Abscisic acid is a key inducer of hydrogen peroxide production in leaves of maize plants exposed to water stress

The histochemical and cytochemical localization of water stress-induced H(2)O(2) production in the leaves of ABA-deficient vp5 mutant and wild-type maize (Zea mays L.) plants were examined, using 3,3-diaminobenzidine and CeCl(3) staining, respectively, and the roles of endogenous ABA in the production of H(2)O(2) induced by water stress were assessed. Water stress induced by polyethylene glycol resulted in the accumulation of H(2)O(2) in mesophyll cells, bundle-sheath cells and vascular bundles of wild-type maize leaves, and the accumulation was substantially blocked in the mutant maize leaves exposed to water stress. Pre-treatments with several apoplastic H(2)O(2) manipulators abolished the majority of H(2)O(2) accumulation induced by water stress in the wild-type leaves. The subcellular localization of H(2)O(2) production was demonstrated in the cell walls, xylem vessels, chloroplasts, mitochondria and peroxisomes in the leaves of wild-type maize plants exposed to water stress, and the accumulation of H(2)O(2) induced by water stress in the cell walls and xylem vessels, but not in the chloroplasts, mitochondria and peroxisomes, was arrested in the leaves of the ABA mutant or the ABA biosynthesis inhibitor (tungstate)-pre-treated maize plants. Pre-treatments with the apoplastic H(2)O(2) manipulators also blocked the apoplastic but not the intracellular H(2)O(2) accumulation induced by water stress in the leaves of wild-type plants. These data indicate that under water stress, the apoplast is the major source of H(2)O(2) production and ABA is a key inducer of apoplastic H(2)O(2) production. These data also suggest that H(2)O(2) generated in the apoplast could not diffuse freely into subcellular compartments.     

一氧化氮参与调节盐胁迫诱导的玉米幼苗脱落酸积累

以三叶一心期的玉米幼苗为实验材料,研究了盐胁迫下玉米幼苗根尖和叶片中一氧化氮（NO）和脱落酸（ABA）积累之间的关系。结果表明,盐胁迫下玉米幼苗NO和ABA的含量均增加,用NO供体硝普钠（Sodium nitroprusside,SNP）处理时,ABA含量亦增加,且累积的时间较盐胁迫下早。用NO合成的抑制剂L-NAME （Nω-nitro-L-arginine methyl ester hydrochloride）和NaN,处理时,可减弱盐胁迫诱导的ABA含量的增加,用NO清除剂cPTIO处理时,这种盐胁迫诱导的ABA增加减少。推测盐胁迫下产生的NO参与调节ABA的积累及逆境下植物的防御反应。     

Involvement of Polyamine Oxidase in Abscisic Acid induced Cytosolic Antioxidant Defense in Leaves of Maize

Using pharmacological and biochemical approaches, the role of maize polyamine oxidase (MPAO) in abscisic acid (ABA)induced antioxidant defense in leaves of maize (Zea mays L.) plants was investigated. Exogenous ABA treatment enhanced the expression of the MPAO gene and the activities of apoplastic MPAO. Pretreatment with two different inhibitors for apoplastic MPAO partly reduced hydrogen peroxide (H2O2) accumulation induced by ABA and blocked the ABA-induced expression of the antioxidant genes superoxide dismutase 4 and cytosolic ascorbate peroxidase and the activities of the cytosolic antioxidant enzymes. Treatment with spermidine, the optimum substrate of MPAO, also induced the expression and the activities of the antioxidant enzymes, and the upregulation of the antioxidant enzymes was prevented by two inhibitors of MPAO and two scavengers of H2O2. These results suggest that MPAO contributes to ABA-induced cytosolic antioxidant defense through H2O2, a Spd catabolic product.     

