Exogenous Abscisic Acid Treatment Research Articles

Abscisic acid enhances alkaline stress tolerance in grapevines: Physiological and transcriptional profiling

Abscisic acid (ABA) is a crucial signaling regulator governing plant growth and survival during adverse conditions. However, the mechanism by which ABA mediates grapevine tolerance to alkali stress has not yet been elucidated. Here, we investigated the role of ABA in regulating physiological characteristics and transcriptome of grapevines under alkali stress through the application of exogenous ABA and fluridone (an ABA biosynthesis inhibitor). The results revealed that alkali stress led to significant reductions in the net photosynthetic rate (Pn), stomatal conductance (Gs), intercellular CO2 concentration (Ci), transpiration rate (Tr), chlorophyll content, the maximum quantum yields of primary photochemistry of PSII (Fv/Fm), nonphotochemical quenching (NPQ), which were greatly alleviated by exogenous ABA (50 and 100 μM) application. Specifically, the application of exogenous ABA (50 μM) increased Pn, chlorophyll content and Fv/Fm by 28.39 %, 30.45 % and 17.29 %, respectively, compared with alkali stress alone. Furthermore, exogenous ABA treatment reduced alkali stress-induced superoxide anions (O2∙−) and hydrogen peroxide (H2O2), malondialdehyde (MDA), and electrolyte leakage, as well as higher proline content and activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). Additionally, exogenous ABA (50 μM) decreased O2∙−, H2O2, MDA, electrolyte leakage by 38.64 %, 28.16 %, 39.29 % and 39.42 %, respectively, respect to alkali stress alone. Moreover, exogenous ABA reduced the Na+, increased K+ and K+/Na+ ratio, as well as the phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI) activities and flavonoid content, and increased zeaxanthin epoxidase (ZEP) and 9-cis-epoxy carotenoid dioxygenase (NCED) activities and endogenous ABA levels in alkali stress-treated grapevines. Exogenous ABA (50 μM) increased flavonoid content and endogenous ABA content by 74.03 % and 20.84 %, compared with alkali stress alone, respectively. Conversely, exogenous fluridone exacerbated alkali stress-induced physiological damage in grapevine plants. Transcriptome analysis revealed that exogenous ABA (50 μM) and fluridone significantly induced the ‘Plant hormone signal transduction (ko04075)’, the ‘MAPK signaling pathway-plant (ko04016)’, ‘ABC transporters (ko02010)’, ‘Photosynthesis-antenna proteins (ko00196)’, ‘Flavonoid biosynthesis (ko00941)’, ‘Phenylpropanoid biosynthesis (ko00940)’ and other biological pathways and key gene involved in chlorophyll metabolism and ion transport. In conclusion, ABA markedly enhanced grapevines tolerance to alkali stress by modulating key biological pathways and physiological characteristics.

Abscisic acid controls sugar accumulation essential to strawberry fruit ripening via the FaRIPK1-FaTCP7-FaSTP13/FaSPT module.

Strawberry is considered as a model plant for studying the ripening of abscisic acid (ABA)-regulated non-climacteric fruits, a process in which sugar plays a fundamental role, while how ABA regulates sugar accumulation remains unclear. This study provides a direct line of physiological, biochemical, and molecular evidence that ABA signaling regulates sugar accumulation via the FaRIPK1-FaTCP7-FaSTP13/FaSPT signaling pathway. Herein, FaRIPK1, a red-initial protein kinase 1 previously identified in strawberry fruit, not only interacted with the transcription factor FaTCP7 (TEOSINTE BRANCHEN 1, CYCLOIDEA, and PCF) but also phosphorylated the critical Ser89 and Thr93 sites of FaTCP7, which negatively regulated strawberry fruit ripening, as evidenced by the transient overexpression (OE) and virus-induced gene silencing transgenic system. Furthermore, the DAP-seq experiments revealed that FvTCP7 bound the motif "GTGGNNCCCNC" in the promoters of two sugar transporter genes, FaSTP13 (sugar transport protein 13) and FaSPT (sugar phosphate/phosphate translocator), inhibiting their transcription activities as determined by the electrophoretic mobility shift assay, yeast one-hybrid, and dual-luciferase reporter assays. The downregulated FaSTP13 and FaSPT transcripts in the FaTCP7-OE fruit resulted in a reduction in soluble sugar content. Consistently, theyeast absorption test revealed that the two transporters had hexose transport activity. Especially, the phosphorylation-inhibited binding of FaTCP7 to the promoters of FaSTP13 and FaSPT could result in the release of their transcriptional activities. In addition, the phosphomimetic form FaTCP7S89D or FaTCP7T93D could rescue the phenotype of FaTCP7-OE fruits. Importantly, exogenous ABA treatment enhanced the FaRIPK1-FaTCP7 interaction. Overall, we found direct evidence that ABA signaling controls sugar accumulation during strawberry fruit ripening via the "FaRIPK1-FaTCP7-FaSTP13/FaSPT" module.

