Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid

Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.

Rice ONAC016 promotes leaf senescence through abscisic acid signaling pathway involving OsNAP

Senescence-induced NAC(senNAC)TFs play a crucial role in senescence during the final stage of leaf development.In this study,we identified a rice senNAC,ONAC016,which functions as a positive regulator of leaf senescence.The expression of ONAC016 increased rapidly in rice leaves during the progression of dark-induced and natural senescence.The onac016-1 knockout mutant showed a delayed leaf yellowing phenotype,whereas the overexpression of ONAC016 accelerated leaf senescence.Notably,ONAC016 expression was upregulated by abscisic acid(ABA),and thus detached leaves of the onac016-1 mutant remained green much longer under ABA treatment.Quantitative RT-PCR analysis showed that ONAC016 upregulates the genes associated with chlorophyll degradation,senescence,and ABA signaling.Yeast one-hybrid and dual-luciferase assays revealed that ONAC016 binds directly to the promoter regions of OsNAP,a key gene involved in chlorophyll degradation and ABA-induced senescence.Taken together,these results suggest that ONAC016 plays an important role in promoting leaf senescence through the ABA signaling pathway involving OsNAP.

Melatonin Alleviates Abscisic Acid Deficiency Inhibition on Photosynthesis and Antioxidant Systems in Rice under Salt Stress

Melatonin and abscisic acid,as major plant hormones,play important roles in the physiological and biochemical activities of crops,but the interaction between the two under salt stress is not yet clear.This study investigated the endogenous levels of melatonin and abscisic acid in rice by using exogenous melatonin,abscisic acid,and their synthetic inhibitors,and examined their interactions under salt stress.The research results indicate that melatonin and abscisic acid can improve rice salt tolerance.Melatonin alleviated the salt sensitivity caused by abscisic acid deficiency,increased antioxidant enzyme activity and antioxidant content in rice treated with abscisic acid synth-esis inhibitors,and reduced total reactive oxygen species content and thiobarbituric acid reactive substance accu-mulation.Melatonin also increased the activity of key photosynthetic enzymes and the content of photosynthetic pigments,maintaining the parameters of photosynthetic gas exchange and chlorophyllfluorescence.In summary,melatonin alleviated the effects of abscisic acid deficiency on photosynthesis and antioxidant systems in rice and improved salt tolerance.This study is beneficial for expanding the understanding of melatonin regulation of crop salt tolerance.

Effects of Abscisic Acid on Cold Resistance in Digitaria sanguinalis(L.)Scop.

[Objectives]This study was conducted to detect the protective effect of abscisic acid on chilling injury of Digitaria sanguinalis(L.)Scop,and whether this effect is related to antioxidant enzymes and osmotic adjustment.[Methods]D.sanguinalis plants were sprayed with abscisic acid solution,and exposed to cold stress at 15℃for 3 d after one day and then at 5℃for 25 to 30 d in a growth chamber.The changes of plant osmotic potential under this treatment were detected.[Results]Under low temperature stress,the osmotic potential of plants in the abscisic acid treatment and the control increased,but the osmotic potential level of the abscisic acid treatment plants was lower.The SOD activity of plants in the ABA treatment and the control decreased under low temperature stress.Under low temperature stress,the activity of catalase and peroxidase in ABA-treated plants was higher than that in control plants.[Conclusions]This study provides a theoretical basis for the impact of abscisic acid on the physiological response of D.sanguinalis to cold injury.

NaCl Facilitates Cell Wall Phosphorus Reutilization in Abscisic Acid Dependent Manner in Phosphorus Deficient Rice Root

Phosphorus(P) starvation in rice facilitates the reutilization of root cell wall P by enhancing the pectin content. NaCl modulates pectin content, however, it is still unknown whether NaCl is also involved in the process of pectin regulated cell wall P remobilization in rice under P starved conditions. In this study, we found that 10 mmol/L NaCl increased the shoot and root biomasses under P deficiency to a remarkable extent, in company with the elevated shoot and root soluble P contents in rice. Further analysis indicated that exogenous NaCl enhanced the root cell wall P mobilization by increasing the pectin methylesterase activity and uronic acid content in pectin suggesting the involvement of NaCl in the process of cell wall P reutilization in P starved rice roots. Additionally, exogenous NaCl up-regulated the expression of P transporter OsPT6, which was induced by P deficiency, suggesting that NaCl also facilitated the P translocation prominently from root to shoot in P starved rice. Moreover, exogenous abscisic acid(ABA) can reverse the NaCl-mediated mitigation under P deficiency, indicating the involvement of ABA in the NaCl regulated root cell wall P reutilization. Taken together, our results demonstrated that NaCl can activate the reutilization of root cell wall P in P starved rice, which is dependent on the ABA accumulation pathway.

