Use of thermography for quantitative studies of spatial and temporal variation of stomatal conductance over leaf surfaces

This paper describes a new approach to the calibration of thermal infrared measurements of leaf temperature for the estimation of stomatal conductance and illustrates its application to thermal imaging of plant leaves. The approach is based on a simple reformulation of the leaf energy balance equation that makes use of temperature measurements on reference surfaces of known conductance to water vapour. The use of reference surfaces is an alternative to the accurate measurement of all components of the leaf energy balance and is of potentially wide application in studies of stomatal behaviour. The resolution of the technique when applied to thermal images is evaluated and some results of using the approach in the laboratory for the study of stomatal behaviour in leaves of Phaseolus vulgaris L. are presented. Conductances calculated from infrared measurements were well correlated with estimates obtained using a diffusion porometer.     

A coupled model of stomatal conductance, photosynthesis and transpiration

A model that couples stomatal conductance, photosynthesis, leaf energy balance and transport of water through the soil–plant–atmosphere continuum is presented. Stomatal conductance in the model depends on light, temperature and intercellular CO₂ concentration via photosynthesis and on leaf water potential, which in turn is a function of soil water potential, the rate of water flow through the soil and plant, and on xylem hydraulic resistance. Water transport from soil to roots is simulated through solution of Richards’ equation. The model captures the observed hysteresis in diurnal variations in stomatal conductance, assimilation rate and transpiration for plant canopies. Hysteresis arises because atmospheric demand for water from the leaves typically peaks in mid‐afternoon and because of uneven distribution of soil matric potentials with distance from the roots. Potentials at the root surfaces are lower than in the bulk soil, and once soil water supply starts to limit transpiration, root potentials are substantially less negative in the morning than in the afternoon. This leads to higher stomatal conductances, CO₂ assimilation and transpiration in the morning compared to later in the day. Stomatal conductance is sensitive to soil and plant hydraulic properties and to root length density only after approximately 10 d of soil drying, when supply of water by the soil to the roots becomes limiting. High atmospheric demand causes transpiration rates, LE, to decline at a slightly higher soil water content, θ_s, than at low atmospheric demand, but all curves of LE versus θ_s fall on the same line when soil water supply limits transpiration. Stomatal conductance cannot be modelled in isolation, but must be fully coupled with models of photosynthesis/respiration and the transport of water from soil, through roots, stems and leaves to the atmosphere.     

Effect of black bean aphid, Aphis fabae, on transpiration, stomatal conductance and crude protein content of faba bean

This study was initiated to investigate effects of damage by 0, 5 and 10 aphids/plant on the physiology of faba bean plants throughout different feeding periods and at two plant development stages. Immediately following removal of Aphis fabae, measurements showed 84–229% increase in transpiration rate. These changes were proportional to the number of aphids and infestation duration. Injury by A. fabae caused the stomatal conductance to be much higher in the leaves of infested plants. Leaf stomatal conductance of the infested plants increased significantly by 51–224% depending on initial aphid densities and feeding intervals. This increase was proportional to the infestation level for each date. Length of infestation period and plant growth stage seemed to have no clear effect on stomatal apertures. Aphid feeding caused a damage of about 7–33% of crude protein levels in the leaf tissue. This reduction increased with increasing infestation levels and time, except for 28‐day‐old plants on 28 days. The physiological effects of aphid feeding on water vapour and chemical composition of damaged leaves are particularly serious when the population is high.     

