Surface wax esters contribute to drought tolerance in Arabidopsis

Waxes are components of the cuticle covering the aerial organs of plants. Accumulation of waxes has previously been associated with protection against water loss, therefore contributing to drought tolerance. However, not much information is known about the function of individual wax components during water deficit. We studied the role of wax ester synthesis during drought. The wax ester load on Arabidopsis leaves and stems was increased during water deficiency. Expression of three genes, WSD1, WSD6 and WSD7 of the wax ester synthase/diacylglycerol acyltransferase (WS/DGAT or WSD) family was induced during drought, salt stress and abscisic acid treatment. WSD1 has previously been identified as the major wax ester synthase of stems. wsd1 mutants have shown reduced wax ester coverage on leaves and stems during normal or drought condition, while wax ester loads of wsd6, wsd7 and of the wsd6wsd7 double mutant were unchanged. The growth and relative water content of wsd1 plants were compromised during drought, while leaf water loss of wsd1 was increased. Enzyme assays with recombinant proteins expressed in insect cells revealed that WSD6 and WSD7 contain wax ester synthase activity, albeit with different substrate specificity compared with WSD1. WSD6 and WSD7 localize to the endoplasmic reticulum (ER)/Golgi. These results demonstrated that WSD1 is involved in the accumulation of wax esters during drought, while WSD6 and WSD7 might play other specific roles in wax ester metabolism during stress.     

Occurrence of Crassulacean acid metabolism in Cissus trifoliata L. (Vitaceae)

Summary Evidence for the operation of CAM in the deciduous climber, Cissus trifoliata L., was obtained in field and laboratory studies. Under natural conditions, diurnal oscillations of titratable acidity and colorimetric measurements of night CO₂ fixation, determined for a period of two and a half years, suggested that acid accumulation was related to plant water status, assessed through the daily courses of stomatal resistance and xylem water potential during dry and rainy seasons. These findings were confirmed by gas exchange studies under controlled conditions which showed that the plant fixed all its CO₂ during the day when it was well irrigated; as water stress increased, dark CO₂ uptake gradually replaced fixation during the day until the plant only performed dark fixation. In severe water stress, even the rate of the latter process decreased until leaves fell.Abbreviations CAM Crassulacean acid metabolism - FW leaf fresh weight - SWC relative soil water content - PAR photosynthetically active radiation - TR total radiation; r, leaf diffusive resistance - WSD water saturation deficit (leaf-air vapour concentration difference) - RWC relative water content of leaves     

Water recycling in leaves of Lithops (Aizoaceae)

Lithops plants consist of a pair of succulent leaves inserted on a short stem; in each growing season, young leaves develop in a cavity formed between the older pair. Young leaves can take up water from the older pair allowing the plant to maintain growth and leaf expansion even without external supply of water. Recycling water between vegetative organs is one of the possible adaptation strategies of plants under drought stress, but it had never been demonstrated experimentally in Lithops. The methodology used to verify the existence of water redistribution from old leaves to young leaves was fluorescence microscopy, using two dyes to follow the water pathway inside the plant: Sulforhodamine G (SRG) and 5(6)-carboxyfluoroscein diacetate (CFDA). In Lithops fluorescent tracers loaded into old leaves were found in young leaves, in 74% of the cases for SRG, in 59% of the cases for CFDA. Our data demonstrate that young leaves take up water from the old ones following both a symplastic and an apoplastic pathway. Water recycling is therefore one of the adaptive responses of these plants allowing them to perform at least a complete growth cycle even during prolonged drought stress periods, using the water stored in the older leaves.     

Leaf cell water and enzyme activity

This work supports further the thesis that under conditions of water stress, cell water content may supersede hormonal regulation in effecting enzyme activity, thus becoming a regulatory factor in cellular metabolism. Addition of NaCl to the root medium of barley plants (Hordeum vulgare L.) markedly increased leaf RNase activity parallel to an increase of leaf water saturation deficit (WSD). Kinetin and abscisic acid, applied to the salinated plants, also modified RNase activity, as well as leaf-WSD. The familiar pattern of effects of these hormones on leaf RNase as well as leaf chlorophyll content was inverted, kinetin effected a relative increase in RNase activity and a decrease in leaf chlorophyll, whereas abscisic acid effected a relative decrease in RNase activity and maintained chlorophyll content. A close relationship between enzyme activity and leaf WSD became evident when leaf RNase and protease activities in the salinated plants were plotted against leaf WSD. This close relationship was maintained irrespective of the hormonal treatments, which in themselves markedly modified leaf WSD. As predicted, high relative humidity which relived the leaves from salt-induced water stress prevented the salt-induced rise in RNase activity.     

