Role of cytokinins in the regulation of stomatal conductance of wheat seedlings under conditions of rapidly changing local temperature

The influence of an air temperature increase by 4°C and nutrient solution cooling down to 5 ± 1°C on stomatal conductance and hormone level of seven-day-old wheat (Triticum durum L., cv. Bezenchukskaya 139) seedlings was studied. An elevated air temperature resulted in a rapid rise of stomatal conductance preceded by the increase in the level of cytokinins in leaves. Cooling of the nutrient solution induced gradual stomatal closure along with a decreasing cytokinin level in leaves. Hormone concentration in the xylem sap of wheat seedlings was determined, and the rate of hormone transport from the roots to shoots was calculated. The role of cytokinins in the regulation of stomatal conductance under conditions of local thermal treatments is discussed.     

Stomatal behaviour and water movement through roots of wheat plants treated with abscisic acid

Abstract Experiments with isolated roots of wheat plants suggested that when water uptake rates are low, low concentrations of abscisic acid (ABA) may increase the flux of water into roots. This increase was recorded despite an ABA-stimulated reduction in the hydraulic conductance of the whole root system. Hydraulic conductances were measured under steady-state conditions. A system is described where the stomatal behaviour and water movement through roots of a single intact plant may be concurrently monitored. Experiments with intact plants confirmed that application of ABA could increase the rate of water movement into roots when uptake rates were low. No such increase was observed at high flux rates. Application of ABA to roots caused partial stomatal closure and caused conductance to oscillate around a reduced mean value. An ABA-stimulated increase in the turgor sensitivity of stomata is postulated and the significance of this effect is discussed.     

Isolation and characterization of variant wheat cultivars for ABA sensitivity

Genetic variants for abscisic acid (ABA) sensitivity are important for investigating the role of ABA sensitivity in conditioning plant response to environmental stress, and especially to those soil conditions that may elicit a root-mediated hormonal signal. This study was performed in order to isolate variation in ABA sensitivity among wheat (Triticum aestivum and T. durum) cultivars, as characterized by two plant responses: (i) shoot growth reduction in response to 5×10^?2mol m^?3 ABA (racemic) in the root medium of hydroponically grown plants, and (ii) changes in transpiration and gas exchange in a bioassay of detached leaves (leaflaminac) infused with 10^?4mol m^?3 ABA. Very significant (P≤0.01) and repeatable differences were found among 36 wheat cultivars and 19 landraces in the growth rate in ABA-containing nutrient solutions, expressed as a percentage of the growth rate in control nutrient solutions (ABA/control ratio). In duplicate experiments, the ABA/control ratio ranged between 60 and 83% for the least sensitive cultivars (V2151-3, Bethlehem, K1056 and Sunstar) and between 9 and 19% for the most sensitive cultivars (Sundor, Comet, Barkaec and V5). In the transpiration bioassay, performed with seven selected cultivars, it was found that the reductions in transpiration of ABA-infused leaves corresponded very well with the reductions in growth in response to ABA in the root media. Measurement of gas exchange in the detached leaves of two cultivars differing in ABA sensitivity (Bethlehem and Sundor) showed that variable ABA sensitivity was expressed very well in the stomatal conductance, carbon exchange rate (CER) and photosynthetic capacity (CER/C_i ratio) of the leaf. These results therefore allowed us to isolate wheat variants for ABA sensitivity and to conclude that, while ABA sensitivity is expressed in the growth of plants challenged by ABA in the root medium, the control of sensitivity resides, at least partly, in the leaf.     

Hormonal dynamics contributes to divergence in seasonal stomatal behaviour in a monsoonal plant community

The plant hormone abscisic acid (ABA) is a primary regulator of plant transpiration, but its influence in determining seasonal stomatal behaviour in natural plant communities is poorly understood. We examined distantly related vascular plants growing together in a seasonally dry, monsoonal environment to determine whether ABA dynamics contributed to contrasting water use patterns in this natural setting. Regular sampling of angiosperm, cycad, conifer and fern species revealed characteristic seasonal patterns in ABA production, but these were highly distinct among species. Although no general relationship was observed between ABA levels, plant hydration or stomatal conductance among species, the seasonal dynamics in stomatal behaviour within species were predictable functions of either ABA or leaf water potential. Strong divergence in the seasonal role of ABA among species suggests that modification in ABA–stomatal interactions represents an important evolutionary pathway for adaptation in plant water use.     

