Senescence of Rice Leaves XVIII. Changes of Stomatal Aperture during Senescence

Changes of stomatal aperture during the course of developmentof rice leaves were directly observed with a scanning electronmicroscope. The stomata reached their maximal aperture sizeafter senescence began in seedling leaves and the flag leafof mature plants. The small stomatal aperture observed priorto senescence seems to be the normal size of stomata in riceleaves, and thus stomata closure does not seem to be the causeof leaf senescence in rice plants. The stomata retain theircapability of movement during senescence, suggesting that guardcells tend to live longer than mesophyll cells. ⁴Present address: Tobacco Taiwan, Republic of China ResearchInstitute, Taichung, Taiwan, Republic of China (Received March 12, 1987; Accepted September 30, 1987)     

Direct Observation of Reversible and Irreversible Stomatal Responses of Attached Sunflower Leaves to SO(2)

The effects of SO₂ on stomatal aperture of attached sunflower leaves were observed with a remote-control light microscope system that permitted continuous observation of stomatal responses over periods of several hours. The relationship between actual stomatal aperture and stomatal conductance, measured with a porometer, also was examined on leaves before and after exposure to SO₂.

A distinction between uninjured and injured regions was clearly visible on leaves after exposure to 1.5 microliters per liter SO₂ for less than an hour. During the exposure, the mean value of apertures for many stomata, which indicates stomatal conductance and transpiration rate, tended to decrease simultaneously in the uninjured and injured regions. However, the rate of decrease in the injured region was slower than that in the uninjured region because of a transient opening induced by water-soaking in the injured region. The transient opening was less common in stomata near veins and veinlets.

There was a good correlation between pore width and stomatal conductance measured with a porometer before exposure to SO₂. This correlation continued in leaves exposed to SO₂ until visible, irreversible injury occurred, but then it disappeared.

The results of these experiments indicate the necessity of continuous observation of individual stomata under the microscope to understand the effects of air pollutants such as SO₂ on stomatal behavior.

     

不同产地何首乌叶表皮结构的解剖学特征与气候因子的关系

在光学显微镜下观察了不同产地何首乌叶表皮结构特征,应用多元回归方法对不同产地何首乌的叶表皮特征与气候因子的关系进行了分析。观察的叶表皮特征指标有气孔密度、气孔指数、气孔器长、气孔器宽、气孔极区角质加厚、气孔器类型、表皮细胞垂周壁性状及叶表面角质条纹。观察结果:上表皮有少量气孔器分布;在下表皮,气孔器类型为非典型不等细胞型和不规则型,有少量腺鳞分布,气孔器密度每1mm2为241.7(64～573)个,气孔指数为17.1(7.5～26.5)%,气孔器长31.1(20～44)μm,气孔器宽23.1(16～38)μm。随着产地的不同,何首乌叶下表皮结构有明显差异。分析结果显示气孔器、气孔器宽度以及气孔密度均与纬度关系密切,随纬度的升高,气孔器长、气孔器宽呈减小的趋势,气孔密度呈增加的趋势,R2值分别为0.619、0.729、0.772。     

Stomatal Responses to Pressure Changes and Interruptions in the Water Supply of Detached Leaves of Zea mays L

Stomata of Zea mays L. respond to changes in hydrostatic pressure in the water supply of the leaves almost instantaneously and in all leaf parts simultaneously. Therefore, the leaf is a hydraulic unit. The stomata are part of it and their aperture is controlled by the water potential in the water-conducting system. Stomatal aperture is not uniquely related to the relative water content of a leaf. The relation depends also on the humidity in the air and is different for the upper and the lower epidermis.     

The CO(2) response of Vicia guard cells acclimates to growth environment

Stomata of growth chamber-grown Vicia faba leaves have an enhanced CO(2) response, measured as change in stomatal aperture, compared to stomata of greenhouse-grown leaves. Reciprocal transfer experiments showed that the stomatal response to CO(2) acclimated to the growing environment. Stomata of growth chamber-grown leaves transferred to a greenhouse lost their high CO(2) sensitivity within 2-3 d while stomata of greenhouse-grown leaves transferred to a growth chamber acquired a high CO(2) sensitivity within 5-7 d. Experiments measuring the CO(2) responses of stomata in detached epidermis showed that growth chamber and greenhouse-grown stomata have the same contrasting CO(2) sensitivity observed in the intact leaf, indicating that the responses reflect intrinsic guard cell properties. The acclimation properties of the CO(2) response of guard cells have implications for the understanding of stomatal function under the predicted increases in atmospheric CO(2).     