ABA controls H₂O₂ accumulation through the induction of OsCATB in rice leaves under water stress

The production of both ABA and H?O? is induced by drought and can act as signals under stress conditions. We investigated the relationships between ABA, H?O? and catalase (CAT) in rice leaves when rice seedlings were treated with polyethylene glycol as water stress treatment. As a key gene in ABA biosynthesis, OsNCED3 was significantly induced in rice by water stress treatment and such induction preceded the rapid increase in ABA. Water stress inhibited the expression of CATA and CATC but substantially enhanced the expression of CATB. Exogenously applied ABA promoted the expression of CATB also and inhibited the expression of CATC in a concentration-dependent manner. When ABA production was inhibited by using ABA biosynthesis inhibitors nordihydroguaiaretic acid and tungstate, expression of CATB was also subdued while CATC was enhanced under the water stress. Accumulation of H?O? was also reduced when endogenous ABA production was inhibited and showed a correlation with the total activity of catalases. Our results suggest that water stress-induced ABA prevents the excessive accumulation of H?O?, through the induction of the expression of CATB gene during water stress.     

Effect of ABA on the contents of proline, polyamines, and cytokinins in the common ice plants under salt stress

The effects of ABA treatment on the contents of proline, polyamines (PA), and cytokinins (CK) in the facultative halophyte the common ice plant (Mesembryanthemum crystallinum L.) subjected to salt stress were studied. Plants grown in the phytotron chamber on Jonson nutrient medium for 6 weeks were subjected to 6-day-long salinity by a single NaCl adding to medium. During first three days of salinity, half plants of each treatment were placed for 30 min on nutrient medium containing 0, 100, or 300 mM NaCl plus ABA in the final concentration of 1 μM. Salinity reduced biomass accumulation and water and chlorophyll contents in plants. This was accompanied by the increase in the levels of MDA, proline, and sodium ions. ABA treatment of salt-stressed plants favored biomass accumulation and photosynthetic pigment protection, reduced the intensity of oxidative stress and the level of NaCl-induced proline accumulation. ABA treatment increased the contents of putrescine (Put) and spermidine (Spd) in the leaves and roots of control plants (not subjected to salt stress), reduced the losses of Put in the leaves and roots and Spd in the roots in the presence of 100 mM NaCl, and suppressed cadaverine (Cad) accumulation in the roots in the presence of 300 mM NaCl. In the presence of NaCl, ABA reduced the contents of zeatin and zeatin riboside and increased the level of zeatin-O-glucoside in the roots and isopentenyladenosine and isopentenyladenine in the leaves. Thus, ABA protective action under salinity can be realized through the weakening of oxidative stress (a decrease in MDA content) and the regulation of PA, proline, and CK metabolism, which has a great significance in plant adaptation to injurious factors.     

Nitric oxide mediates brassinosteroid-induced ABA biosynthesis involved in oxidative stress tolerance in maize leaves

The role of ABA in brassinosteroid (BR)-induced stress tolerance and the relationship between BR, nitric oxide (NO) and ABA under water stress induced by polyethylene glycol (PEG) were investigated in leaves of maize (Zea mays) plants. Water stress led to oxidative damage. Pre-treatment with the BR biosynthetic inhibitor brassinazole (Brz) aggravated the oxidative damage induced by PEG treatment, which was alleviated by the application of BR or ABA. Pre-treatment with the ABA biosynthetic inhibitor fluridone also aggravated the oxidative damage induced by PEG treatment; however, this was barely alleviated by the application of BR. BR treatment increased the content of ABA and up-regulated the expression of the ABA biosynthetic gene vp14 in maize leaves, which was blocked by pre-treatments with the NO scavenger cPTIO (2,4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) and the nitric oxide synthase inhibitor l-NAME (N(G)-nitro-l-arginine methyl ester. Moreover, BR treatment induced increases in the generation of NO in mesophyll cells of maize leaves, and treatment with the NO donor sodium nitroprusside (SNP) up-regulated the content of ABA and the expression of vp14 in maize leaves. Our results suggest that BR-induced NO production and NO-activated ABA biosynthesis are important mechanisms for BR-enhanced water stress tolerance in leaves of maize plants.     

