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We developed and characterized a paper-based microfluidic sensor for the on-site diagnosis of drought stress in plants. Proline was used as a biomarker for analyzing drought stress, which was extracted by a colorimetric method using the proline-ninhydrin reaction. Paper was used as the main sensor material for the on-site detection of proline as it is easily transportable and cost-effective. The paper-based sensor was fabricated using wax-printing and origami methods, and the sensor was precoated with ninhydrin to allow for easy and convenient on-site use. Furthermore, a sample-to-ninhydrin ratio of 1:2 was found to confer optimal sensitivity to the drought diagnosis sensor. The concentration of proline in a sample was quantified by red-green-blue analysis to determine the change in green color intensity levels in response to distinct proline concentrations, which were detected by the sensor. The limit of detection of proline using the devised sensor was 657 µM, and the green color intensity level decreased with increasing proline concentration. In addition, the sensor was validated in an experimental drought stress model with Arabidopsis and subjected to drought stress for 21 days, and the amount of proline detected was 10 mM. The devised paper-based microfluidic sensor highlights the possibility of the on-site evaluation of drought stress in plants with potential to be utilized in various agricultural areas in the future.

This paper integrates dendrochronological, demographic, and experimental perspectives to improve understanding of the response of white spruce ( Picea glauca (Moench) Voss) tree growth to climatic variability in interior Alaska. The dendrochronological analyses indicate that climate warming has led to widespread declines in white spruce growth throughout interior Alaska that have become more prevalent during the 20th century. Similarly, demographic studies show that white spruce tree growth is substantially limited by soil moisture availability in both mid- and late-successional stands. Interannual variability in tree growth among stands within a landscape exhibits greater synchrony than does growth of trees that occupy different landscapes, which agrees with dendrochronological findings that the responses depend on landscape position and prevailing climate. In contrast, the results from 18 years of a summer moisture limitation experiment showed that growth in midsuccessional upland stands was unaffected by moisture limitation and that moisture limitation decreased white spruce growth in floodplain stands where it was expected that growth would be less vulnerable because of tree access to river water. Taken together, the evidence from the different perspectives analyzed in this study clearly indicates that white spruce tree growth in interior Alaska is vulnerable to the effects of warming on plant water balance.

