Abscisic Acid Responsive Element Binding Protein Research Articles

Overexpression of PtrAREB3 improved cadmium enrichment and tolerance in poplar

Cadmium is a toxic heavy metal that has serious impacts on human and ecosystem health. Poplars play an active role in the phytoremediation of heavy metals in the environment; whereas only a few studies have investigated the molecular mechanisms by which trees absorb heavy metals from the environment. This study identified an abscisic acid-responsive element binding protein, PtrAREB3, in Populus trichocarpa. PtrAREB3 was principally expressed in leaves and markedly induced following the ABA and Cd treatments. Moreover, the poplar PtrAREB3 acts as direct regulator of target genes. Under Cd stress, PtrAREB3 further activates target genes associated with stress response and promotes cadmium tolerance of poplar by reducing the production of oxides, reducing chlorophyll degradation, increasing the synthesis of proline, and activating the antioxidant system. Furthermore, PtrAREB3 might indirectly regulate gene expression relating to Cd uptake, transport, and detoxification; ultimately, this increases Cd accumulation in plants. After 28 days of Cd treatment, the PtrAREB3 over-expression transgenic lines showed 18.2%− 25.7% higher dry biomass accumulation with a net photosynthesis rate than WT. In addition, overexpression of PtrAREB3 increased Cd accumulation in plants by 40.4%− 52.1%. Overall, the overexpression of PtrAREB3 enhanced Cd accumulation and tolerance, which making it an ideal molecular target for genetic improvement in Populus plants grown in metal contaminated areas in the future.

The genetic basis of water\u2010use efficiency and yield in lettuce

BackgroundWater supply limits agricultural productivity of many crops including lettuce. Identifying cultivars within crop species that can maintain productivity with reduced water supply is a significant challenge, but central to developing resilient crops for future water-limited climates. We investigated traits known to be related to water-use efficiency (WUE) and yield in lettuce, a globally important leafy salad crop, in a recombinant inbred line (RIL) lettuce mapping population, produced from a cross between the cultivated Lactuca sativa L. cv. Salinas and its wild progenitor L. serriola L.ResultsWild and cultivated lettuce differed in their WUE and we observed transgressive segregation in yield and water-use traits in the RILs. Quantitative trait loci (QTL) analysis identified genomic regions controlling these traits under well-watered and droughted conditions. QTL were detected for carbon isotope discrimination, transpiration, stomatal conductance, leaf temperature and yield, controlling 4–23 % of the phenotypic variation. A QTL hotspot was identified on chromosome 8 that controlled carbon isotope discrimination, stomatal conductance and yield under drought. Several promising candidate genes in this region were associated with WUE, including aquaporins, late embryogenesis abundant proteins, an abscisic acid-responsive element binding protein and glutathione S-transferases involved in redox homeostasis following drought stress were also identified.ConclusionsFor the first time, we have characterised the genetic basis of WUE of lettuce, a commercially important and water demanding crop. We have identified promising candidate genomic regions determining WUE and yield under well-watered and water-limiting conditions, providing important pre-breeding data for future lettuce selection and breeding where water productivity will be a key target.

The Physiological and Molecular Mechanism of Abscisic Acid in Regulation of Fleshy Fruit Ripening.