Contents of abscisic acid and cytokinins in shoots during dehydration of wheat seedlings

Accumulation of ABA in shoots during 30-min dehydration of wheat seedlings was accompanied by the decline in the content of zeatin nucleotide and the accumulation of zeatin 9-N-glucoside. The total content of zeatin derivatives as well as the content of free base of zeatin remained almost constant. This revised version was published online in July 2006 with corrections to the Cover Date.     

Phytochrome is not involved in the red-light-enhancement of the stomatal blue-light-response in wheat seedlings

The stomatal response to blue light (BL) in wheat seedlings ( Triticum aestivum L. cv. Starke II, Weibull) was enhanced by background red light (R). This enhancement was only slightly affected by the addition of background far-red light (FR). Under similar light treatments, the addition of FR induced a 43% transformation from the far-red-absorbing form towards the red-absorbing form of phytochrome from etiolated oat ( Avena sativa L. cv. Sol II), immobilized on phenyl-sepharose. Furthermore, the enhancement of the stomatal BL-response by 15 min R was not reversed by a subsequent irradiation with 5 min FR. It is concluded that the red-light-enhancement of the stomatal blue-light-response in wheat seedlings does not involve a change in the photostationary state of phytochrome.     

Hexokinase mediates stomatal closure

Stomata, composed of two guard cells, are the gates whose controlled movement allows the plant to balance the demand for CO₂ for photosynthesis with the loss of water through transpiration. Increased guard‐cell osmolarity leads to the opening of the stomata and decreased osmolarity causes the stomata to close. The role of sugars in the regulation of stomata is not yet clear. In this study, we examined the role of hexokinase (HXK), a sugar‐phosphorylating enzyme involved in sugar‐sensing, in guard cells and its effect on stomatal aperture. We show here that increased expression of HXK in guard cells accelerates stomatal closure. We further show that this closure is induced by sugar and is mediated by abscisic acid. These findings support the existence of a feedback‐inhibition mechanism that is mediated by a product of photosynthesis, namely sucrose. When the rate of sucrose production exceeds the rate at which sucrose is loaded into the phloem, the surplus sucrose is carried toward the stomata by the transpiration stream and stimulates stomatal closure via HXK, thereby preventing the loss of precious water.     

Epidermal conductance in different parts of durum wheat grown under Mediterranean conditions: the role of epicuticular waxes and stomata

Abstract. Epidermal (non-stomatally-controlled) conductance from the fourth leaf, first node leaf, flag leaf and ear of durum wheat (Triticum turgidum var durum L.) grown under Mediterranean field conditions has been measured, along with leaf stomatal frequency and the amount and distribution of epicuticular waxes. Measurements were carried out on varieties and land-races from the Middle East, North Africa, ‘Institut National de la Recherche Agricole’ (INRA) and ‘Centra Internacional de Mejora de Maiz y Trigo’ (CIMMYT). Significant differences were observed among genotypes in the epidermal conductances (g_e) of the four organs. For each of the four organs tested, genotypes from the Middle East and CIMMYT showed higher g_e. values than those from North Africa and INRA. Ears showed epidermal conductances that were more than four times higher than those of leaves when g_e. values were expressed per unit dry weight. The amount of epicuticular waxes was higher in the fourth leaves, intermediate in the first node and flag leaves and lower in the ears. For each organ, g_e differences among genotypes were unrelated with the amount of epicuticular waxes. Removal of epicuticular waxes by dipping the organs into chloroform significantly increased the epidermal conductance for the fourth and first node leaves and the ear. However, this did not occur for the flag leaf. For the fourth leaf, g_e of intact leaves and g_e of leaves in which epicuticular waxes were removed were unrelated (r = -0.265). The regression coefficient of this relation for the first node and flag leaves showed values of 0.666 and 0.650 (P > 0.05), respectively, and values were even higher in the ear (r > m 0.892, P > 0.01). Scanning electron microscope analysis showed that wax bloom decreased from the fourth leaf to the flag leaf, whereas the extent of amorphous wax increased. Wax bloom in leaves consisted mainly of deposits of thin wax plates. In the ears and the adaxial surface of flag leaves, fibrillar waxes predominated. In the first node and flag leaves, the wax deposits on the adaxial side cover the surface of the leaf more densely and uniformly than those on the abaxial side. There was no significant correlation between g_e and total stomatal density, or between g_e and either adaxial or abaxial stomatal density for any sample of the three different leaves. The contribution of epicuticular waxes plus total stomatal frequency only explained 42.4, 11.8, 28.3 and 16% of g_e (per unit leaf area) variations for the fourth leaf, first node leaf, flag leaf and the combined variation of the three leaves together, respectively. From these results, it is concluded that complex interrelationship between different morphophysiological characteristics probably control g_e differences among genotypes and that these interrelationships differ for each different plant part.     