Aim: The stress-gradient hypothesis (SGH) postulates that species interactions shift from negative to positive with increasing abiotic stress. Interactions between species are increasingly being recognized as important drivers of species distributions, but it is still unclear whether stress-induced changes in interactions affect continental-to- global scale species distributions. Here, we tested whether associations of vascular plant species in dry grasslands in Europe follow the SGH along a climatic water deficit (CWD) gradient across the continent. Location: Dry grasslands in Europe. Time period: Present. Major taxa studied: Vascular plants. Methods: We built a context-dependent joint species distribution model (JSDM) to estimate the residual associations (i.e., associations that are not explained by the abiotic environment) of 161 plant species as a function of the CWD based on community data from 8,660 vegetation plots. We evaluated changes in residual associations between species for pairs and on the community level, and we compared responses for groups of species with different drought tolerances. Results: We found contrasting shifts in associations for drought-sensitive and drought-tolerant species. For drought-sensitive species, 21% of the pairwise associations became more positive with increasing CWD, whereas 17% became more negative. In contrast, only 17% of the pairwise associations involving drought-tolerant species became more positive, whereas 27% became more negative in areas with a high CWD. Additionally, the incidence of positive associations increased with drought for drought-sensitive species and decreased for drought-tolerant species. Main conclusions: We found that associations of drought-sensitive plant species became more positive with drought, in line with the SGH. In contrast, associations of drought-tolerant species became more negative. Additionally, changes in associations of single species pairs were highly variable. Our results indicate that stress-modulated Aim: The stress-gradient hypothesis (SGH) postulates that species interactions shift from negative to positive with increasing abiotic stress. Interactions between species are increasingly being recognized as important drivers of species distributions, but it is still unclear whether stress-induced changes in interactions affect continental-to- global scale species distributions. Here, we tested whether associations of vascular plant species in dry grasslands in Europe follow the SGH along a climatic water deficit (CWD) gradient across the continent. Location: Dry grasslands in Europe. Time period: Present. Major taxa studied: Vascular plants. Methods: We built a context-dependent joint species distribution model (JSDM) to estimate the residual associations (i.e., associations that are not explained by the abiotic environment) of 161 plant species as a function of the CWD based on community data from 8,660 vegetation plots. We evaluated changes in residual associations between species for pairs and on the community level, and we compared responses for groups of species with different drought tolerances. Results: We found contrasting shifts in associations for drought-sensitive and drought-tolerant species. For drought-sensitive species, 21% of the pairwise associations became more positive with increasing CWD, whereas 17% became more negative. In contrast, only 17% of the pairwise associations involving drought-tolerant species became more positive, whereas 27% became more negative in areas with a high CWD. Additionally, the incidence of positive associations increased with drought for drought-sensitive species and decreased for drought-tolerant species. Main conclusions: We found that associations of drought-sensitive plant species became more positive with drought, in line with the SGH. In contrast, associations of drought-tolerant species became more negative. Additionally, changes in associations of single species pairs were highly variable. Our results indicate that stress-modulated Aim: The stress-gradient hypothesis (SGH) postulates that species interactions shift from negative to positive with increasing abiotic stress. Interactions between species are increasingly being recognized as important drivers of species distributions, but it is still unclear whether stress-induced changes in interactions affect continental-to- global scale species distributions. Here, we tested whether associations of vascular plant species in dry grasslands in Europe follow the SGH along a climatic water deficit (CWD) gradient across the continent. Location: Dry grasslands in Europe. Time period: Present. Major taxa studied: Vascular plants. Methods: We built a context-dependent joint species distribution model (JSDM) to estimate the residual associations (i.e., associations that are not explained by the abiotic environment) of 161 plant species as a function of the CWD based on community data from 8,660 vegetation plots. We evaluated changes in residual associations between species for pairs and on the community level, and we compared responses for groups of species with different drought tolerances. Results: We found contrasting shifts in associations for drought-sensitive and drought-tolerant species. For drought-sensitive species, 21% of the pairwise associations became more positive with increasing CWD, whereas 17% became more negative. In contrast, only 17% of the pairwise associations involving drought-tolerant species became more positive, whereas 27% became more negative in areas with a high CWD. Additionally, the incidence of positive associations increased with drought for drought-sensitive species and decreased for drought-tolerant species. Main conclusions: We found that associations of drought-sensitive plant species became more positive with drought, in line with the SGH. In contrast, associations of drought-tolerant species became more negative. Additionally, changes in associations of single species pairs were highly variable. Our results indicate that stress-modulated species associations might influence the distribution of species over large geographical extents, thus leading to unexpected responses under climate change through shifts in species associations

This review focuses on developing high-throughput phenotyping approaches to quantify key physiological traits at high temporal frequency, involving diverse germplasm to incorporate greater heat and drought stress resilience in crops., Flowering and grain-filling stages are highly sensitive to heat and drought stress exposure, leading to significant loss in crop yields. Therefore, phenotyping to enhance resilience to these abiotic stresses is critical for sustaining genetic gains in crop improvement programs. However, traditional methods for screening traits related to these stresses are slow, laborious, and often expensive. Remote sensing provides opportunities to introduce low-cost, less biased, high-throughput phenotyping methods to capture large genetic diversity to facilitate enhancement of stress resilience in crops. This review focuses on four key physiological traits and processes that are critical in understanding crop responses to drought and heat stress during reproductive and grain-filling periods. Specifically, these traits include: (i) time of day of flowering, to escape these stresses during flowering; (ii) optimizing photosynthetic efficiency; (iii) storage and translocation of water-soluble carbohydrates; and (iv) yield and yield components to provide in-season yield estimates. Moreover, we provide an overview of current advances in remote sensing in capturing these traits, and discuss the limitations with existing technology as well as future direction of research to develop high-throughput phenotyping approaches. In the future, phenotyping these complex traits will require sensor advancement, high-quality imagery combined with machine learning methods, and efforts in transdisciplinary science to foster integration across disciplines.