SlPP2C2 interacts with FZY/SAUR and regulates tomato development via signaling crosstalk of ABA and auxin.

Abscisic acid (ABA) signaling interacts frequently with auxin signaling when it regulates plant development, affecting multiple physiological processes; however, to the best of our knowledge, their interaction during tomato development has not yet been reported. Here, we found that type 2C protein phosphatase (SlPP2C2) interacts with both flavin monooxygenase FZY, an indole-3-acetic acid (IAA) biosynthetic enzyme, and small auxin upregulated RNA (SAUR) of an IAA signaling protein and regulates their activity, thereby affecting the expression of IAA-responsive genes. The expression level of SlPP2C2 was increased by exogenous ABA, IAA, NaCl, or dehydration treatment of fruits, leaves, and seeds, and it decreased in imbibed seeds. Manipulating SlPP2C2 with overexpression, RNA interference, and CRISPR/Cas9-mediated genome editing resulted in pleiotropic changes, such as morphological changes in leaves, stem trichomes, floral organs and fruits, accompanied by alterations in IAA and ABA levels. Furthermore, the RNA-seq analysis indicated that SlPP2C2 regulates the expression of auxin-/IAA-responsive genes in different tissues of tomato. The results demonstrate that SlPP2C2-mediated ABA signaling regulates the development of both vegetative and reproductive organs via interaction with FZY/SAUR, which integrates the cross-talk of ABA and auxin signals during development and affects the expressions of development-related genes in tomato.

Metabolome and transcriptome analyses reveal changes of rapeseed in response to ABA signal during early seedling development

Seed germination is an important development process in plant growth. The phytohormone abscisic acid (ABA) plays a critical role during seed germination. However, the mechanism of rapeseed in response to ABA is still elusive. In order to understand changes of rapeseed under exogenous ABA treatment, we explored differentially expressed metabolites (DEMs) and the differentially expressed genes (DEGs) between mock- and ABA-treated seedlings. A widely targeted LC-MS/MS based metabolomics were used to identify and quantify metabolic changes in response to ABA during seed germination, and a total of 186 significantly DEMs were identified. There are many compounds which are involved in ABA stimuli, especially some specific ABA transportation-related metabolites such as starches and lipids were screened out. Meanwhile, a total of 4440 significantly DEGs were identified by transcriptomic analyses. There was a significant enrichment of DEGs related to phenylpropanoid and cell wall organization. It suggests that exogenous ABA mainly affects seed germination by regulating cell wall loosening. Finally, the correlation analysis of the key DEMs and DEGs indicates that many DEGs play a direct or indirect regulatory role in DEMs metabolism. The integrative analysis between DEGs and DEMs suggests that the starch and sucrose pathways were the key pathway in ABA responses. The two metabolites from starch and sucrose pathways, levan and cellobiose, both were found significantly down-regulated in ABA-treated seedlings. These comprehensive metabolic and transcript analyses provide useful information for the subsequent post-transcriptional modification and post germination growth of rapeseed in response to ABA signals and stresses.