Regulation by ABA of osmotic-stress-induced changes in protein synthesis in tomato roots

Polypeptide synthesis and accumulation were examined in the roots of tomato seedlings exposed to a polyethylene glycol‐imposed water deficit stress. In these roots, the synthesis of a number of polypeptides was induced, while that of several others was enhanced or repressed. To examine the role played by abscisic acid (ABA) in co‐ordinating the accumulation of these proteins, water‐deficit‐stress‐responsive polypeptide synthesis was investigated in the roots of the ABA‐deficient mutant flacca. In the roots of this mutant, the ability to accumulate a complete set of water‐deficit‐stress‐responsive polypeptides was impaired, indicating that ABA is required for their synthesis. The role of ABA was further examined by exposing the roots of both genotypes to exogenous ABA, which, with one exception, elicited the accumulation of all water‐deficit‐stress‐responsive proteins. Polyethylene glycol‐induced polypeptide accumulation was accompanied by a 1·6‐fold increase in the level of endogenous ABA in the roots of wild‐type plants and a 5‐fold increase in the roots of flc. Thus, although the absolute level was lower than that of the wild‐type, flc has the capacity to accumulate ABA in its roots. When fluridone was used to prevent the biosynthesis of ABA, the accumulation of several water‐deficit‐stress‐responsive polypeptides was reduced further. The synthesis of polypeptides was also examined in the roots of salt‐treated seedlings. Salt altered the accumulation of several polypeptides, all of which were previously observed in water‐deficit‐stressed roots, indicating that their synthesis was the result of the osmotic component of the salt stress. However, the accumulation of these polypeptides was not impaired in flc roots, indicating that the role played by ABA in regulating their accumulation in salt‐and polyethylene glycol‐treated roots differs. As such, salt‐ and water‐deficit‐stress‐induced changes in gene expression may be effected by different mechanisms, at least at the level of polypeptide accumulation.     

Involvement of Protein Phosphorylation in Water Stress-induced Antioxidant Defense in Maize Leaves

Using pharmacological and biochemical approaches, the role of protein phosphorylation and the interrelationship between water stress-enhanced kinase activity, antioxidant enzyme activity, hydrogen peroxide (H2O2) accumulation and endogenous abscisic acid in maize (Zea mays L.) leaves were investigated. Water-stress upregulated the activities of total protein phosphorylation and Ca2+ -dependent protein kinase, and the upregulation was blocked in abscisic acid-deficient vp5 mutant. Furthermore, pretreatments with a nicotinamide adenine dinucleotide phosphate oxidase inhibitor and a scavenger of H2O2 significantly reduced the increased activities of total protein kinase and Ca2+-dependent protein kinase in maize leaves exposed to water stress. Pretreatments with different protein kinase inhibitors also reduced the water stress-induced H2O2 production and the water stress-enhanced activities of antioxidant enzymes such as superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase. The data suggest that protein phosphorylation and H2O2 generation are required for water stress-induced antioxidant defense in maize leaves and that crosstalk between protein phosphorylation and H2O2 generation may occur.     

外源ABA对楸树幼苗NaCl胁迫的缓解效应及其生长生理响应特征

以2年生楸树(苏楸1号和008-1)扦插苗为材料,采用盆栽试验法,分析盐胁迫(0.5%NaCl)处理下楸树幼苗生长、生理的变化,并分析不同浓度外源ABA(15、25、35 mg/L)对盐胁迫(30 d)楸树幼苗的缓解效应及其生理生化特性,以探索重度盐胁迫下适合楸树幼苗生长的适宜外源ABA浓度,为增强盐碱地楸树的耐盐性、提高盐碱地的利用提供理论依据。结果显示:(1)0.5%NaCl胁迫下,两品种楸树幼苗叶片表现出不同程度的盐害症状,且‘苏楸1号’叶片盐害症状较‘008-1’严重;随胁迫时间延长,两品种楸树幼苗的相对电导率(REC)均呈先上升后下降的变化趋势,叶绿素(Chl)、相对含水量(RWC)均呈降低趋势,可溶性糖(SS)、可溶性蛋白(SP)、脯氨酸(Pro)以及超氧化物歧化酶(SOD)活性均呈先上升后下降趋势,但‘008-1’的REC显著低于‘苏楸1号’,Chl、RWC、SS、SP、Pro、SOD均显著高于苏楸1号,表明‘008-1’的耐盐性较‘苏楸1号’更强。(2)喷施外源ABA使得盐胁迫下‘008-1’楸树的苗高显著增加、新叶提前萌发,表明外源ABA在一定程度上能够缓解盐胁迫对楸树生长的影响;喷施外源ABA降低了盐胁迫下‘008-1’楸树幼苗叶片的REC,提高了Chl、RWC、SS、SP、Pro、SOD、过氧化物酶(POD)以及过氧化氢酶(CAT)活性,促进了内源激素生长素(IAA)、脱落酸(ABA)、赤霉素(GA3)以及玉米素核苷(ZR)的积累。研究表明,楸树品种‘008-1’的耐盐性更强;外源喷施适宜浓度ABA能够缓解盐胁迫对楸树幼苗生长的影响,降低幼苗叶片细胞膜透性,促进幼苗渗透调节物质的积累,增强渗透调节能力,并提高盐胁迫下幼苗的抗氧化酶活性,促进植物对内源激素含量的调节,从而提高楸树的耐盐性,且以25 mg/L ABA处理的效果最好。     