Summary Determining physiological mechanisms and thresholds for climate‐driven tree die‐off could help improve global predictions of future terrestrial carbon sinks. We directly tested for the lethal threshold in hydraulic failure – an inability to move water due to drought‐induced xylem embolism – in a pine sapling experiment.In a glasshouse experiment, we exposed loblolly pine (Pinus taeda) saplings (n = 83) to drought‐induced water stress ranging from mild to lethal. Before rewatering to relieve drought stress, we measured native hydraulic conductivity and foliar color change. We monitored all measured individuals for survival or mortality.We found a lethal threshold at 80% loss of hydraulic conductivity – a point of hydraulic failure beyond which it is more likely trees will die, than survive, and describe mortality risk across all levels of water stress. Foliar color changes lagged behind hydraulic failure – best predicting when trees had been dead for some time, rather than when they were dying.Our direct measurement of native conductivity, while monitoring the same individuals for survival or mortality, quantifies a continuous probability of mortality risk from hydraulic failure. Predicting tree die‐off events and understanding the mechanism involved requires knowledge not only of when trees are dead, but when they begin dying – having passed the point of no return.

International audience; Desert truffles are edible hypogeous fungi forming ectendomycorrhizal symbiosis with plants of Cistaceae family. Knowledge about the reproductive modes of these fungi and the molecular mechanisms driving the ectendomycorrhizal interaction is lacking. Genomes of the highly appreciated edible desert truffles Terfezia claveryi Chatin and Tirmania nivea Trappe have been sequenced and compared with other Pezizomycetes. Transcriptomes of T. claveryi × Helianthemum almeriense mycorrhiza from well-watered and drought-stressed plants, when intracellular colonizations is promoted, were investigated. We have identified the fungal genes related to sexual reproduction in desert truffles and desert-truffles-specific genomic and secretomic features with respect to other Pezizomycetes, such as the expansion of a large set of gene families with unknown Pfam domains and a number of species or desert-truffle-specific small secreted proteins differentially regulated in symbiosis. A core set of plant genes, including carbohydrate, lipid-metabolism, and defence-related genes, differentially expressed in mycorrhiza under both conditions was found. Our results highlight the singularities of desert truffles with respect to other mycorrhizal fungi while providing a first glimpse on plant and fungal determinants involved in ecto to endo symbiotic switch that occurs in desert truffle under dry conditions.

Tree responses to altered water availability range from immediate (e.g. stomatal regulation) to delayed (e.g. crown size adjustment). The interplay of the different response times and processes, and their effects on long‐term whole‐tree performance, however, is hardly understood. - Here we investigated legacy effects on structures and functions of mature Scots pine in a dry inner‐Alpine Swiss valley after stopping an 11‐yr lasting irrigation treatment. Measured ecophysiological time series were analysed and interpreted with a system‐analytic tree model. - We found that the irrigation stop led to a cascade of downregulations of physiological and morphological processes with different response times. Biophysical processes responded within days, whereas needle and shoot lengths, crown transparency, and radial stem growth reached control levels after up to 4 yr only. Modelling suggested that organ and carbon reserve turnover rates play a key role for a tree’s responsiveness to environmental changes. Needle turnover rate was found to be most important to accurately model stem growth dynamics. - We conclude that leaf area and its adjustment time to new conditions is the main determinant for radial stem growth of pine trees as the transpiring area needs to be supported by a proportional amount of sapwood, despite the growth‐inhibiting environmental conditions. ISSN:0028-646X ISSN:1469-8137

Crassulacean acid metabolism (CAM) photosynthesis is an important biological innovation enabling plant adaptation to hot and dry environments. CAM plants feature high water-use efficiency, with potential for sustainable crop production under water-limited conditions. A deep understanding of CAM-related gene function and molecular evolution of CAM plants is critical for exploiting the potential of engineering CAM into C3 crops to enhance crop production on semi-arid or marginal agricultural lands. With the newly emerging genomics resources for multiple CAM species, progress has been made in comparative genomics studies on the molecular basis and subsequently on the evolution of CAM. Here, recent advances in CAM comparative genomics research in constitutive and facultative CAM plants are reviewed, with a focus on the analyses of DNA/protein sequences and gene expression to provide new insights into the path and driving force of CAM evolution and to identify candidate genes involved in CAM-related biological processes. Potential applications of new computational and experimental technologies (e.g. CRISPR/Cas-mediated genome-editing technology) to the comparative and evolutionary genomics research on CAM plants are offered., This review updates the understanding of the molecular mechanisms underlying CAM evolution from C3 photosynthesis and explores the potential of comparative genomics for identifying candidate genes related to CAM.

