestricted plants. Leaf area reduction in restricted plants was due to a combination of smaller and fewer leaves. Root restriction strongly depressed total dry matter production in both root and shoot. Significant differences of treatments in shoot and root growth rates were akpparent 30 days after sowing. RN plants had a 70% lower net photosynthesis (P_n), stomatal conductance (g_s) and transpiration rate (E) measured on day 50, while root restriction had no effect on P_n in fruiting plants, although g_s and E were significantly decreased due to restriction. Respiration capacity of restricted roots decreased sharply after day 24 compared with unrestricted root systems. Initially, O₂ may have been the limiting resource and root respiration capacity a major factor involved in root restriction, since it causes imbalances in root growth substances and related hormones that alter the plant's morphology.     

Hydraulic architecture of Monstera acuminata: evolutionary consequences of the hemiepiphytic growth form

The hydraulic architecture of the secondary hemiepiphyte Monstera acuminata was examined in native plants from Los Tuxtlas, Veracruz, Mexico, to determine how it compared to better-known growth forms such as trees, shrubs, lianas and primary hemiepiphytes. Monstera acuminata starts its life cycle as a prostrate herb. As it ascends a tree or other vertical support, the stem becomes thicker, produces larger leaves, and may die back from the base upwards until only aerial feeding roots serve to connect the stem to the soil. Unlike the pattern of vessel-size distribution along the stems of woody dicotyledons, M. acuminata has its wider vessels at the top of the stem, decreasing in diameter towards the base. Also peculiar is the fact that Huber values (axis area/distal leaf area) tend to increase exponentially at higher positions within the plant. Based on the hydraulic conductivity ( k _h) and leaf-specific conductivity (LSC, k _h/distal leaf area), the base of the stem potentially acts as a severe hydraulic constriction. This constriction is apparently not limiting, as aerial roots are produced further up the stem. The plants have remarkably strong root pressures, up to 225 kPa, which may contribute to the maintenance of functional vessels by refilling them at night or during periods of very high atmospheric humidity, as in foggy weather and rain. In common with dicotyledonous plants, vessel length, vessel diameter, k _h, specific conductivity ( k _s, k _h/axis area) and LSCs were all positively correlated with axis diameter. The features of the hydraulic architecture of M. acuminata may be an evolutionary consequence of an anatomical constraint (lack of vascular cambium and therefore of secondary growth) and the special requirements of the hemiepiphytic growth form.     

Decoupling the influence of leaf and root hydraulic conductances on stomatal conductance and its sensitivity to vapour pressure deficit as soil dries in a drained loblolly pine plantation

The study examined the relationships between whole tree hydraulic conductance ( K _tree) and the conductance in roots ( K _root) and leaves ( K _leaf) in loblolly pine trees. In addition, the role of seasonal variations in K _root and K _leaf in mediating stomatal control of transpiration and its response to vapour pressure deficit ( D ) as soil-dried was studied. Compared to trunk and branches, roots and leaves had the highest loss of conductivity and contributed to more than 75% of the total tree hydraulic resistance. Drought altered the partitioning of the resistance between roots and leaves. As soil moisture dropped below 50%, relative extractable water (REW), K _root declined faster than K _leaf. Although K _tree depended on soil moisture, its dynamics was tempered by the elongation of current-year needles that significantly increased K _leaf when REW was below 50%. After accounting for the effect of D on g _s, the seasonal decline in K _tree caused a 35% decrease in g _s and in its sensitivity to D , responses that were mainly driven by K _leaf under high REW and by K _root under low REW. We conclude that not only water stress but also leaf phenology affects the coordination between K _tree and g _s and the acclimation of trees to changing environmental conditions.     

Leaf hydraulics and drought stress: response, recovery and survivorship in four woody temperate plant species

Efficient conduction of water inside leaves is essential for leaf function, yet the hydraulic-mediated impact of drought on gas exchange remains poorly understood. Here we examine the decline and subsequent recovery of leaf water potential ( Ψ _leaf), leaf hydraulic conductance ( K _leaf), and midday transpiration ( E ) in four temperate woody species exposed to controlled drought conditions ranging from mild to lethal. During drought the vulnerability of K _leaf to declining Ψ _leaf varied greatly among the species sampled. Following drought, plants were rewatered and the rate of E and K _leaf recovery was found to be strongly dependent on the severity of the drought imposed. Gas exchange recovery was strongly correlated with the relatively slow recovery of K _leaf for three of the four species, indicating conformity to a hydraulic-stomatal limitation model of plant recovery. However, there was also a shift in the sensitivity of stomata to Ψ _leaf suggesting that the plant hormone abscisic acid may be involved in limiting the rate of stomatal reopening. The level of drought tolerance varied among the four species and was correlated with leaf hydraulic vulnerability. These results suggest that species-specific variation in hydraulic properties plays a fundamental role in steering the dynamic response of plants during recovery.     

