Protein lysine succinylation is an important post-translational modification and plays a critical regulatory role in almost every aspects of cell metabolism in both eukaryotes and prokaryotes. Common wheat is one of the major global cereal crops. However, to date, little is known about the functions of lysine succinylation in this plant. Here, we performed a global analysis of lysine succinylation in wheat and examined its overlap with lysine acetylation. In total, 330 lysine succinylated modification sites were identified in 173 proteins. Bioinformatics analysis showed that the modified proteins are distributed in multiple subcellular compartments and are involved in a wide variety of biological processes such as photosynthesis and the Calvin-Benson cycle, suggesting an important role for lysine succinylation in these processes. Five putative succinylation motifs were identified. A protein interaction network analysis revealed that diverse interactions are modulated by protein succinylation. Moreover, 21 succinyl-lysine sites were found to be acetylated at the same position, and 33 proteins were modified by both acetylation and succinylation, suggesting an extensive overlap between succinylation and acetylation in common wheat. Comparative analysis indicated that lysine succinylation is conserved between common wheat and Brachypodium distachyon. These results suggest that lysine succinylation is involved in diverse biological processes, especially in photosynthesis and carbon fixation. This systematic analysis represents the first global analysis of lysine succinylation in common wheat and provides an important resource for exploring the physiological role of lysine succinylation in this cereal crop and likely in all plants.

Protein lysine malonylation, a newly discovered post-translational modification (PTM), plays an important role in diverse metabolic processes in both eukaryotes and prokaryotes. Common wheat is a major global cereal crop. However, the functions of lysine malonylation are relatively unknown in this crop. Here, a global analysis of lysine malonylation was performed in wheat. In total, 342 lysine malonylated sites were identified in 233 proteins. Bioinformatics analysis showed that the frequency of arginine (R) in position + 1 was highest, and a modification motif, K^maR, was identified. The malonylated proteins were located in multiple subcellular compartments, especially in the cytosol (45%) and chloroplast (30%). The identified proteins were found to be involved in diverse pathways, such as carbon metabolism, the Calvin cycle, and the biosynthesis of amino acids, suggesting an important role for lysine malonylation in these processes. Protein interaction network analysis revealed eight highly interconnected clusters of malonylated proteins, and 137 malonylated proteins were mapped to the protein network database. Moreover, five proteins were simultaneously modified by lysine malonylation, acetylation and succinylation, suggesting that these three PTMs may coordinately regulate the function of many proteins in common wheat. Our results suggest that lysine malonylation is involved in a variety of biological processes, especially carbon fixation in photosynthetic organisms. These data represent the first report of the lysine malonylome in common wheat and provide an important dataset for further exploring the physiological role of lysine malonylation in wheat and likely all plants.

Drought stress can negatively impact crop yield and quality. Improving wheat yields under drought stress is a major objective of agronomic research. Glutamine synthetase (GS) is a key enzyme of nitrogen metabolism that is critical to plant growth and development in abiotic stress response. However, to date, no systemic characterization of the GS genes has yet been conducted in wheat and its close relatives. We identified a total of 15 GS genes in Triticum aestivum (2n = 6x = 42; AABBDD), as well as 9 GS genes in Triticum dicoccoides (2n = 4x = 28; AABB), 6 in Aegilops tauschii (2n = 2x = 14; DD), and 5 in Triticum urartu (2n = 2x = 14; AA). The 35 GSs were further clustered into five lineages according to the phylogenetic tree. Synteny analysis revealed that the three subgenomes in bread wheat retained extensive synteny between bread wheat and its three relative species. We identified three up-regulated TaGSs (Ta4A.GSe, Ta4B.GSe, and Ta4D.GSe) from transcriptome data after drought and salt stress. Ta4D.GSe was subsequently used for further functional studies, and its subcellular localization were determined in Arabidopsis protoplasts. Its overexpression in Arabidopsis enhanced drought tolerance by increasing the ability of scavenging of reactive oxygen species (ROS) and osmotic adjustment. We identified GS gene family in four wheat species and performed comparative analyses of their relationships, chromosome locations, conserved motif, gene structure, and synteny. The subcellular localization of Ta4D.GSe was detected and its drought tolerance function was demonstrated. Taken together, these findings provide insight into the potential functional roles of the GS genes in abiotic stress tolerance. Key message: This report clearly shows detailed characterization of GS gene family in four wheat species and demonstrates that Ta4D.GSe plays an important role in enhancing drought tolerance by improving the scavenging of ROS and osmotic adjustment ability in Arabidopsis.

