不同类型和强度的干旱胁迫对黑麦草实验种群物质生产与水分利用的影响

对田间条件下不同类型和强度的干旱胁迫对黑麦草 ( L olium perenne)实验种群物质生产与水分利用特征的影响进行了研究。实验结果表明 :在周期性和持续性胁迫条件下 ,轻度与中度胁迫对草群干物质产量的影响相接近并与对照无显著差异 ,重度胁迫导致干物质产量的降低 ;渐进性胁迫条件下轻度胁迫对干物质产量的影响与对照相接近 ,中度和重度胁迫导致产量的降低 ;在每一种胁迫类型下植物地上部绿色部分的干物质含量与其干物质产量呈反向对应关系 ;草群干物质产量与栽植桶水分散失量之间呈极显著的直线关系 ;周期性胁迫条件下中度胁迫提高了草群水分利用效率 ,持续性和渐进性胁迫条件下草群水分利用效率对干旱胁迫强度的反应随生长季节推移而有所改变     

RNAi-mediated disruption of squalene synthase improves drought tolerance and yield in rice

About one-third of the world's rice area is in rain-fed lowlands and most are prone to water shortage. The identification of genes imparting tolerance to drought in the model cereal plant, rice, is an attractive strategy to engineer improved drought tolerance not only rice but other cereals as well. It is demonstrated that RNAi-mediated disruption of a rice farnesyltransferase/squalene synthase (SQS) by maize squalene synthase improves drought tolerance at both the vegetative and reproductive stages. Twenty-day-old seedlings of wild type (Nipponbare) and seven independent events of transgenic RNAi lines showed no difference in morphology. When subjected to water stress for a period of 32 d under growth chamber conditions, transgenic positives showed delayed wilting, conserved more soil water, and improved recovery. When five independent events along with wild-type plants were subjected to drought at the reproductive stage under greenhouse conditions, the transgenic plants lost water more slowly compared with the wild type, through reduced stomatal conductance and the retention of high leaf relative water content (RWC). After 28 d of slow progressive soil drying, transgenic plants recovered better and flowered earlier than wild-type plants. The yield of water-stressed transgenic positive plants ranged from 14-39% higher than wild-type plants. When grown in plates with Yoshida's nutrient solution with 1.2% agar, transgenic positives from three independent events showed increased root length and an enhanced number of lateral roots. The RNAi-mediated inactivation produced reduced stomatal conductance and subsequent drought tolerance.     

不同类型和强度的干旱胁迫对黑麦草实验种群物质生产与水分利用的影响

 对田间条件下不同类型和强度的干旱胁迫对黑麦草(Lolium perenne)实验种群物质生产与水分利用特征的影响进行了研究。实验结果表明：在周期性和持续性胁迫条件下,轻度与中度胁迫对草群干物质产量的影响相接近并与对照无显著差异,重度胁迫导致干物质产量的降低;渐进性胁迫条件下轻度胁迫对干物质产量的影响与对照相接近,中度和重度胁迫导致产量的降低;在每一种胁迫类型下植物地上部绿色部分的干物质含量与其干物质产量呈反向对应关系;草群干物质产量与栽植桶水分散失量之间呈极显著的直线关系;周期性胁迫条件下中度胁迫提高了草群水分利用效率,持续性和渐进性胁迫条件下草群水分利用效率对干旱胁迫强度的反应随生长季节推移而有所改变。     

Effects of Multi-Stage Continuous Drought on Photosynthetic Characteristics,Yield and Water Use Efficiency of Winter Wheat

A drought event can cause entire crops to fail or yield loss. In order to study the effects of continuous drought on photosynthetic characteristics, yield, and water use efficiency (WUE) of winter wheat (Triticum aestivum L.), the winter wheat variety “Aikang 58” was selected as test material with controlling the water of the pot-planted winter wheat under a mobile rainout shelter. Based on foot planting and safe wintering, winter wheat was evaluated under different drought conditions, including light, moderate and severe drought at the jointing (B), heading (C), and filling (G) stages. The soil water content was controlled in a range of 60% to 70%, 50% to 60%, and 40% to 50% of the field capacity, respectively. In the experiment, there were 9 single-stage droughts, 3 three-stage droughts, and 1 test control (totaling 13 trials). The results are as follows: Under a single-stage drought, the change of net photosynthetic rate (Pn) and stomatal conductance (Gs) have similar trends, and they both decrease significantly with the severity of the drought. Under three-stage continuous droughts, the change curve of Gs shows a constant downward trend; the change curve of Pn showed a “valley shape,” and the minimum value of Pn appeared at the heading stage. All droughts will reduce the yield of winter wheat. Under the three-stage continuous drought conditions, except for light drought, moderate drought and severe drought will cause significant yield reduction, mainly due to lack of water at the jointing and heading stages. Continuous drought will reduce the WUE, and the difference will reach a significant level under moderate and severe drought. The present results suggested that when water resources are scarce, it is a better irrigation model to save water and achieve high grain yield by applying appropriate water stress (60%–70% FC) during the critical growth period of winter wheat.     

