Identification and Analysis of Genes Involved in Auxin, Abscisic Acid, Gibberellin, and Brassinosteroid Metabolisms Under Drought Stress in Tender Shoots of Tea Plants.

Endogenous phytohormones auxin (indole-3-acetic acid [IAA]), abscisic acid (ABA), gibberellin (GA3), and brassinosteroid (BR) play a role in responses to drought stress in higher plants. Tea plant is one of the major economic corps worldwide. The tender shoots of tea plants are the main source for tea production. The effects of drought stress on endogenous IAA, ABA, GA3, and BR metabolisms in tender shoots of tea plants need to be illustrated. In this study, a total of 17 IAA-related genes, 17 ABA-related genes, 18 GA3-related genes, and 8 BR-related genes were identified under drought stress in tender shoots of tea plants, respectively. By using a combination of phytohormone determination, phylogenetic tree construction and sequence analysis, gene expression profiles, functional classification, Kyoto encyclopedia of genes and genomes enrichment, and distribution of genes analysis, we have demonstrated that IAA, ABA, GA3, and BR metabolisms might participate in the regulation of the response to drought stress in tender shoots of tea plants. The expression level of CsLYCE negatively correlated with ABA accumulation under drought stress. Our findings could shed new light on the effects of drought stress on the IAA, ABA, GA3, and BR metabolisms in tender shoots of tea plants.

Preharvest UV-C treatment affected postharvest senescence and phytochemicals alternation of strawberry fruit with the possible involvement of abscisic acid regulation.

As an environmentally friendly approach for fruit quality improvement, the effect of preharvest UV-C on the physiology of strawberry fruit during postharvest storage remains to be assessed. Strawberry fruit developed with supplementary UV-C were stored at room temperature for 2 weeks. Preharvest UV-C attenuated fruit postharvest senescence and altered phytochemicals composition. Higher ester titer was found in the treated fruit at harvest, whereas higher terpene and furanone contents were detected after 72 h of storage. At harvest, polyphenolics accumulated to a higher level in UV-C group, but the difference disappeared after 24 h of storage. Meanwhile, the intrinsic level of abscisic acid and the expressions of FaPYR1, SnRK2, and FaASR in the UV-C-treated fruit was enhanced at harvest but returned to a lower level as storage proceeded. This study highlights the time-dependent effect of preharvest UV-C on strawberry fruit postharvest biochemical indexes and the possible involvement of abscisic acid signaling factors.

DRL1, Encoding A NAC Transcription Factor, Is Involved in Leaf Senescence in Grapevine.

The NAC (for NAM, ATAF1,2, and CUC2) proteins family are plant-specific transcription factors, which play important roles in leaf development and response to environmental stresses. In this study, an NAC gene, DRL1, isolated from grapevine Vitis vinifera L. "Yatomi Rose", was shown to be involved in leaf senescence. The quantity of DRL1 transcripts decreased with advancing leaf senescence in grapevine. Overexpressing the DRL1 gene in tobacco plants significantly delayed leaf senescence with respect to chlorophyll concentration, potential quantum efficiency of photosystem II (Fv/Fm), and ion leakage. Moreover, exogenous abscisic acid (ABA) markedly reduced the expression of DRL1, and the ABA and salicylic acid (SA) concentration was lower in the DRL1-overexpressing transgenic plants than in the wild-type plants. The DRL1 transgenic plants exhibited reduced sensitivity to ABA-induced senescence but no significant change in the sensitivity to jasmonic acid-, SA- or ethylene-induced senescence. Transcriptomic analysis and RNA expression studies also indicated that the transcript abundance of genes associated with ABA biosynthesis and regulation, including 9-cis-epoxycarotenoid dioxygenase (NCED1), NCED5, zeaxanthin epoxidase1 (ZEP1), ABA DEFICIENT2 (ABA2), ABA4, and ABA INSENSITIVE 2 (ABI2), was markedly reduced in the DRL1-overexpressing plants. These results suggested that DRL1 plays a role as a negative regulator of leaf senescence by regulating ABA synthesis.