Soil salinization is one of the major abiotic stresses that adversely affect the yield and quality of crops such as wheat, a leading cereal crop worldwide. Excavating the salt-tolerant genes and exploring the salt tolerance mechanism can help breeding salt-tolerant wheat varieties. Thus, it is essential to identify salt-tolerant wheat germplasm resources. In this study, we carried out a salt stress experiment using Qing Mai 6 (QM6), a salt-tolerant wheat variety, and sequenced the miRNAs and mRNAs. The differentially expressed miRNAs and mRNAs in salt stress conditions were compared with the control. As results, a total of eight salt-tolerance-related miRNAs and their corresponding 11 target mRNAs were identified. Further analysis revealed that QM6 enhances salt tolerance through increasing the expression level of genes related to stress resistance, antioxidation, nutrient absorption, and lipid metabolism balance, and the expression of these genes was regulated by the identified miRNAs. The resulting data provides a theoretical basis for future research studies on miRNAs and novel genes related to salt tolerance in wheat in order to develop genetically improved salt-tolerant wheat varieties.

Glutaredoxins (GRXs) are small-molecular-weight proteins present in a wide range of organisms, and they play a key role in maintaining the redox homeostasis of cells. Most studies on GRXs are conducted in animals and humans, and those conducted on plants are scarce. The number and types of GRX genes vary in different plants. According to the active sites, the GRX family can be further divided into the CPYC, CGFS, and CC subfamilies. The CPYC and CGFS subfamilies are present in eukaryotes. The CC subfamily is exclusively present in higher plants and has the highest number of genes. In this study, 85 GRX genes were identified in common wheat (Triticum aestivum L.) using a bioinformatic method, wherein 12, 9, and 64 belonged to the CPYC, CGFS, and CC subfamilies, respectively. All TaGRX genes were homogeneously distributed in the three subgenomes of wheat. The gene structure analysis revealed that TaGRX members had 1–7 introns. The conserved motif analysis revealed that members of the same TaGRX subfamily had similar motifs. An analysis of cis-regulatory elements of promoters demonstrated that most TaGRX members had auxin-responsive elements; cis-regulatory elements, such as methyl jasmonate (MeJA), MYB, and abscisic acid (ABA), were distributed in all subfamilies, and the cell-cycle regulation element was only observed in the CC and CPYC subfamily members. Additionally, the synteny of the GRX genes in wheat, in wheat and Arabidopsis, and in wheat and barley was analyzed to clarify the evolutionary correlation of TaGRXs. The expression characteristics of TaGRXs were investigated, and TaGRX expression in various tissues and its responses to different abiotic stresses were preliminarily determined. This study provides a reference for the functional analysis of TaGRXs and understanding their role in molecular breeding of wheat.

Drought is a persistent and serious threat to crop yield and quality. The identification and functional characterization of drought tolerance-related genes is thus vital for efforts to support the genetic improvement of drought-tolerant crops. Barley is highly adaptable and renowned for its robust stress resistance, making it an ideal subject for efforts to explore genes related to drought tolerance. In this study, two barley materials with different drought tolerance were subjected to soil drought treatment, including a variety with strong drought tolerance (Hindmarsh) and a genotype with weaker drought tolerance (XZ5). Transcriptomic sequencing data from the aboveground parts of these plants led to the identification of 1,206 differentially expressed genes associated with drought tolerance. These genes were upregulated in Hindmarsh following drought stress exposure but downregulated or unchanged in XZ5 under these same conditions, or were unchanged in Hindmarsh but downregulated in XZ5. Pathway enrichment analyses suggested that these genes are most closely associated with defense responses, signal recognition, photosynthesis, and the biosynthesis of various secondary metabolites. Using protein-protein interaction networks, the ankyrin repeat domain-containing protein 17-like isoform X2 was predicted to impact other drought tolerance-related protein targets in Hindmarsh. In MapMan metabolic pathway analyses, genes found to be associated with the maintenance of drought tolerance in Hindmarsh under adverse conditions were predicted to include genes involved in the abscisic acid, cytokinin, and gibberellin phytohormone signaling pathways, genes associated with redox homeostasis related to ascorbate and glutathione S-transferase, transporters including ABC and AAAP, transcription factors such as AP2/ERF and bHLH, the heat shock proteins HSP60 and HSP70, and the sucrose non-fermenting-1-related protein kinase. Heterologous HvSnRK2 (one of the identified genes, which encodes the sucrose non-fermenting-1-related protein kinase) gene expression in yeast conferred significant drought tolerance, highlighting the functional importance of this gene as one linked with drought tolerance. This study revealed the drought tolerance mechanism of Hindmarsh by comparing transcriptomes while also providing a set of candidate genes for genetic efforts to improve drought tolerance in this and other crop species.