ABRE-binding Factor Research Articles

Integrated physiological and transcriptomic analyses of two warm- and cool-season turfgrass species in response to heat stress

Warm- and cool-season turfgrasses were originated from different locations with contrasting heat tolerance. The molecular mechanisms of heat tolerance have not been extensively studied in turfgrass species. In this study, transcriptomic analysis showed that bermudagrass was more tolerant to heat stress as evidenced by lower contents of H2O2, proline and glutathione than those in tall fescue after heat treatment. RNA sequencing analysis revealed that 32.7% and 17.7% more genes were changed in tall fescue than in bermudagrass after 2 and 12h heat treatment, respectively. GO terms of redox were enriched in bermudagrass whereas metabolite transportation ones were over-represented in tall fescue after 2h treatment. Ubiquitin dependent degradation pathways were commonly regulated in both grass species. CdF-box and FaF-box transgenic Arabidopsis exhibited improved tolerance to heat stress. Regulatory elements analysis revealed that four ABA-responsive elements present in CdF-box promoter, indicating CdF-box could be potentially regulated by ABRE binding factors (ABFs). All these findings provide evidences for understanding heat stress response in warm- and cool-season grass species.

Integrative proteomics and phosphoproteomics reveals phosphorylation networks involved in the maintenance and expression of embryogenic competence in sugarcane callus

Plant embryogenic cell culture allows mass propagation and genetic manipulation, but the mechanisms that determine the fate of these totipotent cells in somatic embryos have not yet been elucidated. Here, we performed label-free quantitative proteomics and phosphoproteomics analyses to determine signaling events related to sugarcane somatic embryo differentiation, especially those related to protein phosphorylation. Embryogenic calli were compared at multiplication (EC0, dedifferentiated cells) and after 14 days of maturation (EC14, onset of embryo differentiation). Metabolic pathway analysis showed enriched lysine degradation and starch/sucrose metabolism proteins during multiplication, whereas the differentiation of somatic embryos was found to involve the enrichment of energy metabolism, including the TCA cycle and oxidative phosphorylation. Multiplication-related phosphoproteins were associated with transcriptional regulation, including SNF1 kinase homolog 10 (KIN10), SEUSS (SEU), and LEUNIG_HOMOLOG (LUH). The regulation of multiple light harvesting complex photosystem II proteins and phytochrome interacting factor 3-LIKE 5 were predicted to promote bioenergetic metabolism and carbon fixation during the maturation stage. A motif analysis revealed 15 phosphorylation motifs. The [D-pS/T-x-D] motif was overrepresented during somatic embryo differentiation. A protein-protein network analysis predicted interactions among SNF1-related protein kinase 2 (SnRK2), abscisic acid-responsive element-binding factor 2 (ABF2), and KIN10, which indicated the role of these proteins in embryogenic competence. The predicted interactions between TOPLESS (TPL) and histone deacetylase 19 (HD19) may be involved in posttranslational protein regulation during somatic embryo differentiation. These results reveal the protein regulation dynamics of somatic embryogenesis and new players in somatic embryo differentiation, including their predicted phosphorylation motifs and phosphosites.

Calcium-dependent Protein Kinase 5 (CPK5) positively modulates drought tolerance through phosphorylating ABA-Responsive Element Binding Factors in oilseed rape (Brassica napus L.)

Drought is an environmental stress that causes severe crop loss. Drought stress can induce abscisic acid (ABA) accumulation and cytoplasmic calcium oscillation. Calcium-dependent protein kinases (CPKs) constitute a group of Ser/Thr protein kinases decoding calcium signals. However, the function and molecular mechanisms of most CPKs in oilseed rape (Brassica napus) remain unknown. Here, we report the functional characterization of BnaCPK5 in drought stress tolerance. BnaCPK5 belongs to Group I of the CPK family and was localized at the plasma membrane and nuclei. Overexpression of BnaCPK5 enhanced drought stress tolerance compared with the control. A screening of interacting proteins identified that BnaCPK5 interacted strongly with two ABA-Responsive Element Binding Factors (ABF/AREBs), BnaABF3 and BnaABF4. BnaCPK5 was shown to phosphorylate both BnaABF3 and BnaABF4 in a kinase assay. Further, it was found that the phosphorylation of BnaABF3 and BnaABF4 by BnaCPK5 increased their transcriptional activities against the famous drought stress marker gene, Responsive to Dehydration (RD) 29B and protein stability. Taken together, these data demonstrate that BnaCPK5 acts as a positive regulator of drought tolerance by, at least in part, phosphorylating two core ABA-signaling components to modulate Late-Embryogenesis Abundant (LEA)-like RD29B expression.