Analysis of eight phytohormone concentrations, expression levels of ABA biosynthesis genes, and ripening-related transcription factors during fruit development in strawberry.

The contents of eight phytohormones and the expression levels of genes encoding enzymes related to abscisic acid (ABA) biosynthesis and deactivation/degradation and transcription factors (TFs) related to fruit ripening were studied in the non-climacteric strawberry fruit (Fragaria × ananassa Duch., cv. 'Seolhyang') at six developmental stages. The hormones tested were ABA, indole-3-acetic acid (IAA), gibberellic acid 4 (GA4), jasmonic acid (JA), methyljasmonate (MJ), jasmonoyl isoleucine (JA-Ile), salicylic acid (SA), and ethylene (ET). The developmental and ripening stages studied were small green (S1, 11 days post-anthesis, DPA), green (S2, 20 DPA), breaker (S3, 24 DPA), pink (S4, 27 DPA), red (S5, 31 DPA), and fully red (S6, 40 DPA). IAA and GA4 contents were highest at S1 and gradually decreased after this stage. ABA content was low at S1-S3 and then increased rapidly until peaking at S6. By contrast, MJ content showed no significant changes over time, while SA content gradually increased. JA, JA-Ile, and ET contents were either insufficient for quantification or undetectable. Expression of the ABA biosynthesis genes FaNCED1 and FaABA2 increased during fruit ripening, whereas expression of the ABA deactivation/degradation genes FaUGT75C1 and FaCYP707A1 was high early in development, when ABA content was low, and then decreased. Among four ripening-related TF genes, FaMYB1, FaMYB5, FaMYB10, and FaASR, only the expression of FaMYB10 seemed to be closely related to strawberry fruit ripening. Our study supports the idea that ABA and FaMYB10 appear to be the key hormone and TF regulating strawberry ripening.

ZmASR3 from the Maize ASR Gene Family Positively Regulates Drought Tolerance in Transgenic Arabidopsis.

Abscisic acid (ABA)-, stress-, and ripening-induced (ASR) proteins are reported to be involved in drought stress responses. However, the function of maize ASR genes in enhancing drought tolerance is not known. Here, nine maize ASR members were cloned, and the molecular features of these genes were analyzed. Phenotype results of overexpression of maize ZmASR3 gene in Arabidopsis showed lower malondialdehyde (MDA) levels and higher relative water content (RWC) and proline content than the wild type under drought conditions, demonstrating that ZmASR3 can improve drought tolerance. Further experiments showed that ZmASR3-overexpressing transgenic lines displayed increased stomatal closure and reduced reactive oxygen species (ROS) accumulation by increasing the enzyme activities of superoxide dismutase (SOD) and catalase (CAT) under drought conditions. Moreover, overexpression of ZmASR3 in Arabidopsis increased ABA content and reduced sensitivity to exogenous ABA in both the germination and post-germination stages. In addition, the ROS-related, stress-responsive, and ABA-dependent pathway genes were activated in transgenic lines under drought stress. Taken together, these results suggest that ZmASR3 acts as a positive regulator of drought tolerance in plants.

Differential Function of Endogenous and Exogenous Abscisic Acid during Bacterial Pattern-Induced Production of Reactive Oxygen Species in Arabidopsis.

Abscisic acid (ABA) plays important roles in positively or negatively regulating plant disease resistance to pathogens. Here, we reassess the role of endogenous and exogenous ABA by using: 35S::ABA2, a previously reported transgenic Arabidopsis line with increased endogenous ABA levels; aba2-1, a previously reported ABA2 mutant with reduced endogenous ABA levels; and exogenous application of ABA. We found that bacterial susceptibility promoted by exogenous ABA was suppressed in 35S::ABA2 plants. The 35S::ABA2 and aba2-1 plants displayed elevated and reduced levels, respectively, of bacterial flagellin peptide (flg22)-induced H2O2. Surprisingly, ABA pre-treatment reduced flg22-induced H2O2 generation. Exogenous, but not endogenous ABA, increased catalase activity. Loss of nicotinamide adenine dinucleotide phosphate oxidase genes, RBOHD and RBOHF, restored exogenous ABA-promoted bacterial susceptibility of 35S::ABA2 transgenic plants. In addition, endogenous and exogenous ABA had similar effects on callose deposition and salicylic acid (SA) signaling. These results reveal an underlying difference between endogenous and exogenous ABA in regulating plant defense responses. Given that some plant pathogens are able to synthesize ABA and affect endogenous ABA levels in plants, our results highlight the importance of reactive oxygen species in the dual function of ABA during plant-pathogen interactions.

