Changing Responses of Stomata to Abscisic Acid and CO2 as Leaves and Plants Age

Willmer, C. M., Wilson, A. B. and Jones, H. G. 1988. Changingresponses of stomata to abscisic acid and CO₂ as leaves andplants age.—J. exp. Bot. 39: 401–410. Stomatal conductances were measured in ageing leaves of Commelinacommunis L. as plants developed; stomatal responses to CO₂ andabscisic acid (ABA) in epidermal strips of C. communis takenfrom ageing leaves of developing plants and in epidermal stripsfrom the same-aged leaves (the first fully-expanded leaf) ofdeveloping plants were also monitored. Stomatal behaviour wascorrelated with parallel measurements of photosynthesis andleaf ABA concentrations. Stomatal conductance in intact leavesdecreased from a maximum of 0-9 cm s^{– 1} at full leaf expansionto zero about 30 d later when leaves were very senescent. Conductancesdeclined more slowly with age in unshaded leaves. Photosynthesisof leaf slices also declined with age from a maximum at fullleaf expansion until about 30 d later when no O₂ exchange wasdetectable. Exogenously applied ABA (0.1 mol m^{– 3}) didnot affect respiration or photosynthesis. In epidermal stripstaken from ageing leaves the widest stomatal apertures occurredabout 10 d after full leaf expansion (just before floweringbegan) and then decreased with age; this decrease was less dramaticin unshaded leaves. The inhibitory effects of ABA on stomatalopening in epidermal strips decreased as leaves aged and wasgreater in the presence of CO₂ than in its absence. When leaveswere almost fully-senescent stomata were still able to open.At this stage, guard cells remained healthy-looking with greenchloroplasts while mesophyll cells were senescing and theirchloroplasts were yellow. Similar data were obtained for stomatain epidermal strips taken from the same-aged leaves of ageingplants. The inhibitory effects of ABA on stomatal opening alsodecreased with plant age. In ageing leaves both free and conjugated ABA concentrationsremained low before increasing dramatically about 30 d afterfull leaf expansion when senescence was well advanced. Concentrationsof free and conjugated ABA remained similar to each other atall times. It is concluded that the restriction of stomatal movements inintact leaves as the leaves and plants age is due mainly toa fall in photosynthetic capacity of the leaves which affectsintracellular CO₂ levels rather than to an inherent inabilityof the stomata to function normally. Since stomatal aperturein epidermal strips declines with plant and leaf age and stomatabecome less responsive to ABA (while endogenous leaf ABA levelsremain fairly constant until leaf senescence) it is suggestedthat some signal, other than ABA, is transmitted from the leafor other parts of the plant to the stomata and influences theirbehaviour. Key words: Abscisic acid, CO₂, Commelina, leaf age, senescence, stomatal sensitivity     

Stomata and Mesophyll Characteristics of Barley Leaf as Affected by Light: Stereological Analysis

The anatomical structure of the second leaf blade of barley{Hordeum vulgare L. cv. Koral) was studied in plants exposedto a photosynthetic photon flux density (PPFD) of 200 µmolm^–2 s^–1 compared with those grown under 25µmolm^–2–11. Design-based stereological methods wereused for the estimation of various leaf anatomical characteristicssuch as mesophyll volume, proportion of intercellular spaces,number of mesophyll cells, mean mesophyll cell volume, and internalleaf surface area. The structure of the mesophyll was more affectedby different levels of PPFD than were the stomatal characteristics.Increased PPFD produced thicker leaves with a larger mesophyllvolume having a higher number of less elongated mesophyll cellsand a larger internal leaf surface area. Key words: Hordeum vulgare, light effect, mesophyll, stereology, stomata     

Contrasted Responses of Two Understorey Species to Direct and Indirect Effects of a Canopy Gap

Positive associations between adult trees and understorey species have been explained either by direct or indirect facilitation. We tested both models by comparing the performance of two understorey species with contrasted stress-tolerance abilities Galium odoratum and Deschampsia flexuosa. Individuals of both species were transplanted in the four combinations of two treatments (gap and removal of an herbaceous competitor, Molinia caerulea). Our experiment demonstrated that direct facilitation of adult trees may explain the restricted occurrence of the shade-demanding Galium within closed forest communities. In contrast, the shade-tolerant Deschampsia was subjected to additional competition within the forest, likely because adult trees had a higher negative effect on light availability and a similar negative effect on nitrogen availability within the forest than did Molinia in the gaps.     

