Regulator Of Abscisic Acid Signaling Research Articles

MYB96 recruits the HDA15 protein to suppress negative regulators of ABA signaling in Arabidopsis

Unlike activation of target genes in response to abscisic acid (ABA), how MYB96 transcription factor represses ABA-repressible genes to further enhance ABA responses remains unknown. Here, we show MYB96 interacts with the histone modifier HDA15 to suppress negative regulators of early ABA signaling. The MYB96-HDA15 complex co-binds to the promoters of a subset of RHO GTPASE OF PLANTS (ROP) genes, ROP6, ROP10, and ROP11, and represses their expression by removing acetyl groups of histone H3 and H4 from the cognate regions, particularly in the presence of ABA. In support, HDA15-deficient mutants display reduced ABA sensitivity and are susceptible to drought stress with derepression of the ROP genes, as observed in the myb96-1 mutant. Biochemical and genetic analyses show that MYB96 and HDA15 are interdependent in the regulation of ROP suppression. Thus, MYB96 confers maximal ABA sensitivity by regulating both positive and negative regulators of ABA signaling through distinctive molecular mechanisms.

Differences in drought resistance in nine North American hybrid poplars

The physiological responses of nine commonly planted North-American hybrid poplars to drought was evaluated, resulting in a ranking of drought resistance. Indicator genes for drought response processes were scored for differences in expression levels in the most sensitive and resistant clones, confirming levels of drought stress and differences consistent with roles in drought resistance. Poplar hybrids are cultivated in North America for environmental applications, agroforestry, and the pulp and paper industry primarily because of their fast growth and limited nutrient requirement. For the same reasons, they have been identified as suitable species for carbon sequestration and as a potential feedstock for carbon–neutral production of energy. The clones deployed on the Canadian prairies are generally regarded as drought sensitive, which poses a problem as water availability has steadily decreased in this region over the past century and a severe water crisis has been predicted. To approach this problem, we tested nine commonly deployed North-American hybrid poplars, developed for large-scale cultivation in the Canadian prairies, for their physiological responses to drought, resulting in a ranking of drought resistance. The difference between the clones showing the most and the least response of drought stress was large, and we used these clones to further examine the differences in the expression of genes known to be up-regulated in response to drought stress. This interrogation showed significant differences in transcript abundance that largely reflected the physiological status of the tested clones, but also many genes being down rather than up-regulated in response to drought stress in the drought-tolerant clone. In particular, putative positive and negative regulators of abscisic acid signaling were expressed at levels consistent with a potential role in observed differences in drought resistance.

The non-DNA binding bHLH transcription factor Paclobutrazol Resistances are involved in the regulation of ABA and salt responses in Arabidopsis

Abscisic acid (ABA) is the key hormone that regulating plant responses to abiotic stresses. Several basic helix-loop-helix (bHLH) transcription factors have been reported to regulate ABA signaling in Arabidopsis. Paclobutrazol Resistances (PREs) are non-DNA binding bHLH transcription factors involved in the regulation of plant response to several different plant hormones including gibberellin, brassinosteroid and auxin. Here, we show that PREs are involved in the regulation of ABA and salt responses in Arabidopsis. Quantitative RT-PCR results showed that the expression levels of PRE6 as well as several other PRE genes were reduced in response to ABA treatment, but increased to salt treatment. Seed germination assays indicated that ABA sensitivity is reduced in the pre6 mutants, but increased in transgenic plants overexpressing PRE6. On the other hand, the 35S:PRE6 transgenic plants showed enhanced tolerance to salt, whereas little, if any changes were observed in the pre6 mutants. Similar responses to ABA and salt treatments were observed in the pre2 mutants and the transgenic plants overexpressing PRE2, and a slight increased resistance to ABA in seed germination was observed in the pre2 pre6 double mutants. Taken together, our results suggest that at least some of the PRE genes are ABA responsive genes, and PREs may function redundantly to regulate ABA and salt responses in Arabidopsis.