Fiber-specific increase of carotenoid content promotes cotton fiber elongation by increasing abscisic acid and ethylene biosynthesis

Cotton fiber is a raw material for the global textile industry and fiber quality is essential to its industrial application.Carotenoids are plant secondary metabolites that may serve as dietary components,regulate light harvesting,and scavenge reactive oxygen species.Although carotenoids accumulate predominantly in rapidly elongating cotton fibers,their roles in cotton fiber development remain poorly understood.In this study,a fiber-specific promoter proSCFP was applied to drive the expression of GhOR1Del,a positive regulator of carotenoid accumulation,to upregulate the carotenoid level in cotton fiber in planta.Fiber length,strength,and fineness were increased in proSCFP:GhOR1Del transgenic cotton and abscisic acid(ABA)and ethylene contents were increased in elongating fibers.The ABA downstream regulator GhbZIP27a stimulated the expression of the ethylene synthase gene GhACO3 by binding to its promoter,suggesting that ABA promoted fiber elongation by increasing ethylene production.These findings suggest the involvement of carotenoids and ABA signaling in promoting cotton fiber elongation and provide a strategy for improving cotton fiber quality.

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,PYL,PP2C and SnRK2 gene families,were identified from the hemp reference genome,including 7 CsPYL(pyrab-actin resistance1-like,ABA receptor),8 CsPP2CA(group A protein phosphatase 2c),and 7 CsSnRK2(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 10μM)treatment had a significant regulatory effect on the selected PYL,PP2C,SnRK2 gene families.CsAHG3 and CsHAI1 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 CsPYL5,CsSnRK2.2,CsSnRK2.3 and CsPP2CA7 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.

RNAi-mediated suppression of the abscisic acid catabolism gene Os ABA8ox1 increases abscisic acid content and tolerance to saline–alkaline stress in rice(Oryza sativa L.)

Saline–alkaline(SA) stress is characterized by high salinity and high alkalinity(high p H), which severely inhibit plant growth and cause huge losses in crop yields worldwide. Here we show that a moderate elevation of endogenous abscisic acid(ABA) levels by RNAi-mediated suppression of Os ABA8 ox1(Os ABA8 ox1-kd), a key ABA catabolic gene, significantly increased tolerance to SA stress in rice plants. We produced Os ABA8 ox1-kd lines in two different japonica cultivars, Dongdao 4 and Nipponbare. Compared with nontransgenic control plants(WT), the Os ABA8 ox1-kd seedlings accumulated 25.9%–55.7% higher levels of endogenous ABA and exhibited reduced plasmalemma injury, ROS accumulation and Na;/K;ratio, and higher survival rates, under hydroponic alkaline conditions simulated by 10, 15, and 20 mmol L-1 of Na;CO;. In pot trials using SA field soils of different alkali levels(p H 7.59, 8.86, and 9.29), Os ABA8 ox1–kd plants showed markedly higher seedling survival rates and more vigorous plant growth, resulting in significantly higher yield components including panicle number(85.7%–128.6%), spikelets per panicle(36.9%–61.9%), branches(153.9%–236.7%), 1000–kernel weight(20.0%–28.6%), and percentage of filled spikelets(96.6%–1340.8%) at harvest time. Under severe SA soil conditions(p H = 9.29, EC = 834.4 μS cm-1),Os ABA8 ox1-kd lines showed an 194.5%–1090.8% increase in grain yield per plant relative to WT plants.These results suggest that suppression of Os ABA8 ox1 to increase endogenous ABA levels provides a new molecular approach for improving rice yield in SA paddies.

Exogenous Application of Abscisic Acid (ABA)Enhances Chilling Tolerance in Seedlings ofNapier Grass (Pennisetum purpureum Schum.)