适应不同光环境小蜡叶片气孔导度、气孔开度和气孔密度的时空变化

对生长在强光环境和弱光环境小蜡叶片的气孔参数测定发现 :气孔导度和气孔开度在 4个取样部位存在异质性分布。气孔导度和气孔开度经回归分析呈线性、指数或多项式分布。统计分析表明 :强光下的叶片气孔导度和气孔开度的相关性明显高于弱光环境叶片的数值。无论强光环境还是弱光环境下的叶片 ,在取样部位编号为1和 3,其气孔导度和气孔开度均存在显著的线性关系。弱光环境下叶片的气孔密度要远低于强光环境下的叶片。强光环境下叶片对变化环境的敏感性要大于弱光环境下的叶片 ,这可能与强光环境叶片具有较高的气孔密度有关。     

Effects of experimental warming on stomatal traits in leaves of maize (Zea may L.)

We examined the warming effects on the stomatal frequency, stomatal aperture size and shape, and their spatial distribution pattern of maize (Zea may L.) leaves using a light microscope, an electron scanning microscope, and geostatistic techniques. A field manipulative experiment was conducted to elevate canopy temperature by 2.08°C, on average. We found that experimental warming had little effect on stomatal density, but significantly increased stomatal index due to the reduction in the number of epidermal cells under the warming treatment. Warming also significantly decreased stomatal aperture length and increased stomatal aperture width. As a result, warming significantly increased the average stomatal aperture area and stomatal aperture circumference. In addition, warming dramatically changed the stomatal spatial distribution pattern with a substantial increase in the average nearest neighbor distance between stomata on both adaxial and abaxial surfaces. The spatial distribution pattern of stomata was scale dependent with regular patterns at small scales and random patterns at larger scales on both leaf surfaces. Warming caused the stomatal distribution to become more regular on both leaf surfaces with smaller L(t) values (Ripley's K‐function, L(t) is an expectation of zero for any value of t) in the warming plots than the control plots.     

CYCLING OF STOMATAL APERTURE IN LEAVES OF PLANTS WITH CRASSULACEAN ACID METABOLISM UNDER CONSTANT CONDITIONS

When leaves of plants with C₃ metabolism are detached and held in darkness, they senesce and the stomata close. Because the relation of senescence and stomatal closure is very close, if not actually causal, the question arose as to whether in the leaves of plants with Crassulacean acid metabolism whose stomata open at night the relationship to senescence would be reversed. Detached leaves of four species of Hoya, floated on water in constant darkness or constant light, were found to show no large differences in stomatal aperture (measured as diffusion resistance) between those in the light or dark, but the aperture changed in a regular circadian rhythm. In some leaves the rhythm was simple, in others the peak showed small secondary peaks, but in all cases the values were nearly the same in the light as in the dark, throughout the cycle. Previous culture of the intact plants under normal day/night conditions gave results similar to those with plants that had had prolonged culture under constant light or darkness. In those cases when the stomata were more open in the dark, the chlorophyll content was greater than when the stomata were more open in the light; but when they were more open in the light, the chlorophyll content showed little difference between light and dark. When the leaves had only their petioles in water they showed greater senescence in the light than in the dark, and the stomata were more tightly closed in the light, especially at the apical ends. All four species of Hoya gave similar results. We deduce that senescence of these leaves is modified by stomatal aperture, and generally in the same direction as in C₃ leaves, but that in continuous light or darkness the primary control over the aperture is the endogenous cycle.     

Some Differences in the Responses of Stomata of the Two Leaf Surfaces in Cotton

Some differences in the responses of the upper and lower stomatain cotton (Gossypium hirsutuni) are presented. These differenceswere observed in the course of some studies in which the transpirationof the two leaf surfaces was measured under controlled environmentconditions and the transpiration data used as an estimate ofstomatal response. In darkness the upper stomata were more or less effectivelyclosed while the lower stomata were partially open. Upon illuminationof the leaf with non-saturation or saturation radiation theupper stomata were slower to open than the lower stomata. Thereductions in stomatal aperture which occurs with the increasein age of leaves commenced earlier in the upper stomata andproceeded at a faster rate than the lower stomata. Sudden exposureto saturation radiation caused the stomata of the two leaf surfacesto oscillate. These oscillations were not observed in youngleaves but in older leaves. During ageing of leaves oscillationsof the upper stomata commenced earlier than oscillations ofthe lower stomata. When the petiole was excised in darknessor light the upper stomata showed a transient increase but notthe lower stomata. Gossypium hirsutum, stomatal responses, transpiration     