Photosynthetic and stomatal conductance responses to acid mist of red spruce seedlings

Abstract Two-year-old seedlings of Picea rubens, growing in open-top chambers in Scotland were treated twice weekly from July 1987 to December 1987, with mist containing ammonium sulphate and nitric acid at a pH of either 2.5 or 5.0. The response of photosynthesis and stomatal conductance to light flux density and carbon dioxide concentration were measured in March 1989. Leaf chlorophyll a and b contents were also measured. Acid mist (pH 2.5) resulted in several significant changes. First, both the rate of light saturated photosynthesis (A_max) and CO₂- saturated rate of photosynthesis (J) were substantially increased, when expressed per unit leaf area. Second, the apparent quantum yield and chlorophylls a and b content increased. Third, as a consequence of the greater chlorophyll content of the leaves treated with acid mist, the rate of A_max, and J, expressed per unit chlorophyll, was substantially reduced in pH 2.5 treated branches. Stomatal conductance was enhanced at all but the highest light flux densities, and was independent of the CO₂ concentration, remaining high for all values of CO₂ concentration used. These results show that acid mist caused a number of responses in the gas exchange and photosynthetic properties of red spruce.     

Effect of partial root excision on transpiration, root hydraulic conductance and leaf growth in wheat seedlings.

Removal of four out of five roots did not lower transpiration and stomatal conductivity of wheat (Triticum durum Desf.) seedlings. Water content of mature expanded leaf lamina remained constant at control levels. The results suggest that the only remaining root was capable to supply the shoot with water. This was evidenced by an increase in hydraulic conductivity of the root system following partial root excision measured at low subatmospheric pressures induced by vacuum. In the absence of a hydrostatic gradient, water flow from reduced root system was initially not higher than from an intact system, but increased subsequently. ABA content was increased in roots 1 h after partial root excision, which might contribute to the increase in hydraulic conductivity.     

Genetic dissection of seminal root architecture in elite durum wheat germplasm

Although root architecture has been shown to play an important role in crop performance, particularly under drought conditions, no information is available on the genetic control of root traits in durum wheat, a crop largely grown in rainfed areas with low rainfall. In our study, a panel of 57 elite durum wheat accessions were evaluated under controlled conditions for root and shoot traits at the seedling stage. Significant genetic variability was detected for all the root and shoot traits that were investigated. Correlation analysis suggested that root and shoot features were only partially controlled by common sets of genes. The high linkage disequilibrium (up to 5 cM) present in the germplasm collection herein considered allowed us to use simple sequence repeat‐based association mapping to identify chromosome regions with significant effects on the investigated traits. In total, 15 chromosome regions showed significant effects on one or more root architectural features. A number of these regions also influenced shoot traits and, in some cases, plant height measured in field conditions. Major effects were detected on chromosome arms 2AL (at Xgwm294), 7AL (at Xcfa2257 and Xgwm332) and 7BL (at Xgwm577 and Xcfa2040). The accessions with the most remarkable differences in root features will provide a valuable opportunity to assemble durum wheat mapping populations well suited for ascertaining the effects of root architecture on water use efficiency and grain yield.     