Oscillations in stomatal conductance of hybrid poplar leaves in the light and dark

Cycling of stomatal conductance in three hybrid poplar ( Populus sp.) cultivars was observed under a variety of conditions. Illumination of plants kept previously in the dark induced very large oscillations with a period of about 40 min and large oscillations with a shorter period (< 10 min) were superimposed on the longer cycles. During these oscillations, large changes in conductance could occur very rapidly (1.0 cm s⁻¹ in 3 min). Plants in constant light also displayed both long and short term cycles in conductance, but these were smaller in amplitude than those induced by sudden illumination. Stomatal oscillations were also observed in darkness and after darkening of previously illuminated plants. These oscillations had shorter (< 30 min) and less regular periods than those observed in the light. Such cycling in the dark is rare. Cycling of the two leaf surfaces was sometimes in synchrony in the light, and more so after a perturbation. Little synchrony between the two surfaces was observed in the dark. Stomatal movements of different leaves on a plant were usually relatively independent. Transient stomatal opening occurred following leaf excision in the light or dark, and often after sudden darkening of intact leaves. Also, stomata of intact leaves sometimes transiently closed following illumination.     

黄土高原苹果园蒸腾导度大气驱动规律比较

植物蒸腾导度是表征土壤-植物-大气连续体（SPAC）中植物-大气间水汽传导过程、反映植物水分调控能力的一类重要变量,常见有冠层导度（G_c）、冠层气孔导度（G_s）与叶片气孔导度（g_s）,明确三者在反映冠层蒸腾过程时的异同或关联性对于理解植物水分利用机制具有重要意义。本研究基于对黄土高原果园苹果树生长季内树干液流（J_s）及环境因子的连续观测,计算了G_c、G_s及脱耦联系数（Ω）等变量,并与短期连续观测的叶片气孔导度（g_s）比较,分析了G_c、G_s和g_s在反映冠层蒸腾特征方面的异同及其关系。结果表明,日变化过程中G_s、g_s呈"单峰"型曲线,而G_c则呈"先增后减,午后抬升"的"双峰"型曲线。g_s与G_s存在较紧密的线性关系（R²=0.80）,但与G_c的线性关系较弱（R²=0.02）。G_c、G_s均随大气水汽压亏缺（VPD）的变化呈现确定的规律,其中,上边界函数呈递减的对数函数关系,平均值则符合先增后减的Log-Normal函数关系（R²>0.95）,拐点对应的VPD值分别为1.33和1.16 kPa。在一日内,G_s对VPD变化的响应过程与g_s对VPDL （基于叶片温度计算的水汽压亏缺）变化的响应过程总体一致,其一致性高于G_c对VPD变化的响应。整个生长季（4-10月）中果树的Ω平均值为0.12,随着Ω递减,G_c与G_s的线性相关性愈趋紧密,其斜率呈递增趋势,G_c越来越趋近于G_s。研究结果表明,在北方地区,基于树干液流的监测能较准确的推导整株并估算林分的冠层蒸腾导度。与实测g_s的变化过程比较,G_s比G_c具有更高的一致性,G_s可以作为描述苹果树水分利用过程响应大气驱动的更为恰当的变量。     

整树水力导度协同冠层气孔导度调节森林蒸腾

冠层气孔导度决定森林的蒸腾效率,它对驱动水汽移动的水汽应力的响应受树木水力结构的影响,并随水汽压亏缺上升和水力导度下降而降低,维持水势在最低阈值之上,避免出现水力灾变,调控冠层蒸腾。由于叶形和树冠结构的特点,部分脱耦联反映了湿润地区阔叶林冠层与大气的水汽交换特征,单纯以气孔导度的变化难以完整描述水分通量的调节规律,因而,需要考虑冠层气孔导度与水力导度协同控制冠层蒸腾的潜在机理。通过整合叶片气孔气体交换、树干液流、冠层微气象和其他环境因子的野外观测值,估测不同时间尺度的森林冠层气孔导度与大气的脱耦联系数和变异范围,以基于树干液流的冠层蒸腾,结合叶片/土壤水势梯度计算的水力导度,分析水力导度影响冠层气孔导度响应水汽压亏缺的敏感性,可以揭示和阐明水力导度和冠层气孔导度联合调节森林蒸腾的机理,对准确估测全球变化背景下森林对水资源利用的潜在生态效应有明显的理论意义。     

Dynamics of stomatal patches for a single surface of Xanthium strumarium L. leaves observed with fluorescence and thermal images