ZmPTF1 regulates drought tolerance in maize by promoting root development and ABA synthesis, by binding to the G-box in the promoter and activating the expression of NCEDs, CBF4, NAC081, and NAC30., Drought stress is the most important environmental stress limiting maize production. ZmPTF1, a phosphate starvation-induced basic helix-loop-helix (bHLH) transcription factor, contributes to root development and low-phosphate tolerance in maize. Here, ZmPTF1 expression, drought tolerance, and the underlying mechanisms were studied by using maize ZmPTF1 overexpression lines and mutants. ZmPTF1 was found to be a positive regulator of root development, ABA synthesis, signalling pathways, and drought tolerance. ZmPTF1 was also found to bind to the G-box element within the promoter of 9-cis-epoxycarotenoid dioxygenase (NCED), C-repeat-binding factor (CBF4), ATAF2/NAC081, NAC30, and other transcription factors, and to act as a positive regulator of the expression of those genes. The dramatically upregulated NCEDs led to increased abscisic acid (ABA) synthesis and activation of the ABA signalling pathway. The up-regulated transcription factors hierarchically regulate the expression of genes involved in root development, stress responses, and modifications of transcriptional regulation. The improved root system, increased ABA content, and activated ABA-, CBF4-, ATAF2-, and NAC30-mediated stress responses increased the drought tolerance of the ZmPTF1 overexpression lines, while the mutants showed opposite trends. This study describes a useful gene for transgenic breeding and helps us understand the role of a bHLH protein in plant root development and stress responses.

ADF5 promotes stomatal closure by regulating actin filament dynamics, and members of the ABF/AREB transcription factor family may serve as potential upstream regulators of ADF5 in the drought stress/ABA signaling pathway., Stomatal movement plays an essential role in plant responses to drought stress, and the actin cytoskeleton and abscisic acid (ABA) are two important components of this process. Little is known about the mechanism underlying actin cytoskeleton remodeling and the dynamic changes occurring during stomatal movement in response to drought stress/ABA signaling. Actin-depolymerizing factors (ADFs) are conserved actin severing/depolymerizing proteins in eukaryotes, and in angiosperms ADFs have evolved actin-bundling activity. Here, we reveal that the transcriptional expression of neofunctionalized Arabidopsis ADF5 was induced by drought stress and ABA treatment. Furthermore, we demonstrated that ADF5 loss-of-function mutations increased water loss from detached leaves, reduced plant survival rates after drought stress, and delayed stomatal closure by regulating actin cytoskeleton remodeling via its F-actin-bundling activity. Biochemical assays revealed that an ABF/AREB transcription factor, DPBF3, could bind to the ADF5 promoter and activate its transcription via the ABA-responsive element core motif ACGT/C. Taken together, our findings indicate that ADF5 participates in drought stress by regulating stomatal closure, and may also serve as a potential downstream target of the drought stress/ABA signaling pathway via members of the ABF/AREB transcription factors family.

Drought stress impacts seedling establishment, survival and whole-plant productivity. Molecular responses to drought stress have been most extensively studied in herbaceous species, mostly considering only aboveground tissues. Coniferous tree species dominate boreal forests, which are predicted to be exposed to more frequent and acute drought as a result of ongoing climate change. The associated impact at all stages of the forest tree life cycle is expected to have large-scale ecological and economic impacts. However, the molecular response to drought has not been comprehensively profiled for coniferous species. We assayed the physiological and transcriptional response of Picea abies (L.) H. Karst seedling needles and roots after exposure to mild and severe drought. Shoots and needles showed an extensive reversible plasticity for physiological measures indicative of drought-response mechanisms, including changes in stomatal conductance (gs), shoot water potential and abscisic acid (ABA). In both tissues, the most commonly observed expression profiles in response to drought were highly correlated with the ABA levels. Still, root and needle transcriptional responses contrasted, with extensive root-specific down-regulation of growth. Comparison between previously characterized Arabidopsis thaliana L. drought-response genes and P. abies revealed both conservation and divergence of transcriptional response to drought. In P. abies, transcription factors belonging to the ABA responsive element(ABRE) binding/ABRE binding factors ABA-dependent pathway had a more limited role. These results highlight the importance of profiling both above- and belowground tissues, and provide a comprehensive framework to advance the understanding of the drought response of P. abies. The results demonstrate that a short-term, severe drought induces severe physiological responses coupled to extensive transcriptome modulation and highlight the susceptibility of Norway spruce seedlings to such drought events.