Salinity and ABA Seed Responses in Pepper: Expression and Interaction of ABA Core Signaling Components.

Abscisic acid (ABA) plays an important role in various aspects of plant growth and development, including adaptation to stresses, fruit development and ripening. In seeds, ABA participates through its core signaling components in dormancy instauration, longevity determination, and inhibition of germination in unfavorable environmental conditions such as high soil salinity. Here, we show that seed germination in pepper was delayed but only marginally reduced by ABA or NaCl with respect to control treatments. Through a similarity search, pepper orthologs of ABA core signaling components PYL (PYRABACTIN RESISTANCE1-LIKE), PP2C (PROTEIN PHOSPHATASE2C), and SnRK2 (SUCROSE NONFERMENTING1 (SNF1)-RELATED PROTEIN KINASE2) genes were identified. Gene expression analyses of selected members showed a low abundance of PYL and SnRK2 transcripts in dry seeds compared to other tissues, and an up-regulation at high concentrations of ABA and/or NaCl for both positive and negative regulators of ABA signaling. As expected, in hydroponically-grown seedlings exposed to NaCl, only PP2C encoding genes were up-regulated. Yeast two hybrid assays performed among putative pepper core components and with Arabidopsis thaliana orthologs confirmed the ability of the identified proteins to function in ABA signaling cascade, with the exception of a CaABI isoform cloned from seeds. BiFC assay in planta confirmed some of the interactions obtained in yeast. Altogether, our results indicate that a low expression of perception and signaling components in pepper seeds might contribute to explain the observed high percentages of seed germination in the presence of ABA. These results might have direct implications on the improvement of seed longevity and vigor, a bottleneck in pepper breeding.

Comparative transcriptome analysis of salt-sensitive and salt-tolerant maize reveals potential mechanisms to enhance salt resistance.

Salt stress is a devastating environmental stress that causes plant growth inhibition and yield reduction. The identification of salt-tolerant genes brings hope for the generation of salinity-tolerant crop plants through molecular breeding. In this study, one salt-sensitive and one salt-tolerant maize inbred line were screened from 242 maize inbred lines. Reactive oxygen species (ROS)-related enzyme activities were detected and salt-responsive comparative transcriptome analysis was performed for control and 220 mM NaCl treated maize leaves. Salt-tolerant maize inbred line (L87) showed higher ROS-related enzyme (SOD, POD, APX and CAT) activities and accumulated relatively lower levels of ROS under salt stress. Of the total DEGs, 1856 upregulated DEGs were specific to L87, including stress tolerance-related members of the 70kDa family of heat shock proteins (Hsp70s) and aquaporins. The DEGs involved in the abscisic acid (ABA), ethylene, jasmonic acid (JA) and salicylic acid (SA) signal transduction pathways may determine the difference in salt tolerance between the two varieties, especially one central component SnRK2, that positively regulates ABA signaling and was only upregulated in L87. Analysis of DEGs related to ROS scavenging showed that some peroxidase (POD), glutathione S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) genes specific to L87 probably enhanced its salt tolerance. The analysis of differentially expressed transcription factors (TFs) suggested that WRKY TFs could contribute to the difference in salt tolerance between the two maize lines. Compared with Salt-sensitive maize inbred line (L29), L87 exhibits specific regulatory mechanisms related to salt tolerance, including plant hormone interactions, ROS scavenging and the regulation of TFs. Our study identifies new candidate genes that may regulate maize tolerance to salt stress and provides useful information for breeding maize with high salt resistance.

Correction: Arabidopsis WRKY6 Transcription Factor Acts as a Positive Regulator of Abscisic Acid Signaling during Seed Germination and Early Seedling Development.

[This corrects the article DOI: 10.1371/journal.pgen.1005833.].