Factors that affect protein abundance of a positive regulator of abscisic acid signalling, the basic leucine zipper transcription factor ABRE-binding factor 2 (ABF2).

Most members of basic leucine zipper (bZIP) transcription factor (TF) subgroup A play important roles as positive effectors in abscisic acid (ABA) signaling during germination and/or in vegetative stress responses. In multiple plant species, one member, ABA insensitive 5 (ABI5), is a major TF that promotes seed maturation and blocks early seeding growth in response to ABA. Other members, referred to as either ABRE‐binding factors (ABFs), ABRE‐binding proteins (AREBs), or D3 protein‐binding factors (DPBFs), are implicated as major players in stress responses during vegetative growth. Studies on the proteolytic regulation of ABI5, ABF1, and ABF3 in Arabidopsis thaliana have shown that the proteins have moderate degradation rates and accumulate in the presence of the proteasome inhibitor MG132. Exogenous ABA slows their degradation and the ubiquitin E3 ligase called KEEP ON GOING (KEG) is important for their degradation. However, there are some reported differences in degradation among subgroup A members. The conserved C‐terminal sequences (referred to as the C4 region) enhance degradation of ABI5 but stabilize ABF1 and ABF3. To better understand the proteolytic regulation of the ABI5/ABFs and determine whether there are differences between vegetative ABFs and ABI5, we studied the degradation of an additional family member, ABF2, and compared its in vitro degradation to that of ABI5. As previously seen for ABI5, ABF1, and ABF3, epitope‐tagged constitutively expressed ABF2 degrades in seedlings treated with cycloheximide and is stabilized following treatment with the proteasome inhibitor MG132. Tagged ABF2 protein accumulates when seedlings are treated with ABA, but its mRNA levels do not increase, suggesting that the protein is stabilized in the presence of ABA. ABF2 is also an in vitro ubiquitination substrate of the E3 ligase KEG and recombinant ABF2 is stable in keg lysates. ABF2 with a C4 deletion degrades more quickly in vitro than full‐length ABF2, as previously observed for ABF1 and ABF3, suggesting that the conserved C4 region contributes to its stability. In contrast to ABF2 and consistent with previously published work, ABI5 with C terminal deletions including an analogous C4 deletion is stabilized in vitro compared to full length ABI5. In vivo expression of an ABF1 C4 deletion protein appears to have reduced activity compared to equivalent levels of full length ABF1. Additional group A family members show similar proteolytic regulation by MG132 and ABA. Altogether, these results together with other work on ABI5 regulation suggest that the vegetative ABFs share proteolytic regulatory mechanisms that are not completely shared with ABI5.

Stomatal regulation: Role of H2S-induced persulfidation in ABA signaling

Stomatal regulation: Role of H2S-induced persulfidation in ABA signaling

Cellular Phosphorylation Signaling and Gene Expression in Drought Stress Responses: ABA-Dependent and ABA-Independent Regulatory Systems.

