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DREB transcription factor can significantly improve the plant resistance to drought, high salt, low temperature, high temperature, and other abiotic stress. A previous study showed that DREB transcription factor gene GmDREB3 from soybean can improve drought resistance in tobacco, however, whether it can improve drought resistance in wheat needs to be further investigated. In the present study, GmDREB3 was transferred to a wheat variety, Shi366 and two positive lines of the T5 generation were obtained by additive culture and molecular identification. GmDREB3‐OE (overexpression) lines conferred drought tolerance characteristics, including higher survival rates, lower malondialdehyde (MDA) content, lower relative conductivity, higher chlorophyll content, and higher proline content than Shi366 wheat plants under drought stress. The SPAD, Fv/Fm, panicle number, and grain yield of GmDREB3‐OE lines were increased but the transpiration rate was decreased, compared to the Shi366 plants in the field condition under drought stress. Significant differences in the stress indices were observed between the GmDREB3–OE lines and Shi366 wheat plants, which suggested that the GmDREB3 significantly improved wheat tolerance to drought stress. In addition, RT‐qPCR analysis indicated that GmDREB3 could enhance the expression of drought‐related genes in wheat. Therefore, GmDREB3 significantly increased drought tolerance in wheat plants along with improved physiological and agronomic traits and grain yield. [ABSTRACT FROM AUTHOR]

Summary: Abscisic acid (ABA) receptors are considered as the targeted manipulation of ABA sensitivity and water productivity in plants. Regulation of their stability or activity will directly affect ABA signalling. Mitogen‐activated protein kinase (MAPK) cascades link multiple environmental and plant developmental cues. However, the molecular mechanism of ABA signalling and MAPK cascade interaction remains largely elusive.TaMPK3 overexpression decreases drought tolerance and wheat sensitivity to ABA, significantly weakening ABA's inhibitory effects on growth. Under drought stress, overexpression lines show lower survival rates, shoot fresh weight and proline content, but higher malondialdehyde levels at seedling stage, as well as decreased grain width and 1000 grain weight in both glasshouse and field conditions at the adult stage. TaMPK3‐RNAi increases drought tolerance.TaMPK3 interaction with TaPYL4 leads to decreased TaPYL4 levels by promoting its ubiquitin‐mediated degradation, whereas ABA treatment diminishes TaMPK3–TaPYL interactions. In addition, the expression of ABA signalling proteins is impaired in TaMPK3‐overexpressing wheat plants under ABA treatment. The MPK3‐PYL interaction module was found to be conserved across monocots and dicots.Our results suggest that the MPK3‐PYL module could serve as a negative regulatory mechanism for balancing appropriate drought stress response with normal plant growth signalling in wheat. [ABSTRACT FROM AUTHOR]

DEAD-box RNA helicases constitute the largest subfamily of RNA helicase superfamily 2 (SF2), and play crucial roles in plant growth, development, and abiotic stress responses. Wheat is one of the most important cereal crops in worldwide, and abiotic stresses greatly restrict its production. So far, the DEAD-box RNA helicase family has yet to be characterized in wheat. Here, we performed a comprehensive genome-wide analysis of the DEAD-box RNA helicase family in wheat, including phylogenetic relationships, chromosomal distribution, duplication events, and protein motifs. A total of 141 TaDEAD-box genes were identified and found to be unevenly distributed across all 21 chromosomes. Whole genome/segmental duplication was identified as the likely main driving factor for expansion of the TaDEAD-box family. Expression patterns of the 141 TaDEAD-box genes were compared across different tissues and under abiotic stresses to identify genes to be important in growth or stress responses. TaDEAD-box57-3B was significantly up-regulated under multiple abiotic stresses, and was therefore selected for further analysis. TaDEAD-box57-3B was localized to the cytoplasm and plasma membrane. Ectopic expression of TaDEAD-box57-3B in Arabidopsis improved tolerance to drought and salt stress as measured by germination rates, root lengths, fresh weights, and survival rates. Transgenic lines also showed higher levels of proline and chlorophyll and lower levels of malonaldehyde (MDA) than WT plants in response to drought or salt stress. In response to cold stress, the transgenic lines showed significantly better growth and higher survival rates than WT plants. These results indicate that TaDEAD-box57-3B may increase tolerance to drought, salt, and cold stress in transgenic plants through regulating the degree of membrane lipid peroxidation. This study provides new insights for understanding evolution and function in the TaDEAD-box gene family. [ABSTRACT FROM AUTHOR]

