Diel trends in stomatal response to ozone and water deficit: a unique relationship of midday values to growth and allometry in Pima cotton?

Plant responses to ozone (O3 ) and water deficit (WD) are commonly observed, although less is known about their interaction. Stomatal conductance (gs ) is both an impact of these stressors and a protective response to them. Stomatal closure reduces inward flux of O3 and outward flux of water. Stomatal measurements are generally obtained at midday when gas exchange is maximal, but these may not be adequate surrogates for stomatal responses observed at other times of day, nor for non-stomatal responses. Here, we find in Pima cotton that stomatal responses to O3 observed at midday do not reflect responses at other times. Stomata were more responsive to O3 and WD near midday, despite being at quasi-steady state, than during periods of active opening or closing in morning or evening. Stomatal responsivity to O3 was not coincident with maximum gas exchange or with periods of active regulation, but coincident with plant sensitivity to O3 previously determined in this cultivar. Responses of pigmentation and shoot productivity were more closely related to stomatal responses at midday than to responses at other times of day under well-watered (WW) conditions, reflecting higher stomatal responsivity, sensitivity to O3 , and magnitude of midday gs . Under WD conditions, shoot responses were more closely related to early morning gs. Root responses were more closely related to early morning gs under both WW and WD. Responses of stomata to O3 at midday were not good surrogates for stomatal responses early or late in the day, and may not adequately predicting O3 flux under WD or when maximum ambient concentrations do not occur near midday.

Unusually high soil nitrogen oxide emissions influence air quality in a high-temperature agricultural region.

Fertilized soils have large potential for production of soil nitrogen oxide (NOx=NO+NO2), however these emissions are difficult to predict in high-temperature environments. Understanding these emissions may improve air quality modelling as NOx contributes to formation of tropospheric ozone (O3), a powerful air pollutant. Here we identify the environmental and management factors that regulate soil NOx emissions in a high-temperature agricultural region of California. We also investigate whether soil NOx emissions are capable of influencing regional air quality. We report some of the highest soil NOx emissions ever observed. Emissions vary nonlinearly with fertilization, temperature and soil moisture. We find that a regional air chemistry model often underestimates soil NOx emissions and NOx at the surface and in the troposphere. Adjusting the model to match NOx observations leads to elevated tropospheric O3. Our results suggest management can greatly reduce soil NOx emissions, thereby improving air quality.

Perchlorate content of plant foliage reflects a wide range of species-dependent accumulation but not ozone-induced biosynthesis.

Perchlorate (ClO4(-)) interferes with uptake of iodide in humans. Emission inventories do not explain observed distributions. Ozone (O3) is implicated in the natural origin of ClO4(-), and has increased since pre-industrial times. O3 produces ClO4(-)in vitro from Cl(-), and plant tissues contain Cl(-) and redox reactions. We hypothesize that O3 exposure may induce plant synthesis of ClO4(-). We exposed contrasting crop species to environmentally relevant O3 concentrations. In the absence of O3 exposure, species exhibited a large range of ClO4(-) accumulation but there was no relationship between leaf ClO4(-) and O3, whether expressed as exposure or cumulative flux (dose). Older, senescing leaves accumulated more ClO4(-) than younger leaves. O3 exposed vegetation is not a source of environmental ClO4(-). There was evidence of enhanced ClO4(-) content in the soil surface at the highest O3 exposure, which could be a significant contributor to environmental ClO4(-).

High ozone increases soil perchlorate but does not affect foliar perchlorate content.

