Endogenous Abscisic Acid Research Articles

Functional Analysis of ScABI3 from Syntrichia caninervis Mitt. in Medicago sativa L.

ABI3 (ABSCISIC ACID INSENSITIVE 3) is a family of B3 transcription factors essential for regulating the abscisic acid (ABA) signaling pathway involved in various biological processes and abiotic stress. Our previous studies demonstrated that ectopic expression of ScABI3 from a desiccation-tolerant moss (Syntrichia caninervis) into Arabidopsis thaliana enhanced abiotic stress tolerance. However, studies on plant transformation using the ABI3 gene are limited and other possible functions of ScABI3 are not known. Here, we transformed the ScABI3 into alfalfa (Medicago sativa L.) and analyzed the effects on phenotype, photosynthetic efficiency, and nutritional quality. The results showed that the endogenous ABA content of the transgenic plants was significantly higher than WT, and the leaf-stem ratio, leaf area, and branch number increased with ScABI3 overexpression in alfalfa. Further analysis of the gas exchange parameters showed that the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), and water-use efficiency (WUE) of the transgenic alfalfa were significantly higher than WT; meanwhile, the protein content of the transgenic lines was higher than the WT, but the crude fat content was lower. Thus, these findings suggest that ScABI3 can be used as a potential candidate gene to manipulate alfalfa’s growth and nutritional quality. This study will provide a theoretical basis for breeding alfalfa varieties and assist in forage production and animal husbandry in the future.

Nitric Oxide Is Essential for Melatonin to Enhance Nitrate Tolerance of Cucumber Seedlings

Melatonin (MT) and nitric oxide (NO) in plants can function cooperatively to alleviate salt stress, sodic alkaline stress and immune response, as well as adventitious root formation. The interaction of MT and NO on the nitrate stress tolerance of cucumber seedlings are not well understood. We investigated the effects of exogenous MT, NO donor (SNP) and NO scavenger (cPTIO) on the growth; photosynthesis; characteristics of root morphological; accumulation of mineral elements, endogenous NO, MT, IAA and ABA; and related genes expression in cucumber (Cucumis sativus L. “Jin You No. 1”) seedlings grown under high nitrate condition (HN). The results showed that MT and NO independently alleviated the inhibition of growth and photosynthesis capacity of cucumber seedlings under nitrate stress. NO was required for MT to enhance the root activity, root length, lateral root number and the accumulation of calcium, magnesium and iron in the roots of cucumber seedlings grown under nitrate stress. Consistently, the expression of adventitious rootless 1 gene (CsARL1) was modulated. Furthermore, exogenous MT induced accumulation of endogenous MT, NO, indole-3-acetic acid (IAA) and abscisic acid (ABA), mainly within 24 h after treatment, in which MT and NO were further increased at 48 h and 96 h, IAA and ABA were further increased at 16 h in the presence of SNP. In contrast, the accumulation of endogenous IAA, MT and ABA slightly decreased within 24 h, NO significantly decreased at 192 h in the presence of cPTIO. Correspondingly, the expression levels of genes involved in nitrogen metabolism (CsNR1 and CsNR2), MT metabolism (CsT5H, CsSNAT2 and Cs2-ODD33), auxin carriers and response factors (CsAUX1, CsGH3.5, CsARF17), ABA synthesis and catabolism (CsNCED1, CsNCED3 and CsCYP707A1) were upregulated by MT, in which CsNR1, CsNR2, CsAUX1, CsNCED3 and CsT5H were further induced in the presence of SNP in roots of cucumber seedlings. These observations indicated that NO act as a crucial factor in MT, alleviating nitrate stress through regulating the mechanism of root growth in cucumber seedlings.

At-ore1 Gene Induces Distinct Novel H2O2-NACs Signaling in Regulating the Leaf Senescence in Soybeans (Glycine max L.)