ERECTAs are receptor-like kinases that regulate plant biomass and stress resistance. In this study, the wheat (Triticum aestivum) TaERECTA gene was used as a probe to identify the SbERECTA family genes (SbERs) in the sorghum (Sorghum bicolor) genome, analyse their subcellular localisation and characterise their expression. Results showed that the two SbER members, SbER10 with three copies (SbER10 _ X1 , SbER10 _ X2 , and SbER10 _ X3) and SbER4 with two copies (SbER4 _ X1 and SbER4 _ X2), were found on chromosomes 10 and 4 of sorghum, respectively. SbER10 had the highest expression level in the pedicel tissue and showed a remarkable response under treatment with abscisic acid, brassinolide, gibberellin and indole-3-acetic acid. SbER10 _ X1 , functioning on the cell membrane and chloroplast, exhibited abundant transcript in only a few sorghum varieties that are grown in mountainous areas and receive strong light, heat, and water supply. Expression of SbER10 _ X1 was significantly and positively correlated with plant biomass of 32 sorghum germplasm resources. These results indicate that SbER10 genes have an important regulatory role in sorghum growth, and increasing SbER10 transcription level offers a potential strategic target for breeding or biotechnological approaches to enhance sorghum biomass and environmental adaptability. The SbERECTA (SbERs) gene family has two members, SbER10 with three copies (SbER10 _ X1 , SbER10 _ X2 , and SbER10 _ X3) and SbER4 with two copies (SbER4 _ X1 and SbER4 _ X2), in the sorghum genome. The promoter of the SbER family contains ABA, low temperature, anaerobic induction and other core elements, and SbER10 genes actively respond to hormone induction. SbER10 _ X1 expression was significantly and positively correlated with plant biomass among 32 sorghum germplasm resources. [ABSTRACT FROM AUTHOR]

Bax inhibitor-1 (BI-1) is an endoplasmic reticulum (ER)-resident cell death suppressor evolutionarily conserved in eukaryotes. The ability of BI-1 to inhibit the biotic and abiotic stresses have been well-studied in Arabidopsis, while the functions of wheat BI-1 are largely unknown. In this study, the wheat BI-1 gene TaBI-1.1 was isolated by an RNAseq analysis of Fusarium graminearum (Fg)-treated wheat. TaBI-1.1 expression was induced by a salicylic acid (SA) treatment and down-regulated by an abscisic acid (ABA) treatment. Based on β-glucuronidase (GUS) staining, TaBI-1.1 was expressed in mature leaves and roots but not in the hypocotyl or young leaves. Constitutive expression of TaBI-1.1 in Arabidopsis enhanced its resistance to Pseudomonas syringae pv. Tomato (Pst) DC3000 infection and induced SA-related gene expression. Additionally, TaBI-1.1 transgenic Arabidopsis exhibited an alleviation of damage caused by high concentrations of SA and decreased the sensitivity to ABA. Consistent with the phenotype, the RNA-seq analysis of 35S::TaBI-1.1 and Col-0 plants showed that TaBI-1.1 was involved in biotic stresses. These results suggested that TaBI-1.1 positively regulates SA signals and plays important roles in the response to biotic stresses. In addition, TaBI-1.1 interacted with the aquaporin TaPIP1, and both them were localized to ER membrane. Furthermore, we demonstrated that TaPIP1 was up-regulated by SA treatment and TaPIP1 transgenic Arabidopsis enhanced the resistance to Pst DC3000 infection. Thus, the interaction between TaBI-1.1 and TaPIP1 on the ER membrane probably occurs in response to SA signals and defense response. [ABSTRACT FROM AUTHOR]

Phosphoinositides are involved in regulation of recruitment and activity of signalling proteins in cell membranes. Phosphatidylinositol (PI) 4-kinases (PI4Ks) generate PI4-phosphate the precursor of regulatory phosphoinositides. No type II PI4K research on the abiotic stress response has previously been reported in plants. A stress-inducible type II PI4K gene, named TaPI4KIIγ, was obtained by de novo transcriptome sequencing of drought-treated wheat (Triticum aestivum). TaPI4KIIγ, localized on the plasma membrane, underwent threonine autophosphorylation, but had no detectable lipid kinase activity. Interaction of TaPI4KIIγ with wheat ubiquitin fusion degradation protein (TaUDF1) indicated that it might be hydrolysed by the proteinase system. Overexpression of TaPI4KIIγ revealed that it could enhance drought and salt stress tolerance during seed germination and seedling growth. A ubdkγ7 mutant, identified as an orthologue of TaPI4KIIγ in Arabidopsis, was sensitive to salt, polyethylene glycol (PEG), and abscisic acid (ABA), and overexpression of TaPI4KIIγ in the ubdkγ7 mutant compensated stress sensitivity. TaPI4KIIγ promoted root growth in Arabidopsis, suggesting that TaPI4KIIγ might enhance stress resistance by improving root growth. Overexpression of TaPI4KIIγ led to an altered expression level of stress-related genes and changes in several physiological traits that made the plants more tolerant to stress. The results provided evidence that overexpression of TaPI4KIIγ could improve drought and salt tolerance. [ABSTRACT FROM AUTHOR]