Progress in Fruit Cracking Control of Gibberellic Acid and Abscisic Acid

Fruit cracking or splitting is a severe physiological disease that significantly affects appearance and quality, compromising the commodity value of fruit and causing substantial economic losses to the producers of several fleshy fruit crops. The growth-promoting plant hormone gibberellins (GAs) and growth-inhibiting abscisic acid (ABA) antagonistically regulate numerous processes throughout the plant life cycle. The homeostasis of GA and ABA plays a significant role in the normal growth and development of fruits, and the imbalance of them may lead to the occurrence of cracking or splitting during the process of fruit growth, development, ripening and postharvest storage. The pathways of GA and ABA metabolism and signaling have been studied widely, and the major components are well characterized, including the genes encoding major biosynthesis and catabolism enzymes and the key signaling components. Nevertheless, our knowledge of the mechanisms of GA and ABA governing fruit cracking is not comprehensive enough. In this review, we summarize the advances in understanding the effects of endogenous GAs and ABA contents in fruits and exogenous GAs and ABA treatments on fruit cracking, and we endeavor to provide some genetic cues on the function of GAs and ABA responsible for fruit cracking modulation. The progress in understanding the molecular bases underlying the actions of GAs and ABA in fruit cracking coordination control will facilitate breeding strategies of cracking-resistant ideotypes of fruits, and also carry great theoretical significance in guiding the establishment of integrated prevention and control measures in fruit cracking.

Characterization of PYL gene family and identification of HaPYL genes response to drought and salt stress in sunflower.

In the context of global climate change, drought and soil salinity are some of the most devastating abiotic stresses affecting agriculture today. PYL proteins are essential components of abscisic acid (ABA) signaling and play critical roles in responding to abiotic stressors, including drought and salt stress. Although PYL genes have been studied in many species, their roles in responding to abiotic stress are still unclear in the sunflower. In this study, 19 HaPYL genes, distributed on 15 of 17 chromosomes, were identified in the sunflower. Fragment duplication is the main cause of the expansion of PYL genes in the sunflower genome. Based on phylogenetic analysis, HaPYL genes were divided into three subfamilies. Members in the same subfamily share similar protein motifs and gene exon-intron structures, except for the second subfamily. Tissue expression patterns suggested that HaPYLs serve different functions when responding to developmental and environmental signals in the sunflower. Exogenous ABA treatment showed that most HaPYLs respond to an increase in the ABA level. Among these HaPYLs, HaPYL2a, HaPYL4d, HaPYL4g, HaPYL8a, HaPYL8b, HaPYL8c, HaPYL9b, and HaPYL9c were up-regulated with PEG6000 treatment and NaCl treatment. This indicates that they may play a role in resisting drought and salt stress in the sunflower by mediating ABA signaling. Our findings provide some clues to further explore the functions of PYL genes in the sunflower, especially with regards to drought and salt stress resistance.

Integrated physio-biochemical and transcriptomic analyses reveal the mechanism underlying ABA-mediated alleviation of salt stress in Limonium bicolor seedlings

Abscisic acid (ABA) plays an important role in regulating plant stress responses. However, there are currently no reports on ABA in regulating the salt tolerance of Limonium bicolor. Here, we analyzed the effects of exogenous ABA on growth, photosynthesis, salt secretion, and physiological and transcriptome changes of L. bicolor seedlings under 300 mM NaCl. Exogenous 10 µM ABA treatment in L. bicolor seedlings treated with salt upregulated the expression of genes related to stomatal development, increased the density and number of stomata, and improved plant gas exchange capacity, thereby increasing the net photosynthesis rate. Exogenous ABA treatment in L. bicolor seedlings under salt stress also upregulated the expression of genes related to salt gland development, promoted salt glands development, increased salt glands density, increased the salt secretion capacity of leaves, and reduced the accumulation of Na+ in the plant. ABA application enhanced salt tolerance of L. bicolor seedlings by increasing content of soluble solutes, enhancing antioxidant enzymes activities, decreasing accumulation of reactive oxygen species, and increasing ABA and indole acetic acid contents. Transcriptome sequencing revealed that genes differentially expressed in L. bicolor seedlings during ABA-induced salt tolerance are involved in many metabolic pathways, including plant hormone signal transduction, proline metabolism, endocytosis and protein processing in the endoplasmic reticulum. Furthermore, some transcription factors, MYB, bHLH, and WRKY, might contribute to the improved salt stress tolerance of L. bicolor conferred by ABA. These results provide a basis for further clarifying the salt tolerance mechanism mediated by exogenous ABA in L. bicolor seedlings.