Senescence is modulated by ORESARA1 (ORE1), a NAC transcription factor that interacts with hormones to fully induce senescence. The At-ore1 gene acts as a suppressor of leaf senescence; however, its exact role in this respect has not been clearly defined. In this study, the function of At-ore1 during leaf senescence was analyzed in soybeans. The precocious leaf senescence of the ore1-1 line was associated with greater chlorophyll loss, leaf necrosis, and redox imbalance in the early vegetative stage during the hyper-accumulation of endogenous abscisic acid (ABA) by enhancing the expression of GmNECD3-related ABA synthesis. At-ore1 induced ABA regulation of the H2O2-GmARF2-GmNAC081 signaling circuit, which relays the At-ore1-induced cell death signal mediation to the caspase-1-like vacuolar processing enzyme (VPE) expression, triggering programmed cell death. In contrast, it was found that At-ore1 functions in IAA to delay leaf-senescence-mediated suppression of the expression of ABA, ROS, and senescence-associated gene 39 (GmSAG39). The IAA-induced GmNAC065 expression controls soybean leaves’ longevity, as discovered by screening At-ore1 expression in ore1-6 for a more stay-green leaf phenotype by helping to increase seed yields. These results uncover a mechanism that modulates ore1 plants’ amplitude expression involved in the ABA/IAA balance in the activation of GmNAC081- or GmNAC065-dependent H2O2 levels, which are crucial in the senescence or delayed leaf senescence of soybeans.

SlSNAT2, a chloroplast-localized acetyltransferase, is involved in Rubisco lysine acetylation and negatively regulates drought stress tolerance in tomato

Drought stress seriously inhibits crop growth and yields and constrains photosynthesis, which is a key biological process. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a key enzyme in the carbon assimilation process, and while its activity is reportedly significantly inhibited under drought stress, little is known about the underlying molecular mechanism. We identified a chloroplast-localized acetyltransferase, SlSNAT2, negatively regulates drought tolerance in tomato. The slsnat2 mutants were conduced to endogenous abscisic acid accumulation in tomato and could promote scavenging of reactive oxygen species through the enhanced activity of antioxidant enzymes. Knocking out this gene could therefore improve photosynthesis under drought stress. Moreover, SlSNAT2 could catalyze the Lys acetylation of rbcL, a large Rubisco subunit. And SlSNAT2 knockout lines were found to maintain higher levels of Rubisco activity under drought stress in the absence of SlSNAT2-catalyzed acetylation of rbcL than wild type plants. Our results demonstrate that SlSNAT2 functions as a negative regulator of drought tolerance. Accordingly, SlSNAT2 could be targeted in molecular designs for tomato breeding. • SlSNAT2 acetylates rbcL and negatively regulates Rubisco activity and endogenous ABA content in tomato. SlSNAT2 knockout lines stabilize photosynthesis in tomato under drought stress conditions.

Paclobutrazol Ameliorates Low-Light-Induced Damage by Improving Photosynthesis, Antioxidant Defense System, and Regulating Hormone Levels in Tall Fescue.

Paclobutrazol (PBZ) is a plant-growth regulator (PGR) in the triazole family that enhances plant tolerance to environmental stresses. Low-light (LL) intensity is a critical factor adversely affecting the growth of tall fescue (Festuca arundinacea Schreb.). Therefore, in this study, tall fescue seedlings were treated with PBZ under control and LL conditions to investigate the effects of PBZ on enhancing LL stress resistance by regulating the growth, photosynthesis, oxidative defense, and hormone levels. Our results reveal that LL stress reduced the total biomass, chlorophyll (Chl) content, photosynthetic capacity, and photochemical efficiency of photosystem II (PSII) but increased the membrane lipid peroxidation level and reactive oxygen species (ROS) generation. However, the application of PBZ increased the photosynthetic pigment contents, net photosynthetic rate (Pn), maximum quantum yield of PSII photochemistry (Fv/Fm), ribulose-1,5-bisphosphate carboxylase (RuBisCO) activity, and starch content. In addition, PBZ treatment activated the antioxidant enzyme activities, antioxidants contents, ascorbate acid-glutathione (AsA-GSH) cycle, and related gene expression, lessening the ROS burst (H2O2 and O2∙−). However, the gibberellic acid (GA) anabolism was remarkably decreased by PBZ treatment under LL stress, downregulating the transcript levels of kaurene oxidase (KO), kaurenoic acid oxidase (KAO), and GA 20-oxidases (GA20ox). At the same time, PBZ treatment up-regulated 9-cis-epoxycarotenoid dioxygenase (NCED) gene expression, significantly increasing the endogenous abscisic acid (ABA) concentration under LL stress. Thus, our study revealed that PBZ improves the antioxidation and photosynthetic capacity, meanwhile increasing the ABA concentration and decreasing GA concentration, which ultimately enhances the LL stress tolerance in tall fescue.