Oxidative stress caused by accumulation of reactive oxygen species (ROS) is capable of damaging effects on numerous cellular components. Glutathione peroxidases (GPXs, EC 1.11.1.9) are key enzymes of the antioxidant network in plants. In this study, W69 and W106, two putative GPX genes, were obtained by de novo transcriptome sequencing of salt-treated wheat (Triticum aestivum) seedlings. The purified His-tag fusion proteins of W69 and W106 reduced H2O2 and t-butyl hydroperoxide (t-BHP) using glutathione (GSH) or thioredoxin (Trx) as an electron donor in vitro, showing their peroxidase activity toward H2O2 and toxic organic hydroperoxide. GFP fluorescence assays revealed that W69 and W106 are localized in chloroplasts. Quantitative real-time PCR (Q-RT-PCR) analysis showed that two GPXs were differentially responsive to salt, drought, H2O2, or ABA. Isolation of the W69 and W106 promoters revealed some cis-acting elements responding to abiotic stresses. Overexpression of W69 and W106 conferred strong tolerance to salt, H2O2, and ABA treatment in Arabidopsis. Moreover, the expression levels of key regulator genes (SOS1, RbohD and ABI1/ABI2) involved in salt, H2O2 and ABA signaling were altered in the transgenic plants. These findings suggest that W69 and W106 not only act as scavengers of H2O2 in controlling abiotic stress responses, but also play important roles in salt and ABA signaling. [ABSTRACT FROM AUTHOR]

During the past two decades, large amounts of chemical N fertilizers have been applied in rice (Oryza sativa L.)--wheat (Triticum aestivum L.) double-cropping systems in the Taihu Lake region of southern China to achieve high yield. To understand the N balance and environmental impact, a 3-yr field experiment was conducted to determine the integrated N input and output in a rice--wheat rotation system in this region. The results indicated little N surplus in the soil despite different patterns of N input and output in rice and wheat seasons. Although total N input was higher for rice than wheat, the output was also proportionately higher so that the balance was similar between the two crops. Crop harvest removed 48% of N input for both rice and wheat seasons. Denitrification and NH3 volatilization were the primary paths for N loss (22% each of total output) in the rice season, while runoff and leaching (22%) and denitrification (21%) were the main paths for N loss in the wheat season. Although the total annual N input reached up to 606 kg N ha-1, the annual N output was as high as 599 kg N ha-1, of which 52% was lost into the environment, leaving little N in the soil. These results demonstrated that the overwhelming majority of N input did not remain in the soil but rather was quickly emitted. Proper practices are imperative to optimize the N balance and minimize N loss. [ABSTRACT FROM AUTHOR]

Field undisturbed tension-free monolith lysimeters and 15N-labeled urea were used to investigate the fate of fertilizer nitrogen in paddy soil in the Taihu Lake region under a summer rice-winter wheat rotation system. We determined nitrogen recovered by rice and wheat, N remained in soil, and the losses of reactive N (i.e., NH3, N2O, NO3 −, organic N and NH4 +) to the environment. Quantitative allocation of nitrogen fate varied for the rice and wheat growing seasons. At the conventional application rate of 550 kg N ha−1 y−1 (250 kg N ha−1 for wheat and 300 kg N ha−1 for rice), nitrogen recovery of wheat and rice were 49% and 41%, respectively. The retention of fertilizer N in soil at harvest accounted for 29% in the wheat season and for 22% in the rice season. N losses through NH3 volatilization from flooded rice paddy was 12%, far greater than that in the wheat season (less than 1%), while N leaching and runoff comprised only 0.3% in the rice season and 5% in the wheat season. Direct N2O emission was 0.12% for the rice season and 0.14% for the wheat season. The results also showed that some dissolved organic N (DON) were leached in both crop seasons. For the wheat season, DON contributed 40–72% to the N- leaching, in the rice season leached DON was 64–77% of the total N leaching. With increasing fertilizer application rate, NH3 volatilization in the rice season increased proportionally more than the fertilizer increase, N leaching in the wheat season was proportional to the increase of fertilizer rate, while N2O emission increased less in proportion than fertilizer increase both in the rice season and wheat season. [ABSTRACT FROM AUTHOR]