Fluorescence and thermal imaging were used to examine the dynamics of stomatal patches for a single surface of Xanthium strumarium L. leaves following a decrease in ambient humidity. Patches were not observed in all experiments, and in many experiments the patches were short-lived. In some experiments, however, patches persisted for many hours and showed complex temporal and spatial patterns. Rapidly sampled fluorescence images showed that the measurable variations of these patches were sufficiently slow to be captured by fluorescence images taken at 3-min intervals using a saturating flash of light. Stomatal patchiness with saturating flashes of light was not demonstrably different from that without saturating flashes of light, suggesting that the regular flashes of light did not directly cause the phenomenon. Comparison of simultaneous fluorescence and thermal images showed that the fluorescence patterns were largely the result of stomatal conductance patterns, and both thermal and fluorescence images showed patches of stomatal conductance that propagated coherently across the leaf surface. These nondispersing patches often crossed a given region of the leaf repeatedly at regular intervals, resulting in oscillations in stomatal conductance for that region. The existence of these coherently propagating structures has implications for the mechanisms that cause patchy stomatal behaviour as well as for the physiological ramifications of this phenomenon.     

北京山区侧柏林冠层-大气蒸腾导度模拟及环境因子响应

蒸腾导度模型是衡量冠层-大气界面水汽输出的重要阻力模型,研究其特征及对环境因子的响应,为揭示森林冠层-大气界面水汽输出阻力机制提供理论依据。以首都圈森林生态系统定位观测研究站侧柏林为研究对象,采用TDP热探针法测定侧柏林树干液流密度,同步监测光合有效辐射、饱和水汽压差、气温、风速等主要环境因子,分析冠层导度和空气动力学导度的动态变化,构建冠层-大气蒸腾导度模型并模拟,明确冠层-大气蒸腾导度对各环境因子的响应关系。结果表明:蒸腾导度季节变化表现为非生长季与冠层导度趋势一致,生长季与空气动力学导度趋势一致,全年均为单峰趋势。冬季蒸腾导度与冠层导度保持较稳定差值(45 mol m^(-2 )s^-1左右),其他季节蒸腾导度与冠层导度、空气动力学导度的最大差值,均在各季节冠层导度、空气动力学导度的峰值水平。全年日均蒸腾导度冬季最大(86.92 mol m^(-2 )s^-1),其他季节较小且稳定(40—50 mol m^(-2 )s^-1之间)。在非生长季各环境因子对蒸腾导度的影响与对冠层导度的影响基本一致,温度为主要影响因子(r=-0.198),其他环境因子影响较小(r<0.1);在生长季中风速为主要影响因子(r=0.488),光合有效辐射(r=0.228)和饱和水汽压差(r=-0.299)的影响明显升高,温度的影响降低(r=0.114)。蒸腾导度模型较好的模拟了冠层-大气界面侧柏蒸腾不同季节的变化规律,阐明了各环境因子和冠层导度、空气动力学导度对蒸腾导度的影响机制,证实在生长季应重视空气动力学导度对蒸腾的影响。     

Sensitivity of infrared water vapor analyzers to oxygen concentration and errors in stomatal conductance

Use of infrared analyzers to measure water vapor concentrations in photosynthesis systems is becoming common. It is known that sensitivity of infrared carbon dioxide and water vapor analyzers is affected by the oxygen concentration in the background gas, particularly for absolute analyzers, but the potential for large errors in estimates of stomatal conductance due to effects of oxygen concentration on the sensitivity of infrared water vapor analyzers is not widely recognized. This work tested three types of infrared water vapor analyzers for changes in sensitivity of infrared water vapor analyzers depending on the oxygen content of the background gas. It was found that changing from either 0 or 2% to 21% oxygen in nitrogen decreased the sensitivity to water vapor for all three types of infrared water vapor analyzers by about 4%. The change in sensitivity was linear with oxygen mole fraction. The resulting error in calculated stomatal conductance would depend strongly on the leaf to air vapor pressure difference and leaf temperature, and also on whether leaf temperature was directly measured or calculated from energy balance. Examples of measurements of gas exchange on soybean leaves under glasshouse conditions indicated that changing from 21% to 2% oxygen produced an artifactual apparent increase in stomatal conductance which averaged about 30%. Similar errors occurred for `conductances' of wet filter paper. Such errors could affect inferences about the carbon dioxide dependence of the sensitivity of photosynthesis to oxygen. This revised version was published online in June 2006 with corrections to the Cover Date.     

