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Drought stress is a crucial environmental factor that limits plant growth, development, and productivity. Autophagy of misfolded proteins can help alleviate the damage caused in plants experiencing drought. However, the mechanism of autophagy‐mediated drought tolerance in plants remains largely unknown. Here, we cloned the gene for a maize (Zea mays) selective autophagy receptor, NEXT TO BRCA1 GENE 1 (ZmNBR1), and identified its role in the response to drought stress. We observed that drought stress increased the accumulation of autophagosomes. RNA sequencing and reverse transcription‐quantitative polymerase chain reaction showed that ZmNBR1 is markedly induced by drought stress. ZmNBR1 overexpression enhanced drought tolerance, while its knockdown reduced drought tolerance in maize. Our results established that ZmNBR1 mediates the increase in autophagosomes and autophagic activity under drought stress. ZmNBR1 also affects the expression of genes related to autophagy under drought stress. Moreover, we determined that BRASSINOSTEROID INSENSITIVE 1A (ZmBRI1a), a brassinosteroid receptor of the BRI1‐like family, interacts with ZmNBR1. Phenotype analysis showed that ZmBRI1a negatively regulates drought tolerance in maize, and genetic analysis indicated that ZmNBR1 acts upstream of ZmBRI1a in regulating drought tolerance. Furthermore, ZmNBR1 facilitates the autophagic degradation of ZmBRI1a under drought stress. Taken together, our results reveal that ZmNBR1 regulates the expression of autophagy‐related genes, thereby increasing autophagic activity and promoting the autophagic degradation of ZmBRI1a under drought stress, thus enhancing drought tolerance in maize. These findings provide new insights into the autophagy degradation of brassinosteroid signaling components by the autophagy receptor NBR1 under drought stress. [ABSTRACT FROM AUTHOR]

Summary: Mitogen‐activated protein kinase (MPK) is a critical regulator of the antioxidant defence system in response to various stimuli. However, how MPK directly and exactly regulates antioxidant enzyme activities is still unclear. Here, we demonstrated that a NAC transcription factor ZmNAC49 mediated the regulation of antioxidant enzyme activities by ZmMPK5.ZmNAC49 expression is induced by oxidative stress. ZmNAC49 enhances oxidative stress tolerance in maize, and it also reduces superoxide anion generation and increases superoxide dismutase (SOD) activity. A detailed study showed that ZmMPK5 directly interacts with and phosphorylates ZmNAC49 in vitro and in vivo. ZmMPK5 directly phosphorylates Thr‐26 in NAC subdomain A of ZmNAC49. Mutation at Thr‐26 of ZmNAC49 does not affect the interaction with ZmMPK5 and its subcellular localisation.Further analysis found that ZmNAC49 activates the ZmSOD3 expression by directly binding to its promoter. ZmMPK5‐mediated ZmNAC49 phosphorylation improves its ability to bind to the ZmSOD3 promoter. Thr‐26 of ZmNAC49 is essential for its transcriptional activity. In addition, ZmSOD3 enhances oxidative stress tolerance in maize.Our results show that phosphorylation of Thr‐26 in ZmNAC49 by ZmMPK5 increased its DNA‐binding activity to the ZmSOD3 promoter, enhanced SOD activity and thereby improved oxidative stress tolerance in maize. [ABSTRACT FROM AUTHOR]

Summary: Drought stress seriously limits crop productivity. Although studies have been carried out, it is still largely unknown how plants respond to drought stress. Here we find that drought treatment can enhance the phosphorylation activity of brassinosteroid‐signaling kinase 1 (ZmBSK1) in maize (Zea mays).Our genetic studies reveal that ZmBSK1 positively affects drought tolerance in maize plants. ZmBSK1 localizes in plasma membrane, interacts with calcium/calmodulin (Ca2+/CaM)‐dependent protein kinase (ZmCCaMK), and phosphorylates ZmCCaMK. Ser‐67 is a crucial phosphorylation site of ZmCCaMK by ZmBSK1.Drought stress enhances not only the interaction between ZmBSK1 and ZmCCaMK but also the phosphorylation of Ser‐67 in ZmCCaMK by ZmBSK1. Furthermore, Ser‐67 phosphorylation in ZmCCaMK regulates its Ca2+/CaM binding, autophosphorylation and transphosphorylation activity, and positively affects its function in drought tolerance in maize.Our results reveal an important role for ZmBSK1 in drought tolerance and suggest a direct regulatory mode of ZmBSK1 phosphorylating ZmCCaMK. [ABSTRACT FROM AUTHOR]