Drought is a severe and complex abiotic stress that negatively affects plant growth and crop yields. Numerous genes with various functions are induced in response to drought stress to acquire drought stress tolerance. The phytohormone abscisic acid (ABA) accumulates mainly in the leaves in response to drought stress and then activates subclass III SNF1-related protein kinases 2 (SnRK2s), which are key phosphoregulators of ABA signaling. ABA mediates a wide variety of gene expression processes through stress-responsive transcription factors, including ABA-RESPONSIVE ELEMENT BINDING PROTEINS (AREBs)/ABRE-BINDING FACTORS (ABFs) and several other transcription factors. Seed plants have another type of SnRK2s, ABA-unresponsive subclass I SnRK2s, that mediates the stability of gene expression through the mRNA decay pathway and plant growth under drought stress in an ABA-independent manner. Recent research has elucidated the upstream regulators of SnRK2s, RAF-like protein kinases, involved in early responses to drought stress. ABA-independent transcriptional regulatory systems and ABA-responsive regulation function in drought-responsive gene expression. DEHYDRATION RESPONSIVE ELEMENT (DRE) is an important cis-acting element in ABA-independent transcription, whereas ABA-RESPONSIVE ELEMENT (ABRE) cis-acting element functions in ABA-responsive transcription. In this review article, we summarize recent advances in research on cellular and molecular drought stress responses and focus on phosphorylation signaling and transcription networks in Arabidopsis and crops. We also highlight gene networks of transcriptional regulation through two major regulatory pathways, ABA-dependent and ABA-independent pathways, that ABA-responsive subclass III SnRK2s and ABA-unresponsive subclass I SnRK2s mediate, respectively. We also discuss crosstalk in these regulatory systems under drought stress.

DcABF3, an ABF transcription factor from carrot, alters stomatal density and reduces ABA sensitivity in transgenic Arabidopsis

Abscisic acid-responsive element (ABRE)-binding factors (ABFs) are important transcription factors involved in various physiological processes in plants. Stomata are micro channels for water and gas exchange of plants. Previous researches have demonstrated that ABFs can modulate the stomatal development in some plants. However, little is known about stomata-related functions of ABFs in carrots. In our study, DcABF3, a gene encoding for ABF transcription factor, was isolated from carrot. The open reading frame of DcABF3 was 1329 bp, encoding 442 amino acids. Expression profiles of DcABF3 indicated that DcABF3 can respond to drought, salt or ABA treatment in carrots. Overexpressing DcABF3 in Arabidopsis led to the increase of stomatal density which caused severe water loss. Expression assay indicated that overexpression of DcABF3 caused high expression of stomatal development-related transcription factor genes, SPCH, FAMA, MUTE and SCRMs. Increased antioxidant enzyme activities and higher expression levels of stress-related genes were also found in transgenic lines after water deficit treatment. Changes in expression of ABA synthesis-related genes and AtABIs indicated the potential role of DcABF3 in ABA signaling pathway. Under the treatment of exogenous ABA, DcABF3-overexpression Arabidopsis seedlings exhibited increased root length and germination rate. Our findings demonstrated that heterologous overexpression of DcABF3 positively affected stomatal development and also reduced ABA sensitivity in transgenic Arabidopsis.

ABF3 enhances drought tolerance via promoting ABA-induced stomatal closure by directly regulating ADF5 in Populus euphratica.

Water availability is a main limiting factor for plant growth, development, and distribution throughout the world. Stomatal movement mediated by abscisic acid (ABA) is particularly important for drought adaptation, but the molecular mechanisms in trees are largely unclear. Here, we isolated an ABA-responsive element binding factor, PeABF3, in Populus euphratica. PeABF3 was preferentially expressed in the xylem and young leaves, and was induced by dehydration and ABA treatments. PeABF3 showed transactivation activity and was located in the nucleus. To study its functional mechanism in poplar responsive to drought stress, transgenic triploid white poplars (Populus tomentosa 'YiXianCiZhu B385') overexpressing PeABF3 were generated. PeABF3 overexpression significantly enhanced stomatal sensitivity to exogenous ABA. When subjected to drought stress, PeABF3 overexpression maintained higher photosynthetic activity and promoted cell membrane integrity, resulting in increased water-use efficiency and enhanced drought tolerance compared with wild-type controls. Moreover, a yeast one-hybrid assay and an electrophoretic mobility shift assay revealed that PeABF3 activated the expression of Actin-Depolymerizing Factor-5 (PeADF5) by directly binding to its promoter, promoting actin cytoskeleton remodeling and stomatal closure in poplar under drought stress. Taken together, our results indicate that PeABF3 enhances drought tolerance via promoting ABA-induced stomatal closure by directly regulating PeADF5 expression.