Stomata of Micropropagated DelphiniumPlants Respond to ABA, CO2, Light and Water Potential, but Fail to Close Fully

Morphological and physiological characteristics of micropropagatedplants of Delphinium cv. Princess Caroline were studied. Leavesproduced in vitro showed poor control of water loss which appearsto result from restricted responses by stomata and not frompoor cuticular development. Stomata of leaves produced in vitrowere larger and more frequent than those produced during acclimatization.Despite the fact that stomata from isolated epidermis of leavesproduced in vitro reduced their apertures when exposed to turgor-reducingtreatments, they did not close fully. This, together with highstomatal frequencies might explain the poor control of waterloss shown by intact leaves produced in culture when exposedto dry air. While leaves from acclimatized plants showed almostcomplete closure with ABA, low water potentials, darkness andCO₂, stomata from leaves produced in vitro reduced their apertureswhen exposed to those factors, but only to a limit. Therefore,stomata from leaves cultured in vitro seem to be partially functional,but some physiological or anatomical alteration prevents themfrom closing fully. Stomata from leaves produced in vitro wereparticularly insensitive to ABA which appears to be partly associatedwith the high cytokinin concentration in the culture medium.In the long-term, this stomatal insensitivity to ABA might contributeto plant losses when micropropagated plantlets are transferredto soil. Key words: Micropropagation, stomatal physiology, dehydration, PEG, ABA, BAP, darkness, CO₂, Delphinium     

Modification of the CO2 Responses of Maize Stomata by Abscisic Acid and by Naturally-Occurring and Synthetic Cytokinins

Fragments of maize leaves were incubated at controlled temperatureand irradiance either on distilled water or on one of threeconcentrations of cytokinin (10^–1, 10^–2 and 10^–3mol m^–3). The effects of zeatin or kinetin on stomatalaperture were determined by stripping abaxial epidermis fromthe fragments after incubation and immediately measuring stomatalapertures under the microscope. At each cytokinin concentrationleaf pieces were incubated at 5 or 350 µmol mol^–1CO₂ with or without ABA (10^–1 mol m^–3). At 5.0 µmolmol^–1 CO₂ increasing the concentrations of zeatin hada negligible effect upon stomatal aperture. When air containing350 umol mol^–1 CO₂ was bubbled through the incubationsolutions, apertures of stomata incubated on water were morethan halved. Increasing cytokinin concentrations reduced theeffect of CO₂ on stomata and incubation on 10^–1 mol m^–3zeatin completely removed any CO₂ response. The addition ofABA restored the effect of CO₂, even at the highest cytokininconcentration. Key words: Maize, CO₂ response, ABA, Cytokinins     

Leaf carbohydrate responses to CO2 enrichment at the top of a tropical forest

The accumulation of non-structural leaf carbohydrates is one of the most consistent plant responses to elevated CO₂. It has been found in both fast-and slow-growing plants and is largely independent of the duration of exposure. Changes in leaf quality are thus to be expected, irrespective of other plant responses to atmospheric CO₂ enrichment. However, there is no experimental evidence from tropical forests, the biome with the largest biomass carbon pool. Here we report in situ mesophyll responses of mature tropical trees to a doubling of CO₂. Individually CO₂-enriched leaves on 25 to 35-m-tall forest trees living at 26–35°C can be assumed to experience little sink limitation, and so, may be expected to exhibit no or very little carbohydrate accumulation. We tested this hypothesis using the leaf cup method on leaves accessible via the canopy crane of the Smithsonian Tropical Research Institute in a semi-deciduous tropical forest in Panamá. We also investigated the influence of the leaf-specific light regime, another possible environmental determinant of leaf carbon gain and mobile leaf carbohydrates. Total non-structural carbohydrates (TNC) reached a new steady state concentration after less than 4 days of exposure to twice ambient CO₂ concentration. Against expectation, all four tree species investigated (Anacardium excelsum, Cecropia longipes, C. peltata, Ficus insipida) accumulated significant amounts of TNC (+41 to +61%) under elevated CO₂. The effect was stronger at the end of the daylight period (except for Ficus), but was still significant in all four species at the end of the dark period. In contrast, neither artificial nor natural shading affected leaf TNC. Taken together, these observations suggest that TNC accumulation reflects a mesophyll-bound tissue response specific to elevated CO₂, presumably unrelated to sink limitations. Thus, leaves of tropical forests seem not to be an exception, and will most likely contain more non-structural carbohydrates in a CO₂-rich world. Received: 28 January 1998 / Accepted: 9 April 1998     