Summary: l‐Arabinose (l‐Ara) is a major monosaccharide in plant polysaccharides and glycoproteins, and functions in plant growth and development. However, the potential role of l‐Ara during abscisic acid (ABA)‐mediated seed germination has been largely ignored. Here, our results showed a function of l‐Ara during ABA‐mediated seed germination.ABA slowed down the reduction of l‐Ara in seed cell wall, and exogenous l‐Ara aggravated the inhibition of ABA on germination. We further found that MUR4, encoding URIDINE 5′‐DIPHOSPHATE‐d‐XYLOSE 4‐EPIMERASE 1, played a vital role in ABA‐mediated germination. MUR4 was highly expressed in embryo and induced by ABA in both seeds and seedlings. Overexpression of MUR4 conferred hypersensitive seed germination and early postgermination growth to ABA.Further analysis revealed that ABSCISIC ACID INSENSITIVE4 (ABI4) positively modulated the MUR4 expression by directly binding the Coupling Element1 motif of MUR4 promoter. Consistently, abi4‐1 mutant had a lower l‐Ara content in seed cell wall, while a higher l‐Ara content in seed cell wall was observed in ABI4 overexpressors. Genetic analysis suggested that overexpression of MUR4 in abi4‐1 partly restored the ABA sensitivity of abi4‐1.We established the link between ABA and l‐Ara during ABA‐mediated seed germination and cotyledon greening in Arabidopsis and revealed the potential molecular mechanism. [ABSTRACT FROM AUTHOR]

In maize ( Zea mays), the mitogen-activated protein kinase Zm MPK5 has been shown to be involved in abscisic acid ( ABA)-induced antioxidant defence and to enhance the tolerance of plants to drought, salt stress and oxidative stress. However, the underlying molecular mechanisms are poorly understood. Here, using Zm MPK5 as bait in yeast two-hybrid screening, a protein interacting with Zm MPK5 named Zm ABA2, which belongs to a member of the short-chain dehydrogenase/reductase family, was identified. Pull-down assay and bimolecular fluorescence complementation analysis and co-immunoprecipitation test confirmed that Zm MPK5 interacts with Zm ABA2 in vitro and in vivo. Phosphorylation of Ser173 in Zm ABA2 by Zm MPK5 was shown to increase the activity of Zm ABA2 and the protein stability. Various abiotic stimuli induced the expression of Zm ABA2 in leaves of maize plants. Pharmacological, biochemical and molecular biology and genetic analyses showed that both Zm MPK5 and Zm ABA2 coordinately regulate the content of ABA. Overexpression of Zm ABA2 in tobacco plants was found to elevate the content of ABA, regulate seed germination and root growth under drought and salt stress and enhance the tolerance of tobacco plants to drought and salt stress. These results suggest that Zm ABA2 is a direct target of Zm MPK5 and is involved in ABA biosynthesis and functions. [ABSTRACT FROM AUTHOR]

The influence of the porosity on the static mechanical strength of the carbon fiber fabric reinforced epoxy composites laminates was investigated. The tensile, compressive, bending, and interlaminar strength test on the CFRP laminates with porosity of 0.33% and 1.50% were conducted and simulated by a finite element analysis model. The article proposes the failure criterion of the static mechanical strength of the fabric fiber reinforced composites based on the improved Hashin failure criterion that is suitable for the undirectional composite laminates. The basic composite strength parameters are used to evaluate the mechanical properties of CFRP laminates with different porosities. A finite element analysis model is established by using software ABAQUS™ combined with the sudden stiffness degradation model. The experiment results show that the tensile, compressive, bending, and interlaminar strength decrease with the increasing porosities. The tensile, compressive, bending, and interlaminar strength of the fabric carbon fiber reinforced epoxy composites laminates are simulated accurately by the finite element model. [ABSTRACT FROM AUTHOR]