Tulip transcription factor TgWRKY75 activates salicylic acid and abscisic acid biosynthesis to synergistically promote petal senescence.

WRKY transcription factors play a central role in controlling plant organ senescence; however, it is unclear whether and how they regulate petal senescence in the widely grown ornamental plant tulip (Tulipa gesneriana). In this study, we report that TgWRKY75 promotes petal senescence by enhancing the synthesis of both abscisic acid (ABA) and salicylic acid (SA) in tulip and in transgenic Arabidopsis. The expression level of TgWRKY75 was up-regulated in senescent petals, and exogenous ABA or SA treatment induced its expression. The endogenous contents of ABA and SA significantly increased during petal senescence and in response to TgWRKY75 overexpression. Two SA synthesis-related genes, TgICS1 and TgPAL1, were identified as direct targets of TgWRKY75, which binds to their promoters. In parallel, TgWRKY75 activated the expression of the ABA biosynthesis-related gene TgNCED3 via directly binding to its promoter region. Site mutation of the W-box core motif located in the promoters of TgICS1, TgPAL1, and TgNCED3 eliminated their interactions with TgWRKY75. In summary, our study demonstrates a dual regulation of ABA and SA biosynthesis by TgWRKY75, revealing a synergistic process of tulip petal senescence through feedback regulation between TgWRKY75 and the accumulation of ABA and SA.

A transcriptional regulation of ERF15 contributes to ABA-mediated cold tolerance in tomato.

Cold stress is a major meteorological threat to crop growth and yield. Abscisic acid (ABA) plays important roles in plant cold tolerance by activating the expression of cold-responsive genes; however, the underlying transcriptional regulatory module remains unknown. Here, we demonstrated that the cold- and ABA-responsive transcription factor ETHYLENE RESPONSE FACTOR 15 (ERF15) positively regulates ABA-mediated cold tolerance in tomato. Exogenous ABA treatment significantly enhanced cold tolerance in wild-type tomato plants but failed to rescue erf15 mutants from cold stress. Transcriptome analysis showed that ERF15 was associated with the expression of cold-responsive transcription factors such as CBF1 and WRKY6. Further RT-qPCR assays confirmed that the ABA-induced increased in CBF1 and WRKY6 transcripts was suppressed in erf15 mutants when the plants were subjected to cold treatment. Moreover, yeast one-hybrid assays, dual-luciferase assays and electrophoretic mobility shift assays demonstrated that ERF15 activated the transcription of CBF1 and WRKY6 by binding their promoters. Silencing CBF1 or WRKY6 significantly decreased cold tolerance. Overall, our study identified the role of ERF15 in conferring ABA-mediated cold tolerance in tomato plants by activating CBF1 and WRKY6 expression. This study not only broadens our knowledge of the mechanism of ABA-mediated cold tolerance in plants but also highlights ERF15 as an ideal target gene for cold-tolerant crop breeding.

Exogenous abscisic acid treatment regulates protein secretion in sorghum cell suspension cultures

ABSTRACT Drought stress adversely affects plant growth, often leading to total crop failure. Upon sensing soil water deficits, plants switch on biosynthesis of abscisic acid (ABA), a stress hormone for drought adaptation. Here, we used exogenous ABA application to dark-grown sorghum cell suspension cultures as an experimental system to understand how a drought-tolerant crop responds to ABA. We evaluated intracellular and secreted proteins using isobaric tags for relative and absolute quantification. While the abundance of only ~ 7% (46 proteins) intracellular proteins changed in response to ABA, ~32% (82 proteins) of secreted proteins identified in this study were ABA responsive. This shows that the extracellular matrix is disproportionately targeted and suggests it plays a vital role in sorghum adaptation to drought. Extracellular proteins responsive to ABA were predominantly defense/detoxification and cell wall-modifying enzymes. We confirmed that sorghum plants exposed to drought stress activate genes encoding the same proteins identified in the in vitro cell culture system with ABA. Our results suggest that ABA activates defense and cell wall remodeling systems during stress response. This could underpin the success of sorghum adaptation to drought stress.