Napier grass, an important forage crop with potentials in multi-purpose applications, is widely grown throughout the tropics and subtropics. Low temperature severely limits its productivity and geographical distribution in temperate regions of the world. In this study, we investigated the effect of exogenous abscisic Acid(ABA) on chilling tolerance of napier grass(Pennisetum purpureum Schum.) seedlings. Seven-day-old napier grass seedlings were cultured in dd H_2O or ABA solution at different concentrations and exposed to 1 ℃ for different time durations. The chilling injury, membrane stability index(MSI) and proline content were estimated from leaf samples. The results showed that there was obvious morphological injury of leaf blighting and restrained growth for the seedlings under chilling stress, but this damage can be largely reduced(by 2/3) when the seedlings were treated by 100 μmol/L ABA in the culture solution, and that the application of exogenous ABA can help to maintain a good stability of leaf cell membrane as expressed by a high MSI value and a low level of proline in leaf cells. These results suggested that exogenous ABA can significantly alleviate chilling injury in napier grass seedlings by maintaining the stability of leaf cell membrane during chilling stress, and that the chilling tolerance was not ensured by a proline accumulation although a passive accumulation of proline was observed in the seedlings under chilling stress. Our results lay a preliminary foundation for future investigations on the molecular mechanisms of ABA induced chilling or freezing tolerance in napier grass.

Research and Applicationon Plant Hormone--Abscisic Acid, ABA

Abscisic Acid (ABA), along with ethylene, gibberellins, cytokinins and auxins, is regarded as five kinds of important plant hormone. ABA was first isolated from cotton bud by Addcott Ohhuma’s group in 1963. Until 1965, its plane structure was determined. It was formally named as Abscisic acid in "the International Conference of Plant Regulator" in 1967. Scientists all over the world have made a long-term unremitting effort

Exogenous Application of Abscisic Acid or Gibberellin Acid Has Different Effects on Starch Granule Size Distribution in Grains of Wheat

Granule size distribution of wheat starch is an important characteristic that can affect its chemical composition and the functionality of wheat products. Two high-yield winter wheat cultivars were used to evaluate the effects of the application of exogenous ABA or GA during the reproductive phase of the initial grain filling on starch granule size distribution and starch components in grains at maturity. The results indicated that a bimodal curve was found in the volume and surface area distribution of grain starch granules, and a unimodal curve was observed for the number distribution under all treatments. The exogenous ABA resulted in a significant increase in the proportions （both by volume and by surface area） of B-type （〈9.9 Ixm in diameter） starch granules, with a reduction in those of A-type （〉9.9 ~tm） starch granules, while, the exogenous GA3 led to converse effects on size distribution of those starch granules. The exogenous ABA also increased starch, amylose and amylopectin contents at maturity but significantly reduced the ratio of amylose to amylopectin. Application of GA3 significantly reduced starch content, amylopectin content but increased the ratio of amylose to amylopectin. The ratio of amylose to amylopectin showed a significant and negative relationship with the volume proportion of granules 〈9.9 μm, but was positively related to the volume proportion of granules 22.8-42.8 μm.

A novel impedance immunosensor based on O-phenylenediamine modified gold electrode to analyze abscisic acid

An impedance immunosensor based on O-phenylenediamine modified gold electrode for the determination of phytohormone abscisic acid(ABA) was proposed.The operating pH,absorption time,absorption temperature and concentration of anti-ABA antibody were investigated to optimize the analytical performance.The calibration curve for the determination of ABA was obtained from this impedance immunosensor under optimal conditions.The results showed that the detection limit at about 1 ng/mL in the range of 10-5000 ng/mL...

Effects of Formulated Fertilizer Synergist on Abscisic Acid Accumulation, Proline Content and Photosynthetic Characteristics of Rice under Drought

To investigate the effects of formulated fertilizer synergist on the drought tolerance in rice, pot experiment was conducted to analyze the photosynthetic characteristics and the accumulation of abscisic acid （ABA） and proline in middle-season rice variety Peiliangyou 93. The synergist could improve the net photosynthetic rate, and coordination between the water loss and the CO2 absorption as well as reduce the harmful effect on photosynthetic process under drought conditions. Under drought, the ABA accumulated massively both in roots and leaves, while the ABA content in roots was far higher than that in leaves. The results indicate that synergist could increase the ABA accumulation, but reduce the proline accumulation in rice plant under drought.