Overexpression of a Pea DNA Helicase (PDH45) in Peanut (Arachis hypogaea L.) Confers Improvement of Cellular Level Tolerance and Productivity Under Drought Stress

Peanut, a major edible oil seed crop globally is predominantly grown under rainfed conditions and suffers yield losses due to drought. Development of drought-tolerant varieties through transgenic technology is a valid approach. Besides superior water relation traits like water mining, intrinsic cellular level tolerance mechanisms are important to sustain the growth under stress. To achieve this objective, the focus of this study was to pyramid drought adaptive traits by overexpressing a stress responsive helicase, PDH45 in the background of a genotype with superior water relations. PCR, Southern, and RT-PCR analyses confirmed stable integration and expression of the PDH45 gene in peanut transgenics. At the end of T₃ generation, eight transgenic events were identified as promising based on stress tolerance and improvement in productivity. Several transgenic lines showed stay-green phenotype and increased chlorophyll stability under stress and reduced chlorophyll retardation under etherel-induced simulated stress conditions. Stress-induced root growth was also substantially higher in the case of transformants. This was reflected in increased WUE (low Δ¹³C) and improved growth rates and productivity. The transgenics showed 17.2 and 26.75 % increase in yield under non-stress and stress conditions over wild type ascertaining the feasibility of trait pyramiding strategy for the development of drought-tolerant peanut.     

Effect of Pre-Anthesis Drought Hardening on Post-Anthesis Physiological Characteristics,Yield and WUE in Winter Wheat

A drought event can cause yield loss or entire crops to fail. In order to study the effects of continuous drought on physiological characteristics, yield, and water use efficiency (WUE) of winter wheat (Triticum aestivum L.), the variety “Zhoumai 22” was grown in controlled water conditions of the pot-planted winter wheat under a mobile rainout shelter. Foot planting and safe wintering were used to evaluate, winter wheat under different drought conditions, including light, moderate and severe drought at the jointing, heading, and filling stages. The soil water content was controlled at 60–70%, 50–60%, or 40–50% of field capacity. Experimental trials included 3 pre-anthesis drought hardening, 3 three-stage continuous drought, and 1 test control conditions. Under drought stress conditions, winter wheat leaf water potential, soil plant analysis development (SPAD), photosynthesis parameters, and yield declined due to pre-anthesis drought hardening. And the degree of decline: continuous drought > pre-anthesis drought hardening. Changes in the post-anthesis photosynthetic capacity of winter wheat were mainly related to the pre-anthesis drought level, rather than post-anthesis rehydration. The threshold of non-stomata limiting factors caused by photosynthesis at the filling stage is 40–50%FC, while comprehensive yield and WUE affected, the yield in severe drought treatments was the most significant, B3C3 and B3C3G3 decreased by 55.68% and 55.88%, respectively. Pre-anthesis drought was the main reason for the decreased crop yield. Thus, severe drought should be avoided during planting, while pre-anthesis light drought is a suitable choice for water-saving and crop production, as proper pre-anthesis drought hardening (60–70% FC) is feasible and effective.     

Mechanisms of selection for drought stress tolerance and avoidance in Impatiens capensis (Balsaminaceae)

For longer lived annual plants, high water-use efficiency (WUE) and low stomatal conductance are hypothesized to confer a fitness advantage under drought stress. To directly test the adaptive significance of WUE and stomatal conductance under drought stress, inbred lines of Impatiens capensis were grown in two field environments (watered and not-watered), in a year of unusual early-season drought. In contrast to the results from a previous study of late-season drought in the same system, selection was detected for lower WUE, increased stomatal conductance, and early flowering time. These findings suggest that early-season drought conditions may select for drought avoidance traits such as low WUE and early reproduction, whereas later drought selects for tolerance traits such as high WUE.     