Hormone and RNA-seq analyses reveal the mechanisms underlying differences in seed vigour at different maize ear positions.

KEY MESSAGE: ABA/GA4 ratio, stress resistance, carbon and nitrogen metabolism, and chromatin structure play important roles in vigour differences of seeds located at different maize ear positions. Seed vigour, which ensures rapid and uniform field emergence across diverse environments, differs at different maize ear positions. However, little is known regarding the associated mechanisms. In this study, we determined that seed vigour, stress resistance, and carbon and nitrogen metabolism were higher in seeds from middle and bottom section of the ear, while the ABA/GA4 ratio in the embryos was significantly lower. Compared with the seeds subjected to repeated pollination during silking, less variation in seed vigour and the ABA/GA4 ratio in the embryos was observed in seeds at different ear positions subjected to single pollination after complete silking. This indicated that single pollination can reduce, but not eliminate, the differences in seed vigour at different ear positions. RNA-seq analysis indicated that the seed vigour differences at the different locations of the maize ears of the single pollinated treatment were related to carbon and nitrogen metabolism. In contrast, the differences in seed vigour under repeated pollination were related to chromatin structure. The present study contributes to our understanding of the mechanisms underlying differences in seed vigour at different positions on the maize ear.

Genome-Wide Identification and Characterization of the AREB/ABF/ABI5 Subfamily Members from Solanum tuberosum.

Abscisic acid (ABA) plays crucial roles in plant development and adaption to environmental stresses. The ABA-responsive element binding protein/ABRE-binding factor and ABA INSENSITIVE 5 (AREB/ABF/ABI5) gene subfamily members, which belong to the basic domain/leucine zipper (bZIP) transcription factors family, participate in the ABA-mediated signaling pathway by regulating the expression of their target genes. However, information about potato (Solanum tuberosum) AREB/ABF/ABI5 subfamily members remains scarce. Here, seven putative AREB/ABF/ABI5 members were identified in the potato genome. Sequences alignment revealed that these members shared high protein sequence similarity, especially in the bZIP region, indicating that they might possess overlapping roles in regulating gene expression. Subcellular localization analysis illustrated that all seven AREB/ABF/ABI5 members were localized in the nucleus. Transactivation activity assays in yeast demonstrated that these AREB/ABF/ABI5 members possessed distinct transcriptional activity. Electrophoretic mobility shift assays (EMSA) confirmed that all of these AREB/ABF/ABI5 members could have an affinity to ABRE in vitro. The expression patterns of these AREB/ABF/ABI5 genes showed that they were in response to ABA or osmotic stresses in varying degrees. Moreover, most AREB/ABF/ABI5 genes were induced during stolon swelling. Overall, these results provide the first comprehensive identification of the potato AREB/ABF/ABI5 subfamily and would facilitate further functional characterization of these subfamily members in future work.

The regulatory mechanism of chilling-induced dormancy transition from endo-dormancy to non-dormancy in Polygonatum kingianum Coll.et Hemsl rhizome bud.