Short-term and long-term effects of plant water deficits on stomatal response to humidity in Corylus avellana L.

Short-term (hours) changes in plant water status were induced in hazel (Corylus avellana L.) by changing the evaporative demand on a major portion of the shoot while maintaining a branch in a constant environment. Stomatal conductance of leaves on the branch was influenced little by these short-term changes in water status even with changes in leaf water potential as great as 8 bars. Long-term (days) changes in plant water status were imposed by soil drying cycles. Stomatal conductance progessively decreased with increases in long-term water stress. Stomata still responded to humidity with long-term water stress but the range of the conductance response decreased. Threshold responses of stomata to leaf water potential were not observed with either short-term or long-term changes in plant water status even when leaves wilted. It is suggested that concurrent measurements of plant water status may not be sufficient for explaining stomatal and other plant responses to drought.     

Distribution of hydraulic resistance in seedlings,sprouts and an adult tree ofPasania edulis Makino

Hydraulic resistance is an important factor in predicting water status. Hydraulic resistance of petiols, stems and branches, and roots was measured inPasania edulis Makino in order to compare the distribution of resistance between current seedlings, current stump sprouts and a 16 year old adult tree. Total resistance showed only minor variations despite large variations in plant size. This result is thought to be consistent with allometry between leaf mass and supportive organ mass, and with changes in permeability of conductive organs. Root resistance was low in sprouts and the adult tree due to their mature root systems. Current seedlings with undeveloped root systems had high root resistance. The proportion of petiol resistance in total resistance was high compared to the proportion of their conductive distance, and was thought to be a limiting factor of tree water status. The petiol resistance of the adult tree leaves was higher than for seedling and sprout leaves. From a comparison with the leaf water relation characteristics, the petiol resistance was thought to be provided as low values for intolerant leaves against water stress in order to compensate water inflow, and high values for tolerant leaves to regulate water inflow.     

Optimizing thermal imaging as a technique for detecting stomatal closure induced by drought stress under greenhouse conditions

Temperature of leaves or canopies can be used as an indicator of stomatal aperture, which is considered a sensitive response to soil water deficit. In this paper we analyse the robustness and sensitivity of thermal imaging for detecting changes in stomatal conductance and leaf water status in a range of plant species. Thermal imaging successfully distinguished between irrigated and non-irrigated plants of a variety of species under greenhouse or controlled chamber conditions, with strong correlations between thermal indices and stomatal conductance measured by porometry. Our results also highlighted issues that need to be addressed in order to be confident of always detecting drought stress using this technique. These include variability in leaf angles and the limited reliability of 'wet' and 'dry' leaves to represent leaves with stomata fully open or stomata fully closed. These results should assist the design of protocols for application in crop production or ecosystem monitoring.     

Stomatal action directly feeds back on leaf turgor: new insights into the regulation of the plant water status from non‐invasive pressure probe measurements

Uptake of CO₂ by the leaf is associated with loss of water. Control of stomatal aperture by volume changes of guard cell pairs optimizes the efficiency of water use. Under water stress, the protein kinase OPEN STOMATA 1 (OST1) activates the guard‐cell anion release channel SLOW ANION CHANNEL‐ASSOCIATED 1 (SLAC1), and thereby triggers stomatal closure. Plants with mutated OST1 and SLAC1 are defective in guard‐cell turgor regulation. To study the effect of stomatal movement on leaf turgor using intact leaves of Arabidopsis, we used a new pressure probe to monitor transpiration and turgor pressure simultaneously and non‐invasively. This probe permits routine easy access to parameters related to water status and stomatal conductance under physiological conditions using the model plant Arabidopsis thaliana. Long‐term leaf turgor pressure recordings over several weeks showed a drop in turgor during the day and recovery at night. Thus pressure changes directly correlated with the degree of plant transpiration. Leaf turgor of wild‐type plants responded to CO₂, light, humidity, ozone and abscisic acid (ABA) in a guard cell‐specific manner. Pressure probe measurements of mutants lacking OST1 and SLAC1 function indicated impairment in stomatal responses to light and humidity. In contrast to wild‐type plants, leaves from well‐watered ost1 plants exposed to a dry atmosphere wilted after light‐induced stomatal opening. Experiments with open stomata mutants indicated that the hydraulic conductance of leaf stomata is higher than that of the root–shoot continuum. Thus leaf turgor appears to rely to a large extent on the anion channel activity of autonomously regulated stomatal guard cells.     