Investigations into the Stornatal Physiology of Upland Cotton: I. The Effects of Hour of Day, Solar Radiation, Temperature and Leaf Water-Content on Stomatal Behaviour

Stomatal behaviour was studied under field conditions usingan infiltration method with solutions of iso-butanol dilutedwith ethylene glycol. For mature leaves not suffering waterstrain, stomata in both epidermes commence opening at sunrise(6.30 a.m. local time) and have a maximum aperture between 10a.m. and 3 p.m.; subsequently a slow closure commences and thisis complete by 7 pjn. For leaves suffering marked water stress,i.e. with relative turgidity values falling below 85 per cent.,the period of maximum aperture is curtailed by a varying amountdependent upon the extent of water stress; closure occurs earlier,and no evidence has been found for a second opening later inthe day. For leaves not suffering water strain it is shown by means ofa multiple correlation analysis that stomatal aperture is significantlycorrelated with solar radiation, temperature, and hour of day.It is suggested that the hour-of-day effect may be a true diurnalrhythm controlled by factors within the leaf. The correlationsnoted above are not found for leaves with low values of relativeturgidity, since the effects of water strain tend to over-ridethose due to the other factors.     

适应不同光环境小蜡叶片气孔导度、气孔开度和气孔密度的时空变化

对生长在强光环境和弱光环境小蜡叶片的气孔参数测定发现:气孔导度和气孔开度在4个取样部位存在异质性分布.气孔导度和气孔开度经回归分析呈线性、指数或多项式分布.统计分析表明:强光下的叶片气孔导度和气孔开度的相关性明显高于弱光环境叶片的数值.无论强光环境还是弱光环境下的叶片,在取样部位编号为1和3,其气孔导度和气孔开度均存在显著的线性关系.弱光环境下叶片的气孔密度要远低于强光环境下的叶片.强光环境下叶片对变化环境的敏感性要大于弱光环境下的叶片,这可能与强光环境叶片具有较高的气孔密度有关.     

Boron deficiency affects cell morphology and structure of young leaves of radish

Boron (B) is an essential microelement for the growth and development of plants. B-deficient radish plants grew slowly compared to B-sufficient controls. Soluble B and cell wall-bound B decreased in young leaves on removal of B from culture medium. In old leaves, B deficiency reduced soluble B content but there was no significant effect on cell wall-bound B content compared to controls. The mesophyll cells in the middle of leaves were enlarged abnormally and had greater cell wall thickness under B-deficient conditions. B deficiency reduced the stomata frequency, inhibited the stomata aperture, and guard cells had thickened cell walls. B-starved leaves showed decreased photosynthesis and stomatal conductance. These indicate that B deficiency could interfere with cell wall development, especially irregular guard cell walls as a result of B deficiency severely affected the rhythmic stomatal closing and opening, preventing the normal functioning of stomata. Correspondingly, photosynthesis was indirectly affected, and plant growth decreased.     

Dynamics of spatial heterogeneity of stomatal closure in Tradescantia virginiana altered by growth at high relative air humidity

The spatial heterogeneity of stomatal closure in response to rapid desiccation of excised well-watered Tradescantia virginiana leaves grown at moderate (55%) or high (90%) relative air humidity (RH) was studied using a chlorophyll fluorescence imaging system under non-photorespiratory conditions. Following rapid desiccation, excised leaves grown at high RH had both a greater heterogeneity and a higher average value of PSII efficiency (Phi(PSII)) compared with leaves grown at moderate RH. Larger decreases in relative water content resulted in smaller decreases in water potential and Phi(PSII) of high RH-grown leaves compared with moderate RH-grown leaves. Moreover, the Phi(PSII) of excised high RH-grown leaves decreased less with decreasing water potential, implying that the stomata of high RH-grown leaves are less sensitive to decreases in leaf water potential compared with moderate RH-grown leaves. After desiccation, some non-closing stomata were distributed around the main vein in high RH-grown leaves. Direct measurements of stomatal aperture showed 77% stomatal closure in the margins after 2 h desiccation compared with 40% closure of stomata in the main-vein areas in high RH-grown leaves. Faster closure of stomata in leaf margins compared with main-vein areas of leaves grown at high RH was related to substantially lower relative water content in these areas of the leaves.     