Transcriptomic analysis of poco1, a mitochondrial pentatricopeptide repeat protein mutant in Arabidopsis thaliana

BackgroundFlowering is a crucial stage during plant development. Plants may respond to unfavorable conditions by accelerating reproductive processes like flowering. In a recent study, we showed that PRECOCIOUS1 (POCO1) is a mitochondrial pentatricopeptide repeat (PPR) protein involved in flowering time and abscisic acid (ABA) signaling in Arabidopsis thaliana. Here, we use RNA-seq data to investigate global gene expression alteration in the poco1 mutant.ResultsRNA-seq analysis was performed during different developmental stages for wild-type and poco1 plants. The most profound differences in gene expression were found when wild-type and poco1 plants of the same developmental stage were compared. Coverage analysis confirmed the T-DNA insertion in POCO1, which was concomitant with truncated transcripts. Many biological processes were found to be enriched. Several flowering-related genes such as FLOWERING LOCUS T (FT), which may be involved in the early-flowering phenotype of poco1, were differentially regulated. Numerous ABA-associated genes, including the core components of ABA signaling such as ABA receptors, protein phosphatases, protein kinases, and ABA-responsive element (ABRE) binding proteins (AREBs)/ABRE-binding factors (ABFs) as well as important genes for stomatal function, were mostly down-regulated in poco1. Drought and oxidative stress-related genes, including ABA-induced stress genes, were differentially regulated. RNA-seq analysis also uncovered differentially regulated genes encoding various classes of transcription factors and genes involved in cellular signaling. Furthermore, the expression of stress-associated nuclear genes encoding mitochondrial proteins (NGEMPs) was found to be altered in poco1. Redox-related genes were affected, suggesting that the redox state in poco1 might be altered.ConclusionThe identification of various enriched biological processes indicates that complex regulatory mechanisms underlie poco1 development. Differentially regulated genes associated with flowering may contribute to the early-flowering phenotype of poco1. Our data suggest the involvement of POCO1 in the early ABA signaling process. The down-regulation of many ABA-related genes suggests an association of poco1 mutation with the ABA signaling deficiency. This condition further affects the expression of many stress-related, especially drought-associated genes in poco1, consistent with the drought sensitivity of poco1. poco1 mutation also affects the expression of genes associated with the cellular regulation, redox, and mitochondrial perturbation.

Chromosome doubling mediates superior drought tolerance in Lycium ruthenicum via abscisic acid signaling

Plants are continuously affected by unfavorable external stimuli, which influences their productivity and growth. Differences in gene composition and expression patterns lead homologous polyploid plants to exhibit different physiological phenomena, among which enhanced environmental adaptability is a powerful phenotype conferred by polyploidization. The mechanisms underlying the differences in stress tolerance between diploids and autotetraploids at the molecular level remain unclear. In this research, a full-length transcription profile obtained via the single-molecule real-time (SMRT) sequencing of high-quality single RNA molecules for use as background was combined with next-generation transcriptome and proteome technologies to probe the variation in the molecular mechanisms of autotetraploids. Tetraploids exhibited an increase in ABA content of 78.4% under natural conditions and a superior stress-resistance phenotype under severe drought stress compared with diploids. The substantial differences in the transcriptome profiles observed between diploids and autotetraploids under normal growth conditions were mainly related to ABA biosynthesis and signal transduction pathways, and 9-cis-epoxycarotenoid dioxygenase 1 (NCED1) and NCED2, which encode key synthetic enzymes, were significantly upregulated. The increased expression of the ABRE-binding factor 5-like (ABF5-like) gene was a pivotal factor in promoting the activation of the ABA signaling pathway and downstream target genes. In addition, ABA strongly induced the expression of osmotic proteins to increase the stress tolerance of the plants at the translational level. We consider the intrinsic mechanisms by which ABA affects drought resistance in tetraploids and diploids to understand the physiological and molecular mechanisms that enhance abiotic stress tolerance in polyploid plants.