Reversible Histone H2B Monoubiquitination Fine-Tunes Abscisic Acid Signaling and Drought Response in Rice

Histone H2B monoubiquitination (H2Bub1) plays important roles in several physiological and developmental processes, but its roles in the regulation of plant stress responses remain elusive. Here, we report that H2Bub1 is crucially involved in abscisic acid (ABA) signaling and drought response in rice. We found that rice HISTONE MONOUBIQUITINATION2 (OsHUB2), an E3 ligase for H2Bub1, interacted with OsbZIP46, a key transcription factor regulating ABA signaling and drought response in rice. Genetic analyses suggest that OsHUB2, upregulated by drought and ABA, positively modulates ABA sensitivity and drought resistance. The H2Bub1 levels were increased in the target genes of OsbZIP46 under the drought stress and ABA treatments, which were positively correlated with their increased expression levels. Interestingly, MODD, a reported suppressor of ABA signaling and drought resistance by mediating OsbZIP46 deactivation and degradation, could reduce the H2Bub1 levels in the target genes of OsbZIP46 by recruiting a putative deubiquitinase OsOTLD1. Suppression of OsOTLD1 in vivo resulted in increased H2Bub1 levels and expression of OsbZIP46 target genes. Collectively, these findings established an elaborate mechanism of histone monoubiquitination in the fine-turning of ABA signaling and drought response by balancing H2Bub1 deposition and removal.

Abscisic Acid Inhibits Rice Protein Phosphatase PP45 via H2O2 and Relieves Repression of the Ca2+/CaM-Dependent Protein Kinase DMI3.

In plants, Ca2+/calmodulin-dependent protein kinase (CCaMK) is a positive regulator of abscisic acid (ABA) responses, including root growth, antioxidant defense, and tolerance of both water stress and oxidative stress. However, the underlying molecular mechanisms are poorly understood. Here, we show a direct interaction between DMI3 (Doesn't Make Infections 3), a rice (Oryza sativa) CCaMK and PP45, a type 2C protein phosphatase in rice (PP2C). This interaction involves the CaM binding domain of DMI3 and the PP2C domain of PP45. In the absence of ABA, PP45 directly inactivates DMI3 by dephosphorylating Thr-263 in DMI3. However, in the presence of ABA, ABA-induced H2O2 production by the NADPH oxidases RbohB/E inhibits the activity of PP45 not only by inhibiting the expression of PP45 but also by oxidizing Cys-350 and Cys-428 residues to form PP45 intermolecular dimers. ABA-induced oxidation of Cys-350 and Cys-428 in PP45 blocked the interaction between PP45 and DMI3 and substantially prevented PP45-mediated inhibition in DMI3 activity. Genetic analysis indicated that PP45 is an important negative regulator of ABA signaling. These results reveal important pathways for the inhibition of DMI3 under the basal state and for its ABA-induced activation in rice.

Hydrogen peroxide plays an important role in PERK4-mediated abscisic acid-regulated root growth in Arabidopsis.

Abscisic acid (ABA) is a crucial factor that affects primary root tip growth in plants. Previous research suggests that reactive oxygen species (ROS), especially hydrogen peroxide, are important regulators of ABA signalling in root growth of Arabidopsis. PROLINE-RICH EXTENSIN-LIKE RECEPTOR KINASE 4 (PERK4) plays an important role in ABA responses. Arabidopsis perk4 mutants display attenuated sensitivity to ABA, especially in primary root growth. To gain insights into the mechanism(s) of PERK4-associated ABA inhibition of root growth, in this study we investigated the involvement of ROS in this process. Normal ROS accumulation in the primary root in response to exogenous ABA treatment was not observed in perk4 mutants. PERK4 deficiency prohibits ABA-induced expression of RESPIRATORY BURST OXIDASE HOMOLOGUE (RBOH) genes, therefore the perk4-1 mutant showed decreased production of ROS in the root. The perk4-1/rbohc double mutant displayed the same phenotype as the perk4 and rbohc single mutants in response to exogenous ABA treatment. The results suggest that PERK4-stimulated ROS accumulation during ABA-regulated primary root growth may be mediated by RBOHC.