Distribution profile of stomatal conductance and its interrelations to transpiration rate and water dynamics in young maize laminas under sulfate deprivation

Seven-day-old maize (Zea mays) plants were grown hydroponically for 10 days in S-deprived nutrient solution. The distribution profiles according to the position on the stem of the S-deprived laminas’ stomatal conductance, transpiration rate, photosynthetic rate, dry mass, water content, and specific surface area were monitored relative to control among others. Photochemical efficiency of photosystem II remained unaffected by the deprivation, as well as the specific surface area of all but the embryonic laminas after d2. In S-deficient plants, the embryonic (L0) and the uppermost lamina or the one below it presented mostly significant changes. The response ratios (Rr) of the L0 stomatal conductance oscillated; the oscillation started with an increase at d2. The corresponding Rr values of L0 transpiration and photosynthetic rates started oscillating at d4 in the same fashion. At d8, an increasing gradient appeared in water-content Rr values from L1 to the uppermost lamina. At d10, all but the embryonic laminas presented significantly reduced Rr values in water content. Changes in dry mass and surface area of laminas were synchronized. In control, the transpiration rate expressed per DM unit remained constant during the examined period, while under the deprivation it followed a power function of surface area.     

Epidermal conductance, stomatal density and stomatal size among genotypes of Sorghum bicolor (L.) Moench

Abstract. The ability of a plant to survive severe water deficits depends on its ability to restrict water loss through the leaf epidermis after stomata attain minimum aperture. At this stage, the rate of water loss is regulated by the epidermal conductance (g_c). Low g_c would be a useful selection criterion to identify genotypes with enhanced survival capability. Consequently, variation in g_c among Sorghum bicolor (L.) Moench genotypes was evaluated. Since there is little conclusive evidence linking g _c with leaf waxiness, alternative hypotheses relating g _c to stomatal trails were also examined. Epidermal conductance varied from 6.3 to 17.6mmol m⁻² s⁻¹ among sorghum genotypes. It was unrelated to stomatal pore length which varied with genotype and to pore depth which was similar for all genotypes measured. However, g _c, increased with increasing stomatal density. This indicates that stomatal density plays a direct role in water loss even at very low conductances. The association of low stomatal density with low g _c is consistent with the hypothesis that at the smallest stomata aperture, water loss from the epidermis above guard cell teichodes becomes a significant source of leaf water loss. Since low g _c is directly related to crop survival under severe water deficits, it is recommended that genotypes with low g _c. be selected using the selection criterion of stomatal density.     

黄土高原油松冠层气孔导度和蒸腾变化特征与模拟

油松是黄土高原重要的造林树种,模拟其冠层气孔导度和蒸腾对区域水量平衡计算和人工林可持续经营具有重要意义。基于2015—2018年TDP(Thermal dissipation probes)方法所测得液流数据,分析了黄土高原地区油松冠层平均气孔导度(g_c)与冠层蒸腾(Tr)的变化特征与影响因素,并采用Penman-Monteith公式和Jarvis型气孔导度模型模拟了其g_c和Tr的变化过程,结果表明：(1)该地区油松g_c和Tr日内变化均呈现单峰型,日均蒸腾耗水量为(1.25±0.57) mm/d,生长季(4—10月)总蒸腾耗水量均值为195.47 mm。(2)g_c的日内变化受太阳辐射(Rad)驱动(偏相关系数为0.65),当Rad高于300 W/m²时,驱动作用减弱;g_c的日内变化受水汽压亏缺(VPD)控制(偏相关系数为-0.41),随VPD的增加而降低;g_c的日际变化受土壤水分限制(偏相关系数为0.46),当根区相对有效含水率(RE...     