Effect of increased irradiance on the hormone content,water relations,and leaf elongation in wheat seedlings

In wheat (Triticum durum Desf., cv. Bezenchukskaya 139) seedlings, an increase in irradiance from 20 to 400 μmol/(m² s) PAR enhanced transpiration and increased stomatal conductance by three times on the background of reduced relative water content (RWC). After this treatment, leaves quickly ceased to grow and became even shrunk later. In 40 or 50 min, leaf growth was resumed. At this period, we observed an increase in hydraulic conductivity and RWC and also in leaf extensibility. As soon as 10 min after treatment, some changes in hormone content were noted. In the zones of leaf growth and its mature part, zeatin and zeatin riboside were accumulated, whereas ABA accumulation was observed in the zone of leaf growth and in the roots. The results obtained indicate that leaf expansion at increased irradiance was related to changes in cell-wall extensibility and hydraulic conductivity. The first effect could be due to cytokinin accumulation, whereas the second one, to ABA accumulation.     

Soil salinity effects on transpiration and net photosynthetic rates,stomatal conductance and Na+ and Cl− contents in durum wheat

Leaf gas exchange, plant growth and leaf ion content were measured in wheat (Triticum durum L. cv. HD 4502) exposed to steady- state salinities (1.6, 12.0 and 16.0 dS nr⁻¹) for 8 weeks. Salinity reduced leaf area and number of tillers, and increased Na⁺ and Cl⁻ concentrations in leaves. Leaf- to- leaf gradients of these ions were observed. The oldest leaf contained 6 to 8 times more Na⁺ and Cl⁻ than the flag leaf. Net photosynthetic rate (P_N), transpiration rate (E) and stomatal conductance (g_S) were the highest in flag leaf, declined in the middle and fully expanded leaves, and were minimum in the oldest leaves. These processes were reduced by salinity with similar leaf- to- leaf gradients. Intercellular CO₂ concentrations in the older leaves were higher than in the flag leaf in non-saline plants, and increased similarly with salinity. Leaf age was the major factor in reducing P_N, and senescence processes were promoted by salinity.     

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

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

Stomatal and hydraulic conductance in growing sugarcane: stomatal adjustment to water transport capacity*

Abstract Stomatal conductance per unit leaf area in well-irrigated field- and greenhouse-grown sugarcane increased with leaf area up to 0.2 m² plant ¹, then declined so that maximum transpiration per plant tended to saturate rather than increase linearly with further increase in leaf area. Conductance to liquid water transport exhibited parallel changes with plant size. This coordiantion of vapour phase and liquid phase conductances resulted in a balance between water loss and water transport capacity, maintaining leaf water status remarkably constant over a wide range of plant size and growing conditions. The changes in stomatal conductance were not related to plant or leaf age. Partial defoliation caused rapid increases in stomatal conductance, to re-establish the original relationship with remaining leaf area. Similarly, pruning of roots caused rapid reductions in stomatal conductance, which maintained or improved leaf water status. These results suggest that sugarcane stomata adjusted to the ratio of total hydraulic conductance to total transpiring leaf area. This could be mediated by root metabolites in the transpiration stream, whose delivery per unit leaf area would be a function of the relative magnitudes of root system size, transpiration rate and leaf area.     

Influence of temperature increase on evaportranspiration rate and cytokinin content in wheat seedlings

The content of cytokinins especially zeatin nucleotide decreased in shoots as a result of temperature increase. Simultaneously the cytokinins accumulated in roots. The changes in cytokinins distribution were followed by a decline of evapotranspiration after its initial temperature-induced uprising. This revised version was published online in July 2006 with corrections to the Cover Date.     