Drought effects on photosynthesis and fluorescence in hard wheat cultivars grown in the field

Net photosynthesis of the flag leaf of hard wheat ( Triticum durum L. evs Valforte, Produra, Adamello, Karel, Appulo and El Amel from the collection of the Instituto di Cerealicultura. Foggia, Italy) of different water potential has been studied on three consecutive years. Net photosynthesis was measured in natural conditions with a LI-COR portable instrument and in saturating CO₂ with an oxygen electrode. Net photosynthesis and stomatal conductance were significantly lower in the unirrigated leaves. However, the ratio of intercellular CO₂, concentration (C₁) to ambient CO₃ concentration (C_a) around the stressed plants was similar to the irrigated control. The maximal rate of photosynthesis in saturating CO₂, (P_nmax). measured in the second year of the experiment, was quite close to photosynthesis under natural conditions, indicating that CO₂ supply was not limiting. These results suggest that altered mesophyll photosynthetic capacity, rather than stomatal closure, causes the observed reduction in photosynthesis in the unirrigated plants. The variable fluorescence yield (_v/F_m) in predarkened leaves measured for two consecutive years, did not show differences between treatments or between cultivars. However, the analysis of the slow transients, measured the last year of the experiment, showed a linear relation between the fluorescence decline from the maximum initial level (P) and maximum photosynthesis (P_nmax).     

Stomatal limitation of photosynthesis in winter production of greenhouse tomato plants

In May, greenhouse tomato ( Lycopersicon esculentum Mill.) plants near the end of their winter production cycle were shown to exhibit a diurnal photosynthetic decrease. In order to identify the physiological causes of this decline, we compared in May the photosynthetic characteristics of the fifth youngest leaves from tomato plants of different ages corresponding to a winter production (11-month-old plants) and to a spring production (5-month-old plants). Although the leaves were developed simultaneously under the same environmental conditions, only the ones from the winter production showed a diurnal decline of the in situ CO₂ assimilation rate (A _{CO 2}). This was accompanied by a decline of internal CO₂ and stomatal conductance and by large accumulations of hexoses. When stomatal closure was relieved under saturated CO₂ concentration (5%) using a leaf-disc electrode system, the fifth leaves of both tomato cultures had similar maximum quantum efficiency of O₂ evolution (Φ_max), light-saturated rate of O₂ evolution (P_max) and quantum efficiency of photosystem II (PSII) photochemistry (ΔF/F'_m, q _P and q _N ). We concluded that the diurnal decline of A _{CO 2} observed in winter tomato production during May originates from a stomatal limitation that is not dependent on environmental conditions but rather related to the developmental stage of the plants.     

Does calcium ameliorate the negative effect of NaCl on melon root water transport by regulating aquaporin activity?

The hydraulic conductance ( L ₀) of detached, exuding root systems from melon ( Cucumis melo cv. Amarillo oro) was measured. All plants received a half-strength Hoagland nutrient solution, and plants stressed either solely with NaCl (50 mM) or with NaCl (50 mM) following treatment (2 d) with CaCl₂ (10 mM) were compared with controls and CaCl₂-treated (10 mM) plants. The L ₀ of NaCl-treated plants was markedly decreased when compared to control and CaCl₂-treated plants, but the decrease was smaller when NaCl was added to plants previously treated with CaCl₂. A similar effect was observed when the flux of Ca²⁺ into the xylem and the Ca²⁺ concentration in the plasma membrane of the root cells were determined. In control, CaCl₂- and NaCl + CaCl₂-treated plants, HgCl₂ treatment (50 μM) caused a sharp decline in L ₀ to values similar to those of NaCl-stressed roots, but L ₀ was restored by treatment with 5 mM DTT. However, in NaCl roots only a slight effect of Hg²⁺ and DTT were observed. The effect of all treatments on L ₀ was similar to that on osmotic water permeability ( P _f) of individual protoplasts isolated from roots. The results suggest that NaCl decreased the passage of water through the membrane and roots by reducing the activity of Hg-sensitive water channels. The ameliorative effect of Ca²⁺ on NaCl stress could be related to water-channel function.     

Contrasting effects of simazine on the photosynthetic physiology and leaf morphology of two Populus clones

Differential effects of simazine (2-chloro-4,6-bis(ethylamino)- s -triazine) on the physiology of two Populus clones were investigated in a greenhouse study. Additions of 5 mg/pot simazine to young plants had no deleterious morphological or physiological effects on clone NC 5328 ( P. x euramericana cv. I 45/51; Section Aigeiros), but reduced the rate of CO₂ fixation, increased CO₂ compensation concentrations and lowered the specific leaf weight of clone NE 388 ( P. maximowiczü x P. trichocarpa cv. Kingston; section Tacamahaca). Abaxial leaf conductance to water vapor was not affected in NE 388. Deleterious effects of simazine on NE 388 were detected ca 48 h after exposure of plants to simazine and generally became more pronounced thereafter. Visual symptoms of injury were evident at ca 2 weeks after simazine application.
Toxic responses to simazine in clone NE 388 varied in different portions of the crown. Inhibition of photosynthesis and increased CO₂ compensation concentrations were more pronounced in the region of recently matured leaves, but were somewhat less in the region of expanding leaves. Older mature leaves in the lower crown region showed no visual symptoms of injury and the rate of photosynthesis and CO₂ compensation concentrations were largely unaffected.     