Silicon-Induced Tolerance against Arsenic Toxicity by Activating Physiological, Anatomical and Biochemical Regulation in Phoenix dactylifera (Date Palm).

Arsenic is a toxic metal abundantly present in agricultural, industrial, and pesticide effluents. To overcome arsenic toxicity and ensure safety for plant growth, silicon (Si) can play a significant role in its mitigation. Here, we aim to investigate the influence of silicon on date palm under arsenic toxicity by screening antioxidants accumulation, hormonal modulation, and the expression profile of abiotic stress-related genes. The results showed that arsenic exposure (As: 1.0 mM) significantly retarded growth attributes (shoot length, root length, fresh weight), reduced photosynthetic pigments, and raised reactive species levels. Contrarily, exogenous application of Si (Na2SiO3) to date palm roots strongly influenced stress mitigation by limiting the translocation of arsenic into roots and shoots as compared with the arsenic sole application. Furthermore, an enhanced accumulation of polyphenols (48%) and increased antioxidant activities (POD: 50%, PPO: 75%, GSH: 26.1%, CAT: 51%) resulted in a significant decrease in superoxide anion (O2•−: 58%) and lipid peroxidation (MDA: 1.7-fold), in silicon-treated plants, compared with control and arsenic-treated plants. The Si application also reduced the endogenous abscisic acid (ABA: 38%) under normal conditions, and salicylic acid (SA: 52%) and jasmonic acid levels (JA: 62%) under stress conditions as compared with control and arsenic. Interestingly, the genes; zeaxanthin epoxidase (ZEP) and 9-cis-epoxycarotenoid dioxygenase (NCED-1) involved in ABA biosynthesis were upregulated by silicon under arsenic stress. Likewise, Si application also upregulated gene expression of plant plasma membrane ATPase (PMMA-4), aluminum-activated malate transporter (ALMT) responsible for maintaining cellular physiology, stomatal conductance, and short-chain dehydrogenases/reductases (SDR) involved in nutrients translocation. Hence, the study demonstrates the remarkable role of silicon in supporting growth and inducing arsenic tolerance by increasing antioxidant activities and endogenous hormones in date palm. The outcomes of our study can be employed in further studies to better understand arsenic tolerance and decode mechanism.

Transcriptome analysis revealed the expression levels of genes related to abscisic acid and auxin biosynthesis in grapevine (Vitis vinifera L.) under root restriction.