Protein kinases play crucial roles in response to external environment stress signals. A putative protein kinase, W55a, belonging to SNF1-related protein kinase 2 ( SnRK2) subfamily, was isolated from a cDNA library of drought-treated wheat seedlings. The entire length of W55a was obtained using rapid amplification of 5′ cDNA ends ( 5′-RACE) and reverse transcription-polymerase chain reaction( RT-PCR). It contains a 1 029 -bp open reading frame ( ORF) encoding 342 amino acids. The deduced amino acid sequence of W55a had eleven conserved catalytic subdomains and one Ser/Thr protein kinase active-site that characterize Ser/Thr protein kinases. Phylogenetic analysis showed that W55a was 90.38% homologous with rice SAPK1, a member of the SnRK2 family. Using nullisomic-tetrasomic and ditelocentric lines of Chinese Spring, W55a was located on chromosome 2BS. Expression pattern analysis revealed that W55a was upregulated by drought and salt, exogenous abscisic acid, salicylic acid, ethylene and methyl jasmonate, but was not responsive to cold stress. In addition, W55a transcripts were abundant in leaves, but not in roots or stems, under environmental stresses. Transgenic Arabidopsis plants overexpressing W55a exhibited higher tolerance to drought. Based on these findings, W55a encodes a novel dehydration-responsive protein kinase that is involved in multiple stress signal transductions. [ABSTRACT FROM AUTHOR]

Abstract: Construction of chromosome-specific BAC library plays an important role for simplifying sequencing, physical mapping, and gene cloning of plant with complex genome such as common wheat (Triticum aestivum L.), and identification of sorted chromosomes is a vital step of library construction. Based on previous studies, the identification of chromosomes (arms) 6VS, 3B, and 7BL sorted from ditelosomic and normal wheat were performed through fluorescence in situ hybridization (FISH), primed in situ DNA labeling (C-PRINS), and PCR amplification methods, respectively. The results indicated that all these methods could efficiently identify the flow sorted chromosomes. Chromosome staining before flow sorting and chromosome damage from physical shear force during suspension of chromosome preparation and flow sorting did not impact obviously the results of identification. Amongst the 3 methods, PCR is the fastest one with good repetition, and better for rapid determination of the constitution of chromosomal peaks on the univariate flow karyotype histogram, but there are no visible signals hybridized on the sorted chromosomes and the purity of sorted chromosomes cannot be determined in this method. FISH can provide a visible and repetitive result and is suitable for identifying the purity of the sorted chromosomes, but it is time-consuming, complex, and obligatory for special probes. C-PRINS, combining the advantages of FISH and PCR, has the potential for chromosomes identification, although the hybridization signal was not stable enough and its repetition was not satisfactory at present. If combined with in situ hybridization in suspension, C-PRINS is probably a new way for chromosome flow sorting. [Copyright &y& Elsevier]

The WRKY transcription factor superfamily is known to participate in plant growth and stress response. However, the role of this family in wheat (Triticum aestivum L.) is largely unknown. Here, a salt-induced gene TaWRKY13 was identified in an RNA-Seq data set from salt-treated wheat. The results of RT-qPCR analysis showed that TaWRKY13 was significantly induced in NaCl-treated wheat and reached an expression level of about 22-fold of the untreated wheat. Then, a further functional identification was performed in both Arabidopsisthaliana and Oryza sativa L. Subcellular localization analysis indicated that TaWRKY13 is a nuclear-localized protein. Moreover, various stress-related regulatory elements were predicted in the promoter. Expression pattern analysis revealed that TaWRKY13 can also be induced by polyethylene glycol (PEG), exogenous abscisic acid (ABA), and cold stress. After NaCl treatment, overexpressed Arabidopsis lines of TaWRKY13 have a longer root and a larger root surface area than the control (Columbia-0). Furthermore, TaWRKY13 overexpression rice lines exhibited salt tolerance compared with the control, as evidenced by increased proline (Pro) and decreased malondialdehyde (MDA) contents under salt treatment. The roots of overexpression lines were also more developed. These results demonstrate that TaWRKY13 plays a positive role in salt stress. [ABSTRACT FROM AUTHOR]