The ripening of fleshy fruits is coupled with the degradation of both chlorophyll and cell walls, as well as changes in the metabolism of phenylpropanoids, flavonoids, starch/sucrose, and carotenoids. These processes are controlled by phytohormones and other factors, including abscisic acid (ABA), ethylene, auxin, polyamines, sugar, and reactive oxygen species. The ripening of climacteric fruits is controlled by ethylene and non-climacteric fruit ripening is regulated mainly by ABA. Also, ABA and ethylene may interact in both types of fruit ripening. ABA concentrations in fleshy fruits are regulated in response to developmental and environmental cues and are controlled by the relative rates of ABA biosynthesis and catabolism, the former mainly via 9-cis-epoxycarotenoid dioxygenases (NCEDs) and β-glucosidases and the latter via ABA 8'-hydroxylases (CYP707As) and β-glycosyltransferases. In strawberry fruit ripening, ABA is perceived via at least two receptors, Pyrabactin resistance (PYR)/PYR-like (PYL) and putative abscisic acid receptor (ABAR), which are linked separately to the conserved signaling pathway ABA-FaPYR1-FaABIl-FaSnRK2 and the novel signaling pathway ABA-FaABAR-FaRIPK1-FaABI4. Downstream signaling components include important transcription factors, such as AREB (ABA responsive element binding protein)/ABF (ABRE binding factors ABA responsive factor), ethylene response factor (ERF), and V-myb Myeloblastosis viral oncogene homolog (MYB), as well as ripening-related genes. Finally, a comprehensive model of ABA linked to ethylene, sugar, polyamines, auxin and reactive oxygen species in the regulation of strawberry fruit ripening is proposed. Next, new integrated mechanisms, including two ABA signaling pathways, ABA and ethylene signaling pathways, and ABA/ethylene to other phytohormones are interesting and important research topics in ripening, especially in non-climacteric fruits.

Exogenous Application of Melatonin Improves Drought Tolerance in Coffee by Regulating Photosynthetic Efficiency and Oxidative Damage

The protective role of melatonin in plants under abiotic stress has been reported, but little information is available on its mitigation effect on coffee (Coffea arabica) plants. The objective of this study was to determine the effect of exogenous application of 100 µM melatonin in coffee leaves under 3 months of drought stress treatment. Melatonin was found to alleviate the drought-induced damage in coffee through reducing the rate of chlorophyll degradation, electrolyte leakage, malonaldehyde content, and activating various antioxidant enzymes, such as catalase, guaiacol peroxidase, and superoxide dismutase. Melatonin application suppressed the expression of chlorophyll degradation gene PAO encoding pheophorbide a oxygenase, and upregulated the expression of photosynthetic gene RBCS2 encoding ribulose-1,5-bisphosphate oxygenase (Rubisco) protein, and a drought-related gene AREB encoding abscisic acid-responsive element binding protein. The photosynthetic efficiency of photosystem II under dark adaptation was also improved upon melatonin application in drought-stressed plants. Our results showed that both foliar spray and direct soil application of melatonin could improve drought tolerance by regulating photosynthetic efficiency and oxidative damage in C. arabica seedlings. This study provides insights in application of melatonin as a protective agent against drought stress in improvement of crop yields.

Transcriptional modulation of AREB-1 by CRISPRa improves plant physiological performance under severe water deficit

Plants are sessile organisms, which are vulnerable to environmental stresses. As such, plants have developed multiple molecular, physiological, and cellular mechanisms to cope with natural stressors. However, these environmental adversities, including drought, are sources of the main agribusiness problems since they interfere with plant growth and productivity. Particularly under water deprivation conditions, the abscisic acid-responsive element-binding protein AREB1/ABF2 plays an important role in drought stress response and physiological adaptation. In this investigation, we provide substantial confirmation for the role of AREB1/ABF2 in plant survival under severe water deficit using the CRISPR activation (CRISPRa) technique to enhance the AREB1 gene expression. In our strategy, the inactive nuclease dCas9 was fused with an Arabidopsis histone acetyltransferase 1, which improves gene expression by remodeling chromatin. The AREB1 overexpression promotes an improvement in the physiological performance of the transgenic homozygous plants under drought, which was associated with an increase in chlorophyll content, antioxidant enzyme activity, and soluble sugar accumulation, leading to lower reactive oxygen species accumulation. Finally, we found that the CRISPR-mediated up-regulation of AREB1 changes the abundance of several downstream ABA-inducible genes, allowing us to report that CRISPRa dCas9-HAT is a valuable biotechnological tool to improve drought stress tolerance through the positive regulation of AREB1.

Gaining Insight into Exclusive and Common Transcriptomic Features Linked to Drought and Salinity Responses across Fruit Tree Crops.