KEY MESSAGE: We identified three dormant stages of Polygonatum kingianum and changes that occurred during dormancy transition in the following aspects including cell wall and hormones, as well as interaction among them. Polygonatum kingianum Coll.et Hemsl (P. kingianum) is an important traditional Chinese medicine, but the mechanism of its rhizome bud dormancy has not yet been studied systematically. In this study, three dormancy phases were induced under controlled conditions, and changes occurring during the transition were examined, focusing on phytohormones and the cell wall. As revealed by HPLC-MS (High Performance Liquid Chromatography-Mass Spectrometry) analysis, the endo- to non-dormancy transition was association with a reduced abscisic acid (ABA)/gibberellin (GA3) ratio, a decreased level of auxin (IAA) and an increased level of trans-zeatin (tZR). Transmission electron microscopy showed that plasmodesmata (PDs) and the cell wall of the bud underwent significant changes between endo- and eco-dormancy. A total of 95,462 differentially expressed genes (DEGs) were identified based on transcriptomics, and clustering and principal component analysis confirmed the different physiological statuses of the three types of bud samples. Changes in the abundance of transcripts associated with IAA, cytokinins (CTKs), GA, ABA, brassinolide (BR), jasmonic acid (JA), ethylene, salicylic acid (SA), PDs and cell wall-loosening factors were analysed during the bud dormancy transition in P. kingianum. Furthermore, nitrilase 4 (NIT4) and tryptophan synthase alpha chain (TSA1), which are related to IAA synthesis, were identified as hub genes of the co-expression network, and strong interactions between hormones and cell wall-related factors were observed. This research will provide a good model for chilling-treated rhizome bud dormancy in P. kingianum and cultivation of this plant.

Abscisic acid prevents pollen abortion under high-temperature stress by mediating sugar metabolism in rice spikelets.

Heat stress at the pollen mother cell (PMC) meiotic stage leads to pollen sterility in rice, in which the reactive oxygen species (ROS) and sugar homeostasis are always adversely affected. This damage is reversed by abscisic acid (ABA), but the mechanisms underlying the interactions among the ABA, sugar metabolism, ROS and heat shock proteins in rice spikelets under heat stress are unclear. Two rice genotypes, Zhefu802 (a recurrent parent) and fgl (its near-isogenic line) were subjected to heat stress of 40°C after pre-foliage sprayed with ABA and its biosynthetic inhibitor fluridone at the meiotic stage of PMC. The results revealed that exogenous application of ABA reduced pollen sterility caused by heat stress. This was achieved through various means, including: increased levels of soluble sugars, starch and non-structural carbohydrates, markedly higher relative expression levels of heat shock proteins (HSP24.1 and HSP71.1) and genes related to sugar metabolism and transport, such as sucrose transporters (SUT) genes, sucrose synthase (SUS) genes and invertase (INV) genes as well as increased antioxidant activities and increased content of adenosine triphosphate and endogenous ABA in spikelets. In short, exogenous application of ABA prior to heat stress enhanced sucrose transport and accelerated sucrose metabolism to maintain the carbon balance and energy homeostasis, thus ABA contributed to heat tolerance in rice.

Modulation of ABA responses by the protein kinase WNK8.

Abscisic acid (ABA) regulates growth and developmental processes in response to limiting water conditions. ABA functions through a core signaling pathway consisting of PYR1/PYL/RCAR ABA receptors, type 2C protein phosphatases (PP2Cs), and SnRK2-type protein kinases. Other signaling modules might converge with ABA signals through the modulation of core ABA signaling components. We have investigated the role of the protein kinase WNK8 in ABA signaling. WNK8 interacted with PP2CA and PYR1, phosphorylated PYR1 in vitro, and was dephosphorylated by PP2CA. A hypermorphic wnk8-ct Arabidopsis mutant allele suppressed ABA and glucose hypersensitivities of pp2ca-1 mutants during young seedling development, and WNK8 expression in protoplasts suppressed ABA-induced reporter gene expression. We conclude that WNK8 functions as a negative modulator of ABA signaling.

Arabidopsis thaliana NGATHA1 transcription factor induces ABA biosynthesis by activating NCED3 gene during dehydration stress.