We aimed to determine whether humic acid (HA) can alleviate the injury of millet caused by drought and its potential mechanism. Millet seeds (Jingu 21 and Zhangza 10) were soaked in different concentrations of HA (0, 50, 10, 200, and 300 mg L(−1)) for 12 h. The physiological and photosynthetic characteristics of millet seedlings, including growth parameters, osmotic regulators, antioxidase activity, photosynthesis, chlorophyll fluorescence, and P700 parameters, were determined before and after drought stress. HA significantly promoted the growth of millet seedlings under drought stress. Pretreatment with 100 mg L(−1) or 200 mg L(−1) HA significantly increased free proline, soluble protein, and activity of the antioxidant enzyme system (superoxide dismutase, peroxidase, and catalase) in both Zhangza 10 and Jingu 21. The accumulation of reactive oxygen species ([Image: see text] and H(2)O(2)) was reduced in HA treatments compared with that of the control (P

High-throughput and accurate measurements of plant traits facilitate identification of gene function. Along with recent advances in quantitative genomics, there is a growing need for precise quantification of multiple traits in plants. However, it is difficult continuously to quantify plant adaptive responses to environmental stress responses such as drought because multiple environmental factors are intricately involved in the phenotype. To solve this problem, we developed an automatic phenotyping system for evaluating the growth responses of individual Arabidopsis plants to a wide range of environmental conditions. The RIKEN Integrated Plant Phenotyping System (RIPPS) controls soil moisture for single plants by automatically weighing and watering 120 continuously rotating pots under controlled light, humidity and temperature growth conditions. RIPPS also records individual rosette size and expansion rate by photographing plants every 2 h. We used RIPPS to establish phenotype evaluation methods for Arabidopsis growth response and water use efficiency under various water conditions, and analyzed the involvement of ABA metabolism in determining water use efficiency. We also used RIPPS to analyze salinity tolerance in Arabidopsis plants.

This study synchronously investigated maize growth and physiological responses to progressive soil drying. It indicates that hydraulic and chemical changes may regulate plant development and functioning during onset of drought., A better understanding of physiological responses of crops to drought stress is important for ensuring sustained crop productivity under climate change. Here, we studied the effect on 15-day-old maize (Zea mays L.) plants of a 6 d non-lethal period of soil drying [soil water potential (SWP) decreased from –0.20 MPa to –0.81 MPa]. Root growth was initially stimulated during drying (when SWP decreased from –0.31 MPa to –0.38 MPa, compared with –0.29 MPa in well-watered pots), followed by inhibition during Days 5–6 (SWP from –0.63 MPa to –0.81 MPa). Abscisic acid (ABA) in the root began to accumulate as the root water potential declined during Days 2–3. Leaf elongation was inhibited from Day 4 (SWP less than –0.51 MPa), just after leaf ABA content began to increase, but coinciding with a decline in leaf water potential. The stomatal conductance was restricted earlier in the younger leaf (fourth) (on Day 3) than in the older leaf (third). The ethylene content of leaves and roots decreased during drying, but after the respective increase in ABA contents. This work identified critical timing of hydraulic and chemical changes at the onset of soil drying, which can be important in initiating early stomatal and growth responses to drought.

ZAT18 modulated genes involved in hormone signaling transduction and stress response pathways, resulting in enhanced osmotic stress responses in seed germination and improved drought stress tolerance., Environmental stress poses a global threat to plant growth and reproduction, especially drought stress. Zinc finger proteins comprise a family of transcription factors that play essential roles in response to various abiotic stresses. Here, we found that ZAT18 (At3g53600), a nuclear C2H2 zinc finger protein, was transcriptionally induced by dehydration stress. Overexpression (OE) of ZAT18 in Arabidopsis improved drought tolerance while mutation of ZAT18 resulted in decreased plant tolerance to drought stress. ZAT18 was preferentially expressed in stems, siliques, and vegetative rosette leaves. Subcellular location results revealed that ZAT18 protein was predominantly localized in the nucleus. ZAT18 OE plants exhibited less leaf water loss, lower content of reactive oxygen species (ROS), higher leaf water content, and higher antioxidant enzyme activities after drought treatment when compared with the wild type (WT). RNA sequencing analysis showed that 423 and 561 genes were transcriptionally modulated by the ZAT18 transgene before and after drought treatment, respectively. Pathway enrichment analysis indicated that hormone metabolism, stress, and signaling were over-represented in ZAT18 OE lines. Several stress-responsive genes including COR47, ERD7, LEA6, and RAS1, and hormone signaling transduction-related genes including JAZ7 and PYL5 were identified as putative target genes of ZAT18. Taken together, ZAT18 functions as a positive regulator and plays a crucial role in the plant response to drought stress.