The periodic wetting of leaves enhances water relations and growth of the long‐lived conifer Araucaria angustifolia

The importance of foliar absorption of water and atmospheric solutes in conifers was recognised in the 1970s, and the importance of fog as a water source in forest environments has been recently demonstrated. Araucaria angustifolia (Araucariaceae) is an emergent tree species that grows in montane forests of southern Brazil, where rainfall and fog are frequent events, leading to frequent wetting of the leaves. Despite anatomical evidence in favour of leaf water absorption, there is no information on the existence and physiological significance of a such process. In this study, we test the hypothesis that the use of atmospheric water by leaves takes place and is physiologically relevant for the species, by comparing growth, water relations and nutritional status between plants grown under two conditions of soil water (well‐watered and water‐stressed plants) and three types of leaf spraying (none, water and nutrient solution spray). Leaf spraying had a greater effect in improving plant water relations when plants were under water stress. Plant growth was more responsive to water available to the leaves than to the roots, and was equally increased by both types of leaf spraying, with no interaction with soil water status. Spraying leaves with nutrient solution increased shoot ramification and raised the concentrations of N, P, K, Zn, Cu and Fe in the roots. Our results provide strong indications that water and nutrients are indeed absorbed by leaves of A. angustifolia, and that this process might be as important as water uptake by its roots.     

Genotypic variation in coleus in the ability to accumulate abscisic acid in response to water deficit

Terry, P. H., Krizek, D. T. and Mirecki, R. M. 1988. Genotypic variation in coleus in the ability to accumulate abscisic acid in response to water deficit. - Physiol. Plant. 72: 441–449.
Abscisic acid (ABA) concentration and plant and soil water potentials were determined in leaves of three cultivars of Coleus blumei Benth. cvs. Marty, Buckley Supreme and PI354190, chosen for their differences in sensitivity to drought, SO₂ and/or chilling stress. Plants were subjected to 'gradual' soil moisture stress (SMS) for 0–6 days, during which time the soil dried out slowly and to more 'rapid' SMS for 0–9 days where the soil dried out at a faster rate. Plants were propagated from clonal stock in the greenhouse and transferred to the growth chamber, where they were maintained for 1 week prior to beginning water stress treatment. ABA concentration was determined as the methyl ester using a gas chromatography - electron capture detector method. Samples for ABA determinations were taken from the third pair of leaves from the apex at the same time each day (1430 h). Measurements of stomatal conductance (C₅) and leaf water potential (φ_l) were made on the fourth pair of leaves from the apex, using the same plants as those sampled for ABA. During the more rapid stress portion of the study soil matric potentials (φ_m) were monitored on a daily basis. Despite large cultivar differences in ABA concentration at 0–6 days, by 7–9 days these differences had largely disappeared. Except for drought-insensitive cv. Marty, there was generally little correlation between ABA levels and measurements of plant and soil water status at 7–9 days.     