Ozone (O) is implicated in the natural source inventory of ClO, a hydrophilic salt that migrates to groundwater and interferes with the uptake of iodide in mammals, including humans. Tropospheric O is elevated in many urban and some rural areas in the United States and globally. We previously showed that controlled O exposure at near-ambient concentrations (up to 114 nL L, 12-h mean) did not increase foliar ClO. Under laboratory conditions, O has been shown to oxidize Cl to ClO. Plant tissues contain Cl and exhibit responses to O invoking redox reactions. As higher levels of O are associated with stratospheric incursion and with developing megacities, we have hypothesized that exposure of vegetation to such elevated O may increase foliar ClO. This would contribute to ClO in environments without obvious point sources. At these high O concentrations (up to 204 nL L, 12-h mean; 320 nL L maximum), we demonstrated an increase in the ClO concentration in surface soil that was linearly related to the O concentration. There was no relationship of foliar ClO with O exposure or dose (stomatal uptake). Accumulation of ClO varied among species at low O, but this was not related to soil surface ClO or to foliar ClO concentrations following exposure to O. These data extend our previous conclusions to the highest levels of plausible O exposure, that tropospheric O contributes to environmental ClO through interaction with the soil but not through increased foliar ClO.

Demonstration of a diel trend in sensitivity of Gossypium to ozone: a step toward relating O3 injury to exposure or flux.

Plant injury by ozone (O3) occurs in three stages, O3 entrance through stomata, overcoming defences, and attack on bioreceptors. Concentration, deposition, and uptake of O3 are accessible by observation and modelling, while injury can be assessed visually or through remote sensing. However, the relationship between O3 metrics and injury is confounded by variation in sensitivity to O3. Sensitivity weighting parameters have previously been assigned to different plant functional types and growth stages, or by differentially weighting O3 concentrations, but diel and seasonal variability have not been addressed. Here a plant sensitivity parameter (S) is introduced, relating injury to O3 dose (uptake) using three independent injury endpoints in the crop species, Pima cotton (Gossypium barbadense). The diel variability of S was determined by assessment at 2h intervals. Pulses of O3 (15 min) were used to assess passive (constitutive) defence mechanisms and dose was used rather than concentration to avoid genetic or environmental effects on stomatal regulation. A clear diel trend in S was apparent, with maximal sensitivity in mid-afternoon, not closely related to gas exchange, whole leaf ascorbate, or total antioxidant capacity. This physiologically based sensitivity parameter provides a novel weighting factor to improve modelled relationships between either flux or exposure to O3, and O3 impacts. This represents a substantial improvement over concentration- or phenology-based weighting factors currently in use. Future research will be required to characterize the variability and metabolic drivers of diel changes in S, and the performance of this parameter in prediction of O3 injury.

Root and shoot gas exchange respond additively to moderate ozone and methyl jasmonate without induction of ethylene: ethylene is induced at higher O3 concentrations.

The available literature is conflicting on the potential protection of plants against ozone (O(3)) injury by exogenous jasmonates, including methyl jasmonate (MeJA). Protective antagonistic interactions of O(3) and MeJA have been observed in some systems and purely additive effects in others. Here it is shown that chronic exposure to low to moderate O(3) concentrations (4-114 ppb; 12 h mean) and to MeJA induced additive reductions in carbon assimilation (A (n)) and root respiration (R (r)), and in calculated whole plant carbon balance. Neither this chronic O(3) regime nor MeJA induced emission of ethylene (ET) from the youngest fully expanded leaves. ET emission was induced by acute 3 h pulse exposure to much higher O(3) concentrations (685 ppb). ET emission was further enhanced in plants treated with MeJA. Responses of growth, allocation, photosynthesis, and respiration to moderate O(3) concentrations and to MeJA appear to be independent and additive, and not associated with emission of ET. These results suggest that responses of Pima cotton to environmentally relevant O(3) are not mediated by signalling pathways associated with ET and MeJA, though these pathways are inducible in this species and exhibit a synergistic O(3)×MeJA interaction at very high O(3) concentrations.

No interaction between methyl jasmonate and ozone in Pima cotton: growth and allocation respond independently to both.