Hydrogen Sulfide Positively Regulates Abscisic Acid Signaling through Persulfidation of SnRK2.6 in Guard Cells

The phytohormone abscisic acid (ABA) plays pivotal roles in triggering stomatal closure and facilitating adaptation of plants to drought stress. Hydrogen sulfide (H2S), a small signaling gas molecule, is involved in ABA-dependent stomatal closure. However, how H2S regulates ABA signaling remains largely unclear. Here, we show that ABA induces the production of H2S catalyzed by L-CYSTEINE DESULFHYDRASE1 (DES1) in guard cells, and H2S in turn positively regulates ABA signaling through persulfidation of Open Stomata 1 (OST1)/SNF1-RELATED PROTEIN KINASE2.6 (SnRK2.6). Two cysteine (Cys) sites, Cys131 and Cys137, which are exposed on the surface of SnRK2.6 and close to the activation loop, were identified to be persulfidated, which promotes the activity of SnRK2.6 and its interaction with ABA response element-binding factor 2 (ABF2), a transcription factor acting downstream of ABA signaling. When Cys131, Cys137, or both residues in SnRK2.6 were substituted with serine (S), H2S-induced SnRK2.6 activity and SnRK2.6–ABF2 interaction were partially (SnRK2.6C131S and SnRK2.6C137S) or completely (SnRK2.6C131SC137S) compromised. Introduction of SnRK2.6C131S, SnRK2.6C137S, or SnRK2.6C131SC137S into the ost1-3 mutant could not rescue the mutant phenotype: less sensitivity to ABA- and H2S-induced stomatal closure and Ca2+ influx as well as increased water loss and decreased drought tolerance. Taken together, our study reveals a novel post-translational regulatory mechanism of ABA signaling whereby H2S persulfidates SnRK2.6 to promote ABA signaling and ABA-induced stomatal closure.

Arabidopsis CPK6 positively regulates ABA signaling and drought tolerance through phosphorylating ABA-responsive element-binding factors.

Abscisic acid (ABA) regulates numerous developmental processes and drought tolerance in plants. Calcium-dependent protein kinases (CPKs) are important Ca2+ sensors playing crucial roles in plant growth and development as well as responses to stresses. However, the molecular mechanisms of many CPKs in ABA signaling and drought tolerance remain largely unknown. Here we combined protein interaction studies, and biochemical and genetic approaches to identify and characterize substrates that were phosphorylated by CPK6 and elucidated the mechanism that underlines the role of CPK6 in ABA signaling and drought tolerance. The expression of CPK6 is induced by ABA and dehydration. Two cpk6 T-DNA insertion mutants are insensitive to ABA during seed germination and root elongation of seedlings; in contrast, overexpression of CPK6 showed the opposite phenotype. Moreover, CPK6-overexpressing lines showed enhanced drought tolerance. CPK6 interacts with and phosphorylates a subset of core ABA signaling-related transcription factors, ABA-responsive element-binding factors (ABFs/AREBs), and enhances their transcriptional activities. The phosphorylation sites in ABF3 and ABI5 were also identified through MS and mutational analyses. Taken together, we present evidence that CPK6 mediates ABA signaling and drought tolerance through phosphorylating ABFs/AREBs. This work thus uncovers a rather conserved mechanism of calcium-dependent Ser/Thr kinases in ABA signaling.