The root system is essential for the stable growth of plants. Roots help anchor plants in the soil and play a crucial role in water uptake, mineral nutrient absorption and endogenous phytohormone formation. Root-restriction (RR) cultivation, a powerful technique, confines plant roots to a specific soil space. In the present study, roots of one-year-old “Muscat Hamburg” grapevine under RR and control (nR) treatments harvested at 70 and 125 days after planting were used for transcriptome sequencing, and in total, 2031 (nR7 vs. nR12), 1445 (RR7 vs. RR12), 1532 (nR7 vs. RR7), and 2799 (nR12 vs. RR12) differentially expressed genes (DEGs) were identified. Gene Ontology (GO) enrichment analysis demonstrated that there were several genes involved in the response to different phytohormones, including abscisic acid (ABA), auxin (IAA), ethylene (ETH), gibberellins (GAs), and cytokinins (CTKs). Among them, multiple genes, such as PIN2 and ERF113, are involved in regulating vital plant movements by various phytohormone pathways. Moreover, following RR cultivation, DEGs were enriched in the biological processes of plant-type secondary cell wall biosynthesis, the defense response, programmed cell death involved in cell development, and the oxalate metabolic process. Furthermore, through a combined analysis of the transcriptome and previously published microRNA (miRNA) sequencing results, we found that multiple differentially expressed miRNAs (DEMs) and DEG combinations in different comparison groups exhibited opposite trends, indicating that the expression levels of miRNAs and their target genes were negatively correlated. Furthermore, RR treatment indeed significantly increased the ABA content at 125 days after planting and significantly decreased the IAA content at 70 days after planting. Under RR cultivation, most ABA biosynthesis-related genes were upregulated, while most IAA biosynthesis-related genes were downregulated. These findings lay a solid foundation for further establishing the network through which miRNAs regulate grapevine root development through target genes and for further exploring the molecular mechanism through which endogenous ABA and IAA regulate root architecture development in grapevine.

Improving pre-harvest sprouting resistance in rice by editing OsABA8ox using CRISPR/Cas9.

Knock out OsABA8ox helps improve pre-harvest spouting resistance and do not affect rice yield. Pre-harvest sprouting(PHS) is a phenomenon that the seeds of crops germinate preharvest, which reduces the yield and quality of rice. Abscisic acid(ABA) is one of the phytohormones that promotes seed dormancy. ABA8' hydroxylase is the main enzyme that can catabolism ABA in plant. There are three genes that encode ABA8' hydroxylase in rice, named OsABA8ox1, OsABA8ox2 and OsABA8ox3. In this study, we use CRISPR/Cas9 gene editing technology to target these three genes in Ningjing6 and find that the knockout transgenic lines are all significantly strengthen in seed dormancy and have no effect on the yield. By a series of quantitative experiments, we consider that after knock out OsABA8ox, the high endogenous ABA level will influence the ABA signal which suppress the substantial and energy metabolism in the seeds, and finally led to higher dormancy.

OsWRKY114 Inhibits ABA-Induced Susceptibility to Xanthomonas oryzae pv. oryzae in Rice.

The phytohormone abscisic acid (ABA) regulates various aspects of plant growth, development, and stress responses. ABA suppresses innate immunity to Xanthomonas oryzae pv. oryzae (Xoo) in rice (Oryza sativa), but the identity of the underlying regulator is unknown. In this study, we revealed that OsWRKY114 is involved in the ABA response during Xoo infection. ABA-induced susceptibility to Xoo was reduced in OsWRKY114-overexpressing rice plants. OsWRKY114 attenuated the negative effect of ABA on salicylic acid-dependent immunity. Furthermore, OsWRKY114 decreased the transcript levels of ABA-associated genes involved in ABA response and biosynthesis. Moreover, the endogenous ABA level was lower in OsWRKY114-overexpressing plants than in the wild-type plants after Xoo inoculation. Taken together, our results suggest that OsWRKY114 is a negative regulator of ABA that confers susceptibility to Xoo in rice.

Increased dehydrin level decreases leaf rolling grade by altering the reactive oxygen species homeostasis and abscisic acid content in maize subjected to osmotic stress.