Highlight Osmotic stress enhances the rate of protein glycation and monosaccharide autoxidation is the main pathway., Among the environmental alterations accompanying oncoming climate changes, drought is the most important factor influencing crop plant productivity. In plants, water deficit ultimately results in the development of oxidative stress and accumulation of osmolytes (e.g. amino acids and carbohydrates) in all tissues. Up-regulation of sugar biosynthesis in parallel to the increasing overproduction of reactive oxygen species (ROS) might enhance protein glycation, i.e. interaction of carbonyl compounds, reducing sugars and α-dicarbonyls with lysyl and arginyl side-chains yielding early (Amadori and Heyns compounds) and advanced glycation end-products (AGEs). Although the constitutive plant protein glycation patterns were characterized recently, the effects of environmental stress on AGE formation are unknown so far. To fill this gap, we present here a comprehensive in-depth study of the changes in Arabidopsis thaliana advanced glycated proteome related to osmotic stress. A 3 d application of osmotic stress revealed 31 stress-specifically and 12 differentially AGE-modified proteins, representing altogether 56 advanced glycation sites. Based on proteomic and metabolomic results, in combination with biochemical, enzymatic and gene expression analysis, we propose monosaccharide autoxidation as the main stress-related glycation mechanism, and glyoxal as the major glycation agent in plants subjected to drought.

Highlight We reveal that ABA affects flowering through two independent regulatory mechanisms: the activation of GI and CO functions upstream of the florigen genes and the down-regulation of SOC1 signalling., One strategy deployed by plants to endure water scarcity is to accelerate the transition to flowering adaptively via the drought escape (DE) response. In Arabidopsis thaliana, activation of the DE response requires the photoperiodic response gene GIGANTEA (GI) and the florigen genes FLOWERING LOCUS T (FT) and TWIN SISTER OF FT (TSF). The phytohormone abscisic acid (ABA) is also required for the DE response, by promoting the transcriptional up-regulation of the florigen genes. The mode of interaction between ABA and the photoperiodic genes remains obscure. In this work we use a genetic approach to demonstrate that ABA modulates GI signalling and consequently its ability to activate the florigen genes. We also reveal that the ABA-dependent activation of FT, but not TSF, requires CONSTANS (CO) and that impairing ABA signalling dramatically reduces the expression of florigen genes with little effect on the CO transcript profile. ABA signalling thus has an impact on the core genes of photoperiodic signalling GI and CO by modulating their downstream function and/or activities rather than their transcript accumulation. In addition, we show that as well as promoting flowering, ABA simultaneously represses flowering, independent of the florigen genes. Genetic analysis indicates that the target of the repressive function of ABA is the flowering-promoting gene SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), a transcription factor integrating floral cues in the shoot meristem. Our study suggests that variations in ABA signalling provide different developmental information that allows plants to co-ordinate the onset of the reproductive phase according to the available water resources.

Plant hormones play an important role in plant adaptation to abiotic stress, but hormonal responses of cool-season turfgrass species to drought stress are not well documented. This study was to investigate responses of hormones and photosynthesis to drought stress and examine if drought stress – induced hormone alteration is associated with stress tolerance in kentucky bluegrass (KBG, Poa pratensis L.). The grass was grown in a growth chamber for 6 weeks and then subjected to drought stress [40%-50% evapotranspiration (ET) replacement)] for 28 d. Drought stress caused cell membrane damage, resulting in decline in photosynthetic rate (Pn), chlorophyll content, and visual quality in KBG. Drought stressed grass had higher leaf abscisic acid (ABA), lower leaf trans-zeatin riboside (ZR), isopentenyl adenosine (iPA), and indole-3-acetic acid (IAA), but similar level of leaf gibberellin A4 (GA4) when compared to the control (well-watered). On average, drought stress treatment reduced leaf ZR by 59.1%, iPA by 50.4%, IAA by 26.7%, while increased ABA by 108.5% when compared to the control at the end of drought stress (28 d). The turf quality and photosynthetic rate was positively correlated with cytokinins and IAA, but negatively correlated with ABA and ABA/cytokinins (CK) ratio under drought stress. The results of this study suggest drought stress-induced injury to Kentucky bluegrass may be associated with hormonal alteration, and the plants with higher cytokinins and IAA and less ABA under drought stress may have better photosynthetic function and performance.