Arabidopsis small ubiquitin-related modifier protease ASP1 positively regulates abscisic acid signaling during early seedling development.

The small ubiquitin-related modifier (SUMO) modification plays an important role in the regulation of abscisic acid (ABA) signaling, but the function of the SUMO protease, in ABA signaling, remains largely unknown. Here, we show that the SUMO protease, ASP1 positively regulates ABA signaling. Mutations in ASP1 resulted in an ABA-insensitive phenotype, during early seedling development. Wild-type ASP1 successfully rescued, whereas an ASP1 mutant (C577S), defective in SUMO protease activity, failed to rescue, the ABA-insensitive phenotype of asp1-1. Expression of ABI5 and MYB30 target genes was attenuated in asp1-1 and our genetic analyses revealed that ASP1 may function upstream of ABI5 and MYB30. Interestingly, ASP1 accumulated upon ABA treatment, and ABA-induced accumulation of ABI5 (a positive regulator of ABA signaling) was abolished, whereas ABA-induced accumulation of MYB30 (a negative regulator of ABA signaling) was increased in asp1-1. These findings support the hypothesis that increased levels of ASP1, upon ABA treatment, tilt the balance between ABI5 and MYB30 towards ABI5-mediated ABA signaling.

The Ubiquitin E3 Ligase RHA2b Promotes Degradation of MYB30 in Abscisic Acid Signaling.

The phytohormone abscisic acid (ABA) is critical for plants encountering abiotic stress. We reported previously that the Arabidopsis (Arabidopsis thaliana) transcription factor MYB30 participates in ABA responses via SUMO ligase SAP-MIZ Domain-Containing SIZ1-mediated sumoylation. Here, we show that the RING-type ubiquitin E3 ligase RHA2b, which positively regulates ABA signaling, interacted with and ubiquitinated MYB30 to modulate MYB30 stability through the 26S proteasome pathway. The degradation rate of MYB30 was repressed significantly in the rha2b-1 mutant. Phenotypic analyses showed that MYB30 acts genetically downstream of RHA2b in ABA signaling. Substitutions of lysine-283 (K283) and K165 blocked ubiquitination, suggesting that these residues are sites of ubiquitination. K283 residue substitution significantly inhibited the degradation of MYB30 induced by ABA. The K165 site functioned additively with K283 in ABA-induced MYB30 degradation and ABA responses. At the same time, sumoylation protected MYB30 from degradation under cycloheximide and ABA treatment. Compared with MYB30, overexpression of MYB30-SUMO1 partially recovered the ABA sensitivity of siz1-2 But MYB30-SUMO1 exhibited similar localization with MYB30 in nuclei. Overall, our results suggest that RHA2b targets MYB30 for degradation to modulate ABA signaling. Considering that the K283 residue also is the major site for sumoylation, we propose that sumoylation and ubiquitination act antagonistically in the ABA response to regulate the stability of MYB30 by occupying the same residue.

CARK1 mediates ABA signaling by phosphorylation of ABA receptors

The function of abscisic acid (ABA) is mediated by its receptors termed RCARs/PYR1/PYLs. Modulation of ABA signaling is vital for plant growth and development. The RCAR-PP2C-SnRK2 regulatory modules have been defined as the core components in ABA signaling. However, it is still not clear whether and how the ABA receptors could be modified at the initial post-translational stage to fine-tune ABA transduction pathway. Here we identify and characterize the putative receptor-like cytoplasmic kinase (RLCK) in Arabidopsis named CARK1, which interacts with RCAR3 (PYL8) and RCAR11 (PYR1) in the manner of phosphorylation. Structural studies of CARK1 revealed the critical active site, N204, which accounts for the kinase activity and the direct interaction with RCAR3/RCAR11. CARK1 phosphorylates RCAR3/RCAR11 at one conserved threonine site, T77/T78. Our genetic analyses further demonstrated that CARK1 positively regulates ABA-mediated physiological responses and overexpression of CARK1 in Arabidopsis distinctly promotes the drought resistance. Moreover, the phosphor-mimic form of RCAR11 in the cark1 mutant is able to functionally complement the ABA sensitivity. CARK1 positively regulates ABA-responsive gene expression and enhances RCAR3/RCAR11’s inhibition to Clade A PP2C. Taken together, our studies strongly support the functional significance of CARK1 in positively regulating ABA signaling via phosphorylation on RCAR3/RCAR11 in Arabidopsis.