Dehydrins (DHNs) are stress proteins involved in the development of protective reactions in plants against dehydration. The relationship between DHNs and morphological responses such as leaf rolling in plants exposed to water deficit is not well known. In this study, we detected how variations in DHN levels affect the leaf rolling response in maize exposed to osmotic stress in relation to the antioxidant system and ABA level. In this context, we altered the DHN levels in maize seedlings by treatment with bio-regulators (salicylic acid and abscisic acid) under PEG6000-free and PEG6000-induced osmotic stress. When the DHN levels were increased by the bio-regulators (25µM SA and 100µM ABA), the relative expression level of the Zea mays dehydrin COR410 gene increased in the seedlings, while reactive oxygen species (ROS) and leaf rolling grade decreased. Moreover, induction of DHNs caused increases in the antioxidant enzyme activity and content of antioxidant substances, and very high amounts of endogenous abscisic acid. When DHN level was suppressed by a bio-regulator (200µM SA) in the maize seedlings, dehydrin COR410 expression level decreased, while ROS and the leaf rolling grade increased. Moreover, the antioxidant enzyme activity and content of antioxidant substances decreased in the seedlings, while the amount of abscisic acid increased. Taken all together, an increase in DHN level by bio-regulator treatment can stimulate the antioxidant system, enable abscisic acid regulation, and thus reduce leaf rolling through decreased ROS levels. The results also indicated that DHNs may be involved in the signal pathways inducing expression of some genes related to leaf rolling response, possibly by modulating ROS levels, in maize seedlings exposed to osmotic stress.

Effect of salicylic acid treatment on antioxidant capacity and endogenous hormones in winter jujube during shelf life

This study investigated the effects of exogenous salicylic acid (SA) treatment (0, control; 3 mmol L−1) on the antioxidant and hormone levels of winter jujube during shelf life (20 d) at 4 °C. The results showed that 3 mmol L−1 SA treatment preferably maintained firmness, color, titratable acidity, and total soluble solids, and effectively reduced the respiratory intensity and TSS/TA value (13.08%) of the fruit. Compared with the control group, the SA group had a higher content of sucrose (14.03%) and malic acid (29.13%). Meanwhile, SA reduced the accumulation of H2O2 (27.73%) and O2− (45.44%) by enhancing the activity of antioxidant enzymes (superoxide dismutase, peroxidase, catalase, ascorbate peroxidase) and the content of antioxidant substances (ascorbic acid, total phenols, total flavonoids, and glutathione) in the fruit. In addition, 3 mmol L−1 SA treatment led to higher levels of endogenous abscisic acid (18.49%) and SA (20.47%) in fruit, and lower concentration of jasmonic acid (42.68%), but had a weak effect on indole acetic acid levels.

Suppression of GhGLU19 encoding β-1,3-glucanase promotes seed germination in cotton

BackgroundIn eudicots, germination begins with water uptake by the quiescent dry seed and is greatly related to the permeability of micropyle enriched callose layers. Once imbibition starts, seeds undergo a cascade of physiological, biochemical, and molecular events to initiate cellular activities. However, the effects of callose on water uptake and following seed metabolic events during germination are largely unknown. Cotton (Gossypium hirsutum) is a eudicot plant with natural fiber and edible oil production for humans. Here, we addressed this question by examining the role of GhGLU19, a gene encoding β-1,3-glucanase, in cotton seed germination.ResultsGhGLU19 belongs to subfamily B and was expressed predominately in imbibed seeds and early seedlings. Compared to wild type, GhGLU19-suppressing and GhGLU19-overexpressing transgenic cotton lines showed the higher and lower seed germination percentage, respectively. Callose was enriched more at inner integument (ii) than that in embryo and seed coat in cotton seeds. In GhGLU19-suppressing lines, callose at ii of cotton seeds was greatly increased and brought about a prolonged water uptake process during imbibition. Both proteomic and transcriptomic analysis revealed that contrary to GhGLU19-overexpressing lines, the glycolysis and pyruvate metabolism was decreased, and abscisic acid (ABA) biosynthesis related genes were downregulated in imbibed seeds of GhGLU19-suppressing lines. Also, endogenous ABA was significantly decreased in GhGLU19-suppressing line while increased in GhGLU19-overexpressing line.ConclusionsOur results demonstrate that suppression of GhGLU19 improves cotton seed germination via accumulating callose of inner integument, modulating glycolysis and pyruvate metabolism, and decreasing ABA biosynthesis. This study provides a potential way for improving germination percentage in cotton seed production, and other eudicot crops.

SbWRKY55 regulates sorghum response to saline environment by its dual role in abscisic acid signaling.