Humidity Pretreatment Affects the Responses of Stomata and CO2 Assimilation to Vapor Pressure Difference in C3 and C4 Plants

Experiments were carried out to investigate the long-term influenceof humidity on the short-term responses of stomata and CO₂ assimilationto vapor pressure difference in Oryza sativa (rice, C₃ species)and Panicum maximum (green panic, C₄ species). Plants were grownfor four weeks in growth chambers set at 35% and 85% relativehumidity at 25C air temperature, 38+2 Pa CO₂ partial pressureand 1,700µmol m^-2s^-1 photon flux density. Soil was saturatedwith water in both humidity treatments. Low humidity pretreatmentscaused low leaf conductance and low rates of transpiration andCO₂ assimilation in O. sativa, but small changes in stomatalresponses to humidity and in CO₂ assimilation were found inP. maximum. From the short-term gas exchange experiments, itwas noted that the responsiveness of leaf conductance to vaporpressure difference were affected by humidity pretreatmentsin O. sativa, whereas unaffected in P. maximum. In O. sativameasurements of CO₂ assimilation as a function of internal CO₂partial pressure (A-Ci curve) indicated that low humidity pretreatmentsreduced the CO₂ assimilation at high internal CO₂ partial pressure,but the initial slope of the A-Ci curve was unaffected. Furthermore,plant characteristics such as total dry weight and leaf areaof plants subjected to low umidity were lower than plants subjectedto high humidity. The reductions in O. sativa, however, werelarger than in P. maximum. Stomatal frequency from low humiditygrown plant was higher than that from high humidity grown plantsin both species although there is no significant difference.The data indicated that if the short term inhibition of netCO₂ assimilation at a high vapor pressure difference was imposedduring vegetative growth, the photosynthetic biochemistry andthe resultant plant growth were largely depressed in O. sativa,a C₃ species. (Received May 26, 1992; Accepted November 2, 1992)     

Responses of Stomata to IAA and Fusicoccin at the Opposite Phases of an Entrained Rhythm

The stomata of Commelma communis showed reduced opening responsesto light and low CO₂ concentrations during the night phase oftheir entrained circadian rhythm. Increased supplies of potassiumions, and treatments with indol-3-ylacetic acid and fusicoccin,failed to promote opening during the night phase to a levelequivalent to that in the day phase. The inability of fusiccocinto overcome the suppression of opening during the night phasecontrasts with its ability to counteract the closure inducedby agents such as CO₂, darkness and abscisic acid. It is concludedthat there are at least two basic mechanisms by which the turgorof guard cells can be regulated, one which is susceptible tooverriding control by fusicoccin and another which is unaffectedby fusicoccin. Several previous studies had shown a positive correlation betweenmalate in the epidermis (mainly located in guard cells) andstomatal opening. In the present experiments the aperture/malatecorrelation was broken in epidermis treated with fusicoccinduring the night phase of the rhythm. The amount of malate presentexceeded that associated with the same stomatal aperture inthe day phase. Possible explanations are (1) that fusicoccinstimulates similar proton fluxes out of the guard cells duringboth phases of the rhythm, but an unknown factor imposes a restrictionon stomatal opening during the night phase; (2) that there arelower proton fluxes in the night phase (limited, for example,by a reduced supply of ATP) but chloride availability or transportis reduced to an even greater extent so that a larger productionof malate in the guard cells is required. Key words: Stomata, IAA, Fusicoccin, Rhythms     

Primary Productivity and Water Balance of Grassland Vegetation on Three Soils in a Continuous CO2 Gradient: Initial Results from the Lysimeter CO2 Gradient Experiment