Abscisic acid accumulation maintains maize primary root elongation at low water potentials by restricting ethylene production

Previous work showed that primary root elongation in maize (Zea mays L.) seedlings at low water potentials (psi(w)) requires the accumulation of abscisic acid (ABA) (R.E. Sharp, Y. Wu, G.S. Voetberg, I.N. Saab, M.E. LeNoble [1994] J Exp Bot 45: 1743-1751). The objective of the present study was to determine whether the inhibition of elongation in ABA-deficient roots is attributable to ethylene. At a psi(w) of -1.6 MPa, inhibition of root elongation in dark-grown seedlings treated with fluridone to impose ABA deficiency was largely prevented with two inhibitors of ethylene synthesis (aminooxyacetic acid and aminoethoxyvinylglycine) and one inhibitor of ethylene action (silver thiosulfate). The fluridone treatment caused an increase in the rate of ethylene evolution from intact seedlings. This effect was completely prevented with aminooxyacetic acid and also when ABA was supplied at a concentration that restored the ABA content of the root elongation zone and the root elongation rate. Consistent results were obtained when ABA deficiency was imposed using the vp5 mutant. Both fluridone-treated and vp5 roots exhibited additional morphological symptoms of excess ethylene. The results demonstrate that an important role of ABA accumulation in the maintenance of root elongation at low psi(w) is to restrict ethylene production.     

Wounding stress induces alcohol dehydrogenase in maize and lettuce seedlings

To understand the effect of wounding stress on alcohol dehydrogenase(ADH, EC 1.1.1.1) in monocotyledonous and dicotyledonous plants, maize(Zea mays L.) and lettuce (Lactucasativa L.) seedlings were subjected to wounding stress and ADHactivity and abscisic acid (ABA) concentration were determined. In response tothe stress, the ADH activity in seedlings of both species increased rapidly asaresult of increased synthesis of the ADH. At 12 h after thestress,the activities in the wounded lettuce and maize seedlings, respectively,increased to 1.7- and 1.5-fold of that in non-stressed seedlings. Woundingstress also increased the concentration of endogenous ABA during the first 6h in both seedlings. The maximum increased levels of ABA in thelettuce and maize seedlings were 4.9- and 4.7-fold of that in the non-stressedseedlings, respectively.     

水分胁迫诱导玉米叶片质外体产生H2O2机理

通过组织化学染色、电镜观察、酶活性分析对水分胁迫诱导玉米叶片质外体产生H2O2进行了研究。结果表明:水分胁迫能够诱导玉米叶片内源ABA的积累,ABA参与了水分胁迫诱导的玉米叶片H2O2的产生,质膜NADPH氧化酶、细胞壁过氧化物酶(POD)以及质外体多胺氧化酶(PAO)是水分胁迫诱导玉米细胞在质外体产生H2O2的来源,其中质膜NADPH氧化酶是主要来源;内源ABA的积累参与了水分胁迫激活的质膜NADPH氧化酶、细胞壁POD和质外体PAO活性的提高。研究认为,水分胁迫诱导玉米细胞在质外体产生H2O2可能是由于水分胁迫下内源ABA的积累通过激活质膜NADPH氧化酶、细胞壁POD以及质外体PAO的活性而实现的。     