Melatonin (N-acetyl-5-methoxytryptamine) is involved in many developmental processes and responses to various abiotic stresses in plants. Most of the studies on melatonin focus on its functions and physiological responses in plants, while its regulation mechanism remains unknown. Caffeic acid 3-O-methyltransferase (COMT) functions at a key step of the biosynthesis process of melatonin. In this study, a COMT-like gene, TaCOMT (Traes_1AL_D9035D5E0.1) was identified in common wheat (Triticum aestivum L.). Transient transformation in wheat protoplasts determined that TaCOMT is localized in cytoplasm. TaCOMT in wheat was induced by drought stress, gibberellin (GA)3 and 3-Indoleacetic acid (IAA), but not by ABA. In TaCOMT transgenic Arabidopsis, melatonin contents were higher than that in wild type (WT) plants. Under D-Mannitol treatment, the fresh weight of the transgenic Arabidopsis was significantly higher than WT, and transgenic lines had a stronger root system compared to WT. Drought tolerance assays in pots showed that the survival rate of TaCOMT-overexpression lines was significantly higher than that of WT lines. this phenotype was similar to that the WT lines treated with melatonin under drought condition. In addition, the TaCOMT transgenic lines had higher proline content and lower malondialdehyde (MDA) content compared to WT lines after drought treatment. These results indicated that overexpression of the wheat TaCOMT gene enhances drought tolerance and increases the content of melatonin in transgenic Arabidopsis. It could be one of the potential genes for agricultural applications. [ABSTRACT FROM AUTHOR]

Adaptation to drought and salt stresses is a fundamental part of plant cell physiology and is of great significance for crop production under environmental stress. Heat shock proteins (HSPs) are molecular chaperones that play a crucial role in folding, assembling, translocating, and degrading proteins. However, their underlying mechanisms and functions in stress tolerance remain elusive. Here, we identified the HSP TaHSP17.4 in wheat by analyzing the heat stress-induced transcriptome. Further analysis showed that TaHSP17.4 was significantly induced under drought, salt, and heat stress treatments. Intriguingly, yeast-two-hybrid analysis showed that TaHSP17.4 interacts with the HSP70/HSP90 organizing protein (HOP) TaHOP, which plays a significant role in linking HSP70 and HSP90. We found that TaHSP17.4 - and TaHOP -overexpressing plants have a higher proline content and a lower malondialdehyde content than wild-type plants under stress conditions and display strong tolerance to drought, salt, and heat stress. Additionally, qRT-PCR analysis showed that stress-responsive genes relevant to reactive oxygen species scavenging and abscisic acid signaling pathways were significantly induced in TaHSP17.4 - and TaHOP -overexpressing plants under stress conditions. Together, our findings provide insight into HSP functions in wheat and two novel candidate genes for improvement of wheat varieties. [ABSTRACT FROM AUTHOR]

Oxidative stress caused by accumulation of reactive oxygen species (ROS) is capable of damaging effects on numerous cellular components. Glutathione peroxidases (GPXs, EC 1.11.1.9) are key enzymes of the antioxidant network in plants. In this study, W69 and W106, two putative GPX genes, were obtained by de novo transcriptome sequencing of salt-treated wheat (Triticum aestivum) seedlings. The purified His-tag fusion proteins of W69 and W106 reduced H2O2 and t-butyl hydroperoxide (t-BHP) using glutathione (GSH) or thioredoxin (Trx) as an electron donor in vitro, showing their peroxidase activity toward H2O2 and toxic organic hydroperoxide. GFP fluorescence assays revealed that W69 and W106 are localized in chloroplasts. Quantitative real-time PCR (Q-RT-PCR) analysis showed that two GPXs were differentially responsive to salt, drought, H2O2, or ABA. Isolation of the W69 and W106 promoters revealed some cis-acting elements responding to abiotic stresses. Overexpression of W69 and W106 conferred strong tolerance to salt, H2O2, and ABA treatment in Arabidopsis. Moreover, the expression levels of key regulator genes (SOS1, RbohD and ABI1/ABI2) involved in salt, H2O2 and ABA signaling were altered in the transgenic plants. These findings suggest that W69 and W106 not only act as scavengers of H2O2 in controlling abiotic stress responses, but also play important roles in salt and ABA signaling. [ABSTRACT FROM AUTHOR]