The present study aimed at identifying and mapping key genes expressed in root tissues involved in drought and salinity tolerance/resistance conserved among different fruit tree species. Twenty-six RNA-Seq samples were analyzed from six published studies in five plant species (Olea europaea, Vitis riparia Michx, Prunus mahaleb, Prunus persica, Phoenix dactylifera). This meta-analysis used a bioinformatic pipeline identifying 750 genes that were commonly modulated in three salinity studies and 683 genes that were commonly regulated among three drought studies, implying their conserved role in resistance/tolerance/response to these environmental stresses. A comparison was done on the genes that were in common among both salinity and drought resulted in 82 genes, of which 39 were commonly regulated with the same trend of expression (23 were upregulated and 16 were downregulated). Gene set enrichment and pathway analysis pointed out that pathways encoding regulation of defense response, drug transmembrane transport, and metal ion binding are general key molecular responses to these two abiotic stress responses. Furthermore, hormonal molecular crosstalk plays an essential role in the fine-tuning of plant responses to drought and salinity. Drought and salinity induced a different molecular “hormonal fingerprint”. Dehydration stress specifically enhanced multiple genes responsive to abscisic acid, gibberellin, brassinosteroids, and the ethylene-activated signaling pathway. Salt stress mostly repressed genes encoding for key enzymes in signaling proteins in auxin-, gibberellin-(gibberellin 2 oxidase 8), and abscisic acid-related pathways (aldehyde oxidase 4, abscisic acid-responsive element-binding protein 3). Abiotic stress-related genes were mapped into the chromosome to identify molecular markers usable for the improvement of these complex quantitative traits. This meta-analysis identified genes that serve as potential targets to develop cultivars with enhanced drought and salinity resistance and/or tolerance across different fruit tree crops in a biotechnological sustainable way.

Overexpression of the Soybean NAC Gene GmNAC109 Increases Lateral Root Formation and Abiotic Stress Tolerance in Transgenic Arabidopsis Plants

NACs are plant-specific transcription factors that have crucial roles in plant development and biotic and/or abiotic stress responses. This study characterized the functions of the soybean NAC gene GmNAC109 using an overexpression construct in Arabidopsis lines. Sequence analysis revealed that GmNAC109 is highly homologous to ATAF1 (Arabidopsis Transcription Activation Factor 1), which regulates biotic and abiotic stress responses. GmNAC109 protein localized to the nucleus and its C-terminal domain exhibited transcriptional activation activity. Salt, dehydration, and cold stresses significantly increased expression of GmNAC109 in soybean. Similarly, Arabidopsis plants overexpressing GmNAC109 were more tolerant to drought and salt stress than wild-type Col-0 plants. Stress response-related genes, such as DREB1A (drought-responsive element-binding 1A), DREB2A, AREB1 (ABSCISIC ACID-RESPONSIVE ELEMENT BINDING PROTEIN 1), AREB2, RD29A (RESPONSIVE TO Desiccation 29A), and COR15A (COLD REGULATED 15A) were upregulated in GmNAC109-overexpressing transgenic Arabidopsis lines. The transgenic lines showed upregulation of the ABA-responsive genes ABI1 (ABA INSENSITIVE 1) and ABI5 and hypersensitivity to ABA. However, GmNAC109 did not increase expression of the ABA-biosynthetic gene NCED3 (NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3) and endogenous ABA content in the transgenic lines. Overexpression of GmNAC109 significantly increased lateral root formation in transgenic Arabidopsis lines. Expression of AIR3 (AUXIN-INDUCED IN ROOT CULTURES 3) and ARF2 (AUXIN RESPONSE FACTOR 2) was increased and decreased in these transgenic lines, respectively, indicating that GmNAC109 is involved in the auxin signaling pathway and thereby helps to regulate hairy root formation. Our results provide a basis for development of soybean lines with improved tolerance to abiotic stresses via genetic manipulation.