The plant hormone abscisic acid (ABA) is accumulated after drought stress and plays critical roles in the responses to drought stress in plants, such as gene regulation, stomatal closure, seed maturation, and dormancy. Although previous reports revealed detailed molecular roles of ABA in stress responses, the factors that contribute to the drought-stress responses-in particular, regulation of ABA accumulation-remain unclear. The enzyme NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3) is essential for ABA biosynthesis during drought stress, and the NCED3 gene is highly induced by drought stress. In the present study, we isolated NGATHAs (NGAs) as candidate transcriptional regulators of NCED3 through a screen of a plant library harboring the transcription factors fused to a chimeric repressor domain, SRDX. The NGA proteins were directly bound to a cis-element NGA-binding element (NBE) in the 5' untranslated region (5' UTR) of the NCED3 promoter and were suggested to be transcriptional activators of NCED3 Among the single-knockout mutants of four NGA family genes, we found that the NGATHA1 (NGA1) knockout mutant was drought-stress-sensitive with a decreased expression level of NCED3 during dehydration stress. These results suggested that NGA1 essentially functions as a transcriptional activator of NCED3 among the NGA family proteins. Moreover, the NGA1 protein was degraded under nonstressed conditions, and dehydration stress enhanced the accumulation of NGA1 proteins, even in ABA-deficient mutant plants, indicating that there should be ABA-independent posttranslational regulations. These findings emphasize the regulatory mechanisms of ABA biosynthesis during early drought stress.

Identification and Expression Analyses of SBP-Box Genes Reveal Their Involvement in Abiotic Stress and Hormone Response in Tea Plant (Camellia sinensis).

The SQUAMOSA promoter binding protein (SBP)-box gene family is a plant-specific transcription factor family. This family plays a crucial role in plant growth and development. In this study, 20 SBP-box genes were identified in the tea plant genome and classified into six groups. The genes in each group shared similar exon-intron structures and motif positions. Expression pattern analyses in five different tissues demonstrated that expression in the buds and leaves was higher than that in other tissues. The cis-elements and expression patterns of the CsSBP genes suggested that the CsSBP genes play active roles in abiotic stress responses; these responses may depend on the abscisic acid (ABA), gibberellic acid (GA), and methyl jasmonate (MeJA) signaling pathways. Our work provides a comprehensive understanding of the CsSBP family and will aid in genetically improving tea plants.

Transcriptomic analysis of drought stress responses of sea buckthorn (Hippophae rhamnoidessubsp. sinensis) by RNA-Seq.

Sea buckthorn is one of the most important eco-economic tree species in China due to its ability to grow and produce acceptable yields under limited water and fertilizer availability. In this study, the differentially expressed genes under drought stress (DS) of sea buckthorn were identified and compared with control (CK) by RNA-Seq. A total of 122,803 unigenes were identified in sea buckthorn, and 70,025 unigenes significantly matched a sequence in at least one of the seven databases. A total of 24,060 (19.59%) unigenes can be assigned to 19 KEGG pathways, and 1,644 unigenes were differentially expressed between DS and CK, of which 519 unigenes were up-regulated and 1,125 unigenes down-regulated. Of the 47 significantly enriched GO terms, 14, 7 and 26 items were related to BP, CC and MF, respectively. KEGG enrichment analysis showed 398 DEGs involved in 97 different pathways, of which 119 DEGs were up-regulated and 279 DEGs were down-regulated under drought stress. In addition, we found 4438 transcriptor factors (TFs) in sea buckthorn, of which 100 were differentially expressed between DS and CK. These results lay a first foundation for further investigations of the very specific functions of these unigenes in sea buckthorn in response to drought stress.

Genetic and Physio-Biochemical Characterization of a Novel Premature Senescence Leaf Mutant in Rice (Oryza sativa L.).