LEAF DEVELOPMENT IN ISOPHYLLOUS AND FACULTATIVELY ANISOPHYLLOUS SPECIES OF PENTADENIA (GESNERIACEAE)

Shoots of Pentadenia orientandina exhibit varying degrees of anisophylly, ranging from pairs of equal-sized leaves to pairs of large ventral and small dorsal leaves. In this study we compare phyllotaxis, leaf expansion, and accompanying histological changes in extremely anisophyllous shoots of this species and in isophyllous shoots of the related species, P. crassicaulis. In P. orientandina, decussate phyllotaxis is modified at leaf initiation, and angles of leaf insertion appear to be further changed during leaf expansion. In both species, leaf primordia of a pair are not distinguishable at inception, suggesting an equivalent developmental potential. In P. orientandina, size differences between ventral and dorsal leaves become significant at the P2 or P3 stage, coincident with lamina initiation. Minute dorsal leaves are arrested in their development at the P3 stage and mature without differentiation of multiple epidermis, stomata, mesophyll and most vascular tissue. Variation in dorsal leaf structure in P. orientandina emphasizes the plasticity of leaf development in this facultatively anisophyllous species.     

Effect of water-deficit stress on cotton plants expressing the Bacillus thuringiensis toxin

Bacillus thuringiensis (Bt) crops require a high dosage of Bt toxin to delay development of insect resistance, in particular, when the refuge strategy is applied. This strategy is threatened by plant developmental and environmental factors that might reduce Bt toxin concentration and Bt efficacy in Bt crops. Growth of Bt (Cry1Ac) cotton under prolonged, moderate water deficit as a single stress factor was evaluated. Bt cotton plants were analysed for physiological performance, Bt toxin concentration and Bt efficacy. For performance analysis, leaf and total plant dry weight and leaf area were measured. Bt toxin concentration was determined by an immuno‐assay. Effects of Bt toxin on growth and mortality of African cotton bollworm, Helicoverpa armigera, larvae were measured in different plant organs. Leaves from young plants exposed for 30 days to moderate water deficit had both higher Bt toxin concentrations and were more effective against larvae than leaves, flowers or bolls from mature flowering plants exposed to 60 days of moderate water deficit. Although growth of Bt cotton plants under moderate water‐deficit conditions decreased Bt concentrations in leaves, flowers and bolls, this had no effect on efficacy against first‐instar cotton bollworm larvae. No significant evidence was found that moderate water deficit, as a single stress factor, decreases Bt efficacy in Bt cotton.     

Effects of Age and Water Stress on Endogenous Levels of Plant Growth Regulators in Euphorbia lathyrus L.

The effect of droughting and waterlogging on changes in endogenouslevels of hormones have been studied in Euphorbia lathyrus L.with particular reference to plant age and leaf age. Younger tissues (i.e. immature leaves and young plants) synthesizedsignificantly greater amounts of ABA per unit fresh weight thandid corresponding older tissue. The increased ABA levels correlatedwell with corresponding increases in water saturation deficit(WSD) and stomatal resistance in a given treatment but the relationshipbetween the absolute values of these three parameters variedgreatly between seedlings of different ages. Endogenous ethylenelevels in leaves and stems were unaffected by droughting irrespectiveof age. Imposition of waterlogging led to marked increases in ABA levelsin young plants though this was not strictly related to changesin WSD or stomatal resistance. Endogenous levels of ethyleneincreased in response to waterlogging, this being marked inthe older seedlings which also had a higher incidence of senescenceand abscission. The more complex effects of waterlogging, the significance ofage of tissue on hormonal responses to stress, and the adaptivevalue of such responses to younger tissues are discussed.     

Stem and leaf hydraulic properties are finely coordinated in three tropical rain forest tree species

Coordination of stem and leaf hydraulic traits allows terrestrial plants to maintain safe water status under limited water supply. Tropical rain forests, one of the world's most productive biomes, are vulnerable to drought and potentially threatened by increased aridity due to global climate change. However, the relationship of stem and leaf traits within the plant hydraulic continuum remains understudied, particularly in tropical species. We studied within‐plant hydraulic coordination between stems and leaves in three tropical lowland rain forest tree species by analyses of hydraulic vulnerability [hydraulic methods and ultrasonic emission (UE) analysis], pressure‐volume relations and in situ pre‐dawn and midday water potentials (Ψ). We found finely coordinated stem and leaf hydraulic features, with a strategy of sacrificing leaves in favour of stems. Fifty percent of hydraulic conductivity (P₅₀) was lost at ?2.1 to ?3.1 MPa in stems and at ?1.7 to ?2.2 MPa in leaves. UE analysis corresponded to hydraulic measurements. Safety margins (leaf P₅₀ – stem P₅₀) were very narrow at ?0.4 to ?1.4 MPa. Pressure‐volume analysis and in situ Ψ indicated safe water status in stems but risk of hydraulic failure in leaves. Our study shows that stem and leaf hydraulics were finely tuned to avoid embolism formation in the xylem.     