Maize ABP2 enhances tolerance to drought and salt stress in transgenic Arabidopsis

Abiotic stresses, especially drought and salt, severely affect maize production, which is one of the most important cereal crops in the world. Breeding stress-tolerant maize through biotechnology is urgently needed to maintain maize production. Therefore, it is important to identify new genes that can enhance both drought and salt stress tolerance for molecular breeding. In this study, we identified a maize ABA (abscisic acid)-responsive element (ABRE) binding protein from a 17-day post-pollination (dpp) maize embryo cDNA library by yeast one-hybrid screen using the ABRE2 sequence of the maize Cat1 gene as bait. This protein, designated, ABRE binding protein 2 (ABP2), belongs to the bZIP transcription factor family. Endogenous expression of ABP2 in maize can be detected in different tissues at various development stages, and can be induced by drought, salt, reactive oxygen species (ROS)-generating agents, and ABA treatment. Constitutive expression of ABP2 in transgenic Arabidopsis plants enhanced tolerance to drought and salt stress, and increased sensitivity to ABA. In exploring the mechanism by which ABP2 can stimulate abiotic stress tolerance, we found that ROS levels were reduced and expression of stress-responsive and carbon metabolism-related genes was enhanced by constitutive ABP2 expression in transgenic plants. In short, we identified a maize bZIP transcription factor which can enhance both drought and salt tolerance of plants.

Abscisic Acid Is a Major Regulator of Grape Berry Ripening Onset: New Insights into ABA Signaling Network

Grapevine is a world-wide cultivated economically relevant crop. The process of berry ripening is non-climacteric and does not rely on the sole ethylene signal. Abscisic acid (ABA) is recognized as an important hormone of ripening inception and color development in ripening berries. In order to elucidate the effect of this signal at the molecular level, pre-véraison berries were treated ex vivo for 20 h with 0.2 mM ABA and berry skin transcriptional modulation was studied by RNA-seq after the treatment and 24 h later, in the absence of exogenous ABA. This study highlighted that a small amount of ABA triggered its own biosynthesis and had a transcriptome-wide effect (1893 modulated genes) characterized by the amplification of the transcriptional response over time. By comparing this dataset with the many studies on ripening collected within the grapevine transcriptomic compendium Vespucci, an extended overlap between ABA- and ripening modulated gene sets was observed (71% of the genes), underpinning the role of this hormone in the regulation of berry ripening. The signaling network of ABA, encompassing ABA metabolism, transport and signaling cascade, has been analyzed in detail and expanded based on knowledge from other species in order to provide an integrated molecular description of this pathway at berry ripening onset. Expression data analysis was combined with in silico promoter analysis to identify candidate target genes of ABA responsive element binding protein 2 (VvABF2), a key upstream transcription factor of the ABA signaling cascade which is up-regulated at véraison and also by ABA treatments. Two transcription factors, VvMYB143 and VvNAC17, and two genes involved in protein degradation, Armadillo-like and Xerico-like genes, were selected for in vivo validation by VvABF2-mediated promoter trans-activation in tobacco. VvNAC17 and Armadillo-like promoters were induced by ABA via VvABF2, while VvMYB143 responded to ABA in a VvABF2-independent manner. This knowledge of the ABA cascade in berry skin contributes not only to the understanding of berry ripening regulation but might be useful to other areas of viticultural interest, such as bud dormancy regulation and drought stress tolerance.

Evaluation of genetic diversity of cis-acting elements of Abscisic acid responsive element binding protein (ABRE-BP) in selected Sri Lankan rice varieties