The role of abscisic acid in disturbed stomatal response characteristics of Tradescantia virginiana during growth at high relative air humidity

In this study, the role of abscisic acid (ABA) in altered stomatal responses of Tradescantia virginiana leaves grown at high relative air humidity (RH) was investigated. A lower ABA concentration was found in leaves grown at high RH compared with leaves grown at moderate RH. As a result of a daily application of 20 microM ABA to leaves for 3 weeks during growth at high RH, the stomata of ABA-treated leaves grown at high RH showed the same behaviour as did the stomata of leaves grown at moderate RH. For example, they closed rapidly when exposed to desiccation. Providing a high RH around a single leaf of a plant during growth at moderate RH changed the stomatal responses of this leaf. The stomata in this leaf grown at high RH did not close completely in response to desiccation in contrast to the stomata of the other leaves from the same plant. The ABA concentration on a fresh weight basis, though not on a dry weight basis, of this leaf was significantly lower than that of the others. Moreover, less closure of stomata was found in the older leaves of plants grown at high RH in response to desiccation compared with younger leaves. This was correlated with a lower ABA concentration in these leaves on a fresh weight basis, though not on a dry weight basis. Stomata of leaves grown at moderate RH closed in response to short-term application of ABA or sodium nitroprusside (SNP), while for leaves grown at high RH there was a clear difference in stomatal responses between the leaf margins and main-vein areas. The stomatal aperture in response to short-term application of ABA or SNP at the leaf margins of leaves grown at high RH remained significantly wider than in the main-vein areas. It was concluded that: (i) a long-term low ABA concentration in well-watered plants during growth at high RH could be a reason for less or no stomatal closure under conditions of drought stress; and (ii) the long-term ABA concentration on a fresh weight basis rather than on a dry weight basis is likely to be responsible for structural or physiological changes in stomata during leaf growth.     

Electrical Currents at the Leaf Surface of Commelina communis and their Relationship to Stomatal Activity

A vibrating probe was used to detect and measure electricalcurrents at the surface of excised leaves and isolated leafepidermis from Commelina commnunis. Currents of up to 4.0 µAcm^–2 moving out from the leaf surface were observed whenthe stomata were open. When the stomata were almost closed nocurrent was detected and when they were fully closed a smallinwardly directed current was observed. There appeared to bea linear relationship between current and stomatal aperture.The current was stimulated by fusicoccin and eliminated by increasingthe external pH suggesting that it was brought about by a flowof H⁺ from the leaf surface. Key words: Electrical currents, leaves, stomata, vibrating probe     

The nature of heterogeneity in the stomatal behaviour of Phaseolus vulgaris L. primary leaves

The aim of this research was to investigate the nature of heterogeneity in stomatal conductance and, in particular, to determine whether the characteristic 'patchy' pattern of water infiltration is reflected in measurements on individual stomata. Silicone rubber replicas were made of primary leaves of glasshouse-grown Phaseolus vulgaris L. plants, and the leaves were then infiltrated with water at controlled sub-atmospheric gas pressures according to their estimated or measured stomatal conductance. Seven leaves examined in detail all showed patchy infiltration, and the mean sire of infiltrated areas was negatively correlated with the prevailing stomatal conductance. In four of the leaves, a one millimetre wide transect across the leaf was selected for further detailed study. Measurements of mean peristomatal groove distance (PGD) and stomatal frequency were made along the transect and related to the state of infiltration. Analysis of variance indicated that, in all four cases, variation in PGD among patches was highly significant, but there was no significant difference between patches of different infiltration categories. Thus, local (patch-level) variation in stomatal aperture appeared to bear no relation to the infiltration status of the patches. The dominant source of stomatal variability was between individual pores in the same locality, which accounted for 82% or more of the total variability. Taking into account variation in stomatal frequency, correlations between predicted stomatal conductance and the extent of infiltration were significant in only one out of the seven leaves studied. Possible reasons for these results are discussed. It is suggested that the infiltration method misrepresents the underlying state of the stomata as being either open or closed, when there is little evidence for this from measurements of stomatal dimensions. For these unstressed plants under relatively stable conditions, it is concluded that the 'unit of variability' in stomatal heterogeneity may rest at the individual pore ('micro') scale, rather than at the areolar patch ('macro') scale, or above.     