Field studies of atmospheric CO₂ effects on ecosystems usually include few levels of CO₂ and a single soil type, making it difficult to ascertain the shape of responses to increasing CO₂ or to generalize across soil types. The Lysimeter CO₂ Gradient (LYCOG) chambers were constructed to maintain a linear gradient of atmospheric CO₂ (~250 to 500 μl l⁻¹) on grassland vegetation established on intact soil monoliths from three soil series. The chambers maintained a linear daytime CO₂ gradient from 263 μl l⁻¹ at the subambient end of the gradient to 502 μl l⁻¹ at the superambient end, as well as a linear nighttime CO₂ gradient. Temperature variation within the chambers affected aboveground biomass and evapotranspiration, but the effects of temperature were small compared to the expected effects of CO₂. Aboveground biomass on Austin soils was 40% less than on Bastrop and Houston soils. Biomass differences between soils resulted from variation in biomass of Sorghastrum nutans, Bouteloua curtipendula, Schizachyrium scoparium (C₄ grasses), and Solidago canadensis (C₃ forb), suggesting the CO₂ sensitivity of these species may differ among soils. Evapotranspiration did not differ among the soils, but the CO₂ sensitivity of leaf-level photosynthesis and water use efficiency in S. canadensis was greater on Houston and Bastrop than on Austin soils, whereas the CO₂ sensitivity of soil CO₂ efflux was greater on Bastrop soils than on Austin or Houston soils. The effects of soil type on CO₂ sensitivity may be smaller for some processes that are tightly coupled to microclimate. LYCOG is useful for discerning the effects of soil type on the CO₂ sensitivity of ecosystem function in grasslands. Author Contributions: PF conceived study, analyzed data, and wrote the paper. AK, AP analyzed data. DH, VJ, RJ, HJ, and WP conceived study, and conducted research.     

Temperature and CO2Responses of Leaf and Canopy Photosynthesis: a Clarification using the Non-rectangular Hyperbola Model of Photosynthesis

The responses of C₃leaf and canopy gross photosynthesis to increasingtemperature and CO₂can be readily understood in terms of thetemperature and CO₂dependencies of quantum yield (     

Direct capture and conversion of CO2 from air by growing a cyanobacterial consortium at pH up to 11.2

Bioenergy with carbon capture and storage (BECCS) is recognized as a potential negative emission technology, needed to keep global warming within safe limits. With current technologies, large-scale implementation of BECCS would compromise food production. Bioenergy derived from phototrophic microorganisms, with direct capture of CO₂ from air, could overcome this challenge and become a sustainable way to realize BECCS. Here we present an alkaline capture and conversion system that combines high atmospheric CO₂ transfer rates with high and robust phototrophic biomass productivity (15.2 ± 1.0 g/m ²/d). The system is based on a cyanobacterial consortium, that grows at high alkalinity (0.5 mol/L) and a pH swing between 10.4 and 11.2 during growth and harvest cycles.     

不同尺度上植物叶气孔导度对升高CO2的响应

植物叶气孔导度对大气CO2浓度升高的响应可表现在以下几个层面：在叶水平上,叶气孔导度和气孔密度下降;在植物个体水平上,单位叶面积蒸腾下降,植株的水分利用率升高;在生态系统水平上,蒸散降低,土壤泾流和土壤水分含量增加;在全球尺度上,扩大了温室气体的增温效应,同时也降低了全球降雨量增加的趋势。正是因为植物叶气孔导度的变化会影响全球水循环,所以它在全球变化中起着非常重要的作用。但目前的研究结果还不能外推到更大的尺度上去。     

Environmental Responses of the Post-lower Illumination CO(2) Burst as Related to Leaf Photorespiration

As leaf irradiance is decreased in increments, a single transient CO₂ burst is exhibited by C₃ plant leaves. This post-lower illumination CO₂ burst (PLIB) is sensitive to changes in irradiance, to changes in the concentrations of O₂ and CO₂, and to temperature. Increasing O₂ concentrations above ambient produces a progressively larger PLIB while increasing CO₂ concentrations above ambient produces a progressively smaller PLIB. The PLIB, which exhibits many responses to environment common with other methods for measuring photorespiration and photosynthesis, is proposed as a measure of photorespiration in illuminated leaves of C₃ plants. Although the PLIB cannot be used as a quantitative measurement of photorespiration, we propose that the PLIB is a rapid, easy, relatively inexpensive, nondestructive method for evaluating photorespiration in intact illuminated C₃ leaves in air.     