The maize Viviparous10/Viviparous13 locus encodes the Cnx1 gene required for molybdenum cofactor biosynthesis

Abscisic acid (ABA), auxin and nitrate are important signaling molecules that affect plant growth responses to the environment. The synthesis or metabolism of these compounds depends on the molybdenum cofactor (MoCo). We show that maize (Zea mays) viviparous10 (vp10) mutants have strong precocious germination and seedling lethal phenotypes that cannot be rescued with tissue culture. We devised a novel PCR-based method to clone a transposon-tagged allele of vp10, and show that Vp10 encodes the ortholog of Cnx1, which catalyzes the final common step of MoCo synthesis. The seedling phenotype of vp10 mutants is consistent with disruptions in ABA and auxin biosynthesis, as well as a disruption in nitrate metabolism. Levels of ABA and auxin are reduced in vp10 mutants, and vp10 seedlings lack MoCo-dependent enzyme activities that are repairable with exogenous molybdenum. vp10 and an Arabidopsis cnx1 mutant, chlorate6 (chl6), have similar defects in aldehyde oxidase (AO) enzyme activity, which is required for ABA synthesis. Surprisingly, chl6 mutants do not show defects in abiotic stress responses. These observations confirm an orthologous function for Cnx1 and Vp10, as well as defining a characteristic viviparous phenotype to identify other maize cnx mutants. Finally, the vp10 mutant phenotype suggests that cnx mutants can have auxin- as well as ABA-biosynthesis defects, while the chl6 mutant phenotype suggests that low levels of AO activity are sufficient for normal abiotic stress responses.     

Effects of abscisic acid on the contents of polyamines and proline in common bean plants under salt stress

The effects of ABA treatment on the contents of polyamines (PAs) and proline (Pro) in the glycophyte Phaseolus vulgaris L. during plant adaptation to salt stress were studied. Two-week-old common bean seedlings grown in the phytotron chamber on the Jonson nutrient medium were subjected to salinity for 6 days by one-time NaCl addition to medium up to final concentrations of 50 and 100 mM. During first three days of salinity, the root system was daily treated with ABA (1, 5, 10, or 50 μM) for 30 min. Salt stress (100 mM NaCl) elevated the level of endogenous ABA, increased the content of Pro 14-fold, reduced sharply the content of free PAs (putrescine, spermidine, spermine, and cadaverine), and the accumulation of 1,3-diaminopropan, a product of oxidation of high-molecular PAs. Common bean plant treatment with 1 μM ABA weakened the adverse effects of salt stress (100 mM NaCl), which was manifested in the maintenance of plant growth, stimulation of chlorophyll (a and b) and carotenoid accumulation, a stabilization of water and Na⁺ balance. Seedling treatment with ABA suppressed NaCl-induced Pro and intracellular ABA accumulation and restored the levels of putrescine and spermidine. The content of spermine in the leaves of plants subjected to salt stress and treated with ABA was approximately threefold higher than in control plants, whereas the content of cadaverine increased under similar conditions more than fivefold. Simultaneously, the contents of 1,3-diaminopropan and malondialdehyde as well as activity of superoxide dismutase were reduced, which indicates a weakening of oxidative stress, one of the possible causes of defensive ABA effects against salt stress. In addition, the suppression by exogenous ABA of Pro accumulation and stimulation of PA content under salt stress confirm indirectly our hypothesis that ABA is involved in the coordinated regulation of two biosynthetic pathways, Pro and PA formation, which use a common precursor, glutamate, and play an important protective role during stress in plants.     

Exogenous Melatonin Improves the Growth of Rice Seedlings by Regulating Redox Balance and Ion Homeostasis Under Salt Stress