A Tonoplast-Associated Calcium-Signaling Module Dampens ABA Signaling during Stomatal Movement.

Stomatal movement, critical for photobiosynthesis, respiration, and stress responses, is regulated by many factors, among which abscisic acid (ABA) is critical. Early events of ABA signaling involve Ca2+ influx and an increase of cytoplasmic calcium ([Ca2+]cyt). Positive regulators of this process have been extensively studied, whereas negative regulators are obscure. ABA-induced stomatal closure involves K+ flux and vacuolar convolution. How these processes are connected with Ca2+ is not fully understood. We report that pat10-1, a null mutant of Arabidopsis (Arabidopsis thaliana) PROTEIN S-ACYL TRANSFERASE10 (PAT10), is hypersensitive to ABA-induced stomatal closure and vacuolar convolution. A similar phenotype was observed in cbl2;cbl3, the double mutant of CBL2 and CBL3, whose tonoplast association depends on PAT10. Functional loss of the PAT10-CBL2/CBL3 system resulted in enhanced Ca2+ influx and [Ca2+]cyt elevation. Promoting vacuolar K+ accumulation by overexpressing NHX2 suppressed ABA-hypersensitive stomatal closure and vacuolar convolution of the mutants, suggesting that PAT10-CBL2/CBL3 positively mediates vacuolar K+ accumulation. We have identified CBL-interacting protein kinases (CIPKs) that mediate CBL2/CBL3 signaling during ABA-induced stomatal movement. Functional loss of the PAT10-CBL2/3-CIPK9/17 system in guard cells enhanced drought tolerance. We propose that the tonoplast CBL-CIPK complexes form a signaling module that negatively regulates ABA signaling during stomatal movement.

Arabidopsis subtilase SASP is involved in the regulation of ABA signaling and drought tolerance by interacting with OPEN STOMATA 1.

Arabidopsis Senescence-Associated Subtilisin Protease (SASP) has previously been reported to participate in leaf senescence and in the development of inflorescences and siliques. Here, we describe a new role of SASP in the regulation of abscisic acid (ABA) signaling. SASP encodes a subtilase and its expression was considerably induced by darkness, ABA, and ethylene treatments. sasp knockout mutants displayed obvious developmental phenotypes such as early flowering and smaller leaves. In particular, the sasp mutants exhibited enhanced ABA sensitivity during seed germination and seedling growth, heightened ABA-mediated leaf senescence, and increased production of reactive oxygen species (ROS). Importantly, the sasp mutants also showed remarkably increased tolerance to drought, with expression of six ABA signaling-related genes being either up- or down-regulated following ABA treatment. Interaction assays demonstrated that SASP physically interacts with OPEN STOMATA 1 (OST1) at the cell periphery. Co-expression of SASP and OST1 led to degradation of OST1, whereas this degradation was impaired in sasp-1 protoplasts. ROS attenuation assays demonstrated that in sasp-1 mutant guard cells the attenuation rate markedly decreased. Taken together, the results demonstrate that SASP plays an important role in regulating ABA signaling and drought tolerance through interaction with OST1.