Effects of Exogenous Trehalose on the Metabolism of Sugar and Abscisic Acid in Tomato Seedlings Under Salt Stress

Salt stress a ects the growth and development of plants, which results in a decrease in crop quality and yield. In this study, we used tomato seedlings treated with salt and trehalose as experimental materials and analyzed them using the technique for order preference by similarity to ideal solution analysis to select the optimal trehalose concentration for treatment. We also determined the contents of sugar and abscisic acid (ABA) and detected the expression of genes involved in the metabolism of sugar and ABA by quantitative real-time PCR. Results showed that the optimal trehalose concentration was 2 mmol/L for tomato seedlings under salt stress. Exogenous trehalose decreased the starch content and increased the soluble sugar con- tent by a ecting the expression of genes related to the metabolism of starch and soluble sugar. Exogenous trehalose altered the accumulation and distribution of sugar by inducing the upregulation of sugar transporter genes. Furthermore, trehalose increased the ABA content to induce salt stress response by regulating the expression of genes related to the synthesis and metabolism of ABA. In conclusion, trehalose can e ectively alleviate salt stress and enhance salt tolerance of tomato. These ndings provide a novel perspective and a better resource to investigate the salt tolerance mechanism and a new method for alleviating salt stress in tomato.

Abscisic acid and jasmonic acid are involved in drought priming-induced tolerance to drought in wheat

Drought stress is a limiting factor for wheat production and food security.Drought priming has been shown to increase drought tolerance in wheat.However,the underlying mechanisms are unknown.In the present study,the genes encoding the biosynthesis and metabolism of abscisic acid(ABA)and jasmonic acid(JA),as well as genes involved in the ABA and JA signaling pathways were up-regulated by drought priming.Endogenous concentrations of JA and ABA increased following drought priming.The interplay between JA and ABA in plant responses to drought priming was further investigated using inhibitors of ABA and JA biosynthesis.Application of fluridone(FLU)or nordihydroguaiaretic acid(NDGA)to primed plants resulted in lower chlorophyll-fluorescence parameters and activities of superoxide dismutase and glutathione reductase,and higher cell membrane damage,compared to primed plants(PD)under drought stress.NDGA+ABA,but not FLU+JA,restored priming-induced tolerance,as indicated by a finding of no significant difference from PD under drought stress.Under drought priming,NDGA induced the suppression of ABA accumulation,while FLU did not affect JA accumulation.These results were consistent with the expression of genes involved in the biosynthesis of ABA and JA.They suggest that ABA and JA are required for priming-induced drought tolerance in wheat,with JA acting upstream of ABA.

Abscisic acid induces differential expression of genes involved in wound-induced suberization in postharvest tomato fruit

Fruit wounding occurred at harvest and transportation requires rapid suberization as a major part of the healing process to prevent infection and desiccation. The focus of this work was to explore the mediation of abscisic acid(ABA) on woundinduced suberization and to determine expression profiles of specific genes involved in wound-induced suberization in tomato fruit. The measurements of weight loss and fruit firmness suggested wound-induced suberization started at 2 d after wounding. The suberization process with the accumulation of suberin polyphenolics(SPP) and polyaliphatics(SPA) observed through autofluorescence microscopy and Sudan IV staining was accelerated by ABA. Expressions of SlPAL5 and Sl4 CL involved in the synthesis of SPP reached the highest at 4 and 8 d after wounding following ABA application, respectively. Associated with SPA biosynthesis, SlLACS1 and SlLACS2 showed the most abundant transcripts at 8 and 6 d in ABA group, respectively. Transcript levels including SlKCSs, SlCYP86 B1, SlFAR3, and Sl GPATs were up-regulated at 2 d after wounding by ABA. Activities of polyphenol oxidase and lipoxygenase were also enhanced during wound-induced suberization following ABA application. The results in this study proved that ABA accelerated the wound-induced suberization progress by increasing the transcript levels of relevant genes in postharvest tomato fruit.