Drought and chlorophyll fluorescence in field-grown potato (Solanum tuberosum)

Light interception, stomatal conductance and chlorophyll fluorescence were measured in potato ( Solanum tuberosum L.) grown either irrigated, or droughted from the time of plant emergence. Compared with the irrigated treatment, drought reduced both light interception and stomatal conductance. In both treatments, the yields of variable fluorescence in the dark- and light-adapted states (F_y/F_m and F'_v/F'_m, respectively) were negatively correlated with photosynthetic photon flux density (PPFD) and mirrored daytime changes in PPFD. Photochemical quenching was positively correlated with PPFD, but the dominant effect of F'_v/F'_m resulted in a decrease in the quantum yield of photosystem II (PSII) electron transport with increasing PPFD.
Drought had no significant effect on the functioning of PSII and the balance between photochemical and non-photochemical quenching was unaffected. Non-photochemical quenching was not increased by drought and the quantum yield of PSII electron transport was unaffected. It is concluded that, in leaves of droughted plants, excess energy, resultant of stomatal limitation of photosynthesis, was dissipated by photochemical quenching such as increased photorespiration.     

Correlations of stomatal conductance with hydraulic and chemical factors in several deciduous tree species in a natural habitat

Recent research in whole-plant stomatal physiology, conducted largely with potted plants in controlled environments, suggests that stomatal conductance ( g _s) might be more closely linked to plant chemical variables than to hydraulic variables. To test this in a field situation, seasonal g _s was examined in relation to a number of plant and environmental variables in 11 temperate, deciduous forest tree species. Stomatal conductance was generally better correlated with environmental variables (air temperature, vapor pressure deficit, PPFD) than with plant variables, and slightly better correlated with plant hydraulic variables (shoot water and osmotic potentials) than with plant chemical variables (xylem sap ABA concentration, xylem sap pH). We examined a model, developed previously for maize, which describes regulation of g _s by xylem sap ABA concentration with leaf water status acting to modify stomatal sensitivity to the ABA signal. This model explained slightly more variation in seasonal g _s in the forest trees than did single plant variables but not more variation than most single environmental variables. Response surface models, especially those incorporating environmental variables, were more consistently successful at explaining g _s across species.     

Nitrate reductase activity, nitrogenase activity and photosynthesis of black alder exposed to chilling temperatures

Actinorhizal ( Frankia -nodulated) black alder [ Alnus glutinosa (L.) Gaertn.] seedlings fertilized with 0.36 m M nitrate (low nitrate fertilizer treatment) or 7.14 m M nitrate (high nitrate fertilizer treatment) and acclimated in a growth chamber for 2 weeks were exposed to 2.5 h of night-time chilling temperatures of −1 to 4°C. Cold treatment decreased nitrogenase activity (acetylene reduction activity) 33% for low nitrate fertilized plants and 41% for high nitrate fertilized plants. Recovery of nitrogenase activity occurred within 7 days after chilling treatment. In contrast, in vivo nitrate reductase (NR) activities of leaves and fine roots increased immediately after chilling then decreased as nitrogenase activities recovered. Fine roots of alder seedlings exhibited NR activities proportional to the amounts of nitrate in the rooting medium. In contrast, the NR activities of leaves were independent of substrate and tissue nitrate levels and corresponded to nitrogenase activity in the root nodules. In a separate experiment, net photosynthesis (PS) of similarly treated black alder seedlings was measured before and after chilling treatments. Net PS declined in response to chilling by 17% for plants receiving low nitrate fertilizer and 19% for plants receiving high nitrate fertilizer. After chilling, stomatal conductance (g_s) decreased by 39% and internal CO₂ concentration (c_i) decreased by 5% in plants receiving the high nitrate fertilizer, whereas plants receiving the low nitrate fertilizer showed no change in g_s and a 13% increase in c_i. Results indicate that chilling stimulates stomatal closure only at the high nitrate level and that interference with biochemical functions is probably the major impact of chilling on PS.     

Effects of manganese toxicity on leaf CO2 assimilation of contrasting common bean genotypes

Parameters related to leaf photosynthesis were evaluated in three genotypes of common bean ( Phaseolus vulgaris L.) with contrasting tolerance to Mn toxicity. Two short-term studies in solution culture were used to assess the effect of excess Mn on CO₂ assimilation in mature and immature leaves. Mn toxicity decreased total chlorophyll content only in immature leaves, with a consequent reduction of leaf CO₂ assimilation. Mature leaves that showed brown speckles characteristic of Mn toxicity, did not suffer any detriment in their capacity to assimilate CO₂, at least in a 4-day experiment. Stomatal conductance and transpiration were not affected by the presence of high levels of Mn in leaf tissue. Lower stomatal conductance and transpiration rates were observed only in leaves with advanced chlorosis. Differences among genotypes were detected as increased chlorosis in the more sensitive genotype ZPV-292, followed by A-283 and less chlorosis in the tolerant genotype CALIMA. Since CO₂ assimilation expressed per unit of chlorophyll was not different between high-Mn plants and control plants, we conclude that the negative effect of Mn toxicity on CO₂ assimilation can be explained by a reduction in leaf chlorophyll content.