Premature senescence greatly affects the yield production and the grain quality in plants, although the molecular mechanisms are largely unknown. Here, we identified a novel rice premature senescence leaf 85 (psl85) mutant from ethyl methane sulfonate (EMS) mutagenesis of cultivar Zhongjian100 (the wild-type, WT). The psl85 mutant presented a distinct dwarfism and premature senescence leaf phenotype, starting from the seedling stage to the mature stage, with decreasing level of chlorophyll and degradation of chloroplast, declined photosynthetic capacity, increased content of malonaldehyde (MDA), upregulated expression of senescence-associated genes, and disrupted reactive oxygen species (ROS) scavenging system. Moreover, endogenous abscisic acid (ABA) level was significantly increased in psl85 at the late aging phase, and the detached leaves of psl85 showed more rapid chlorophyll deterioration than that of WT under ABA treatment, indicating that PSL85 was involved in ABA-induced leaf senescence. Genetic analysis revealed that the premature senescence leaf phenotype was controlled by a single recessive nuclear gene which was finally mapped in a 47 kb region on the short arm of chromosome 7, covering eight candidate open reading frames (ORFs). No similar genes controlling a premature senescence leaf phenotype have been identified in the region, and cloning and functional analysis of the gene is currently underway.

Comparative Transcriptomic Profiling to Understand Pre- and Post-Ripening Hormonal Regulations and Anthocyanin Biosynthesis in Early Ripening Apple Fruit.

The 'Hongyu' apple is an early ripening apple cultivar and usually used for fresh marketing. Due to the short ripening period, most of the fruit are harvested at the commercial maturity stage for proper marketing distribution and a longer shelf life. Fruit ripening involves delicate changes to its metabolic and physiological traits through well-organized synchronization of several hormones and regulatory steps. A clear understanding of these hormonal alterations is crucial for extending the period from commercial to physiological ripening. This study was intended to clarify the hormonal alterations and anthocyanin biosynthesis process prior to and immediate after, the harvesting of apple fruit considering the commercial maturity stage. Fruits harvested at 120 Days after flowering (DAF) (HY_4th) was considered as commercially ripened, 110 DAF (HY_3rd) as pre-ripening and 120 DAF followed by five days storage at 20 °C (HY_20 °C_5) as post-ripening samples. Three different stages of fruit were used for transcriptome assembly using RNA-Seq. Results revealed 9187 differentially expressed genes (DEGs) in the post-ripening samples, which was comparatively lower (922 DEGs) in the pre-ripening fruits. DEGs were subjected to Gene Ontology analysis and 31 categories were significantly enriched in the groups 'biological process,' 'molecular function' and 'cellular component.' The DEGs were involved in hormonal signaling pathways like ethylene, abscisic acid (ABA), auxin, gibberellin (GA), brassinosteroid (BR) and anthocyanin biosynthesis pathways such as PAL, 4CL, CHI, DFR, F3H, UFGT. Several transcription factors like the MADS-box gene, MYB, bHLH, NAC, WRKY and HSF were differentially expressed between the pre- and post-ripening fruits. Selected DEGs were subjected to gene expression analysis using quantitative RT-PCR (qRT-PCR) and the results were consistent with those of RNA-Seq. Our data suggested that in addition to ethylene, ABA and other hormones also play key roles in regulating apple fruit ripening and may interact with the ethylene signaling process. Additionally, our data provided an exhibition of the expression pattern of genes in the anthocyanin biosynthesis pathway.

ABA Receptor Subfamily III Enhances Abscisic Acid Sensitivity and Improves the Drought Tolerance of Arabidopsis.

The phytohormone abscisic acid (ABA) regulates plant growth, the developmental process, and abiotic stresses. ABA signaling is induced in response to mediate plant acclimation to environmental challenges, including high salinity and drought. The ABA-binding receptors (RCAR/PYR1/PYL), composing of 14 members, are the core components of the ABA-signaling pathway. Here, we observed that the three subfamilies within the RCARs showed different expression patterns at the basal and exogenous ABA levels. Subsequently, we generated transgenic plants overexpressing subfamily III, RCAR11⁻RCAR14, respectively. The transgenic plants showed increased ABA sensitivity in seed germination and post-germination seedling establishment and root length. Further studies revealed that the overexpressing subfamily III transgenic plants enhanced drought resistance, increased water-use efficiency, and accelerated stress-responsive gene expression compared with the wild-type plants. These findings confirm that the subfamily III plays a key role in ABA-mediated developmental processes and, more importantly, is involved in drought tolerance in the ABA-dependent pathway.

Sprouting of paradormant and endodormant grapevine buds under conditions of forced growth: similarities and differences.