北京山区油松和元宝槭冠层气孔导度特征及其环境响应

叶片气孔是植物进行水汽交换的通道, 影响着植物的蒸腾和光合作用。然而叶片气孔行为受环境条件和树种类型的影响, 不同树种冠层气孔导度对环境因子响应的差异性, 以及在生长季不同时期叶片气孔对冠层蒸腾的调节作用是否会发生改变, 仍不清楚。该研究目的是通过探究各环境因子对不同树种冠层气孔导度的相对贡献率以及叶片气孔对冠层蒸腾的调节作用, 为深入了解植物水分利用状况和山区森林经营提供参考依据。于2018年生长季以北京八达岭国家森林公园内的58年生油松(Pinus tabuliformis)和39年生元宝槭(Acer truncatum)为研究对象, 利用热扩散技术对其树干液流进行连续监测, 并同步监测环境因子。利用彭曼公式计算冠层气孔导度(G_s)。主要结果: (1)油松和元宝槭日间G_s在日、月时间尺度上存在明显差异。5-7月油松和元宝槭日动态G_s均随饱和水汽压差(VPD)和太阳辐射(GR)的增加呈上升趋势, 上升持续时间比8月和9月长; 在月尺度上, 随着VPD、GR的降低和土壤湿度(VWC)的升高, G_s从5月到9月整体上升。(2)利用增强回归树法分析得到VWC和VPD对G_s的贡献率最大, 其次是GR、气温和风速。VWC和VPD对油松G_s的贡献率分别为66.4%和17.4%, 对元宝槭G_s的贡献率分别为54.8%和21.0%。(3)油松和元宝槭的dG_s/dlnVPD值与参考冠层气孔导度之间的斜率均显著高于0.6, 气孔调节作用相对较强。综上所述, 气孔对环境因子的响应在树种以及生长季不同时期之间存在差异, 为防止水分过度散失, 两树种在不同土壤水分条件下均通过严格的气孔调节控制蒸腾量。     

Limitation of transpiration by hydraulic conductance and xylem cavitation in Betula occidentalis

The extent to which stomatal conductance (g_s) was capable of responding to reduced hydraulic conductance (k)and preventing cavitation-inducing xylem pressures was evaluated in the small riparian tree, Betula occidentalis Hook. We decreased k by inducing xylem cavitation in shoots using an air-injection technique. From 1 to 18 d after shoot injection we measured midday transpiration rate (E), g_s, and xylem pressure (Ψ_p-xylem) on individual leaves of the crown. We then harvested the shoot and made direct measurements of k from the trunk (2–3 cm diameter) to the distal tip of the petioles of the same leaves measured for E and g_s. The k measurement was expressed per unit leaf area (k_l, leaf-specific conductance). Leaves measured within 2 d of shoot injection showed reduced g_s and E relative to non-injected controls, and both parameters were strongly correlated with k_l At this time, there was no difference in leaf Ψ_p-xylem between injected shoots and controls, and leaf Ψ_p-xylem was not significantly different from the highest cavitation-inducing pressure (Ψ_p-cav) in the branch xylem (-1.43 ± 0.029 MPa, n=8). Leaves measured 7–18 d after shoots were injected exhibited a partial return of g_s and E values to the control range. This was associated with a decrease in leaf Ψ_p-xylem below Ψ_p-cav and loss of foliage. The results suggest the stomata were incapable of long-term regulation of E below control values and that reversion to higher E caused dieback via cavitation.     

