Enhanced Drought Stress Research Articles

Raffinose synthase enhances drought tolerance through raffinose synthesis or galactinol hydrolysis in maize and Arabidopsis plants

Raffinose and its precursor galactinol accumulate in plant leaves during abiotic stress. RAFFINOSE SYNTHASE (RAFS) catalyzes raffinose formation by transferring a galactosyl group of galactinol to sucrose. However, whether RAFS contributes to plant drought tolerance and, if so, by what mechanism remains unclear. In this study, we report that expression of RAFS from maize (or corn, Zea mays) (ZmRAFS) is induced by drought, heat, cold, and salinity stresses. We found that zmrafs mutant maize plants completely lack raffinose and hyper-accumulate galactinol and are more sensitive to drought stress than the corresponding null-segregant (NS) plants. This indicated that ZmRAFS and its product raffinose contribute to plant drought tolerance. ZmRAFS overexpression in Arabidopsis enhanced drought stress tolerance by increasing myo-inositol levels via ZmRAFS-mediated galactinol hydrolysis in the leaves due to sucrose insufficiency in leaf cells and also enhanced raffinose synthesis in the seeds. Supplementation of sucrose to detached leaves converted ZmRAFS from hydrolyzing galactinol to synthesizing raffinose. Taken together, we demonstrate that ZmRAFS enhances plant drought tolerance through either raffinose synthesis or galactinol hydrolysis, depending on sucrose availability in plant cells. These results provide new avenues to improve plant drought stress tolerance through manipulation of the raffinose anabolic pathway.

Coordinated regulation of carbon and nitrogen assimilation confers drought tolerance in maize (Zea mays L.)

Drought is a major limiting factor affecting crop productivity. To gain a comprehensive understanding of transcriptomic reprogramming under drought stress in maize (Zea mays L.), we performed physiological and transcriptomic analysis of leaf tissues of two maize cultivars, LY66 and LY99, subjected to a progressively severe drought and subsequent recovery. LY66 exhibited greater drought tolerance and recovery than LY99. RNA-Seq analysis showed that differentially expressed genes mainly enriched in pathways involved in carbon (C) and nitrogen (N) metabolism, and plant hormone signal transduction. As judged from RNA-Seq data, enzyme activity, and metabolite contents, LY66 maintained a higher relatively stable C and N metabolism than LY99, which were related to higher photosynthesis and sucrose biosynthesis capacity, higher N assimilation and protein biosynthesis capacity under drought stress, respectively. Interestingly, the expression of several genes encoding key enzymes involved in auxin biosynthesis and signaling were activated to higher levels in LY66 than in LY99 under drought stress. Our results indicate that LY66 exhibits coordinated regulation of C and N assimilation, possibly involved in phytohormone signaling pathways, which enhances drought stress tolerance.

CAM plant expansion favored indirectly by asymmetric climate warming and increased rainfall variability.

Recent observational evidence suggests that nighttime temperatures are increasing faster than daytime temperatures, while in some regions precipitation events are becoming less frequent and more intense. The combined ecological impacts of these climatic changes on crassulacean acid metabolism (CAM) plants and their interactions with other functional groups (i.e., grass communities) remain poorly understood. Here we developed a growth chamber experiment to investigate how two CAM-grass communities in desert ecosystems of the southwestern United States and northern Mexico respond to asymmetric warming and increasing rainfall variability. Grasses generally showed competitive advantages over CAM plants with increasing rainfall variability under ambient temperature conditions. In contrast, asymmetric warming caused mortality of both grass species (Bouteloua eriopoda and Bouteloua curtipendula) in both rainfall treatments due to enhanced drought stress. Grass mortality indirectly favored CAM plants even though the biomass of both CAM species Cylindropuntia imbricata and Opuntia phaeacantha significantly decreased. The stem's volume-to-surface ratio of C. imbricata was significantly higher in mixture than in monoculture under ambient temperature (both P < 0.0014); however, the difference became insignificant under asymmetric warming (both P > 0.1625), suggesting that warming weakens the negative effects of interspecific competition on CAM plant growth. Our findings suggest that while the increase in intra-annual rainfall variability enhances grass productivity, asymmetric warming may lead to grass mortality, thereby indirectly favoring the expansion of co-existing CAM plants. This study provides novel experimental evidence showing how the ongoing changes in global warming and rainfall variability affect CAM-grass growth and interactions in dryland ecosystems.