Overexpression of HVA1 gene from barley generates tolerance to salinity and water stress in transgenic mulberry (Morus indica)

Late embryogenesis abundant (LEA) proteins are members of a large group of hydrophilic proteins found primarily in plants. The barley hva1 gene encodes a group 3 LEA protein and is induced by ABA and water deficit conditions. We report here the over expression of hva1 in mulberry under a constitutive promoter via Agrobacterium-mediated transformation. Molecular analysis of the transgenic plants revealed the stable integration and expression of the transgene in the transformants. Transgenic plants were subjected to simulated salinity and drought stress conditions to study the role of hva1 in conferring tolerance. The transgenic plants showed better cellular membrane stability (CMS), photosynthetic yield, less photo-oxidative damage and better water use efficiency as compared to the non-transgenic plants under both salinity and drought stress. Under salinity stress, transgenic plants show many fold increase in proline concentration than the non-transgenic plants and under water deficit conditions proline is accumulated only in the non-transgenic plants. Results also indicate that the production of HVA1 proteins helps in better performance of transgenic mulberry by protecting membrane stability of plasma membrane as well as chloroplastic membranes from injury under abiotic stress. Interestingly, it was observed that hva1 conferred different degrees of tolerance to the transgenic plants towards various stress conditions. Amongst the lines analysed for stress tolerance transgenic line ST8 was relatively more salt tolerant, ST30, ST31 more drought tolerant, and lines ST11 and ST6 responded well under both salinity and drought stress conditions as compared to the non-transgenic plants. Thus hva1 appears to confer a broad spectrum of tolerance under abiotic stress in mulberry.     

Osmotin-expressing transgenic tea plants have improved stress tolerance and are of higher quality

Drought is a major stress that affects the yield and quality of tea, a widely consumed beverage crop grown in more than 20 countries of the world. Therefore, osmotin gene-expressing transgenic tea plants produced using earlier optimized conditions were evaluated for their tolerance of drought stress and their quality. Improved tolerance of polyethylene glycol-induced water stress and faster recovery from stress were evident in transgenic lines compared with the normal phenotype. Significant improvements in growth under in-vitro conditions were also observed. Besides enhanced reactive oxygen species-scavenging enzyme activity, the transgenic lines contained significantly higher levels of flavan-3-ols and caffeine, key compounds that govern quality and commercial yield of the beverage. The selected transgenic lines have the potential to meet the demands of the tea industry for stress-tolerant plants with higher yield and quality. These traits of the transgenic lines can be effectively maintained for generations because tea is commercially cultivated through vegetative propagation only.     

Field evaluation of transgenic wheat plants stably expressing the HVA1 gene for drought tolerance

The barley HVA1 gene, encoding a member of the group 3 late embryogenesis abundant (LEA) proteins, has previously been introduced into spring wheat cv. Hi-Line to determine its effect on drought tolerance (Sivamani E, Bahieldin A, Wraith JM, Al-Niemi T, Dyer WE, Ho T-HD, Qu R (2000) Improved biomass productivity and water use efficiency under water deficit conditions in transgenic wheat constitutively expressing the barley HVA1 gene. Plant Sci 155, 1–9). T4 progeny from six independent transgenic events (lines 111/1, 1/1, 11/2, 84, 765 and 1201) were tested in nine field experiments over six cropping seasons. In the first two seasons, the total biomass per plot and the grain yield per plot of line 111/1 were higher than those of line 1/1, and higher than those of the wild-type control in the second season. The grain yield per plot of line 11/2 was significantly lower than that of the transgenic lines 111/1 and 1/1 in the third season, and this line was not tested further. In the fourth season, the plant height and grain yield per plot of line 111/1 were significantly higher than those of the wild-type control. Under dryland conditions in the fifth season, line 111/1 showed significantly greater plant height, total biomass per plot and grain yield per plot than the wild-type control in at least two of the four locations, as well as across locations. In the sixth season, newly developed transgenic lines 1201 and 765 significantly overyielded the two original transgenic lines 111/1 and 1/1, the non-expressing transgenic line 84 as well as the wild-type control in the three yield attributes and leaf water measurement, namely relative water content (RWC). This result coincided with the rate of HVA1 transgene expression of the different genotypes. Differences in total seed storage protein concentrations between the transgenic lines and the wild-type control within or across environmental conditions were insignificant. These field trials show that the HVA1 gene has the potential to confer drought stress protection in transgenic spring wheat.     