Variation in stomatal aperture in leaves of Avena fatua L. observed by low-temperature scanning electron microscopy

Abstract. A novel technique to record the variability of stomatal aperture over the leaf surface is described. This combines observations of leaf surfaces using low-temperature scanning electron microscopy (LTSEM), with digital image analysis to produce the most accurate aperture measurements obtained to date. Leaf samples are rapidly immobilized by cryo-fixation in liquid nitrogen and stored in a purpose-built cryo-storage system. Specimens can be collected in the field, remote from the cryopreparation system, and stored for up to several weeks before being examined on the LTSEM. The advantages of this method are that the time frame of the measurements is accurately known, and is identical for all stomatal apertures in a sample, and the precision of the measurements is not limited by the resolving power of the microscope. Measurements of stomatal aperture were obtained from leaves of field grown Avena fatua using the above procedure. Leaf surface conductance (g_sur) was determined by porometry immediately before cryo-fixation of the same region of the leaf. Measurements of aperture size showed a high degree of variability within each specimen, with coefficients of variation similar to those found in previous studies. Stomatal conductance (g_s) was calculated from stomatal dimensions using formulae derived elsewhere. A linear regression between the computed values of g_s and porometric estimates of g_sur showed good agreement with the regression line passing through the origin with a slope of 1.0 (R²=0.96). Applications of the experimental system are discussed.     

The effect of competition from neighbours on stomatal conductance in lettuce and tomato plants

Competition decreased transpiration from young lettuce plants after 2 days, before any reductions in leaf area became apparent, and stomatal conductance (g(s) ) of lettuce and tomato plants was also reduced. Stomatal closure was not due to hydraulic signals or competition for nutrients, as soil water content, leaf water status and leaf nitrate concentrations were unaffected by neighbours. Competition-induced stomatal closure was absent in an abscisic acid (ABA)-deficient tomato mutant, flacca, indicating a fundamental involvement of ABA. Although tomato xylem sap ABA concentrations were unaffected by the presence of neighbours, ABA/pH-based stomatal modulation is still likely to underlie the response to competition, as soil and xylem sap alkalization was observed in competing plants. Competition also modulated leaf ethylene production, and treatment of lettuce plants with an ethylene perception inhibitor (1-methylcyclopropene) diminished the difference in g(s) between single and competing plants grown in a controlled environment room, but increased it in plants grown in the greenhouse: ethylene altered the extent of the stomatal response to competition. Effects of competition on g(s) are discussed in terms of the detection of the absence of neighbours: increases in g(s) and carbon fixation may allow faster initial space occupancy within an emerging community/crop.     

Responses of stomatal conductance to light, humidity and temperature in winter wheat and barley grown at three concentrations of carbon dioxide in the field

Responses of leaf stomatal conductance to light, humidity and temperature were characterized for winter wheat and barely grown at ambient (about 350 μmol mol^?1 in the daytime), ambient + 175 and ambient + 350 μmol mol^?1 concentrations of carbon dioxide in open‐topped chambers in field plots over a three year period. Stomatal responses to environment were determined by direct manipulation of single environmental factors, and those results were compared with responses derived from natural day to day variation in mid‐day stomatal conductance. The purpose of these experiments was to determine the magnitude of reduction in stomatal conductance at elevated [CO₂], and to assess whether the relative response of conductance to elevated [CO₂] was constant across light, humidity and temperature conditions. The results indicated that light, humidity and temperature all significantly affected the relative decrease in stomatal conductance at elevated [CO₂]. The relative decrease in conductance with elevated [CO₂] was greater at low light, low water vapour pressure difference, and high temperature in both species. For measurements made at saturating light near mid‐day, the ratio of mid‐day stomatal conductances at doubled [CO₂] to that at ambient [CO₂] ranged from 0.42 to 0.86, with a mean of 0.66 in barley, and from 0.33 to 0.80, with a mean of 0.56 in wheat. Day‐to‐day variation in the relative effect of elevated [CO₂] on conductance was correlated with the relative stimulation of [CO₂] assimilation rate and with temperature. Some limitations of multiple linear regression, multiplicative, and ‘Ball–Berry' models as summaries of the data are discussed. In barley, a better fit to the models occurred in individual years than for the combined data, and in wheat a better fit to the models occurred when data from near the end of the season were removed.