High Nitrogen Enhance Drought Tolerance in Cotton through Antioxidant Enzymatic Activities, Nitrogen Metabolism and Osmotic Adjustment.

Drought is one of the most important abiotic stresses and hampers many plant physiological processes under suboptimal nitrogen (N) concentration. Seedling tolerance to drought stress is very important for optimum growth and development, however, the enhancement of plant stress tolerance through N application in cotton is not fully understood. Therefore, this study investigates the role of high N concentration in enhancing drought stress tolerance in cotton. A hydroponic experiment supplying low (0.25 mM) and high (5 mM) N concentrations, followed by 150 g L−1 polyethylene glycol (PEG)-induced stress was conducted in a growth chamber. PEG-induced drought stress inhibited seedling growth, led to oxidative stress from excessive malondialdehyde (MDA) generation, and reduced N metabolism. High N concentrations alleviated oxidative damage and stomatal limitation by increasing antioxidant enzymatic activities, leaf relative water content, and photosynthesis in cotton seedlings under drought stress. The results revealed that the ameliorative effects of high N concentration may be ascribed to the enhancement of N metabolizing enzymes and an increase in the amounts of osmoprotectants like free amino acids and total soluble protein. The present data suggest that relatively high N concentrations may contribute to drought stress tolerance in cotton through N metabolism, antioxidant capacity, and osmotic adjustment.

Study the effect of 24-epibrassinolide application on the Cu/Zn-SOD expression and tolerance to drought stress in common bean.

To study the possibility of increasing the drought tolerance of common bean with the exogenous application of 24-epibrassinolide (EBL), an experiment was conducted in 2016 and 2017. In this experiment, two irrigation levels (optimal irrigation and drought stress) were applied to the main plots and two common bean genotypes (Kusha cultivar and COS16 genotype) and four EBL concentrations (0, 2, 4, and 6μM) were allocated to sub-plots as factorial. In the flowering stage, drought stress was applied and plants were sprayed with EBL. The results showed that drought stress reduced relative water content (RWC) and increased proline content, malondialdehyde (MDA) content, and antioxidant enzymes activity. However, exogenous application of EBL reduced the seed yield loss and increased the drought stress tolerance in both common bean genotypes by decreasing the MDA content and increasing the RWC, proline content, antioxidant enzymes activity, and nitrate reductase activity. It can beconcluded that foliar spray of 4µM EBL as the best concentration may increase the seed yield and enhance the drought stress tolerance of common bean. Also, Cu/Zn-SOD was up-regulated in response to the drought stress and exogenous EBL. The COS16 genotype showed better response to the drought stress and exogenous EBL than the Kusha cultivar, because of the higher up-regulation of Cu/Zn-SOD in this genotype compared to the Kusha cultivar. Therefore, EBL can be used as a plant growth regulator to enhance drought stress tolerance and minimize the seed yield loss of common bean caused by water deficit.

Vitis vinifera bZIP14 functions as a transcriptional activator and enhances drought stress resistance via suppression of reactive oxygen species

Background: Drought stress affects grapevine growth and development and reduces berry yield and quality. Identifying genes that are involved in the plant response to drought stress will enable the development of new grape strains that are tolerant to drought. Objective: We cloned the VvibZIP14 gene from Vitis vinifera and analyzed its role in drought resistance. Methods: Gene expression was analyzed by quantitative real-time PCR. Subcellular localization was assessed with a transient expression assay. The transactivation activity of the protein was evaluated in yeast. The physiologic role of VvibZIP14 was analyzed by overexpressing VvibZIP14 in Arabidopsis following drought stress. Hydrogen peroxide accumulation in Arabidopsis was visualized by diaminobenzidine staining. Results: Drought stress caused the accumulation of VvibZIP14, which was localized in the nucleus and had transcriptional activity. Transgenic plants showed improved resistance to drought stress and reduced electrolyte leakage compared to plants overexpressing empty vector, whereas chlorophyll content, photosystem II maximal photochemical efficiency, and net photosynthetic rate were higher. Catalase, peroxidase, and superoxide dismutase activities were also increased in VvibZIP14-overexpressing plants subjected to drought stress. Conclusions: VvibZIP14 functions as a transcription factor that confers resistance to drought stress in grape by enhancing the antioxidant response.

Overexpression of soybean DREB1 enhances drought stress tolerance of transgenic wheat in the field.

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.