MAIN CONCLUSION: Bud-break assays under forced growth conditions suggest that a drop in ABA content and an increase in sugars are common features in the sprouting of paradormant (PD) and endodormant (ED) grapevine buds. However, increases in cell division and in respiration are unique characteristics of the ED budding. In tropical and subtropical regions where the variations in day length and temperatures are minor throughout the year, the rupture of grapevine buds can be achieved during the current growing season given rise to a double-cropping system annually. However, it is unknown whether the breaking buds are in the paradormancy (PD) or endodormancy (ED) stage. In this study, we compared the breakage of PD and ED buds under conditions of forced growth. To do this, the expression of genes related to the metabolism of phytohormones and sugars, and of relevant physiological functions such as respiration and cell division was analyzed temporally throughout the incubation period in both types of buds. An early fall in the expression of the ABA biosynthesis gene (VvNCED1) and increases in genes related to sugar metabolism and transports were observed during the incubation period in both types of buds. However, while in the PD buds, the genes related to respiration and the cell cycle did not undergo significant changes in their expression during the incubation period, in the ED buds, the expression of these genes together with those related to auxin and cytokinin biosynthesis experienced a large increase. The results suggest that a drop in ABA content and an increase in sugars are early signals for the onset of bud break in both PD and ED vines, while the increase in respiration and cell division are unique characteristics of the ED buds, which reflect its transition from a resting state to a state of active growth.

The 6xABRE Synthetic Promoter Enables the Spatiotemporal Analysis of ABA-Mediated Transcriptional Regulation.

The water stress-associated hormone abscisic acid (ABA) acts through a well-defined signal transduction cascade to mediate downstream transcriptional events important for acclimation to stress. Although ABA signaling is known to function in specific tissues to regulate root growth, little is understood regarding the spatial pattern of ABA-mediated transcriptional regulation. Here, we describe the construction and evaluation of an ABSCISIC ACID RESPONSIVE ELEMENT (ABRE)-based synthetic promoter reporter that reveals the transcriptional response of tissues to different levels of exogenous ABA and stresses. Genome-scale yeast one-hybrid screens complemented these approaches and revealed how promoter sequence and architecture affect the recruitment of diverse transcription factors (TFs) to the ABRE. Our analysis also revealed ABA-independent activity of the ABRE-reporter under nonstress conditions, with expression being enriched at the quiescent center and stem cell niche. We show that the WUSCHEL RELATED HOMEOBOX5 and NAC DOMAIN PROTEIN13 TFs regulate QC/SCN expression of the ABRE reporter, which highlights the convergence of developmental and DNA-damage signaling pathways onto this cis-element in the absence of water stress. This work establishes a tool to study the spatial pattern of ABA-mediated transcriptional regulation and a repertoire of TF-ABRE interactions that contribute to the developmental and environmental control of gene expression in roots.

A stress recovery signaling network for enhanced flooding tolerance in Arabidopsis thaliana.

Abiotic stresses in plants are often transient, and the recovery phase following stress removal is critical. Flooding, a major abiotic stress that negatively impacts plant biodiversity and agriculture, is a sequential stress where tolerance is strongly dependent on viability underwater and during the postflooding period. Here we show that in Arabidopsis thaliana accessions (Bay-0 and Lp2-6), different rates of submergence recovery correlate with submergence tolerance and fecundity. A genome-wide assessment of ribosome-associated transcripts in Bay-0 and Lp2-6 revealed a signaling network regulating recovery processes. Differential recovery between the accessions was related to the activity of three genes: RESPIRATORY BURST OXIDASE HOMOLOG D, SENESCENCE-ASSOCIATED GENE113, and ORESARA1, which function in a regulatory network involving a reactive oxygen species (ROS) burst upon desubmergence and the hormones abscisic acid and ethylene. This regulatory module controls ROS homeostasis, stomatal aperture, and chlorophyll degradation during submergence recovery. This work uncovers a signaling network that regulates recovery processes following flooding to hasten the return to prestress homeostasis.