Genome-Wide Analysis of the Universal Stress Protein Gene Family in Blueberry and Their Transcriptional Responses to UV-B Irradiation and Abscisic Acid

Universal stress proteins (USPs) play essential roles in plant development, hormonal regulation, and abiotic stress responses. However, the characteristics and functional divergence of USP family members have not been studied in blueberry (Vaccinium corymbosum). In this study, we identified 72 VcUSP genes from the Genome Database for Vaccinium. These VcUSPs could be divided into five groups based on their phylogenetic relationships. VcUSPs from groups Ⅰ, Ⅳ, and Ⅴ each possess one UspA domain; group Ⅰ proteins also contain an ATP-binding site that is not present in group Ⅳ and Ⅴ proteins. Groups Ⅱ and Ⅲ include more complex proteins possessing one to three UspA domains and UspE or UspF domains. Prediction of cis-regulatory elements in the upstream sequences of VcUSP genes indicated that their protein products are likely involved in phytohormone signaling pathways and abiotic stress responses. Analysis of RNA deep sequencing data showed that 21 and 7 VcUSP genes were differentially expressed in response to UV-B radiation and exogenous abscisic acid (ABA) treatments, respectively. VcUSP41 and VcUSP68 expressions responded to both treatments, and their encoded proteins may integrate the UV-B and ABA signaling pathways. Weighted gene co-expression network analysis revealed that VcUSP22, VcUSP26, VcUSP67, VcUSP68, and VcUSP41 were co-expressed with many transcription factor genes, most of which encode members of the MYB, WRKY, zinc finger, bHLH, and AP2 families, and may be involved in plant hormone signal transduction, circadian rhythms, the MAPK signaling pathway, and UV-B-induced flavonoid biosynthesis under UV-B and exogenous ABA treatments. Our study provides a useful reference for the further functional analysis of VcUSP genes and blueberry molecular breeding.

A 14-3-3 Protein-Encoding Gene, BdGF14g, Confers Better Drought Tolerance by Regulating ABA Biosynthesis and Signaling

Abscisic acid (ABA), a phytohormone, enacts a cardinal function in coping with abiotic stress. 14-3-3 proteins can interact with ABA-responsive-element-binding transcription factors (ABFs), a chief constituent of ABA signaling, and play critical roles in the dehydration response involving ABA signaling. Meanwhile, whether and how 14-3-3 proteins regulate ABA signaling to respond to aridity stress is yet to be fully investigated. Herein, BdGF14g, a 14-3-3 gene induced by ABA, H2O2, and PEG treatments, was identified in Brachypodium distachyon (B. distachyon). Overexpression of BdGF14g improved drought stress tolerance in tobacco plants, with a higher survival rate, longer root length, enhanced cell membrane stability, and increased antioxidase activity compared with non-transgenic controls in coping with dehydration. Both drought and exogenous ABA treatments resulted in smaller stomatal apertures in BdGF14g-transgenic lines. Additionally, when an ABA biosynthesis inhibitor was added, the better growth statuses, less H2O2 accumulation, and higher activities of catalase and superoxide dismutase under mannitol stress disappeared. Moreover, BdGF14g interacted with NtABF2, upregulated the endogenous ABA content, and enhanced the transcription of ABA-related genes, including NtNCED1, a crucial ABA biosynthesis gene, under drought conditions. In conclusion, BdGF14g acts as a positive factor in the water deficiency response by affecting ABA biosynthesis and signaling in tobacco plants.