Rhizobacteria-induced systemic tolerance against drought stress in Sorghum bicolor (L.) Moench

Induction of systemic tolerance in sorghum [Sorghum bicolor (L.) Moench] against drought stress was studied by screening a large collection of rhizobacterial isolates for their potential to exhibit this essential plant growth-promoting trait. This was done by means of a greenhouse assay that measured the relative change in both plant height and -biomass (roots and shoots) between rhizobacteria-primed versus non-primed (naïve) plants under drought stress conditions. In order to elucidate the metabolomic changes in S. bicolor that conferred the drought stress tolerance after treatment (priming) with selected isolates, untargeted ultra-high performance liquid chromatography-high definition mass spectrometry (UHPLC-HDMS)-based metabolomics was carried out. Intracellular metabolites were methanol-extracted from rhizobacteria-primed and naïve S. bicolor roots and shoots. Extracts were analysed on a UHPLC-HDMS system and the generated data were chemometrically mined to determine signatory metabolic profiles and bio-markers related to induced systemic tolerance. The metabolomic results showed significant treatment-related differential metabolic reprogramming between rhizobacteria-primed and naïve plants, correlating to the ability of the selected isolates to protect S. bicolor against drought stress. The selected isolates, identified by means of 16S rRNA gene sequencing as members of the genera Bacillus and Pseudomonas, were screened for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity by means of an in vitro assay and the presence of the acdS gene was subsequently confirmed by PCR for strain N66 (Pseudomonas sp.). The underlying key metabolic changes in the enhanced drought stress tolerance observed in rhizobacteria-primed S. bicolor plants included (1) augmented antioxidant capacity; (2) growth promotion and root architecture modification as a result of the upregulation of the hormones gibberellic acid, indole acetic acid and cytokinin; (3) the early activation of induce systemic tolerance through the signalling hormones brassinolides, salicylic acid and jasmonic acid and signalling molecules sphingosine and psychosine; (4) the production of the osmolytes proline, glutamic acid and choline; (5) the production of the epicuticular wax docosanoic acid and (6) ACC deaminase activity resulting in lowered ethylene levels. These results unravelled key molecular details underlying the PGPR-induced systemic tolerance in sorghum plants, providing insights for the plant priming for abiotic stress.

Induction of drought tolerance by inoculation of Bacillus aryabhattai on sugarcane seedlings

In twenty-first century, free-living endophytic and rhizosphere-competent microbes have become breakthrough strategies to meet global demands for sustainable foods and renewable fuelstocks owing to their great potential to produce stress-tolerant food and energy crops. Here, we investigate how Bacillus aryabhattai could mitigate water stress by drought in sugarcane seedlings. Briefly, the sugarcane genotypes, namely IAC91-1099 and RB85-5156, both exposed to cells-centrifuged B. aryabhattai suspension at 1x10 8 CFU mL -1 and non-treated were developed under irrigation regimes of 0, 7, 14 and 21 days to simulate different degrees of soil moisture content. The osmotolerant bacterium remarkably enhanced development of aboveground structures and root system, mainly in irrigated seedlings of IAC91-1099. Clearly noted that microbial metabolism depends on genotype and soil water potential to promote plant growth. This bacterium probably enabled sugarcane plants in early phenological stages to cope with water deficit by regulation of plant growth hormones along with solubilization of nutrients. Of particular importance, the bacterium exerted more pronounced effects by conferring drought tolerance at 7 and 14 days irrigation interval, irrespective of sugarcane genotype, which also translated into increased biomass of shoots and roots. Microbial degradation of ethylene precursors exuded in rhizosphere could reasonably explain why B. aryabhattai -associated seedlings developed deeper root systems to uptake water and nutrients and, conse­quently, allocating larger amounts of dry matter to shoots. Collectively, our findings provide relevant insights on the beneficial effects of the cacti-associated B. aryabhattai in alleviating the harmful effects of drought stress in seedlings and contribute to increasing our understanding of the phenotypic outcomes from the interaction be­tween sugarcane genotypes and a beneficial rhizobacterium. Hence, this bacterial inoculant forms a low-cost and ecologically sound strategy to enhancing drought stress tolerance in sugarcane crops planted in water-limited zones in Brazil.