Ozone (O3) is damaging to plants, inducing signalling pathways involving antagonism between jasmonates and ethylene. These pathways mediate O3 responses, particularly to acute exposure, and their manipulation protected several species against acute and chronic O3. We use chronic daily exposure of up to 163 ppb O3, and twice weekly application of up to 320 microg plant(-1) methyl jasmonate (MeJA) to test two hypothesizes: 1) a low rate of MeJA does not affect growth but increases O3 sensitivity; 2) a high rate inhibits growth but reduces O3 sensitivity. Both hypotheses were rejected. Growth declined with increases in both MeJA and O3. MeJA at 40 microg plant(-1) caused no direct effect, and at 160 microg plant(-1) reduced growth similarly at all O3. Neither rate altered O3 sensitivity. These additive responses are not consistent with protection by MeJA in this system. They may reflect inter-specific differences in signalling, since O3 concentrations used here exceeded some reported acute exposures. Alternatively, parallel responses to O3 and MeJA may suggest that O3-induced jasmonates play a developmental role in chronic response but no protective role in the absence of lesions characteristic of acute exposure. MeJA appears useful as a probe of these mechanisms.

O3 impacts on plant development: a meta-analysis of root/shoot allocation and growth.

The mechanism of O3 action on plants remains poorly characterized. Symptoms include visible lesions on the leaf surface, reduced growth and a hypothesized reduction in allocation of carbohydrate to roots. The generality of this latter phenomenon has not been demonstrated. Here, a meta-analysis is performed of all available experimental data, to test the hypotheses that O3 exposure of the shoot inhibits biomass allocation below ground (the root/shoot allometric coefficient, k) and inhibits whole-plant growth rate [relative growth rate (RGR)]. Both k and RGR were significantly reduced by O3 (5.6 and 8.2%, respectively). Variability in k was greater than in RGR, and both exhibited some positive as well as mostly negative responses. The effects on k were distinct from the effects on RGR. In some cases, k was reduced while RGR was unaffected. Slow-growing plants (small RGR) exhibited the largest declines in k. These observations may have mechanistic implications regarding O3 phytotoxicity. There were no effects of type of exposure chamber on sensitivity to O3. The analyses indicate that the O3 inhibition of allocation to roots is real and general, but variable. Further experiments are needed for under-represented plant groups, to characterize exceptions to this generalization and to evaluate O3--environment interactions.

Ozone impacts on cotton: towards an integrated mechanism.

Vegetation removes tropospheric ozone (O(3)) mainly through uptake by stomata. O(3) reduces growth, photosynthesis, and carbohydrate allocation. Effects on mesophyll photosynthesis, may reducing carbohydrate source strength and, indirectly, carbohydrate translocation. Alternatively direct translocation, itself, could explain all of these observations. O(3)-reduced root proliferation inhibits exploitation of soil resources and interferes with underground carbon sequestration. Simulations with cotton suggest O(3)-disrupted root development could indirectly reduce shoot photosynthesis. Strong evidence for O(3) impacts on both carbon assimilation and carbon translocation exists, but data determining the primacy of direct or indirect O(3) effects on either or both processes remain inconclusive. Phloem loading may be particularly sensitive to O(3). Further research on metabolic feedback control of carbon assimilation and phloem loading activity as affected by O(3) exposure is required.

Ozone increases root respiration but decreases leaf CO2 assimilation in cotton and melon.

It is well established that exposure of plant foliage to tropospheric ozone (O3) inhibits photosynthetic gas exchange in leaves and the translocation of current photosynthate to sink tissues. It is less clear what impact O3-reduced source strength has on the physiological responses of sink tissue such as fine roots. The responses were investigated of carbon acquisition in leaves and carbon utilization in the respiration of fine roots, following chronic (weeks) and acute (hours) exposures to O3 in open top chambers. Previous reports indicate increased, decreased, and unchanged rates of root respiration following exposure to O3. A decline in source activity is confirmed, but an increase in sink respiration is reported in fine roots of Pima cotton (cv. S-6) and muskmelon (cv. Ambrosia hybrid). Leaf source strength and root sink activity changed in opposing directions, thus there was no positive correlation that might indicate direct substrate control of root function. Additional linkages between shoot and root following exposure to O3 may be involved.