Overexpression of a C3HC4‐type E3‐ubiquitin ligase contributes to salinity tolerance by modulating Na+ homeostasis in rice

AbstractSoil salinity has a negative effect on crop yield. Therefore, plants have evolved many strategies to overcome decreases in yield under saline conditions. Among these, E3‐ubiquitin ligase regulates salt tolerance. We characterized Oryza sativa Really Interesting New Gene (RING) Finger C3HC4‐type E3 ligase (OsRFPHC‐4), which plays a positive role in improving salt tolerance. The expression of OsRFPHC‐4 was downregulated by high NaCl concentrations and induced by abscisic acid (ABA) treatment. GFP‐fused OsRFPHC‐4 was localized to the plasma membrane of rice protoplasts. OsRFPHC‐4 encodes a cellular protein with a C3HC4‐RING domain with E3 ligase activity. However, its variant OsRFPHC‐4C161A does not possess this activity. OsRFPHC‐4‐overexpressing plants showed enhanced salt tolerance due to low accumulation of Na+ in both roots and leaves, low Na+ transport in the xylem sap, high accumulation of proline and soluble sugars, high activity of reactive oxygen species (ROS) scavenging enzymes, and differential regulation of Na+/K+ transporter expression compared to wild‐type (WT) and osrfphc‐4 plants. In addition, OsRFPHC‐4‐overexpressing plants showed higher ABA sensitivity under exogenous ABA treatment than WT and osrfphc‐4 plants. Overall, these results suggest that OsRFPHC‐4 contributes to the improvement of salt tolerance and Na+/K+ homeostasis via the regulation of changes in Na+/K+ transporters.

The responses of genotypes with contrasting NUtE to exogenous ABA during the flowering stage in Brassica napus

A large amount of nitrogen (N) fertilizer is applied to maintain a stable yield because of the low N utilization efficiency (NUtE) in rapeseed (Brassica napus L.), which increases production costs and has a negative impact on the environment. Abscisic acid (ABA) has been reported to have crosstalk with nitrate. In this study, exogenous ABA treatment and RNA-seq were used to explore the response patterns of rapeseed with contrasting NUtE to ABA signals. Our results showed that ABA application on leaf promoted the N allocation from vegetative organs to grain and increased the number of seeds per silique, grain yield, N harvest index (NHI), and the NUtE in N-efficient rapeseed. Transcriptome analysis showed the DEGs were mainly enriched in N metabolism, ABC transporters, and MAPK signaling pathways. Among them, the Glutamate dehydrogenase (GS) genes and the Glutamine synthetase gene were more highly expressed in the N-efficient genotype leaf than the N-inefficient genotype after ABA treatment. In contrast, the opposite expression level was observed in seven genes, including nitrate transporter 2.1, Nitrate Reductase genes, and GS genes in the root. In conclusion, the shoot and root of rapeseed may have different response mechanisms to ABA stimulation, and N-efficient rapeseed is more sensitive to ABA treatment, which may be one of the reasons for the difference in NUtE

Genome-wide identification of the AOMT gene family in wax apple and functional characterization of SsAOMTs to anthocyanin methylation.

Anthocyanins are major pigments in the peels of red-series wax apple fruits, and two principal components of them, namely, the cyanin and the peonidin, are non-methoxylated and methoxylated anthocyanins, respectively. Anthocyanin O-methyltransferases (AOMTs) are an important group of enzymes that have the ability to catalyze anthocyanins methylation to promote the solubility, stability, and bioactivity of anthocyanins. Although AOMT genes have been studied in a variety of plants, the function of them in wax apple is generally not well understood. The anthocyanin composition in peels of two wax apple cultivars was determined by High Performance Liquid Chromatography Tandem Mass Spectrometry (HPLS-MS). The genome-wide analysis of the AOMT genes was performed with bioinformatics technology, and the expression patterns of different plant tissues, cultivars, fruit ripening stages, and exogenous abscisic acid (ABA) treatments were analyzed by transcriptome sequencing analysis and real-time quantitative PCR verification. An initial functional evaluation was carried out in vitro using recombinant the Anthocyanin O-methyltransferase Gene 5 of S. samarangense (SsAOMT5) protein. Only two main compositions of anthocyanin were found in peels of two wax apple cultivars, and it was worth noting that Tub Ting Jiang cultivar contained non-methoxylated anthocyanin (Cy3G) only, whereas Daye cultivar contained both non-methoxylated and methoxylated (Pn3G) anthocyanins. A total of six SsAOMT genes were identified in the whole genome of wax apple, randomly distributing on three chromosomes. A phylogenic analysis of the protein sequences divided the SsAOMT gene family into three subgroups, and all SsAOMTs had highly conserved domains of AOMT family. In total, four types of stress- related and five types of hormone- related cis-elements were discovered in the promoter region of the SsAOMTs. Expression pattern analysis showed that SsAOMT5 and SsAOMT6 were expressed in all tissues to varying degrees; notably, the expression of SsAOMT5 was high in the flower and fruit and significantly higher in Daye peels than those of other cultivars in the fruit ripening period. Exogenous ABA treatment significantly increased anthocyanin accumulation, but the increase of methoxylated anthocyanin content did not reach significant level compared with those without ABA treatment, whereas the expression of SsAOMT5 upregulated under ABA treatment. We identified two homologous SsAOMT5 genes from Daye cultivar (DSsAOMT5) and Tub Ting Jiang cultivar (TSsAOMT5); the results of functional analyses to two SsAOMT5 recombinant proteins in vitro demonstrated that DSsAOMT5 showed methylation modification activity, but TSsAOMT5 did not. In conclusion, SsAOMT5 was responsible for methylated anthocyanin accumulation in the peels of wax apple and played an important role in red coloration in wax apple peels.