Stem cycle analyses help decipher the nonlinear response of trees to concurrent warming and drought

High-resolution analysis of stem radius variation can quantify the impact of warming and drought on stem water balance and stem growth in black spruce [ Picea mariana (Mill.) B.S.P.)]. Drought affected plant water status and stem growth. However, warming affects the components of the circadian stem cycle differently if the impacts occur in the daytime or nighttime. The interactive effect of abiotic stresses had less impact on the circadian stem cycle than when the stresses occurred independently. Warming and recent droughts in boreal regions reflect the multiple dimensions of climate change. How these climate-related stresses will affect the stem growth of trees remains to be described. Plant water relations can detect the dynamics of stem depletion and replenishment under conditions of climate-forced stress. This study aimed to verify the impacts of a combination of asynchronous warming (nighttime versus daytime warming) and drought on stem water balance and stem growth in black spruce [Picea mariana (Mill.) B.S.P.)]. We investigated the water status and variations in stem radius of black spruce saplings growing in a controlled environment from May through August. We grew four-year-old saplings in warmer conditions either during the day (DW) or night (NW) at temperatures ca. 6 °C warmer than the ambient air temperature (CT). We then simulated a one-month drought in June. Automatic point dendrometers provided a high-resolution analysis of variations in stem radius, and we also monitored leaf water potentials and volumetric soil water content during the entire experimental period. We detected significant reductions in stem radius variation under water deficit conditions. In the daytime warming scenario, we observed a significant increase in the duration of contraction and a decrease in expansion of the stems. The amplitude of this contraction and expansion was reduced under the nighttime warming conditions. The main effect of warming was to enhance drought stress by accelerating soil water depletion. Changes in predawn water potential drove the duration of stem circadian cycles under conditions of daytime warming, whereas irreversible growth dynamics drove these cycles under nighttime warming conditions due to the midday water potential. The interaction of night/daytime asynchronous warming and drought reduced the amplitude rather than the duration of stem contraction and expansion. Water deficit decreased stem growth during the growing season. Asymmetric warming (as a single independent treatment) affected the timing and magnitude of stem circadian cycles. Under daytime warming scenarios, the duration of contraction and expansion were regulated mainly by predawn water potential, inducing longer (shorter) durations of contraction (expansion). Under nighttime warming, the smaller amplitudes of stem contraction and expansion were associated with midday water potential. Therefore, the interaction of abiotic stresses had less of an impact on the circadian stem cycle components than when these stresses were applied independently.

Calcium Application Enhances Drought Stress Tolerance in Sugar Beet and Promotes Plant Biomass and Beetroot Sucrose Concentration.

Numerous studies have demonstrated the potential of sugar beet to lose the final sugar yield under water limiting regime. Ample evidences have revealed the important role of mineral nutrition in increasing plant tolerance to abiotic stresses. Despite the vital role of calcium (Ca2+) in plant growth and development, as well as in stress responses as an intracellular messenger, its role in alleviating drought stress in sugar beet has been rarely addressed. Here, an attempt was undertaken to investigate whether, and to what extent, foliar application of Ca2+ confers drought stress tolerance in sugar beet plants exposed to drought stress. To achieve this goal, sugar beet plants, which were grown in a high throughput phenotyping platform, were sprayed with Ca2+ and submitted to drought stress. The results showed that foliar application of Ca2+ increased the level of magnesium and silicon in the leaves, promoted plant growth, height, and leaf coverage area as well as chlorophyll level. Ca2+, in turn, increased the carbohydrate levels in leaves under drought condition and regulated transcriptionally the genes involved in sucrose transport (BvSUC3 and BvTST3). Subsequently, Ca2+ enhanced the root biomass and simultaneously led to induction of root (BvSUC3 and BvTST1) sucrose transporters which eventually supported the loading of more sucrose into beetroot under drought stress. Metabolite analysis revealed that the beneficial effect of Ca2+ in tolerance to drought induced-oxidative stress is most likely mediated by higher glutathione pools, increased levels of free polyamine putrescine (Put), and lower levels of amino acid gamma-aminobutyric acid (GABA). Taken together, this work demonstrates that foliar application of Ca2+ is a promising fertilization strategy to improve mineral nutrition efficiency, sugar metabolism, redox state, and thus, drought stress tolerance.