Ecological effects of particulate matter.

Atmospheric particulate matter (PM) is a heterogeneous material. Though regulated as un-speciated mass, it exerts most effects on vegetation and ecosystems by virtue of the mass loading of its chemical constituents. As this varies temporally and spatially, prediction of regional impacts remains difficult. Deposition of PM to vegetated surfaces depends on the size distribution of the particles and, to a lesser extent, on the chemistry. However, chemical loading of an ecosystem may be determined by the size distribution as different constituents dominate different size fractions. Coating with dust may cause abrasion and radiative heating, and may reduce the photosynthetically active photon flux reaching the photosynthetic tissues. Acidic and alkaline materials may cause leaf surface injury while other materials may be taken up across the cuticle. A more likely route for metabolic uptake and impact on vegetation and ecosystems is through the rhizosphere. PM deposited directly to the soil can influence nutrient cycling, especially that of nitrogen, through its effects on the rhizosphere bacteria and fungi. Alkaline cation and aluminum availability are dependent upon the pH of the soil that may be altered dramatically by deposition of various classes of PM. A regional effect of PM on ecosystems is linked to climate change. Increased PM may reduce radiation interception by plant canopies and may reduce precipitation through a variety of physical effects. At the present time, evidence does not support large regional threats due to un-speciated PM, though site-specific and constituent-specific effects can be readily identified. Interactions of PM with other pollutants and with components of climate change remain important areas of research in assessment of challenges to ecosystem stability.

Ozone impacts on allometry and root hydraulic conductance are not mediated by source limitation nor developmental age.

O(3)could reduce growth and carbohydrate allocation to roots by direct inhibition of photosynthesis and source strength. Alternatively, O(3) could reduce growth indirectly by inhibition of root hydraulic development through a primary lesion in carbohydrate translocation. Another alternative is that O(3) could slow the rate of plant development, only apparently altering carbohydrate allocation at a given plant age. Pima cotton (Gossypium barbadense L.) is used to address these possibilities, and four hypotheses were tested/ (1) O(3) exposure reduces leaf pools of soluble sugars; (2) pruning leaf area and reducing source strength to match that of O(3)-treated plants reproduces O(3)-effects; (3) pruning lower leaf area more closely reproduces O(3) effects than pruning upper leaf area; and (4) manipulating plant age and thereby plant size to match O(3)-treated plants reproduces O(3)-effects. All were falsified. Soluble sugars did not decline. Pruning upper and lower leaves and manipulating plant age all reduced biomass and leaf area similarly to O(3)-exposure, but neither reproduced O(3) effects on biomass allocation nor root function. It is concluded that O(3) induces an allometric shift in carbohydrate allocation that is not mediated by photosynthetic inhibition nor by alteration of developmental age. Effects of O(3) could be mediated by direct effects on phloem loading, with consequent inhibition of translocation to roots and root system development.

Genotypic variability in vulnerability of leaf xylem to cavitation in water-stressed and well-irrigated sugarcane.

Genotypic variability in vulnerability of leaf xylem to water-stress-induced cavitation was determined in four sugarcane (Saccharum sp.) clones using detached leaf segments in a hydraulic conductivity apparatus. Vulnerability curves were constructed by plotting the percentage of maximum conductivity versus leaf water potential (psi(I)) and fitting curves using a Weibull function. The psi(I) at which each clone lost 10, 50, and 80% of maximum conductivity was determined. Maximum conductivity per unit of leaf width was positively associated with metaxylem vessel diameter. The commercial clone H65-7052 exhibited the highest and the nondomesticated S. spontaneum exhibited the lowest conductivity. All four clones lost substantial conductivity at values of psi(I) less negative than -1.4 MPa, but H65-7052 was able to maintain 50% conductivity to lower psi(I) than the other clones. S. spontaneum sustained the most negative psi(I) (-1.99 MPa) before reaching the 80% conductivity loss point. Clone H69-8235 was consistently the most vulnerable to initial loss of conductivity. These vulnerability functions were used in conjunction with field measurements of psi(I) to estimate diurnal losses in leaf hydraulic conductivity under irrigated and droughted conditions. H69-8235 lost up to 50% of its conductivity during the day, even when well irrigated, and more than 80% when subjected to drought. The other clones exhibited lower conductivity losses. These losses are apparently reversed overnight by root pressure. Despite their close genetic relationships, these clones exhibited large differences in conductivity, in the vulnerability of their xylem to cavitation, and in gas exchange behavior. The potential for altering water relations by selecting for particular hydraulic characteristics is discussed.