SbWRKY55 functions as a key component of the ABA-mediated signaling pathway; transgenic sorghum regulates plant responses to saline environments and will help save arable land and ensure food security. Salt tolerance in plants is triggered by various environmental stress factors and endogenous hormonal signals. Numerous studies have shown that WRKY transcription factors are involved in regulating plant salt tolerance. However, the underlying mechanism for WRKY transcription factors regulated salt stress response and signal transduction pathways remains largely unknown. In this study, the SbWRKY55 transcription factor was found to be the key component for reduced levels of salt and abscisic acid in SbWRKY55 overexpression significantly reduced salt tolerance in sorghum and Arabidopsis. Mutation of the homologous gene AtWRKY55 in A. thaliana significantly enhanced salt tolerance, and SbWRKY55 supplementation in the mutants restored salt tolerance. In the transgenic sorghum with SbWRKY55 overexpression, the expression levels of genes involved in the abscisic acid (ABA) pathway were altered, and the endogenous ABA content decreased. Yeast one-hybrid assays and dual-luciferase reporter assay showed that SbWRKY55 binds directly to the promoter of SbBGLU22 and inhibits its expression level. In addition, both in vivo and in vitro biochemical analyses showed that SbWRKY55 interacts with the FYVE zinc finger protein SbFYVE1, blocking the ABA signaling pathway. This could be an important feedback regulatory pathway to balance the SbWRKY55-mediated salt stress response. In summary, the results of this study provide convincing evidence that SbWRKY55 functions as a key component in the ABA-mediated signaling pathway, highlighting the dual role of SbWRKY55 in ABA signaling. This study also showed that SbWRKY55 could negatively regulate salt tolerance in sorghum.

OsPP65 Negatively Regulates Osmotic and Salt Stress Responses Through Regulating Phytohormone and Raffinose Family Oligosaccharide Metabolic Pathways in Rice

Although type 2C protein phosphatases (PP2Cs) have been demonstrated to play important roles in regulating plant development and various stress responses, their specific roles in rice abiotic stress tolerance are still largely unknown. In this study, the functions of OsPP65 in rice osmotic and salt stress tolerance were investigated. Here, we report that OsPP65 is responsive to multiple stresses and is remarkably induced by osmotic and salt stress treatments. OsPP65 was highly expressed in rice seedlings and leaves and localized in the nucleus and cytoplasm. OsPP65 knockout rice plants showed enhanced tolerance to osmotic and salt stresses. Significantly higher induction of genes involved in jasmonic acid (JA) and abscisic acid (ABA) biosynthesis or signaling, as well as higher contents of endogenous JA and ABA, were observed in the OsPP65 knockout plants compared with the wild-type plants after osmotic stress treatment. Further analysis indicated that JA and ABA function independently in osmotic stress tolerance conferred by loss of OsPP65. Moreover, metabolomics analysis revealed higher endogenous levels of galactose and galactinol but a lower content of raffinose in the OsPP65 knockout plants than in the wild-type plants after osmotic stress treatment. These results together suggest that OsPP65 negatively regulates osmotic and salt stress tolerance through regulation of the JA and ABA signaling pathways and modulation of the raffinose family oligosaccharide metabolism pathway in rice. OsPP65 is a promising target for improvement of rice stress tolerance using gene editing.

Vapour pressure deficit and endogenous ABA level modulate stomatal responses of tomato plants to soil water deficit