This study evaluated the effects of foliar spraying melatonin (MT) on the growth of salt-stressed rice. Seedlings were treated with 50 and 100 mM of NaCl and different concentrations of MT (25, 50, 100, 200, 300, and 400 μM) for 14 days. Different concentrations of MT could promote plant growth significantly under salt stress, particularly at concentrations of 200, 300, and 400 μM. A concentration of 200 μM MT was considered as optimal and used in a subsequent experiment on biomass, water content, antioxidation, mineral nutrition, salt absorption, and distribution of salt-stressed rice seedlings. Results showed that MT’s promoting effect on plant growth under salt stress was evident with time, particularly under high salt stress. MT improved the activities of antioxidant enzymes, reduced membrane lipid peroxidation, alleviated cell injury in plant leaves, and increased N content and Si accumulation in the leaves and roots under salt stress, particularly under high salinity. This compound also inhibited Na uptake and upward transport, but it promoted or maintained the uptake and upward transport of K and Ca in salt-stressed rice. Thus, MT improved the ion homeostasis of K/Na and Ca/Na in plants, particularly in the leaves. Foliar spraying of MT alleviated salt stress on rice by promoting nutrient accumulation or translocation, improving ion homeostasis, which is evident in the leaves, and consequently enhancing its salt resistance. The antioxidative improvement caused by MT might also be related to the improved ion homeostasis.

     

Water stress-induced abscisic acid accumulation triggers the increased generation of reactive oxygen species and up-regulates the activities of antioxidant enzymes in maize leaves

The interrelationship among water-stress-induced abscisic acid (ABA) accumulation, the generation of reactive oxygen species (ROS), and the activities of several antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) was investigated in leaves of detached maize (Zea mays L.) plants exposed to -0.7 MPa water stress induced by polyethylene glycol (PEG 6000). Time-course analyses of ABA content, the production of ROS, and the activities of antioxidant enzymes in water-stressed leaves showed that a significant increase in the content of ABA preceded that of ROS, which was followed by a marked increase in the activities of these antioxidant enzymes. Pretreatment with an ABA biosynthesis inhibitor, tungstate, significantly suppressed the accumulation of ABA, and also reduced the increased generation of ROS and the up-regulation of these antioxidant enzymes in water-stressed leaves. A mild oxidative stress induced by paraquat, which generates O(2)(-) and then H(2)O(2), resulted in a significant enhancement in the activities of antioxidant enzymes in non-water-stressed leaves. Pretreatment with some ROS scavengers, such as Tiron and dimethylthiourea (DMTU), and an inhibitor of NAD(P)H oxidase, diphenyleneiodonium (DPI), almost completely arrested the increase in ROS and the activities of these antioxidant enzymes induced by water stress or ABA treatment. These data suggest that water stress-induced ABA accumulation triggers the increased generation of ROS, which, in turn, leads to the up-regulation of the antioxidant defence system.     

Role of nitric oxide in abscisic acid-induced subcellular antioxidant defense of maize leaves

The sources of nitric oxide (NO) production in response to abscisic acid (ABA) and the role of NO in ABA-induced hydrogen peroxide (H(2)O(2)) accumulation and subcellular antioxidant defense in leaves of maize (Zea mays L.) plants were investigated. ABA induced increases in generation of NO and activity of nitric oxide synthase (NOS) in maize leaves. Such increases were blocked by pretreatment with each of the two NOS inhibitors. Pretreatments with a NO scavenger or NR inhibitors inhibited ABA-induced increase in production of NO, but did not affect the ABA-induced increases in activity of NOS, indicating that ABA-induced NO production originated from sources of NOS and NR. ABA- and H(2)O(2)-induced increases in expression of the antioxidant genes superoxide dismutase 4 (SOD4), cytosolic ascorbate peroxidase (cAPX), and glutathione reductase 1 (GR1) and the activities of the chloroplastic and cytosolic antioxidant enzymes were arrested by pretreatments with the NO scavenger, inhibitors of NOS and NR, indicating that NO is involved in the ABA- and H(2)O(2)-induced subcellular antioxidant defense reactions. On the other hand, NO donor sodium nitroprusside (SNP) reduced accumulation of H(2)O(2) induced by ABA, and c-PTIO reversed the effect of SNP in decreasing the accumulation of H(2)O(2). SNP induced increases in activities of subcellular antioxidant enzymes, and the increases were substantially prevented from occurring by the pretreatment with c-PTIO. These results suggest that ABA induces production of H(2)O(2) and NO, which can up-regulate activities of the subcellular antioxidant enzymes, to prevent overproduction of H(2)O(2) in maize plants. There is a negative feedback loop between NO and H(2)O(2) in ABA signal transduction in maize plants.