Stem and leaf hydraulics of congeneric tree species from adjacent tropical savanna and forest ecosystems

Leaf and stem functional traits related to plant water relations were studied for six congeneric species pairs, each composed of one tree species typical of savanna habitats and another typical of adjacent forest habitats, to determine whether there were intrinsic differences in plant hydraulics between these two functional types. Only individuals growing in savanna habitats were studied. Most stem traits, including wood density, the xylem water potential at 50% loss of hydraulic conductivity, sapwood area specific conductivity, and leaf area specific conductivity did not differ significantly between savanna and forest species. However, maximum leaf hydraulic conductance (K _leaf) and leaf capacitance tended to be higher in savanna species. Predawn leaf water potential and leaf mass per area were also higher in savanna species in all congeneric pairs. Hydraulic vulnerability curves of stems and leaves indicated that leaves were more vulnerable to drought-induced cavitation than terminal branches regardless of genus. The midday K _leaf values estimated from leaf vulnerability curves were very low implying that daily embolism repair may occur in leaves. An electric circuit analog model predicted that, compared to forest species, savanna species took longer for their leaf water potentials to drop from predawn values to values corresponding to 50% loss of K _leaf or to the turgor loss points, suggesting that savanna species were more buffered from changes in leaf water potential. The results of this study suggest that the relative success of savanna over forest species in savanna is related in part to their ability to cope with drought, which is determined more by leaf than by stem hydraulic traits. Variation among genera accounted for a large proportion of the total variance in most traits, which indicates that, despite different selective pressures in savanna and forest habitats, phylogeny has a stronger effect than habitat in determining most hydraulic traits.     

Developmental expression of β-glucosidase in olive leaves

Plant β-glucosidases catalyze the hydrolysis of glycosidic linkages and play a vital role in defense against pathogens and stress. The present work investigated the relationship between leaf development and β-glucosidase protein content in Olea europea L. (cv. Picual) leaves. The total chlorophyll content increased with leaf age in current-season leaves. Immunoblot analysis revealed that the content of 61 kD protein of β-glucosidase also increased with leaf age, and that the enzyme existed in three isoforms (pI 5.8–6.2). Statistical analysis indicated a strong correlation between chlorophyll and β-glucosidase protein contents.     

Evaluation of leaf water status by means of permittivity at terahertz frequencies

We present an electromagnetic model of plant leaves which describes their permittivity at terahertz frequencies. The complex permittivity is investigated as a function of the water content of the leaf. Our measurements on coffee leaves (Coffea arabica L.) demonstrate that the dielectric material parameters can be employed to determine the leaf water status and, therefore, to monitor drought stress in plant leaves. The electromagnetic model consists of an effective medium theory, which is implemented by a third order extension of the Landau, Lifshitz, Looyenga model. The influence of scattering becomes important at higher frequencies and is modeled by a Rayleigh roughness factor.     

玉米叶片水分利用效率的保守性

水分利用效率是植物个体或生态系统水分利用过程的重要特征参数,可表征不同时空尺度的植物碳-水耦合关系,对植物适应气候变化研究具有重要意义。以玉米为例,利用中国气象局固城农业气象野外科学试验基地2013—2014年玉米不同灌溉方案模拟试验资料,对不同叶位叶片的水分利用效率特征及其影响因素进行分析。结果表明:植株顶部第1片叶片水分利用效率在拔节期和乳熟期呈现明显的峰值,反映出明显的周期变化规律及其与叶片生理生态特征的紧密相关。在相同环境条件下,不同叶位叶片的水分利用效率不存在显著性差异,即玉米叶片水分利用效率具有空间稳定性与叶龄保守性。同时,研究指出叶片光合速率和蒸腾速率在叶位之间的协调变化是导致空间稳定性和叶龄保守性的主要原因。研究结果可为植物水分关系研究提供参考,也可为水分利用效率的尺度化研究提供依据。