Overexpression of soybean GmDREB1 in wheat confers drought tolerance in the field and is associated with increased melatonin content, an increased root system, and delayed leaf senescence., Drought-response-element binding (DREB)-like transcription factors can significantly enhance plant tolerance to water stress. However, most research on DREB-like proteins to date has been conducted in growth chambers or greenhouses, so there is very little evidence available to support their practical use in the field. In this study, we overexpressed GmDREB1 from soybean in two popular wheat varieties and conducted drought-tolerance experiments across a range of years, sites, and drought-stress regimes. We found that the transgenic plants consistently exhibited significant improvements in yield performance and a variety of physiological traits compared with wild-type plants when grown under limited water conditions in the field, for example showing grain yield increases between 4.79–18.43%. Specifically, we found that the transgenic plants had reduced membrane damage and enhanced osmotic adjustment and photosynthetic efficiency compared to the non-transgenic controls. Three enzymes from the biosynthetic pathway of the phytohormone melatonin were up-regulated in the transgenic plants, and external application of melatonin was found to improve drought tolerance. Together, our results demonstrate the utility of transgenic overexpression of GmDREB1 to improve the drought tolerance of wheat in the field.

Glasshouse-based metabolic markers predict yield gap-based drought tolerance determined from multiseason and site field trials which could serve as a potential breeding tool for selecting drought-resilient crops., Water limits crop productivity, so selecting for a minimal yield gap in drier environments is critical to mitigate against climate change and land-use pressure. We investigated the responses of relative water content (RWC), stomatal conductance, chlorophyll content, and metabolites in flag leaves of commercial wheat (Triticum aestivum L.) cultivars to three drought treatments in the glasshouse and in field environments. We observed strong genetic associations between glasshouse-based RWC, metabolites, and yield gap-based drought tolerance (YDT; the ratio of yield in water-limited versus well-watered conditions) across 18 field environments spanning sites and seasons. Critically, RWC response to glasshouse drought was strongly associated with both YDT (r2=0.85, P

In plant cells, calcium-dependent protein kinases (CDPKs) are important sensors of Ca2+ flux resulting from various environmental stresses like cold, drought or salt stress. Previous genome sequence analysis and comparative studies in Arabidopsis (Arabidopsis thaliana L.) and rice (Oryza sativa L.) defined a multi-gene family of CDPKs. Here, we identified and characterised the CDPK gene complement of the model plant, barley (Hordeum vulgare L.). Comparative analysis encompassed phylogeny reconstruction based on newly available barley genome sequence, as well as established model genomes (e.g. O. sativa, A. thaliana, Brachypodium distachyon). Functional gene copies possessed characteristic CDPK domain architecture, including a serine/threonine kinase domain and four regulatory EF-hand motifs. In silico verification was followed by measurements of transcript abundance via real-time polymerase chain reaction (PCR). The relative expression of CDPK genes was determined in the vegetative growth stage under intensifying drought stress conditions. The majority of barley CDPK genes showed distinct changes in patterns of expression during exposure to stress. Our study constitutes evidence for involvement of the barley CDPK gene complement in signal transduction pathways relating to adaptation to drought. Our bioinformatics and transcriptomic analyses will provide an important foundation for further functional dissection of the barley CDPK gene family. Electronic supplementary material The online version of this article (doi:10.1007/s13353-016-0357-2) contains supplementary material, which is available to authorized users.