High vapour pressure deficit (VPD) in the air and water deficits in the soil are considered two key stress factors limiting crop growth and production. To explore their interactive effects on stomatal behaviour and plant water use and the role of abscisic acid (ABA) in mediating these responses, two tomato (Solanum lycopersicum) genotypes, Ailsa Craig (AC) and its ABA-deficient mutant (flacca), were grown at two VPD levels (VPD1, 0.71 kPa and VPD2, 1.25 kPa) and were exposed to progressive soil drying. The results showed that the relatively higher VPD, i.e. VPD2, decreased stomatal conductance only in AC, and increased transpiration rate and water consumption in both genotypes, which was associated with altered stomatal morphological traits, including lowered stomatal density, reduced stomatal size and pore aperture. During progressive soil drying, both AC and flacca grown at VPD2 closed stomata earlier than VPD1-plants with a low leaf ABA concentration, indicating that rising VPD advanced stomatal drought response in an ABA-independent way. At the end of soil drying, the stomata of VPD1-AC plants remained widely opened resulting in fast desiccation of the leaves excised from the plants upon exposure to VPD2, which implied an interaction of VPD and ABA in mediating short-term stomatal response. VPD2 increased the intrinsic water use efficiency (WUE) in AC plants but decreased the whole-leaf and plant WUE in both genotypes, as affirmed by the corresponding changes in leaf δ13C and δ 18O. Compared to δ 18O, δ13C was a more robust indicator of stomatal movement when plants were exposed to combined atmospheric and soil droughts. Collectively, these findings provide some novel insights into the stomatal regulation of tomato plants confronting combined atmospheric and edaphic droughts.

The phytochrome-interacting factor DcPIF3 of carrot plays a positive role in drought stress by increasing endogenous ABA level in Arabidopsis

The phytochrome-interacting factor (PIF) subfamily of basic helix-loop-helix (bHLH) transcription factors plays a critical role in plant growth and development. However, there has been no detailed report on the PIFs in carrot. In this study, we present the identification and characterization of DcPIF gene family in carrot (Daucus carota L.). Phylogenetic analysis indicated that PIFs from carrot and other five plant species could be divided into four groups supported by similar gene structure and motif analysis. Expression profiles showed that all DcPIF genes were tissue-specific and could be induced by drought or abscisic acid (ABA) treatment except DcPIF7.1, among which DcPIF3 was the most responsive. The DcPIF3-overexpressed Arabidopsis plants exhibited more tolerance to drought stress, with higher antioxidant capacity and lower malondialdehyde content after drought treatment than wild type plants. Further stress tolerance assays revealed that DcPIF3 plays a positive role in drought stress by increasing endogenous ABA level and promoting the expression of ABA-related genes. Our results can enrich the understanding of DcPIF family genes and lay a foundation for further investigation of DcPIF3 function to defend against drought stress in carrot.

ABA signaling mediates 5-aminolevulinic acid-induced anthocyanin biosynthesis in red pear fruits

Peel color is an important quality trait of pears and directly affects its commercial value. Although several studies have reported that exogenous 5-aminolevulinic acid (ALA) induces anthocyanin biosynthesis, the mechanisms involved remain unclear. The purpose of this study was to confirm the effects of ALA on anthocyanin biosynthesis in the red pear ‘Nanhong’ and elucidate the effect of abscisic acid (ABA) on ALA-induced anthocyanin accumulation. Application of ALA solution (100 or 300 mg L−1) promoted arbutin, flavonol, and anthocyanin accumulation, but decreased chlorogenic acid and flavanol content, especially at 300 mg L−1. The structural gene transcriptional levels and transcription factors (PuMYB10 and PubHLH33) associated with anthocyanin synthesis were also enhanced by ALA treatment. Furthermore, ALA treatment increased the endogenous ABA content and upregulated the expression of ABA biosynthesis-related genes. Additionally, the influence of ALA on anthocyanin biosynthesis was removed by the ABA biosynthesis inhibitor fluridone, while the expression of PuMYB10, PuUFGT, and PuGSTF12, which were induced by exogenous ABA and ALA treatments, was suppressed. These results suggest that ABA signaling plays a crucial role in ALA-induced anthocyanin accumulation in red pear fruits. The results can provide a theoretical basis for improving the color and fruit quality of red pears.

Biocompatible Abscisic Acid-Sensing Supramolecular Hybridization Probe for Spatiotemporal Fluorescence Imaging in Plant Tissues.