Effects of water deficit on flower opening in coffee (Coffea arabica L.).

The response of coffee (Coffea arabica L.) floral buds to different water deficits followed by re-irrigation was investigated. Flower opening was stimulated by irrigation after one period of water deficit if predawn leaf water potential declined below -0.8 MPa. Similar stimulation of flowering was observed when less severe but more prolonged water deficits (ca. -0.3 to -0.5 MPa for two weeks) were imposed, even if water deficit was relieved by re-irrigation several times during this period. Consistent results were obtained in the field and in two greenhouse locations. Stimulation of flower opening by water deficit followed by re-irrigation was restricted to buds at the "open white cluster" stage of development (Stage 4). Only buds at this stage exhibited development of secondary xylem. Split-root experiments indicated that a root signal stimulated flower opening, independently of predawn or midday leaf water status. Frequent irrigation to prevent flowering, followed by a controlled water deficit and re-irrigation to stimulate flowering, may represent a practical method to synchronize flowering and shorten the harvest period in leeward coffee production areas in Hawaii.

Leaf water relations and maintenance of gas exchange in coffee cultivars grown in drying soil.

Plant water status, leaf tissue pressure-volume relationships, and photosynthetic gas exchange were monitored in five coffee (Coffea arabica L.) cultivars growing in drying soil in the field. There were large differences among cultivars in the rates at which leaf water potential (Psi(L)) and gas exchange activity declined when irrigation was discontinued. Pressure-volume curve analysis indicated that increased leaf water deficits in droughted plants led to reductions in bulk leaf elasticity, osmotic potential, and in the Psi(L) at which turgor loss occurred. Adjustments in Psi(L) at zero turgor were not sufficient to prevent loss or near loss of turgor in three of five cultivars at the lowest values of midday Psi(L) attained. Maintenance of protoplasmic volume was more pronounced than maintenance of turgor as soil drying progressed. Changes in assimilation and stomatal conductance were largely independent of changes in bulk leaf turgor, but were associated with changes in relative symplast volume. It is suggested that osmotic and elastic adjustment contributed to maintenance of gas exchange in droughted coffee leaves probably through their effects on symplast volume rather than turgor.

The magnitude of the stomatal response to blue light : modulation by atmospheric humidity.

The effect of leaf-air vapor pressure difference (VPD) on the magnitude of the stomatal response to blue light was investigated in soybean (Glycine max) by administering blue light pulses (22 seconds by 120 micromoles per square meter per second) at different levels of VPD and temperature. At 20 degrees C and 25 degrees C, the magnitude of the integrated conductance response decreased with increasing VPD (0.4 to 2.6 kiloPascals), due to an earlier onset of stomatal closure that terminated the pulse response. In contrast, at 30 degrees C this magnitude increased with rising VPD (0.9 to 3.5 kiloPascals), due to an increasing maximum excursion of the conductance response despite the accelerated onset of stomatal closure. When the feedforward response of stomata to humidity caused steady state transpiration to decrease with increasing VPD, the magnitude of the pulse-induced conductance response correlated with VPD rather than with transpiration. This suggests that water relations or metabolite movements within epidermal rather than bulk leaf tissue interacted with guard cell photobiological properties in regulating the magnitude of the blue light response. VPD modulation of pulse magnitude could reduce water loss during stomatal responses to transient illumination in natural light environments.