Ecological interactions and the fitness effect of water‐use efficiency: Competition and drought alter the impact of natural MPK12 alleles in Arabidopsis

The presence of substantial genetic variation for water‐use efficiency (WUE) suggests that natural selection plays a role in maintaining alleles that affect WUE. Soil water deficit can reduce plant survival, and is likely to impose selection to increase WUE, whereas competition for resources may select for decreased WUE to ensure water acquisition. We tested the fitness consequences of natural allelic variation in a single gene (MPK12) that influences WUE in Arabidopsis, using transgenic lines contrasting in MPK12 alleles, under four treatments; drought/competition, drought/no competition, well‐watered/competition, well‐watered/no competition. Results revealed an allele × environment interaction: Low WUE plants performed better in competition, resulting from increased resource consumption. Contrastingly, high WUE individuals performed better in no competition, irrespective of water availability, presumably from enhanced water conservation and nitrogen acquisition. Our findings suggest that selection can influence MPK12 evolution, and represents the first assessment of plant fitness resulting from natural allelic variation at a single locus affecting WUE.     

Overexpression of an Arabidopsis thaliana galactinol synthase gene improves drought tolerance in transgenic rice and increased grain yield in the field

Drought stress has often caused significant decreases in crop production which could be associated with global warming. Enhancing drought tolerance without a grain yield penalty has been a great challenge in crop improvement. Here, we report the Arabidopsis thaliana galactinol synthase 2 gene (AtGolS2) was able to confer drought tolerance and increase grain yield in two different rice (Oryza sativa) genotypes under dry field conditions. The developed transgenic lines expressing AtGolS2 under the control of the constitutive maize ubiquitin promoter (Ubi:AtGolS2) also had higher levels of galactinol than the non‐transgenic control. The increased grain yield of the transgenic rice under drought conditions was related to a higher number of panicles, grain fertility and biomass. Extensive confined field trials using Ubi:AtGolS2 transgenic lines in Curinga, tropical japonica and NERICA4, interspecific hybrid across two different seasons and environments revealed the verified lines have the proven field drought tolerance of the Ubi:AtGolS2 transgenic rice. The amended drought tolerance was associated with higher relative water content of leaves, higher photosynthesis activity, lesser reduction in plant growth and faster recovering ability. Collectively, our results provide strong evidence that AtGolS2 is a useful biotechnological tool to reduce grain yield losses in rice beyond genetic differences under field drought stress.     

Adjusting Nitrogen Application in Accordance with Soil Water Availability Enhances Yield and Water Use by Regulating Physiological Traits of Maize under Drip Fertigation

Knowledge of the interactive effects of water and nitrogen (N) on physio-chemical traits of maize (Zea mays L.) helps to optimize water and N management and improve productivity. A split-plot experiment was conducted with three soil water conditions (severe drought, moderate drought, and fully water supply referring to 45%–55%, 65%–75%, and 85%–95% field capacity, respectively) and four N application rates (N₀, N₁₅₀, N₂₄₀, and N₃₃₀ referring to 0, 150, 240, 330 kg N ha^–1 respectively) under drip fertigation in 2014 and 2015 in the Huang-Huai-Hai Plain of China. The results indicated that drought stress inhibited physiological activity of plants (leaf relative water content, root bleeding sap, and net photosynthetic rate), resulting in low dry matter accumulation after silking, yield, and N uptake, whereas increased WUE and NUE. N application rates over than 150 kg ha^–1 aggravated the inhibition of physiological activity under severe drought condition, while it was offset under moderate drought condition. High N application rates (N₃₃₀) still revealed negative effects under moderate drought condition, as it did not consistently enhance plant physiological activity and significantly reduced N uptake as compared to the N₂₄₀ treatment. With fully water supply, increasing N application rates synergistically enhanced physiological activity, promoted dry matter accumulation after silking, and increased yield, WUE, and N uptake. Although the N₂₄₀ treatment reduced yield by 5.4% in average, it saved 27.3% N under full water supply condition as compared with N₃₃₀ treatment. The results indicated that N regulated growth of maize in aspects of physiological traits, dry matter accumulation, and yield as well as water and N use was depended on soil water status. The appropriate N application rates for maize production was 150 kg ha^–1 under moderate drought or 240 kg ha^–1 under fully water supply under drip fertigation, and high N supply (>150 kg ha^–1) should be avoided under severe drought condition.     