Achieving detection of the phytohormone abscisic acid (ABA) is of critical importance for understanding plant growth and development. We report a hybrid supramolecular fluorescent probe that uses bovine serum albumin (BSA) as a host. Aggregation-induced emission of fluorescent chromophores (AIEgens) enables luminescence in the presence of BSA. ABA and its aptamer act as a switch to trigger this fluorescent system, the strategy that exhibits high sensitivity to abscisic acid with a detection limit of 0.098 nM. The probe test strip also enables visualization of ABA content from plants by colorimetric observation with the naked eye. In particular, the high biocompatibility and small molecular size of the prepared fluorescent probe allow for effective monitoring of ABA in plant tissues by fluorescence imaging. This strategy provides a new perspective to achieve the detection of endogenous and exogenous ABA in plants and has important implications for plant biology research.

E3 ligase AtAIRP5/GARU regulates drought stress response by stimulating SERINE CARBOXYPEPTIDASE-LIKE1 turnover.

Ubiquitination is a major mechanism of eukaryotic posttranslational protein turnover that has been implicated in abscisic acid (ABA)-mediated drought stress response. Here, we isolated T-DNA insertion mutant lines in which ABA-insensitive RING protein 5 (AtAIRP5) was suppressed, resulting in hyposensitive ABA-mediated germination compared to wild-type Arabidopsis (Arabidopsis thaliana) plants. A homology search revealed that AtAIRP5 is identical to gibberellin (GA) receptor RING E3 ubiquitin (Ub) ligase (GARU), which downregulates GA signaling by degrading the GA receptor GID1, and thus AtAIRP5 was renamed AtAIRP5/GARU. The atairp5/garu knockout progeny were impaired in ABA-dependent stomatal closure and were markedly more susceptible to drought stress than wild-type plants, indicating a positive role for AtAIRP5/GARU in the ABA-mediated drought stress response. Yeast two-hybrid, pull-down, target ubiquitination, and in vitro and in planta degradation assays identified serine carboxypeptidase-like1 (AtSCPL1), which belongs to the clade 1A AtSCPL family, as a ubiquitinated target protein of AtAIRP5/GARU. atscpl1 single and atairp5/garu-1 atscpl1-2 double mutant plants were more tolerant to drought stress than wild-type plants in an ABA-dependent manner, suggesting that AtSCPL1 is genetically downstream of AtAIRP5/GARU. After drought treatment, the endogenous ABA levels in atscpl1 and atairp5/garu-1 atscpl1-2 mutant leaves were higher than those in wild-type and atairp5/garu leaves. Overall, our results suggest that AtAIRP5/GARU RING E3 Ub ligase functions as a positive regulator of the ABA-mediated drought response by promoting the degradation of AtSCPL1.

Quantitative Analysis of ABA and SA in Rice (Oryza sativa L.) Grown Under Drought Stress

Rice cultivation requires plenty of water for its proper growth, development, and productivity. The regular life cycle of rice plants is disrupted by moderate to severe drought. Both abscisic acid (ABA) and salicylic acid (SA) have involvement in rice physiology under drought. With the limited information, we aimed to study the relationship between ABA and SA concentration in leaves of rice plants under drought. The experiment was performed on a drought-sensitive variety of Swarna MTU 7029 rice. The HPLC method was used to analyse the endogenous ABA and SA content. This study provided data on ABA and SA content in 0.5 mM SA treated and untreated 56 days old rice plants at 7, 14 and 28 days of drought. The result showed that the concentration of ABA was enhanced by 74.6%, 82.8%, and 99.4% during 7, 14, and 28 days of drought, respectively, while it was increased by 64.6%, 74.3% and 78.5% in SA treated plants under 7, 14 and 28 days drought, respectively. The concentration of SA enhanced by 132.2%, 19.7%, and 3.0% during 7, 14, and 28 days of drought, while it was increased to 137.5%, 54.8%, and 23.2% in SA treated plants under 7, 14, and 28 days drought, respectively. This explains that on the 7th day of drought stress, SA may suppress the formation of ABA but not much on the 14th and 28th days of drought stress in treated rice plants. This outcome helps study the interaction of ABA and SA at the gene level under drought stress.