Salinity is a major abiotic stress that affects rice cultivation. Osmotic stress caused by salinity activates tolerance mechanisms in rice. The Abscisic Acid Responsive Element Binding Protein (ABRE-BP), transcribes for a bZIP family transcription factor that binds to the cis-regulatory element Abscisic Acid Responsive Element (ABRE) at the promoter of downstream salinity responsive genes and regulates their expression. Hence, the study was focused on analyzing the nucleotide diversity of a region in ABRE-BP promoter. Analysis of the ABRE-BP sequences of 47 Sri Lankan rice varieties and two reference varieties retrieved from a public database revealed many insertions and deletions (INDELs) and single nucleotide polymorphisms (SNPs) in the promoter. Eleven, abiotic stress related cis-elements were identified, but none of the INDELs spanned over them. However, SNPs either deleted existing cis-elements or created new cis-elements. The salinity stress responsive elements, MYBCORE and GT1GMSCAM4 were detected in few varieties, and were not associated with salinity tolerance based on current available salinity ratings. The presence of SNPs in cis-elements clustered them to nine groups at 68% similarity. The DNA polymorphisms on stress responsive cis-acting elements did not show a strong association with the known salinity ratings of many varieties. Nevertheless, the presence of INDELs affects the relative distance between elements, and thus may alter the expression of the ABRE-BP that regulates downstream stress responsive genes.

Overexpression of the activated form of the AtAREB1 gene (AtAREB1ΔQT) improves soybean responses to water deficit.

Abscisic acid-responsive element binding protein (AREB1) is a basic domain/leucine zipper transcription factor that binds to the abscisic acid (ABA)-responsive element motif in the promoter region of ABA-inducible genes. Because AREB1 is not sufficient to direct the expression of downstream genes under non-stress conditions, an activated form of AREB1 (AREB1ΔQT) was created. Several reports claim that plants overexpressing AREB1 or AREB1ΔQT show improved drought tolerance. In our studies, soybean plants overexpressing AREB1ΔQT were characterized molecularly, and the phenotype and drought response of three lines were accessed under greenhouse conditions. Under conditions of water deficit, the transformed plants presented a higher survival rate (100%) than those of their isoline, cultivar BR 16 (40%). Moreover, the transformed plants displayed better water use efficiency and had a higher number of leaves than their isoline. Because the transgenic plants had higher stomatal conductance than its isoline under well-watered conditions, it was suggested that the enhanced drought response of AREB1ΔQT soybean plants might not be associated with altered transpiration rates mediated by ABA-dependent stomatal closure. However, it is possible that the smaller leaf area of the transgenic plants reduced their transpiration and water use, causing delayed stress onset. The difference in the degree of wilting and percentage of survival between the 35S-AREB1ΔQT and wildtype plants may also be related to the regulation of genes that protect against dehydration because metabolic impairment of photosynthesis, deduced by an increasing internal CO2 concentration, was not observed in the transgenic plants.

The transcription factor SlAREB1 confers drought, salt stress tolerance and regulates biotic and abiotic stress‐related genes in tomato

Members of the abscisic acid-responsive element binding protein (AREB)/abscisic acid-responsive element binding factor (ABF) subfamily of basic leucine zipper (bZIP) transcription factors have been implicated in abscisic acid (ABA) and abiotic stress responses in plants. Here we describe two members identified in cultivated tomato (Solanum lycopersicum), named SlAREB1 and SlAREB2. Expression of SlAREB1 and SlAREB2 is induced by drought and salinity in both leaves and root tissues, although that of SlAREB1 was more affected. In stress assays, SlAREB1-overexpressing transgenic tomato plants showed increased tolerance to salt and water stress compared to wild-type and SlAREB1-down-regulating transgenic plants, as assessed by physiological parameters such as relative water content (RWC), chlorophyll fluorescence and damage by lipoperoxidation. In order to identify SlAREB1 target genes responsible for the enhanced tolerance, microarray and cDNA-amplified fragment length polymorphism (AFLP) analyses were performed. Genes encoding oxidative stress-related proteins, lipid transfer proteins (LTPs), transcription regulators and late embryogenesis abundant proteins were found among the up-regulated genes in SlAREB1-overexpressing lines, especially in aerial tissue. Notably, several genes encoding defence proteins associated with responses to biotic stress (e.g. pathogenesis-related proteins, protease inhibitors, and catabolic enzymes) were also up-regulated by SlAREB1 overexpression, suggesting that this bZIP transcription factor is involved in ABA signals that participate in abiotic stress and possibly in response to pathogens.