Effects of Abscisic Acid Treatment on Berry Coloration and Expression of Flavonoid Biosynthesis Genes in Grape

In order to enhance berry coloration of bright-red grape cultivars, the effects of abscisic acid (ABA) treatment on the quantity and composition of anthocyanins as well as the expression of genes related to flavonoid biosynthesis in the berry were examined. Exogenous ABA treatment increased anthocyanin content, especially petunidin- and malvidin-type anthocyanins. Quantitative real-time PCR analysis revealed that ABA treatment around véraison resulted in the upregulation of genes encoding enzymes responsible for both general flavonoid and anthocyanin biosynthesis. On the other hand, the gene expressions of enzymes involved in proanthocyanidin synthesis were drastically decreased at véraison and remained extremely low even with ABA treatment. Thus, increases in the total amount and composition ratios of petunidin- and malvidin-type anthocyanins were mainly caused by ABA-induced upregulation of uridine diphosphate glucose flavonoid glucosyl transferase, glutathione S-transferase 4, O-methyl transferase and flavonoid 3’, 5’ hydroxylase expression, resulting in the deep coloration of berry of skin.

Phenotype and mechanism analysis of plant dwarfing in pear regulated by abscisic acid

Close planting of dwarf varieties is currently the main cultivation direction for pear trees,and the screening of excellent dwarf varieties is an important goal for breeders.In this study,the dwarfing pear variety‘601D’and its vigorous mutant‘601T’were used to show their biological characteristics and further explore the dwarfing mechanism in‘601D’.The biological characteristics showed that‘601D’had a shorter internode length,a shorter and more compact tree body,thicker and broader leaves,lower stomata density,larger stomata size(dimension),and higher photosynthetic capacity.The biological characteristics of‘601T’showed notable contrasts.The results of endogenous hormone tests indicated that the contents of abscisic acid(ABA),ABA-glucosyl ester,and GA_(4) were higher in‘601D’,but the trans-zeatin content was lower.By transcriptomic analysis,significant differences were found in the biosynthetic and metabolic pathways of ABA.Related transcription factors such as bHLH,WRKY,and homeobox also participated in the regulation of plant dwarfing.We therefore examined three hormones with obvious differences with‘601T’,and found that only ABA could induce‘601T’to return to a dwarfing plant phenotype.Therefore,we conclude that the dwarfing of‘601D’is caused by an excessive accumulation of ABA.This study provides a new theoretical basis for breeding dwarf varieties.

OsbZIP72 Is Involved in Transcriptional Gene-Regulation Pathway of Abscisic Acid Signal Transduction by Activating Rice High-Affinity Potassium Transporter OsHKT1;1

We created CRISPR-Cas9 knock-out and overexpressing OsbZIP72 transgenic rice plants to gain a better understanding of the role and molecular mechanism of OsbZIP72 gene in stress tolerance,which has remained largely elusive.OsbZIP72 was expressed and integrated into rice transgenic plant genomes,and the OsbZIP72 transcript in overexpression lines was elicited by salinity,abscisic acid(ABA)and drought stresses.OsbZIP72 overexpressing plants showed higher tolerance to drought and salinity stresses,while knock-out transgenic lines showed higher sensitivity to these stresses.The differentially expressed genes(DEGs)from RNA-sequencing data encompassed several abiotic stress genes,and the functional classification of these DEGs demonstrated the robust transcriptome diversity in OsbZIP72.Yeast one-hybrid,along with luciferase assay,indicated that OsbZIP72 acted as a transcriptional initiator.Remarkably,electrophoresis mobility assay revealed that OsbZIP72 bound directly to the ABAresponsive element in the OsHKT1;1 promoter region and activated its transcription.Overall,our findings revealed that OsbZIP72 can act as a transcriptional modulator with the ability to induce the expression of OsHKT1;1 in response to environmental stress through an ABA-dependent regulatory pathway,indicating that OsbZIP72 can play a crucial role in the ABA-mediated salt and drought tolerance pathway in rice.

Purification and Analysis of Abscisic Acid-Specifically-Inducible Proteins from Rice Callus

Two ABA-specifically-inducible proteins from rice callus were isolated and purified by precipitation with 65-100% saturated （NH4）2SO4, followed by the DEAE-sepharose, TSK-gel, and two-dimension electrophoresis. Iso-electric points （pl） of the proteins with the same molecular mass （24.5 kD） were 6.1 and 6.9, respectively. The Western blot analysis indicated that the proteins expressed in different tissues were obviously different. The A1 （pl 6.1） protein was only detected in calli treated with ABA and seed embryos （SE）. However, the A2 （pl 6.9） protein was found not only in the calli treated with ABA and SE, but also in the white dry callus. Thus it suggested that the two proteins might play some important roles in the processes of seed embryo （or somatic embryo） formation.