Carbon Isotope Discrimination in Coffee Genotypes Grown under Limited Water Supply.

Photosynthetic gas exchange, plant-water relations characteristics, and stable carbon isotope discrimination (Delta) were evaluated for five Coffea arabica L. genotypes growing under two soil moisture regimes in the field. The Delta of leaf tissue was strongly correlated (r = -0.95) with inherent water use efficiency (ratio of assimilation to stomatal conductance; A/g). The variation in inherent water use efficiency (WUE) among genotypes was 30% for plants irrigated weekly. The higher WUE exhibited by some of these plants resulted from reduced g rather than increased photosynthetic capacity at a given g. Withholding irrigation for 1 month caused Delta to decline substantially in expanding leaf tissue of all genotypes. A strong correlation (r = 0.92) was found between Delta and plant hydraulic efficiency estimated as the ratio of g to the diurnal range in leaf water potential (Psi(l)). The Delta values for plants irrigated weekly adequately predicted drought-induced changes in Delta (r = 0.99) and midday Psi(l) (r = 0.95). The results indicated that Delta might be used to evaluate several aspects of plant performance and response to specific environmental conditions, once suitable background physiological data have been gathered.

Calcium Effects on Stomatal Movement in Commelina communis L. : Use of EGTA to Modulate Stomatal Response to Light, KCl and CO(2).

Stomatal movements depend on both ion influx and efflux; attainment of steady state apertures reflects modulation of either or both processes. The role of Ca(2+) in those two processes was investigated in isolated epidermal strips of Commelina communis, using the Ca(2+) chelator EGTA to reduce apoplastic [Ca(2+)]. The results suggest that a certain concentration of Ca(2+) is an absolute requirement for salt efflux and stomatal closure. EGTA (2 millimolar) increased KCl-dependent stomatal opening in darkness and completely inhibited the dark-induced closure of initially open stomata. Closure was inhibited even in a KCl-free medium. Thus, maintenance of stomata in the open state does not necessarily depend on continued K(+) influx but on the inhibition of salt efflux. Opening in the dark was stimulated by IAA in a concentration-dependent manner, up to 15.4 micrometer without reaching saturation, while the response to EGTA leveled off at 9.2 micrometer. IAA did not inhibit stomatal closure to the extent it stimulated opening. The response to IAA is thus consistent with a primary stimulation of opening, while EGTA can be considered a specific inhibitor of stomatal closing since it inhibits closure to a much larger degree than it stimulates opening. CO(2) causes concentration-dependent reduction in the steady state stomatal aperture. EGTA completely reversed CO(2)-induced closing of open stomata but only partially prevented the inhibition of opening.

Guard cells of Commelina communis L. do not respond metabolically to osmotic stress in isolated epidermis: Implications for stomatal responses to drought and humidity.

We investigated the hypothesis that stomatal aperture is regulated by epidermal water status. Detached epidermal peels of Commelina communis L. or leaf disks with epidermis attached were incubated in graded solutions of mannitol (0-1.2 M) containing KCl. In isolated epidermis, guard-cell solute content of open stomata did not decrease in response to desiccation. Guard cells of closed stomata accumulated solutes to the same extent in all levels of mannitol tested. There was no evidence of stress-induced hydroactive closure nor of inhibition of hydroactive opening, even when guard cells of closed stomata were initially plasmolyzed. Hydropassive, osmometer-like, changes in stomatal aperture in the isolated epidermis were induced by addition or removal of mannitol, but these did not involve changes in guard-cell solute content. In leaf disks, stomata exhibited clear hydroactive stomatal responses. Steady-state guard-cell solute content of initially open and initially closed stomata decreased substantially with increasing mannitol. Stomata were completely closed above approx. 0.4 M mannitol, near the turgor-loss point for the bulk leaf tissue. Stomata of Commelina did not exhibit direct hydroactive responses to environmental or epidermal water status. Stomatal responses to water deficit and low humidity may be indirect, mediated by abscisic acid or other signal metabolite(s) from the mesophyll.