Overexpression of NtabDOG1L promotes plant growth and enhances drought tolerance in Nicotiana tabacum

Drought is one of the major environmental stresses limiting crop growth and production. It is very important to exploit and utilize drought-tolerance genes to improve crop drought-resistance. In this study, we identified two homoeologs of a Nicotiana tabacum (Ntab) DELAY OF GERMINATION (DOG) 1 like gene, named as NtabDOG1L-T and NtabDOG1L-S, respectively. The NtabDOG1L genes were preferentially expressed in roots and their expression levels were induced by polyethylene glycol, high salt, cold, and abscisic acid treatments. Subcellular localization results indicated that NtabDOG1L-T was localized in the nucleus, cytoplasm and cell membrane. Overexpression of NtabDOG1L-T in tobacco resulted in roots growth enhancement in transgenic plants. Furthermore, overexpression of NtabDOG1L-T enhanced drought stress tolerance in transgenic tobacco. The transgenic tobacco lines exhibited lower leaf water loss and electrolyte leakage, lower content of malondialdehyde and reactive oxygen species (ROS), and higher antioxidant enzymes activities after drought treatment when compared with wild type (WT) plants. In addition, the expression levels of several genes encoding key antioxidant enzymes and drought-related proteins were higher in the transgenic plants than in the WT plants under drought stress. Taken together, our results showed that NtabDOG1L functions as a novel regulator that improves plant growth and drought tolerance in tobacco.

The rice SUMO conjugating enzymes OsSCE1 and OsSCE3 have opposing effects on drought stress

Posttranslational modification of proteins by the small ubiquitin-related modifier (SUMO) protein is involved in diverse cellular processes. In sumoylation, SUMO-conjugating enzyme (SCE) conjugates SUMO to substrate proteins. Similarly to yeast and animals, Arabidopsis encodes a single SCE gene, but other plants encode at least two SCE genes. In this study, we report the molecular characterization of three Oryza sativa SCE genes. Their levels of expression are commonly upregulated by drought stress but are differentially regulated by hormones and sugars. Only the OsSCE1 gene showed photoperiod- and light-dependent diurnal oscillations in the leaves. Yeast two-hybrid assays showed that OsSCEs do not show SUMO isoform specificity. Three rice OsSCE proteins localize primarily to the nucleus. Interestingly, OsSCE1 is distributed in specific parts of the nucleus and shows sumoylation activities in the absence of a SUMO ligase in E. coli. In addition, overexpression of the OsSCE1 gene alters the biomass and grain yield parameters in transgenic rice plants. Overexpression of the OsSCE3 gene in transgenic rice plants enhances drought stress tolerance. In contrast, OsSCE1-OX transgenic rice plants are hypersensitive to drought stress. Our results suggest that these genes may be involved in different cellular processes.

An apple (Malus domestica) NAC transcription factor enhances drought tolerance in transgenic apple plants

Plant NAC proteins constitute one of the largest transcription factor families. They play pivotal functions during responses to various abiotic stresses. However, knowledge on roles of NAC proteins in abiotic stress tolerance as well as corresponding mechanisms has not been fully studied in perennial woody plants, including domesticated apple (Malus domestica). In the present study, we characterized the role of apple MdNAC1 transcription factor in response to drought stress. Apple plants overexpressing MdNAC1 gene exhibited promoted tolerance to drought stress, as evident by reduced water loss and electrolyte leakage in leaves, and maintenance of photosynthesis and photosynthetic pigments content under drought conditions. In addition, the levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were significantly lower for transgenic apple lines than those for nontransgenic plants under drought conditions. This was accompanied by higher activities of several antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as increased expression of the associated genes in transgenic lines. Together, our results indicate that overexpression of the apple MdNAC1 gene enhances drought stress tolerance in apple plants by promoting higher photosynthesis and activities of ROS-scavenging enzymes.

Knockdown of Cytochrome P450 Genes Gh_D07G1197 and Gh_A13G2057 on Chromosomes D07 and A13 Reveals Their Putative Role in Enhancing Drought and Salt Stress Tolerance in Gossypium hirsutum.