In this study, the role of the rice (Oryza sativa L.) histidine kinase OsHK3 in abscisic acid (ABA)-induced antioxidant defense was investigated. Treatments with ABA, H2O2, and polyethylene glycol (PEG) induced the expression of OsHK3 in rice leaves, and H2O2 is required for ABA-induced increase in the expression of OsHK3 under water stress. Subcellular localization analysis showed that OsHK3 is located in the cytoplasm and the plasma membrane. The transient expression analysis and the transient RNA interference test in rice protoplasts showed that OsHK3 is required for ABA-induced upregulation in the expression of antioxidant enzymes genes and the activities of antioxidant enzymes. Further analysis showed that OsHK3 functions upstream of the calcium/ calmodulin-dependent protein kinase OsDMI3 and the mitogen- activated protein kinase OsMPK1 to regulate the activities of antioxidant enzymes in ABA signaling. Moreover, OsHK3 was also shown to regulate the expression of nicotinamide adenine dinucleotide phosphate oxidase genes, OsrbohB and OsrbohE, and the production of H2O2 in ABA signaling. Our data indicate that OsHK3 play an important role in the regulation of ABA-induced antioxidant defense and in the feedback regulation of H2O2 production in ABA signaling. [ABSTRACT FROM AUTHOR]

In rice, the Ca2+/calmodulin ( CaM)-dependent protein kinase ( CCaMK) OsDMI3 has been shown to be required for abscisic acid ( ABA)-induced antioxidant defence. However, it is not clear how OsDMI3 participates in this process in rice. In this study, the cross-talk between OsDMI3 and the major ABA-activated MAPK OsMPK1 in ABA-induced antioxidant defence was investigated. ABA treatment induced the expression of OsDMI3 and OsMPK1 and the activities of OsDMI3 and OsMPK1 in rice leaves. In the mutant of OsDMI3, the ABA-induced increases in the expression and the activity of OsMPK1 were substantially reduced. But in the mutant of OsMPK1, the ABA-induced increases in the expression and the activity of OsDMI3 were not affected. Pretreatments with MAPKK inhibitors also did not affect the ABA-induced activation of OsDMI3. Further, a transient expression analysis in combination with mutant analysis in rice protoplasts showed that OsMPK1 is required for OsDMI3-induced increases in the activities of antioxidant enzymes and the production of H2O2. Our data indicate that there exists a cross-talk between OsDMI3 and OsMPK1 in ABA signalling, in which OsDMI3 functions upstream of OsMPK1 to regulate the activities of antioxidant enzymes and the production of H2O2 in rice. [ABSTRACT FROM AUTHOR]

C2H2-type zinc finger proteins (ZFPs) are thought to play important roles in modulating the responses of plants to drought, salinity and oxidative stress. However, direct evidence is lacking for the involvement of these ZFPs in abscisic acid (ABA)-induced antioxidant defense in plants. In this study, the role of the rice ( Oryza sativa L. sub. japonica cv. Nipponbare) C2H2-type ZFP ZFP182 in ABA-induced antioxidant defense and the relationship between ZFP182 and two rice MAPKs, OsMPK1 and OsMPK5 in ABA signaling were investigated. ABA treatment induced the increases in the expression of ZFP182, OsMPK1 and OsMPK5, and the activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) in rice leaves. The transient gene expression analysis and the transient RNA interference (RNAi) analysis in protoplasts showed that ZFP182, OsMPK1 and OsMPK5 are involved in ABA-induced up-regulation in the activities of SOD and APX. Besides, OsMPK1 and OsMPK5 were shown to be required for the up-regulation in the expression of ZFP182 in ABA signaling, but ZFP182 did not mediate the ABA-induced up-regulation in the expression of OsMPK1 and OsMPK5. These results indicate that ZFP182 is required for ABA-induced antioxidant defense and the expression of ZFP182 is regulated by rice MAPKs in ABA signaling. [ABSTRACT FROM AUTHOR]