The TaDREB3 transgene transferred by conventional crossings to different genetic backgrounds of bread wheat improves drought tolerance

Drought tolerance of the wheat cultivar Bobwhite was previously enhanced by transformation with a construct containing the wheat DREB3 gene driven by the stress‐inducible maize Rab17 promoter. Progeny of a single T₂ transgenic line were used as pollinators in crosses with four elite bread wheat cultivars from Western Australia: Bonnie Rock, IGW‐2971, Magenta and Wyalkatchem, with the aim of evaluating transgene performance in different genetic backgrounds. The selected pollinator line, BW8‐9‐10‐3, contained multiple transgene copies, had significantly improved drought tolerance compared with wild‐type plants and showed no growth and development penalties or abnormalities. A single hybrid plant was selected from each cross‐combination for three rounds of backcrossing with the corresponding maternal wheat cultivar. The transgene was detected in all four F₁BC₃ combinations, but stress‐inducible transgene expression was found in only three of the four combinations. Under well‐watered conditions, the phenotypes and grain yield components of the F₂BC₃ transgene‐expressing lines were similar to those of corresponding recurrent parents and null‐segregants. Under severe drought conditions, the backcross lines demonstrated 12–18% higher survival rates than the corresponding control plants. Two from four F₃BC₃ transgenic lines showed significantly higher yield (18.9% and 21.5%) than control plants under limited water conditions. There was no induction of transgene expression under cold stress, and therefore, no improvement of frost tolerance observed in the progenies of drought‐tolerant F₃BC₃ lines.     

Molecular tailoring of farnesylation for plant drought tolerance and yield protection

Protecting crop yield under drought stress is a major challenge for modern agriculture. One biotechnological target for improving plant drought tolerance is the genetic manipulation of the stress response to the hormone abscisic acid (ABA). Previous genetic studies have implicated the involvement of the beta-subunit of Arabidopsis farnesyltransferase (ERA1) in the regulation of ABA sensing and drought tolerance. Here we show that molecular manipulation of protein farnesylation in Arabidopsis, through downregulation of either the alpha- or beta-subunit of farnesyltransferase enhances the plant's response to ABA and drought tolerance. To test the effectiveness of tailoring farnesylation in a crop plant, transgenic Brassica napus carrying an ERA1 antisense construct driven by a drought-inducible rd29A promoter was examined. In comparison with the non-transgenic control, transgenic canola showed enhanced ABA sensitivity, as well as significant reduction in stomatal conductance and water transpiration under drought stress conditions. The antisense downregulation of canola farnesyltransferase for drought tolerance is a conditional and reversible process, which depends on the amount of available water in the soil. Furthermore, transgenic plants were more resistant to water deficit-induced seed abortion during flowering. Results from three consecutive years of field trial studies suggest that with adequate water, transgenic canola plants produced the same amount of seed as the parental control. However, under moderate drought stress conditions at flowering, the seed yields of transgenic canola were significantly higher than the control. Using protein farnesyltransferase as an effective target, these results represent a successful demonstration of engineered drought tolerance and yield protection in a crop plant under laboratory and field conditions.     

Whole plant responses, key processes, and adaptation to drought stress: the case of rice

Most high-yielding rice cultivars developed for irrigated conditions, including the widely grown lowland variety IR64, are highly susceptible to drought stress. This limits their adoption in rainfed rice environments where there is a risk of water shortage during the growing season. Mapping studies using lowland-by-upland rice populations have provided limited information about the genetic basis of variation in yield under drought. One approach to simultaneously improve and understand rice drought tolerance is to generate backcross populations, select superior lines in managed stress environments, and then evaluate which features of the selected lines differ from the recurrent parent. This approach was been taken with IR64, using a range of tolerant and susceptible cultivars as donor parents. Yields of the selected lines measured across 13 widely contracting water environments were generally greater than IR64, but genotype-by-environment effects were large. Traits expected to vary between IR64 and selected lines are plant height, because many donors were not semi-dwarf types, and maturity, because selection in a terminal stress environment is expected to favour earliness. In these experiments it was found that some lines that performed better under upland drought were indeed taller than IR64, but that shorter lines with good yield under drought could also be identified. In trials where drought stress developed in previously flooded (lowland) fields, height was not associated with performance. There was little change in maturity with selection. Other notable differences between IR64 and the selected backcross lines were in their responses to applied ABA and ethylene in greenhouse experiments at the vegetative stage and in leaf rolling observed under chronic upland stress in the field. These observations are consistent with the hypothesis that adaptive responses to drought can effectively allow for improved performance across a broad range of water environments. The results indicate that the yield of IR64 under drought can be significantly improved by backcrossing with selection under stress. In target environments where drought is infrequent but significant in certain years, improved IR64 with greater drought tolerance would be a valuable option for farmers.     