We identified 672, 374, and 379 CYPs proteins encoded by the CYPs genes in Gossypium hirsutum, Gossypium raimondii, and Gossypium arboreum, respectively. The genes were found to be distributed in all 26 chromosomes of the tetraploid cotton, with chrA05, chrA12, and their homeolog chromosomes harboring the highest number of genes. The physiochemical properties of the proteins encoded by the CYP450 genes varied in terms of their protein lengths, molecular weight, isoelectric points (pI), and even grand hydropathy values (GRAVY). However, over 99% of the cotton proteins had GRAVY values below 0, which indicated that the majority of the proteins encoded by the CYP450 genes were hydrophilic in nature, a common property of proteins encoded by stress-responsive genes. Moreover, through the RNA interference (RNAi) technique, the expression levels of Gh_D07G1197 and Gh_A13G2057 were suppressed, and the silenced plants showed a higher concentration of hydrogen peroxide (H2O2) with a significant reduction in the concentration levels of glutathione (GSH), ascorbate peroxidase (APX), and proline compared to the wild types under drought and salt stress conditions. Furthermore, the stress-responsive genes 1-Pyrroline–5-Carboxylate Synthetase (GhP5CS), superoxide dismutase (GhSOD), and myeloblastosis (GhMYB) were downregulated in VIGS plants, but showed upregulation in the leaf tissues of the wild types under drought and salt stress conditions. In addition, CYP450-silenced cotton plants exhibited a high level of oxidative injury due to high levels of oxidant enzymes, in addition to negative effects on CMS, ELWL, RLWC, and chlorophyll content The results provide the basic foundation for future exploration of the proteins encoded by the CYP450 genes in order to understand the physiological and biochemical mechanisms in enhancing drought and salt stress tolerance in plants.

Cucumber Phospholipase D alpha gene overexpression in tobacco enhanced drought stress tolerance by regulating stomatal closure and lipid peroxidation

BackgroundPlant phospholipase D (PLD), which can hydrolyze membrane phospholipids to produce phosphatidic acid (PA), a secondary signaling molecule, has been proposed to function in diverse plant stress responses. Both PLD and PA play key roles in plant growth, development, and cellular processes. PLD was suggested to mediate the regulation of stomatal movements by abscisic acid (ABA) as a response to water deficit. In this research, we characterized the roles of the cucumber phospholipase D alpha gene (CsPLDα, GenBank accession number EF363796) in the growth and tolerance of transgenic tobacco (Nicotiana tabacum) to drought stress.ResultsThe CsPLDα overexpression in tobacco lines correlated with the ABA synthesis and metabolism, regulated the rapid stomatal closure in drought stress, and reduced the water loss. The NtNCED1 expression levels in the transgenic lines and wild type (WT) were sharply up-regulated after 16 days of drought stress compared with those before treatment, and the expression level in the transgenic lines was significantly higher than that in the WT. The NtAOG expression level evidently improved after 8 and 16 days compared with that at 0 day of treatment and was significantly lower in the transgenic lines than in the WT. The ABA content in the transgenic lines was significantly higher than that in the WT. The CsPLDα overexpression could increase the osmolyte content and reduce the ion leakage. The proline, soluble sugar, and soluble protein contents significantly increased. By contrast, the electrolytic leakage and malondialdehyde accumulation in leaves significantly decreased. The shoot and root fresh and dry weights of the overexpression lines significantly increased. These results indicated that a significant correlation between CsPLDα overexpression and improved resistance to water deficit.ConclusionsThe plants with overexpressed CsPLDα exhibited lower water loss, higher leaf relative water content, and heavier fresh and dry matter accumulation than the WT. We proposed that CsPLDα was involved in the ABA-dependent pathway in mediating the stomatal closure and preventing the elevation of intracellular solute potential.

Miscanthus NAC transcription factor MlNAC12 positively mediates abiotic stress tolerance in transgenic Arabidopsis

NAC (NAM, ATAF1/2 and CUC2) transcription factors play critical roles in plant abiotic stress responses. However, knowledge regarding the functional roles of NACs in abiotic stress tolerance and its underlying mechanisms is relatively limited in Miscanthus. In this study, we functionally characterized a novel Miscanthus NAC gene MlNAC12 by ectopic expression in Arabidopsis. MlNAC12 was localized in the nucleus. It could specifically binds to the NAC recognition sequence (NACRS) and has a transactivation activity in the C-terminus. Overexpression of MlNAC12 in Arabidopsis conferred hypersensitivity to exogenous Abscisic acid (ABA) at seed germination and root elongation stages. In addition, MlNAC12 overexpression enhanced germination and root growth under salt (NaCl) stress. Furthermore, MlNAC12 overexpression lines exhibited significantly enhanced drought stress tolerance, which was evidenced by a higher survival rate and a lower water loss rate compared to the wild type (WT). Accordingly, the stomata aperture was remarkably reduced in MlNAC12 overexpression lines in comparison to the WT under drought stress. Furthermore, the accumulation of the reactive oxygen species (ROS) and malondialdehyde (MDA) under abiotic stresses was significantly decreased, accompanied by dramatically enhanced activities of several antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in the transgenic plants. Correspondingly, the expression of six stress-responsive genes was significantly up-regulated in MlNAC12 overexpression lines. Together, our results indicate that MlNAC12 is a positive regulator of drought and salt stress tolerance through activating ROS scavenging enzymes.