Involvement of abscisic acid-responsive element-binding factors in cassava (Manihot esculenta) dehydration stress response

Cassava (Manihot esculenta) is a major staple food, animal feed and energy crop in the tropics and subtropics. It is one of the most drought-tolerant crops, however, the mechanisms of cassava drought tolerance remain unclear. Abscisic acid (ABA)-responsive element (ABRE)-binding factors (ABFs) are transcription factors that regulate expression of target genes involved in plant tolerance to drought, high salinity, and osmotic stress by binding ABRE cis-elements in the promoter regions of these genes. However, there is little information about ABF genes in cassava. A comprehensive analysis of Manihot esculenta ABFs (MeABFs) described the phylogeny, genome location, cis-acting elements, expression profiles, and regulatory relationship between these factors and Manihot esculenta betaine aldehyde dehydrogenase genes (MeBADHs). Here we conducted genome-wide searches and subsequent molecular cloning to identify seven MeABFs that are distributed unevenly across six chromosomes in cassava. These MeABFs can be clustered into three groups according to their phylogenetic relationships to their Arabidopsis (Arabidopsis thaliana) counterparts. Analysis of the 5′-upstream region of MeABFs revealed putative cis-acting elements related to hormone signaling, stress, light, and circadian clock. MeABF expression profiles displayed clear differences among leaf, stem, root, and tuberous root tissues under non-stress and drought, osmotic, or salt stress conditions. Drought stress in cassava leaves and roots, osmotic stress in tuberous roots, and salt stress in stems induced expression of the highest number of MeABFs showing significantly elevated expression. The glycine betaine (GB) content of cassava leaves also was elevated after drought, osmotic, or salt stress treatments. BADH1 is involved in GB synthesis. We show that MeBADH1 promoter sequences contained ABREs and that MeBADH1 expression correlated with MeABF expression profiles in cassava leaves after the three stress treatments. Taken together, these results suggest that in response to various dehydration stresses, MeABFs in cassava may activate transcriptional expression of MeBADH1 by binding the MeBADH1 promoter that in turn promotes GB biosynthesis and accumulation via an increase in MeBADH1 gene expression levels and MeBADH1 enzymatic activity. These responses protect cells against dehydration stresses by preserving an osmotic balance that enhances cassava tolerance to dehydration stresses.

A Cryophyte Transcription Factor, CbABF1, Confers Freezing, and Drought Tolerance in Tobacco.

Abscisic acid responsive element binding factors (ABFs) play crucial roles in plant responses to abiotic stress. However, little is known about the roles of ABFs in alpine subnival plants, which can survive under extreme environmental conditions. Here, we cloned and characterized an ABF1 homolog, CbABF1, from the alpine subnival plant Chorispora bungeana. Expression of CbABF1 was induced by cold, drought, and abscisic acid. Subcellular localization analysis revealed that CbABF1 was located in the nucleus. Further, CbABF1 had transactivation activity, which was dependent on the N-terminal region containing 89 residues. A Snf1-related protein kinase, CbSnRK2.6, interacted with CbABF1 in yeast two-hybrid analysis and bimolecular fluorescence complementation assays. Transient expression assay revealed that CbSnRK2.6 enhanced the transactivation of CbABF1 on ABRE cis-element. We further found that heterologous expression of CbABF1 in tobacco improved plant tolerance to freezing and drought stress, in which the survival rates of the transgenic plants increased around 40 and 60%, respectively, compared with wild-type plants. Moreover, the transgenic plants accumulated less reactive oxygen species, accompanied by high activities of antioxidant enzymes and elevated expression of stress-responsive genes. Our results thus suggest that CbABF1 is a transcription factor that plays an important role in cold and drought tolerance and is a candidate gene in molecular breeding of stress-tolerant crops.

Reducing expression of a nitrate-responsive bZIP transcription factor increases grain yield and N use in wheat.

SummaryNitrogen (N) plays critical role in plant growth; manipulating N assimilation could be a target to increase grain yield and N use. Here, we show that ABRE‐binding factor (ABF)‐like leucine zipper transcription factor TabZIP60 mediates N use and growth in wheat. The expression of TabZIP60 is repressed when the N‐deprived wheat plants is exposed to nitrate. Knock down of TabZIP60 through RNA interference (RNAi) increases NADH‐dependent glutamate synthase (NADH‐GOGAT) activity, lateral root branching, N uptake and spike number, and improves grain yield more than 25% under field conditions, while overexpression of TabZIP60‐6D had the opposite effects. Further investigation shows TabZIP60 binds to ABRE‐containing fragment in the promoter of TaNADH‐GOGAT‐3B and negatively regulates its expression. Genetic analysis reveals that TaNADH‐GOGAT‐3B overexpression overcomes the spike number and yield reduction caused by overexpressing TabZIP60‐6D. As such, TabZIP60‐mediated wheat growth and N use is associated with its negative regulation on TaNADH‐GOGAT expression. These findings indicate that TabZIP60 and TaNADH‐GOGAT interaction plays important roles in mediating N use and wheat growth, and provides valuable information for engineering N use efficiency and yield in wheat.