A mulberry 9-cis-epoxycarotenoid dioxygenase gene MaNCED1 is involved in plant growth regulation and confers salt and drought tolerance in transgenic tobacco.

The phytohormone abscisic acid (ABA) is vital in regulating root elongation, seed germination, and abiotic stress responses in plants. Conversely, the mechanisms of ABA in mulberry root growth, seed germination, and abiotic stress responses are poorly understood. Here, we reported that exogenous ABA and drought treatment inhibited the growth of mulberry seedlings but significantly increased the ratio of root/stem. Inhibition of ABA synthesis by fluridone and sodium tungstate resulted in the decrease of root/stem ratio. We also showed that the expression of MaNCED1 in the root was strongly induced by drought and salt stress. Increasing the expression of MaNCED1 in tobacco using overexpression leads to increased root elongation and reduced seed germination. Compared with the wild type, the accumulation of H2O2 and MDA was reduced, while the POD activity and proline content was increased in the transgenic plants after drought and salt treatment. Further studies revealed increased resistance to drought and salt stress in MaNCED1 overexpressed tobaccos. Meanwhile, the auxin and ethylene signal pathway-related gene expression levels increased in MaNCED1 overexpressed tobaccos. This study demonstrated the roles of mulberry MaNCED1 in regulating plant development and abiotic stress responses. It gave further insights into the coordinated regulation of ABA, auxin, and ethylene in seed growth and germination.

Quality analysis and metabolomic profiling of the effects of exogenous abscisic acid on rabbiteye blueberry.

Blueberry is a characteristic berry fruit shrub of the genus Vaccinium in the Rhododendron family. The fruit is rich in anthocyanins and has a variety of nutritional and health functions. This study aimed to systematically study the effect of exogenous abscisic acid (ABA) application on ripening and metabolites in blueberry fruits. Blueberry fruit ripening was divided into six stages for further analysis. In this study, nontarget metabolomics was performed to demonstrate the effect on metabolite levels. The results showed that 1000 mg/L ABA significantly promoted fruit ripening and increased anthocyanin content. Moreover, exogenous ABA treatment can affect endogenous ABA levels and improve its antioxidant capacity. Important metabolites of the flavonoid pathway were detected, and the results showed that anthocyanin synthesis increased, and some other bioactive metabolite levels decreased. After comprehensive assessments, we believe that 1000 mg/L exogenous ABA application will have positive impacts on blueberry fruit quality and economic benefits.

ABA-triggered ROS burst in rice developing anthers is critical for tapetal programmed cell death induction and heat stress-induced pollen abortion.