冬小麦叶片气孔导度模型水分响应函数的参数化

植物气孔导度模型的水分响应函数用来模拟水分胁迫对气孔导度的影响过程, 是模拟缺水环境下植物与大气间水、碳交换过程的关键算法。水分响应函数包括空气湿度响应函数和土壤湿度(或植物水势)响应函数, 该研究基于田间实验观测, 分析了冬小麦(Triticum aestivum)叶片气孔导度对不同空气饱和差和不同土壤体积含水量或叶水势的响应规律。一个土壤水分梯度的田间处理在中国科学院禹城综合试验站实施, 不同水分胁迫下的冬小麦叶片气体交换过程和气孔导度以及其他的温湿度数据被观测, 同时观测了土壤含水量和叶水势。实验数据表明, 冬小麦叶片气孔导度对空气饱和差的响应呈现双曲线规律, 变化趋势显示大约1 kPa空气饱和差是一个有用的阈值, 在小于1 kPa时, 冬小麦气孔导度对空气饱和差变化反应敏感, 而大于1 kPa后则反应缓慢; 分析土壤体积含水量与中午叶片气孔导度的关系发现, 中午叶片气孔导度随土壤含水量增加大致呈现线性增加趋势, 但在平均土壤体积含水量大于大约25%以后, 气孔导度不再明显增加, 而是维持在较高导度值上下波动; 冬小麦中午叶片水势与相应的气孔导度之间, 随着叶水势的增加, 气孔导度呈现增加趋势。根据冬小麦气孔导度对空气湿度、土壤湿度和叶水势的响应规律, 研究分别采用双曲线和幂指数形式拟合了水汽响应函数, 用三段线性方程拟合了土壤湿度响应函数和植物水势响应函数, 得到的参数可以为模型模拟冬小麦的各类水、热、碳交换过程采用。     

Differential requirement for NO during ABA-induced stomatal closure in turgid and wilted leaves

Abscisic acid (ABA)-induced stomatal closure is mediated by a complex, guard cell signalling network involving nitric oxide (NO) as a key intermediate. However, there is a lack of information concerning the role of NO in the ABA-enhanced stomatal closure seen in dehydrated plants. The data herein demonstrate that, while nitrate reductase (NR)1-mediated NO generation is required for the ABA-induced closure of stomata in turgid leaves, it is not required for ABA-enhanced stomatal closure under conditions leading to rapid dehydration. The results also show that NO signalling in the guard cells of turgid leaves requires the ABA-signalling pathway to be both capable of function and active. The alignment of this NO signalling with guard cell Ca²⁺-dependent/independent ABA signalling is discussed. The data also highlight a physiological role for NO signalling in turgid leaves and show that stomatal closure during the light-to-dark transition requires NR1-mediated NO generation and signalling.     

Effects of patchy stomatal closure on gas exchange measurements following abscisic acid treatment

Gas exchange data and images of leaf fluorescence were collected concurrently as stomata responded to abscisic acid (ABA) application. When 10^?5kmolm^?3 ABA was applied to the transpiration stream in a short pulse, stomatal conductance (g_s), photosynthesis (A) and intercellular CO2 concentration (C_i) decreased rapidly after a short lag period and became approximately constant after 2h. There was an apparent reduction in the A versus c1 relationship as stomata closed, but the data returned to the A versus C1 curve while stomatal conductance was constant or slowly rising during the second hour after ABA treatment. Larger amounts of ABA administered during the pulse caused larger deviations from the A versus c1 relationship. When 10^?7kmolm^?3 ABA was applied continuously through the transpiration stream, gs, A and Cⁱ decreased, but there was no substantial deviation from the A versus c{ curve. Fluorescence images were patchy as stomata closed for all experiments, but became slowly more uniform during the time that gas exchange was returning to the A versus Cj curve. The distribution of con-ductance among patches was not bimodal, and larger devi-ations from the A versus ct curve had greater ranges of pixel values and more pixel values representing low values of Cj during stomatal closure than did experiments show-ing small or no deviation. Estimates of A and gs from fluo-rescence images compared favourably with measured val-ues in most cases, suggesting that the patchy distributions of fluorescence were caused by patchy distributions of stomatal conductance and that apparent reductions in the A versus ct relationship were the result of these patchy stomatai distributions and not direct effects of ABA on mesophyll functioning. The data show that stomatal patches can be temporary and that patchiness may not be reflected in gas exchange data if the range of stomatal con-ductances is not large. These observations may explain some of the discrepancies among previous studies concerning the effect of ABA on the A versus Cⁱ relationship.