In situ observations of stomatal movements in different light-dark regimes: the influence of endogenous rhythmicity and long-term adjustments

The technical device for continuous microscopic observationsof stomatal movements in a gas exchange chamber using digitalimage analysis, earlier described by Kappenet al. (1994), wastechnically improved. By electronic remote control, it is nowpossible repeatedly to record over a period of several daysidentical stomata on a 25 50 mm leaf area. The responses ofindividual stomata to various light-dark sequences in the lightand in the dark phase were investigated on attached leaves ofViciafaba (L.). The amplitude of stomatal oscillations caused byalternatinglight was highest on the first day of an experimental seriesand decreased with repeated application, indicating a long-termadjustment to changing light conditions by decreasing sensitivityto intermittent darkening. In the usual dark phase light-darksequences had little effect on the stomatal aperture, whichremained small. These results were widely convergent with thestomatal conductance calculated from the gas exchange measurements.Very prominent is the role of an endogenous rhythmicity of thestomatal response. It could be demonstrated (1) by autonomousstomatal opening before the light phase started; (2) by a suppressionof dark response in the early morning hours; (3) by a decreasinglight stimulation in the afternoon; (4) by further increaseof aperture for several hours if no light was provided in thelight phase. Closely adjacent stomata could show divergent dark-openingmovements pointing to an autonomous control mechanism locatedin the guard cells. The endogenously controlled morning openingprovides full assimilation capacity in the usually humid morninghours when transpiratory water loss associated with C02-uptakeis comparatively small. Key words: Stomata, changing PPFD, desensitization, endogenous rhythmicity, diurnal courses, Vicia faba     

黄土高原丘陵沟壑区抗侵蚀植物叶表皮的生态适应性

采用叶表皮临时装片法研究了黄土高原丘陵沟壑区不同土壤侵蚀环境沟间地、沟谷地和沟间人工刺槐林地6种抗侵蚀植物叶表皮组成细胞的形态特征.结果表明: 沟间地抗侵蚀植物叶的上表皮气孔开度、气孔密度、气孔指数、气孔器长宽比可塑性、气孔器面积可塑性、表皮毛〖JP2〗密度和表皮细胞密度分别比沟谷地提高93.8%、66.8%、17.9%、36.4%、42.3%、199.4%和46.5%,下表皮分别提高90.4%、76.6%、9.8%、47.1%、43.9%、98.2%和50.1%;沟间人工刺槐林地叶上表皮各形态指标分别比沟谷地提高66.7%、20.5%、11.9%、37.9%、19.8%、113.1%和10.8%,叶下表皮分别提高106.7%、45.8%、11.9%、41.3%、21.2%、52.2%和28.1%.沟间地植物叶上、下叶表皮毛长度分别比沟谷地缩短58.8%和29.7%,表皮细胞面积分别比沟谷地减少40.3%和37.0%.沟间人工刺槐林地叶上、下表皮毛长度分别比沟谷地缩短25.0%和23.6%,表皮细胞面积分别比沟谷地减少22.2%和19.2%.抗侵蚀植物通过增加叶表皮气孔开度、气孔密度、气孔指数、气孔器长宽比可塑性、气孔器面积可塑性、表皮毛密度、表皮细胞密度和减少表皮毛长度、表皮细胞面积来适应较强的土壤侵蚀环境.     

An Investigation into the Stomatal Behaviour of a Wilty Mutant of Pisum sativum

The water relations and stomatal behaviour of a wilty line ofpea (JI 1069) were investigated and compared with those of severalnon-wilty lines (JI 1180, JI 1194, and JI 74). The leaves ofthe wilty line were found to have a lower percent water content,water potential and diffusive resistance and the dimensionsof the stomatal cells were larger than those of the non-wiltytypes. The aperture of stomata on epidermal samples taken from plantsin the light or dark period of a diurnal rhythm was consistentlylarger for the wilty pea than for the non-wilty lines, however,their stomatal responses on detached epidermis to light, CO₂and KC1 concentration were similar. There was no differencein response to ABA of stomata on detached epidermis of wiltyor non-wilty types of pea. Key words: Pisum sativum, Wilty mutant, Water relations, Stomatal behaviour