Genome-wide identification and expression analysis of calmodulin-binding transcription activator genes in banana under drought stress

Banana (Musa acuminata) is an important economical food crop, cultivated worldwide depending on suitable climatic conditions. Its production is hindered because of several biotic and abiotic stresses. Drought stress is one of the main factors, which affect banana yield. To survive under water deficit conditions, plants respond by change in expression of many genes. Calmodulin-binding transcription activator (CAMTA) is a small transcription factor (TF) family with broad range of functions involved in different environmental stress responses including drought. Despite the importance of this family, no comprehensive data is available about this TF family in banana. In this study, five Musa acuminata CAMTA (MuCAMTA) TFs were identified in banana genome. One-month old banana plants cv. Basrai, were subjected to drought stress for twelve days by complete withholding of water. Plants kept for control were watered regularly. Photosynthetic pigments and total RNA was extracted from leaves of drought stressed and control plants. The expression pattern of MuCAMTA genes was identified by qRT-PCR. Drought resulted decrease in chlorophyll pigments. All MuCAMTA genes except MuCAMTA2 showed upregulation under drought stress. MuCAMTA1 showed highest expression (40 fold) under drought stress. MuCAMTA1 could be an ideal candidate to enhance drought stress tolerance in banana plants.

Transgenic creeping bentgrass overexpressing Osa-miR393a exhibits altered plant development and improved multiple stress tolerance.

SummaryMicroRNA393 (miR393) has been implicated in plant growth, development and multiple stress responses in annual species such as Arabidopsis and rice. However, the role of miR393 in perennial grasses remains unexplored. Creeping bentgrass (Agrostis stolonifera L.) is an environmentally and economically important C3 cool‐season perennial turfgrass. Understanding how miR393 functions in this representative turf species would allow the development of novel strategies in genetically engineering grass species for improved abiotic stress tolerance. We have generated and characterized transgenic creeping bentgrass plants overexpressing rice pri‐miR393a (Osa‐miR393a). We found that Osa‐miR393a transgenics had fewer, but longer tillers, enhanced drought stress tolerance associated with reduced stomata density and denser cuticles, improved salt stress tolerance associated with increased uptake of potassium and enhanced heat stress tolerance associated with induced expression of small heat‐shock protein in comparison with wild‐type controls. We also identified two targets of miR393, AsAFB2 and AsTIR1, whose expression is repressed in transgenics. Taken together, our results revealed the distinctive roles of miR393/target module in plant development and stress responses between creeping bentgrass and other annual species, suggesting that miR393 would be a promising candidate for generating superior crop cultivars with enhanced multiple stress tolerance, thus contributing to agricultural productivity.

Exogenous nitric oxide donor and arginine provide protection againstshort-term drought stress in wheat seedlings.

Nitric oxide (NO) is an important plant signaling molecule that has a vital role in abiotic stress tolerance. In the present study, we assesseddrought-induced (15 and 30% PEG, polyethylene glycol) damage in wheat (Triticum aestivum L. cv. Prodip) seedlings and mitigation by the synergistic effect of exogenous Arg (0.5mM l-Arginine) and an NO donor (0.5mM sodium nitroprusside, SNP). Drought stress sharply decreased the leaf relative water content (RWC) but markedly increased the proline (Pro) content in wheat seedlings. Drought stress caused overproduction of reactive oxygen species (ROS) and methylglyoxal (MG) due to the inefficiency of antioxidant enzymes, theglyoxalase system, and theascorbate-glutathione pool. However, supplementation with theNO donor and Arg enhanced the antioxidant defense system (both non-enzymatic and enzymatic components) in drought-stressed seedlings. Application of theNO donor and Arg also enhanced the glyoxalase system and reduced the MG content by increasing the activities of the glyoxalase system enzymes (Gly I and Gly II), which restored the leaf RWC and further increased the Pro content under drought stress conditions. Exogenous NO donor and Arg application enhanced the endogenous NO content, which positively regulated the antioxidant system and reduced ROS production. Thus, the present study reveals the crucial roles of Arg and NO in enhancing drought stress tolerance in wheat seedlings by upgrading their water status and reducing oxidative stress and MG toxicity.