水稻幼苗不同叶位的叶片叶绿素荧光和类胡萝卜素的生物合成对高光胁迫的响应

研究了水稻(Oryza sativa L.)幼苗叶片生长过程中叶绿素荧光和类胡萝卜素各组分含量的变化以及它们对高光胁迫的响应.结果表明:随着叶片的衰老,光合速率、类胡萝卜素不同组分及总的类胡萝卜素含量和叶黄素循环库下降;不同叶龄的叶片经高光胁迫后,第5叶(成熟叶)qN增加的幅度比第6叶(幼嫩叶)和老叶(第3和4叶)大;与高光胁迫前相比,第3、4、5和6叶光系统Ⅱ激发压(1-qP)分别增加了44%、57%、19%和45%;第5叶具有高胡萝卜素含量和高紫黄质到玉米黄质的转化,这与其呈现较强的抗高光胁迫相一致.水稻叶片抵御光抑制的能力与类胡萝卜素水平和类胡萝卜素的生物合成能力以及叶片所处的生长时期相关.     

水稻幼苗不同叶位的叶片叶绿素荧光和类胡萝卜素的生物合成对高光胁迫的响应

研究了水稻 (OryzasativaL .)幼苗叶片生长过程中叶绿素荧光和类胡萝卜素各组分含量的变化以及它们对高光胁迫的响应。结果表明 :随着叶片的衰老 ,光合速率、类胡萝卜素不同组分及总的类胡萝卜素含量和叶黄素循环库下降 ;不同叶龄的叶片经高光胁迫后 ,第 5叶 (成熟叶 )qN增加的幅度比第 6叶 (幼嫩叶 )和老叶 (第 3和 4叶 )大 ;与高光胁迫前相比 ,第 3、4、5和 6叶光系统Ⅱ激发压 (1-qP)分别增加了 4 4 %、5 7%、19%和 4 5 % ;第 5叶具有高胡萝卜素含量和高紫黄质到玉米黄质的转化 ,这与其呈现较强的抗高光胁迫相一致。水稻叶片抵御光抑制的能力与类胡萝卜素水平和类胡萝卜素的生物合成能力以及叶片所处的生长时期相关。     

Responses of Chlorophyll Fluorescence and Carotenoids Biosynthesis to High Light Stress in Rice Seedling Leaves at Different Leaf Position

In the present study, we investigated the changes of photosynthesis, chlorophyll fluorescence and the content of carotenoid pigments in rice ( Oryza sativa L.) seedling leaves and their responses to high light. The results showed that the rate of photosynthesis, the contents of individual and total carotenoids and the pool size of xanthophyll cycle decreased with age increasing of the leaf. When the leaves were exposed to high light for 2 h, the qN of mature leaf (5th leaf) increased more significantly than that of younger (6th leaf) and older leaves (3rd and 4th leaf). Comparing with the leaves before exposure to high light, the excitation pressure on PSⅡ (1-qP) increased by 44%, 57%, 19% and 45% in the 3rd, 4th, 5th and 6th leaf under high light, respectively. The highest content of carotenoids and the greatest conversion of violaxanthin to zeaxanthin were found in the 5th leaf, and it was consistent with the 5th leaf exhibiting the strongest resistance to high light. Our results suggested that the ability of rice leaf to resist photoinhibition is related to the level of carotenoids and the ability of carotenoids biosynthesis.      