Highlight VaNAC26, a member of the NAC gene family from the wild species Vitis amurensis, plays an important role in drought tolerance by positively modulating jasmonic acid synthesis and enhancing the antioxidant system., The growth and fruit quality of grapevines are widely affected by abnormal climatic conditions such as water deficits, but many of the precise mechanisms by which grapevines respond to drought stress are still largely unknown. Here, we report that VaNAC26, a member of the NAC transcription factor family, was upregulated dramatically during cold, drought and salinity treatments in Vitis amurensis, a cold and drought-hardy wild Vitis species. Heterologous overexpression of VaNAC26 enhanced drought and salt tolerance in transgenic Arabidopsis. Higher activities of antioxidant enzymes and lower concentrations of H2O2 and O2 − were found in VaNAC26-OE lines than in wild type plants under drought stress. These results indicated that scavenging by reactive oxygen species (ROS) was enhanced by VaNAC26 in transgenic lines. Microarray-based transcriptome analysis revealed that genes related to jasmonic acid (JA) synthesis and signaling were upregulated in VaNAC26-OE lines under both normal and drought conditions. VaNAC26 showed a specific binding ability on the NAC recognition sequence (NACRS) motif, which broadly exists in the promoter regions of upregulated genes in transgenic lines. Endogenous JA content significantly increased in the VaNAC26-OE lines 2 and 3. Our data suggest that VaNAC26 responds to abiotic stresses and may enhance drought tolerance by transcriptional regulation of JA synthesis in Arabidopsis.

Highlight The relationship between an adenine isopentenyltransferase transgene, reactive oxygen species (ROS) content and the ROS-scavenging system, mechanistically contributing to improved root growth under drought stress in creeping bentgrass, is discussed., Drought stress limits root growth and inhibits cytokinin (CK) production. Increases in CK production through overexpression of isopentenyltransferase (ipt) alleviate drought damages to promote root growth. The objective of this study was to investigate whether CK-regulated root growth was involved in the alteration of reactive oxygen species (ROS) production and ROS scavenging capacity under drought stress. Wild-type (WT) creeping bentgrass (Agrostis stolonifera L. ‘Penncross’) and a transgenic line (S41) overexpressing ipt ligated to a senescence-activated promoter (SAG12) were exposed to drought stress for 21 d in growth chambers. SAG12-ipt transgenic S41 developed a more extensive root system under drought stress compared to the WT. Root physiological analysis (electrolyte leakage and lipid peroxidation) showed that S41 roots exhibited less cellular damage compared to the WT under drought stress. Roots of SAG12-ipt transgenic S41 had significantly higher endogenous CK content than the WT roots under drought stress. ROS (hydrogen peroxide and superoxide) content was significantly lower and content of total and free ascorbate was significantly higher in S41 roots compared to the WT roots under drought stress. Enzymatic assays and transcript abundance analysis showed that superoxide dismutase, catalase, peroxidase, and dehydroascorbate reductase were significantly higher in S41 roots compared to the WT roots under drought stress. S41 roots also maintained significantly higher alternative respiration rates compared to the WT under drought stress. The improved root growth of transgenic creeping bentgrass may be facilitated by CK-enhanced ROS scavenging through antioxidant accumulation and activation of antioxidant enzymes, as well as higher alternative respiration rates when soil water is limited.

Highlight MLP43 interacts with SnRK2.6 and ABF1 and functions as a positive regulator in ABA and drought stress responses through modulating gene expression, ROS production and primary metabolite profiles., Drought stress is one of the disadvantageous environmental conditions for plant growth and reproduction. Given the importance of abscisic acid (ABA) to plant growth and abiotic stress responses, identification of novel components involved in ABA signalling transduction is critical. In this study, we screened numerous Arabidopsis thaliana mutants by seed germination assay and identified a mutant mlp43 (major latex protein-like 43) with decreased ABA sensitivity in seed germination. The mlp43 mutant was sensitive to drought stress while the MLP43-overexpressed transgenic plants were drought tolerant. The tissue-specific expression pattern analysis showed that MLP43 was predominantly expressed in cotyledons, primary roots and apical meristems, and a subcellular localization study indicated that MLP43 was localized in the nucleus and cytoplasm. Physiological and biochemical analyses indicated that MLP43 functioned as a positive regulator in ABA- and drought-stress responses in Arabidopsis through regulating water loss efficiency, electrolyte leakage, ROS levels, and as well as ABA-responsive gene expression. Moreover, metabolite profiling analysis indicated that MLP43 could modulate the production of primary metabolites under drought stress conditions. Reconstitution of ABA signalling components in Arabidopsis protoplasts indicated that MLP43 was involved in ABA signalling transduction and acted upstream of SnRK2s by directly interacting with SnRK2.6 and ABF1 in a yeast two-hybrid assay. Moreover, ABA and drought stress down-regulated MLP43 expression as a negative feedback loop regulation to the performance of MLP43 in ABA and drought stress responses. Therefore, this study provided new insights for interpretation of physiological and molecular mechanisms of Arabidopsis MLP43 mediating ABA signalling transduction and drought stress responses.