Genome Analysis of the Ancient Tracheophyte Selaginella tamariscina Reveals Evolutionary Features Relevant to the Acquisition of Desiccation Tolerance

Resurrection plants, which are the “gifts” of natural evolution, are ideal models for studying the genetic basis of plant desiccation tolerance. Here, we report a high-quality genome assembly of 301 Mb for the diploid spike moss Selaginella tamariscina, a primitive vascular resurrection plant. We predicated 27 761 protein-coding genes from the assembled S. tamariscina genome, 11.38% (2363) of which showed significant expression changes in response to desiccation. Approximately 60.58% of the S. tamariscina genome was annotated as repetitive DNA, which is an almost 2-fold increase of that in the genome of desiccation-sensitive Selaginella moellendorffii. Genomic and transcriptomic analyses highlight the unique evolution and complex regulations of the desiccation response in S. tamariscina, including species-specific expansion of the oleosin and pentatricopeptide repeat gene families, unique genes and pathways for reactive oxygen species generation and scavenging, and enhanced abscisic acid (ABA) biosynthesis and potentially distinct regulation of ABA signaling and response. Comparative analysis of chloroplast genomes of several Selaginella species revealed a unique structural rearrangement and the complete loss of chloroplast NAD(P)H dehydrogenase (NDH) genes in S. tamariscina, suggesting a link between the absence of the NDH complex and desiccation tolerance. Taken together, our comparative genomic and transcriptomic analyses reveal common and species-specific desiccation tolerance strategies in S. tamariscina, providing significant insights into the desiccation tolerance mechanism and the evolution of resurrection plants.

A DEAD-box RNA helicase, RH8, is critical for regulation of ABA signalling and the drought stress response via inhibition of PP2CA activity.

Abscisic acid (ABA) is major plant hormone involved in regulating abiotic stress responses. Several studies have established that an ABA-signalling transduction pathway-from ABA perception to response-functions in plant cells. The group A PP2Cs constitute core components of ABA signalling, and they negatively regulate ABA signalling and stress responses. Recent studies have identified and functionally analysed regulators of PP2C activity; however, the precise regulatory mechanisms remain unclear. In the present study, we used a yeast 2-hybrid (Y2H) screening analysis to identify the DEAD-box RNA helicase RH8, which interacted with PP2CA in the nucleus. rh8 knockout mutants exhibited ABA hyposensitivity and drought-susceptible phenotypes characterized by high levels of transpirational water loss via reduced stomatal closure and decreased leaf temperatures. However, rh8/pp2ca double mutants showed ABA hypersensitivity and drought-tolerant phenotypes, indicating that RH8 and PP2CA function in the same ABA-signalling pathway in the drought stress response; moreover, RH8 functions upstream of PP2CA. In vitro phosphatase and kinase assays revealed that RH8 inhibits PP2CA phosphatase activity. Our data indicate that RH8 and its interacting partner PP2CA modulate the drought stress response via ABA-dependent signalling.

Banana MaABI5 is involved in ABA-induced cold tolerance through interaction with a RING E3 ubiquitin ligase, MaC3HC4-1

Abstract Abscisic acid (ABA) is a phytohormone essential for the adaptation of plants to environmental stresses, and transcription factor abscisic acid-insensitive 5 (ABI5) is a key regulator of ABA signaling and response. Although exogenous application of ABA is effective in retarding chilling injury (CI) symptoms of banana fruit, the possible roles of ABI5 in ABA-induced chilling tolerance in banana fruit remain largely unknown. In this study, ABA treatment significantly induced chilling tolerance in banana fruit, as indicated by the changes in the CI index, peel color, and relative electrolyte leakage. Moreover, MaABI5, a gene encoding banana ABI5, was induced by ABA treatment during low-temperature storage. Importantly, through yeast two-hybrid (Y2H) screening, MaABI5 was found to interact with MaC3HC4-1, a C3HC4-type RING finger E3 ubiquitin ligase. Expression of MaC3HC4-1 was downregulated by ABA application. Furthermore, MaC3HC4-1 possessed intrinsic ubiquitin ligase activity in vitro. The predicted MaABI5–MaC3HC4-1 network will help decipher the possible role of ABA in the cold stress signaling pathway of banana fruit.