Enhancement of the Stomatal Response to Blue Light by Red Light, Reduced Intercellular Concentrations of CO(2), and Low Vapor Pressure Differences

The effects of environmental parameters on the blue light response of stomata were studied by quantifying transient increases in stomatal conductance in Commelina communis following 15 seconds by 0.100 millimole per square meter per second pulses of blue light. Because conductance increases were not observed following red light pulses of the same or greater (30 seconds by 0.200 millimole per square meter per second) fluences, the responses observed could be reliably attributed to the specific blue light response of the guard cells, rather than to guard cell chlorophyll. In both Paphiopedilum harrisianum, which lacks guard cell chloroplasts, and Commelina, the blue light response was enhanced by 0.263 millimole per square meter per second continuous background red light. Thus, the blue light response and its enhancement do not require energy derived from red-light-driven photophosphorylation by the guard cell chloroplasts. In Commelina, reduction of the intercellular concentration of CO₂ by manipulation of ambient CO₂ concentrations resulted in an enhanced blue light response. In both Commelina and Paphiopedilum, the blue light response was decreased by an increased vapor pressure difference. The magnitude of blue-light-specific stomatal opening thus appears to be sensitive to environmental conditions that affect the carbon and water status of the plant.     

Response of Photosynthesis and Dark-CO2-Fixation to Light, CO2 and Temperature in Leaf Slices under Osmotic Stress

Photosynthesis and dark-CO₂-fixation were measured in vacuum-infiltratedleaf slices from the mesophyte Spinacia oleracea and the Mediterraneanxerophyte Arbutus unedo under hypertonic stress as a functionof light-intensity, CO₂-concentration and temperature, in theabsence of stomatal control. Under hypertonic stress, photosynthesis and dark-CO₂-fixationwere inhibited in leaf tissue from both plants. 50% inhibitionof photosynthesis in spinach occurred at about –3.0 MPa,and of dark-CO₂-fixation at about –3.5 MPa. 50% inhibitionof photosynthesis in Arbutus unedo was reached at about –4.0MPa (sorbitol as osmoticum). In both plants, osmotic dehydration decreased the slope andthe maximum of the CO₂- and light-response curves. The slopeof the CO₂-response curve of dark-CO₂-fixation was also decreasedunder hypertonic stress, but the inhibition of the maximal fixationrate was less obvious than for photosynthesis. Photosynthesis and dark-CO₂-fixation differed significantlyin their response to high temperature: under light- and CO₂-saturation,photosynthesis of spinach leaf slices had a temperature optimumat about 37 °C, and it was nearly completely inhibited at45 °C. The rate of dark-CO₂-fixation, however, increasedcontinuously up to 45 °C. Osmotic dehydration increasedthe resistance of photosynthesis to high temperatures. Key words: CO₂ response, Heat stress, Light response, Photosynthesis, Water stress     

Leaf and canopy responses to elevated CO2 in a pine forest under free-air CO2 enrichment

Physiological responses to elevated CO₂ at the leaf and canopy-level were studied in an intact pine (Pinus taeda) forest ecosystem exposed to elevated CO₂ using a free-air CO₂ enrichment (FACE) technique. Normalized canopy water-use of trees exposed to elevated CO₂ over an 8-day exposure period was similar to that of trees exposed to current ambient CO₂ under sunny conditions. During a portion of the exposure period when sky conditions were cloudy, CO₂-exposed trees showed minor (7%) but significant reductions in relative sap flux density compared to trees under ambient CO₂ conditions. Short-term (minutes) direct stomatal responses to elevated CO₂ were also relatively weak (5% reduction in stomatal aperture in response to high CO₂ concentrations). We observed no evidence of adjustment in stomatal conductance in foliage grown under elevated CO₂ for nearly 80 days compared to foliage grown under current ambient CO₂, so intrinsic leaf water-use efficiency at elevated CO₂ was enhanced primarily by direct responses of photosynthesis to CO₂. We did not detect statistical differences in parameters from photosynthetic responses to intercellular CO₂ (A _net-C _i curves) for Pinus taeda foliage grown under elevated CO₂ (550 mol mol^–1) for 50–80 days compared to those for foliage grown under current ambient CO₂ from similar-sized reference trees nearby. In both cases, leaf net photosynthetic rate at 550 mol mol^–1 CO₂ was enhanced by approximately 65% compared to the rate at ambient CO₂ (350 mol mol^–1). A similar level of enhancement under elevated CO₂ was observed for daily photosynthesis under field conditions on a sunny day. While enhancement of photosynthesis by elevated CO₂ during the study period appears to be primarily attributable to direct photosynthetic responses to CO₂ in the pine forest, longer-term CO₂ responses and feedbacks remain to be evaluated.