High temperatures (HT) cause pollen abortion and poor floret fertility in rice, which is closely associated with excessive accumulation of reactive oxygen species (ROS) in the developing anthers. However, the relationships between accumulation of abscisic acid (ABA) and ROS, and their effects on tapetum-specific programmed cell death (PCD) in HT-stressed anthers are poorly characterised. Here, we determined the spatiotemporal changes in ABA and ROS levels, and their relationships with tapetal PCD under HT exposure. Mutants lacking ABA-activated protein kinase 2 (SAPK2) functions and exogenous ABA treatments were used to explore the effects of ABA signalling on the induction of PCD and ROS accumulation during pollen development. HT-induced pollen abortion was tightly associated with ABA accumulation and oxidative stress. The higher ABA level in HT-stressed anthers resulted in the earlier initiation of PCD induction and subsequently abnormal tapetum degeneration by activating ROS accumulation in developing anthers. Interactions between SAPK2 and DEAD-box ATP-dependent RNA helicase elF4A-1 (RH4) were required for ABA-induced ROS generation in developing anthers. The OsSAPK2 knockout mutants showed the impaired PCD responses in the absence of HT. However, the deficiency of SAPK2 functions did not suppress the ABA-mediated ROS generation in HT-stressed anthers.

Abscisic acid alleviates mercury toxicity in wheat (Triticum aestivum L.) by promoting cell wall formation

Mercury (Hg) is a heavy metal (HM) that affects crop growth and productivity. In a previous study, we found that application of exogenous abscisic acid (ABA) alleviated growth inhibition in Hg-stressed wheat seedlings. However, the physiological and molecular mechanisms underlying ABA-mediated Hg detoxification remained unclear. In this study, Hg exposure reduced the plant fresh and dry weights and root numbers. Exogenous ABA treatment significantly resumed the plant growth, increased the plant height and weight, and enriched the roots numbers and biomass. The application of ABA enhanced Hg absorption and raised the Hg levels in the roots. In addition, exogenous ABA decreased Hg-induced oxidative damage and significantly brought down the activities of antioxidant enzymes, such as SOD, POD and CAT. Global gene expression patterns in the roots and leaves exposed to HgCl2 and ABA treatments were examined via RNA-Seq. The data showed that genes related to ABA-mediated Hg detoxification were enriched in functions related to cell wall formation. Weighted gene co-expression network analysis (WGCNA) further indicated that the genes implicated in Hg detoxification were related to cell wall synthesis. Under Hg stress, ABA significantly induced expression of the genes encoding cell wall synthesis enzymes, regulated the activity of hydrolase, and increased the concentrations of cellulose and hemicellulose, hence promoting cell wall synthesis. Taken together, these results suggest that exogenous ABA could alleviate Hg toxicity in wheat by promoting cell wall formation and suppressing translocation of Hg from roots to shoots.

Identification and Expression Analysis of Abscisic Acid Signal Transduction Genes in Hemp Seeds

Abscisic acid (ABA) is involved in regulating diverse biological processes, but its signal transduction genes and roles in hemp seed germination are not well known. Here, the ABA signaling pathway members, <i>PYL</i>, <i>PP2C</i> and <i>SnRK2</i> gene families, were identified from the hemp reference genome, including 7 <i>CsPYL</i> (pyrab-actin resistance1-like, ABA receptor), 8 <i>CsPP2CA</i> (group A protein phosphatase 2c), and 7 <i>CsSnRK2</i> (sucrose nonfermenting1-related protein kinase 2). The content of ABA in hemp seeds in germination stage is lower than that in non-germination stage. Exogenous ABA (1 or 10 μM) treatment had a significant regulatory effect on the selected <i>PYL</i>, <i>PP2C</i>, <i>SnRK2</i> gene families. <i>CsAHG3</i> and <i>CsHAI1</i> were most significantly affected by exogenous ABA treatment. Yeast two-hybrid experiments were performed to reveal that CsPYL5, CsSnRK2.2, and CsSnRK2.3 could interact with CsPP2CA7 and demonstrate that this interaction was ABA-independent. Our results indicated that <i>CsPYL5</i>, <i>CsSnRK2.2</i>, <i>CsSnRK2.3</i> and <i>CsPP2CA7</i> might involve in the ABA signaling transduction pathway of hemp seeds during the hemp seed germination stages. This study suggested that novel genetic views can be brought into investigation of ABA signaling pathway in hemp seeds and lay the foundation for further exploration of the mechanism of hemp seed germination.