Global metabolite profiling based on GC\u2013MS and LC\u2013MS/MS analyses in ABF3-overexpressing soybean with enhanced drought tolerance

Abscisic acid (ABA) is a phytohormone that plays an important role in the adaptive responses to abiotic stresses. We examined the metabolic changes in transgenic soybean that over-expressed Arabidopsis ABA responsive element-binding factor 3 (ABF3), which participates in drought tolerance. Transgenic and non-transgenic plants were exposed to a water deficit, and their metabolic differences were verified by untargeted GC‒MS and LC‒MS/MS analyses. A total of 64 and 476 primary and secondary metabolites from leaf extracts were identified based on GC‒MS and LC‒MS/MS platforms, respectively. Principal component analysis derived from both GC‒MS and LC‒MS/MS data showed a clearly greater separation in the metabolite profiles among three different degrees of drought stress. However, no discrimination of metabolites between transgenic and non-transgenic plants was apparent. Furthermore, except for some free amino acids, quantitative differences in relative levels of those metabolites were less than 50% between genotypes. These results suggest that, during periods of drought, overexpression of ABF3 in transgenic soybean might result in a negligible variance in primary and secondary metabolism when compared with its non-transgenic counterpart.

OsbZIP42 is a positive regulator of ABA signaling and confers drought tolerance to rice.

OsbZIP42 is a positive regulator of ABA signaling and drought stress tolerance. The activation of OsbZIP42 depends on stress-/ABA-activated protein kinase 4 (SAPK4) and an additional ABA-dependent modification of OsbZIP42. Basic leucine zipper transcription factors (bZIP TFs) play important roles in the ABA signaling pathway in plants. Rice OsbZIP42 is a member of the group E bZIP, which is an ortholog of Arabidopsis group A bZIP. This latter group includes abscisic acid-responsive element (ABRE)-binding factors (ABFs) involved in abiotic stress tolerance. The expression of OsbZIP42 was induced by ABA treatment, although it was not induced by drought and salt stresses. Unlike other bZIP TFs, OsbZIP42 contained two transcriptional activation domains. Although the full-length OsbZIP42 protein did not, the N-terminus of the protein interacted with SAPK4. Our results suggest that the activation of OsbZIP42 by SAPK4 requires another ABA-dependent modification of OsbZIP42. Transgenic rice overexpressing OsbZIP42 (OsbZIP42-OX) exhibited a rapidly elevated expression of the ABA-responsive LEA3 and Rab16 genes and was hypersensitive to ABA. Analyses of the OsbZIP42-OX plants revealed enhanced tolerance to drought stress. These results suggest that OsbZIP42 is a positive regulator of ABA signaling and drought stress tolerance depending on its activation, which is followed by an additional ABA-dependent modification. We propose that OsbZIP42 is an important player in rice for conferring ABA-dependent drought tolerance.

Gene Expression Profiling in Sugarcane Genotypes During Drought Stress and Rehydration