EAR1 Negatively Regulates ABA Signaling by Enhancing 2C Protein Phosphatase Activity.

The reversible phosphorylation of proteins by kinases and phosphatases is an antagonistic process that modulates many cellular functions. Protein phosphatases are usually negatively regulated by inhibitor proteins. During abscisic acid (ABA) signaling, these inhibitor proteins comprise PYR1/PYL/RCAR ABA receptors, which inhibit the core negative regulators, the clade A type 2C protein phosphatases (PP2Cs). However, it is not known whether these PP2Cs are positively regulated by other proteins. Here, we identified an Arabidopsis thaliana ear1 (enhancer of aba co-receptor1) mutant that exhibits pleiotropic ABA-hypersensitive phenotypes. EAR1 encodes an uncharacterized protein that is conserved in both monocots and dicots. EAR1 interacts with the N-terminal inhibition domains of all six PP2Cs, ABA INSENSITIVE1 (ABI1), ABI2, HYPERSENSITIVE TO ABA1 (HAB1), HAB2, ABA-HYPERSENSITIVE GERMINATION1 (AHG1), and AHG3, during ABA signaling and enhances the activity of PP2Cs both in vitro and in vivo. ABA treatment caused EAR1 to accumulate in the nucleus. These results indicate that EAR1 is a negative regulator of ABA signaling that enhances the activity of PP2Cs by interacting with and releasing the N-terminal autoinhibition of these proteins.

Chemical Control of ABA Receptors to Enable Plant Protection Against Water Stress.

This chapter describes procedures to find small molecules that bind to abscisic acid (ABA) receptors and improve plant tolerance to water stress. Abscisic acid promotes the interaction between ABA receptors and protein phosphatase 2Cs (PP2Cs), which are negative regulators of ABA signaling. This receptor-mediated ABA-dependent inhibition of PP2C activity is required for ABA action in vivo. ABA agonists can be identified by high-throughput screening for molecules that promote agonist-induced ABA receptor-PP2C interactions using yeast two-hybrid assays. In addition to in vitro assays, an in vivo test to evaluate ABA agonist activity has been devised in which an ABA-inducible GUS reporter strain is used to evaluate the effect of each ABA agonist. The effects of ABA agonists can also be determined using thermal imaging analyses and a water loss assay of stomata. Finally, the ability of each ABA agonist to confer drought stress tolerance upon plants grown in soil is determined. These methods can be used to identify various ABA agonists that protect plants against water stress.

The sHSP22 Heat Shock Protein Requires the ABI1 Protein Phosphatase to Modulate Polar Auxin Transport and Downstream Responses.

The phytohormones abscisic acid (ABA) and indole-3-acetic acid (IAA) response pathways interact synergistically or antagonistically to regulate plant development and environmental adaptation. Here, we show that ABI1, a key negative regulator of ABA signaling, is essential for auxin-modulated root development. We performed a microarray analysis using the loss-of-function mutant abi1-3 and Col-0 seedlings treated with IAA. For sHSP22, an endoplasmic reticulum (ER) small heat shock protein-encoding gene, the induction by IAA was dependent on ABI1shsp22 displayed enhanced sensitivity to ABA in primary root growth. In contrast, overexpression of full-length, but not truncated sHSP22 lacking signal peptide and ER-retention sequences, resulted in decreased ABA sensitivity. Overexpressed (OX) sHSP22 partially rescued the ABA hypersensitivity of abi1-3 In addition, sHSP22 is involved in auxin-regulated hypocotyl elongation at high temperature treatment. sHSP22 also affected accumulation of auxin efflux carrier PIN proteins due to potentiated intracellular trafficking. And sHSP22 OX lines initiated more lateral roots after auxin application. Our results suggest that sHSP22 regulates auxin response through modulating auxin polar transport, and ABI1-sHSP22 provides a novel module orchestrating ABA and auxin signaling crosstalk in Arabidopsis (Arabidopsis thaliana).