Analysis of early responses to drought associated with field drought adaptation in four Sahelian groundnut (Arachis hypogaea L.) cultivars

Groundnut (Arachis hypogaea L.) is the most important oil and cash crop in the sub-Sahelian tropics. Plant adaptation to drought, i.e. cultivars (cvs) that can maintain yield when water is limited, is a complex phenomenon which is not yet fully understood. This study aimed to identify traits expressed at the early stages of the cycle that could reveal cv differences in drought adaptation in the field. The field productivity of four Sahelian groundnut cvs was assessed during three crop seasons in Bambey (Senegal). The same cvs grown in rhizotrons were subjected to early drought stress and to a desiccation test to assess cell membrane tolerance. Between-cv differences were found with respect to pod yield, biomass production, water use efficiency (WUE), stomatal regulation and cell membrane tolerance. Two strategies to cope with water deficit were identified. The first behaviour was characterised by high rapid water loss, late stomatal closure and low cell membrane damage during drought. These traits are all found in the semi-late Virginia cv 57-422 and, into a lesser extent, in the early Spanish cv Fleur 11. For both cvs, biomass production was boosted under favourable conditions in rhizotrons but the semi-late cv had poor pod yield under end-of-season water deficit conditions. The second strategy involved opposite characters, leading to the maintenance of a higher water status, resulting in lower photosynthesis and yield. This characterised the early Spanish cv 73-30, and also, to some extent, the early Spanish cv 55-437. Earliness associated with high WUE, stomatal conductance and cell membrane tolerance, were the main traits of Fleur 11, a cv derived from a Virginia × Spanish cross, which was able to maintain acceptable yield under varying drought patterns in the field. These traits, as they were detectable at an early stage, could therefore be efficiently integrated in groundnut breeding programmes for drought adaptation.     

Drought tolerance through biotechnology: improving translation from the laboratory to farmers' fields

Water availability is a significant constraint to crop production, and increasing drought tolerance of crops is one step to gaining greater yield stability. Excellent progress has been made using models to identify pathways and genes that can be manipulated through biotechnology to improve drought tolerance. A current focus is on translation of results from models in controlled environments to crops in the field. Field testing to demonstrate improved yields under water-limiting conditions is challenging and expensive. More extensive phenotyping of transgenic lines in the greenhouse may contribute to improved predictions about field performance. It is possible that multiple mechanisms of drought tolerance may be needed to provide benefit across the diversity of water stress environments relevant to economic yield.     

PdEPF1 regulates water‐use efficiency and drought tolerance by modulating stomatal density in poplar

Water deficiency is a critical environmental condition that is seriously reducing global plant production. Improved water‐use efficiency (WUE) and drought tolerance are effective strategies to address this problem. In this study, PdEPF1, a member of the EPIDERMAL PATTERNING FACTOR (EPF) family, was isolated from the fast‐growing poplar clone NE‐19 [Populus nigra × (Populus deltoides × Populus nigra)]. Significantly, higher PdEPF1 levels were detected after induction by dehydration and abscisic acid. To explore the biological functions of PdEPF1, transgenic triploid white poplars (Populus tomentosa ‘YiXianCiZhu B385’) overexpressing PdEPF1 were constructed. PdEPF1 overexpression resulted in increased water deficit tolerance and greater WUE. We confirmed that the transgenic lines with greater instantaneous WUE had approximately 30% lower transpiration but equivalent CO₂ assimilation. Lower transpiration was associated with a 28% reduction in abaxial stomatal density. PdEPF1 overexpression not only strongly enhanced WUE, but also greatly improved drought tolerance, as measured by the leaf relative water content and water potential, under limited water conditions. In addition, the growth of these oxPdEPF1 plants was less adversely affected by reduced water availability than plants with a higher stomatal density, indicating that plants with a low stomatal density may be well suited to grow in water‐scarce environments. Taken together, our data suggest that PdEPF1 improves WUE and confers drought tolerance in poplar; thus, it could be used to breed drought‐tolerant plants with increased production under conditions of water deficiency.