The study aimed to assess the molecular responses of sugarcane genotypes exposed to water stress and rehydration. Six sugarcane varieties that show differential tolerance to drought were planted in the pots with three replications and subjected to gradual drought by withholding water for 2, 6 and 10 days and rehydration for 2, 6 and 10 days after the onset of treatment (DAT) at 60 days after planting. To analyze the physiological status, relative water content and cellular membrane injury were measured. Among the six varieties, two varieties, viz. Co 06022 and Co 99004, retained high relative water content and cell membrane injury recorded was also low during severe drought stress. Sugarcane variety Co 8021 was the most vulnerable to drought stress. The relative water content reduced drastically at 2 days of stress and also maximum membrane injury was observed as stress progressed. Signs of wilting were observed in 2 days, while Co 06022 and Co 99004 showed slight wilting only at 10 days of stress. Examination of differentially expressed genes and transcription factors indicated that ABRE-binding factor, trehalose 6-phosphate synthase, DREB2, SNAC1, dehydrin and early responsive to dehydration (ERD4) showed high-level transcript accumulation during initial stress and gradually declined as stress progressed to 10 days in most genotypes. Invertase and calmodulin functional proteins were overexpressed at both stress and recovery stages. Expression levels of most genes reverted back to their control levels during recovery. Results were validated in shoots and leaves by quantitative real-time PCR.

Arabidopsis ABF3 and ABF4 Transcription Factors Act with the NF-YC Complex to Regulate SOC1 Expression and Mediate Drought-Accelerated Flowering

The drought-escape response accelerates flowering in response to drought stress, allowing plants to adaptively shorten their life cycles. Abscisic acid (ABA) mediates plant responses to drought, but the role of ABA-responsive element (ABRE)-binding factors (ABFs) in the drought-escape response is poorly understood. Here, we show that Arabidopsis thaliana ABF3 and ABF4 regulate flowering in response to drought through transcriptional regulation of the floral integrator SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1). The abf3 abf4 mutant displayed ABA-insensitive late flowering under long-day conditions. Ectopic expression of ABF3 or ABF4 in the vasculature, but not in the shoot apex, induced early flowering, whereas expression of ABF3 fused with the SRDX transcriptional repressor domain delayed flowering. We identified SOC1 as a direct downstream target of ABF3/4, and found that SOC1 mRNA levels were lower in abf3 abf4 than in wild-type plants. Moreover, induction of SOC1 by ABA was hampered in abf3 abf4 mutants. ABF3 and ABF4 were enriched at the −1028- to −657-bp region of the SOC1 promoter, which does not contain canonical ABF-ABRE-binding motifs but has the NF-Y binding element. We found that ABF3 and ABF4 interact with nuclear factor Y subunit C (NF-YC) 3/4/9 in vitro and in planta, and induction of SOC1 by ABA was hampered in nf-yc3 yc4 yc9 mutants. Interestingly, the abf3 abf4, nf-yc3 yc4 yc9, and soc1 mutants displayed a reduced drought-escape response. Taken together, these results suggest that ABF3 and ABF4 act with NF-YCs to promote flowering by inducing SOC1 transcription under drought conditions. This mechanism might contribute to adaptation by enabling plants to complete their life cycles under drought stress.

Expression of the Arabidopsis ABF4 gene in potato increases tuber yield, improves tuber quality and enhances salt and drought tolerance.

In this study we show that expression of the Arabidopsis ABF4 gene in potato increases tuber yield under normal and abiotic stress conditions, improves storage capability and processing quality of the tubers, and enhances salt and drought tolerance. Potato is the third most important food crop in the world. Potato plants are susceptible to salinity and drought, which negatively affect crop yield, tuber quality and market value. The development of new varieties with higher yields and increased tolerance to adverse environmental conditions is a main objective in potato breeding. In addition, tubers suffer from undesirable sprouting during storage that leads to major quality losses; therefore, the control of tuber sprouting is of considerable economic importance. ABF (ABRE-binding factor) proteins are bZIP transcription factors that regulate abscisic acid signaling during abiotic stress. ABF proteins also play an important role in the tuberization induction. We developed transgenic potato plants constitutively expressing the Arabidopsis ABF4 gene (35S::ABF4). In this study, we evaluated the performance of 35S::ABF4 plants grown in soil, determining different parameters related to tuber yield, tuber quality (carbohydrates content and sprouting behavior) and tolerance to salt and drought stress. Besides enhancing salt stress and drought tolerance, constitutive expression of ABF4 increases tuber yield under normal and stress conditions, enhances storage capability and improves the processing quality of the tubers.