In search for components of mitogen-activated protein kinase (MAPK) cascades in maize ( Zea mays) involved in response to abscisic acid (ABA) stimulus, a novel MAPK gene, ZmMPK3, from ABA-treated maize leaves cDNA was isolated and characterized. The full length of the ZmMPK3 gene is 1 520 bp and encodes a 376 amino acid protein with a predicted molecular mass of 43.5 kD and a pI of 5.83. ZmMPK3 contains all 11 MAPK conserved subdomains and the phosphorylation motif TEY. Amino acid sequence alignment revealed that ZmMPK3 shared high identity with group-A MAPK in plants. A time course (30–360 min) experiment using a variety of signal molecules and stresses revealed that the transcripts level of ZmMPK3 accumulated markedly and rapidly when maize seedlings were subjected to exogenous signaling molecules: ABA, H2O2, jasmonic acid and salicylic acid, various abiotic stimuli such as cold, drought, ultraviolet light, salinity, heavy metal and mechanical wounding. Its transcription was also found to be tissue-specific regulated. Here, we show that ABA and H2O2 induced a significant increase in the ZmMPK3 activity using immunoprecipitation and in-gel kinase assay. Furthermore, the results showed that the ZmMPK3 protein is localized mainly to the nucleus. These results suggest that the ZmMPK3 may play an important role in response to environmental stresses. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Nitric oxide (NO) is a bioactive molecule involved in many biological events, and has been reported as pro-oxidant as well as anti-oxidant in plants. In the present study, the sources of NO production under water stress, the role of NO in water stress-induced hydrogen peroxide (H2O2) accumulation and subcellular activities of anti-oxidant enzymes in leaves of maize ( Zea mays L.) plants were investigated. Water stress induced defense increases in the generation of NO in maize mesphyll cells and the activity of nitric oxide synthase (NOS) in the cytosolic and microsomal fractions of maize leaves. Water stress-induced defense increases in the production of NO were blocked by pretreatments with inhibitors of NOS and nitrate reductase (NR), suggesting that NO is produced from NOS and NR in leaves of maize plants exposed to water stress. Water stress also induced increases in the activities of the chloroplastic and cytosolic anti-oxidant enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR), and the increases in the activities of anti-oxidant enzymes were reduced by pretreatments with inhibitors of NOS and NR. Exogenous NO increases the activities of water stress-induced subcellular anti-oxidant enzymes, which decreases accumulation of H2O2. Our results suggest that NOS and NR are involved in water stress-induced NO production and NOS is the major source of NO. The potential ability of NO to scavenge H2O2 is, at least in part, due to the induction of a subcellular anti-oxidant defense. [ABSTRACT FROM AUTHOR]

• The role of nitric oxide (NO) and the relationship between NO, hydrogen peroxide (H2O2) and mitogen-activated protein kinase (MAPK) in abscisic acid (ABA)-induced antioxidant defense in leaves of maize ( Zea mays) plants were investigated. • Both ABA and H2O2 induced increases in the generation of NO in mesophyll cells of maize leaves, and H2O2 was required for the ABA-induced generation of NO. Pretreatment with NO scavenger and nitric oxide synthase (NOS) inhibitor substantially reduced the ABA-induced production of NO, and partly blocked the activation of a 46 kDa MAPK and the expression and the activities of several antioxidant enzymes induced by ABA. Treatment with the NO donor sodium nitroprusside (SNP) also induced the activation of the MAPK, and enhanced the antioxidant defense systems. • Conversely, SNP treatment did not induce the production of H2O2, and pretreatments with NO scavenger and NOS inhibitor did not affect ABA-induced H2O2 production. • Our results suggest that ABA-induced H2O2 production mediates NO generation, which, in turn, activates MAPK and results in the upregulation in the expression and the activities of antioxidant enzymes in ABA signaling. [ABSTRACT FROM AUTHOR]