This study systematically investigated the physiological and molecular responses of the wheat mutant ‘XC-MU201’ under high-temperature stress through comprehensive transcriptome analysis and physiological measurements. RNA sequencing of 21 samples across seven different treatment groups revealed, through Weighted Gene Co-expression Network Analysis (WGCNA), 13 modules among 9071 genes closely related to high-temperature treatments. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed significant enrichment of lignin biosynthesis-related modules under high-temperature conditions, especially at the H-10DAT, H-20DAT, and H-30DAT time points. Experimental results demonstrated a significant increase in lignin content in high-temperature-treated samples, confirmed by tissue staining methods, indicating wheat’s adaptation to heat damage through lignin accumulation. The phenylalanine ammonia-lyase gene (TaPAL33) was significantly upregulated under high-temperature stress, peaking at H-30DAT, suggesting its critical role in cellular defense mechanisms. Overexpression of TaPAL33 in the wheat variety ‘Xinchun 11’ enhanced lignin synthesis but inhibited growth. Subcellular localization of GFP-labeled TaPAL33 in tobacco cells showed its distribution mainly in the cytoplasm and cell membrane. Transgenic wheat exhibited higher PAL enzyme activity, enhanced antioxidant defense, and reduced oxidative damage under high-temperature stress, outperforming wild-type wheat. These results highlight TaPAL33’s key role in improving wheat heat tolerance and provide a genetic foundation for future research and applications.

Wheat (Triticum aestivum L.) production is increasingly constrained by drought, heat, and their combination, especially in arid regions like Xinjiang, China. While previous studies have focused on single-stress responses, the molecular mechanisms underlying wheat adaptation to combined drought and heat stress (HD) remain elusive. Here, we performed transcriptome profiling at three key spike developmental stages-Jointing Stage (JS), Stamen and Pistil Primordial Differentiation Stage (SPDS), and Tetrad Stage (TS)-under drought (DS), heat (HS), and combined HD conditions. Comparative analysis revealed that HD triggered the most extensive transcriptional reprogramming, with 3,884, 5,230, and 2,793 stage-specific upregulated genes across JS, SPDS, and TS, respectively. Notably, the overlap between HS- and HD-responsive genes was higher than that between DS and HD during early stages, indicating stress-specific transcriptional shifts. Functional enrichment highlighted pathways related to osmotic adjustment, oxidative defense, and phytohormone signaling, particularly ABA and JA biosynthesis. Among stress-responsive transcription factors, the AP2/ERF family showed prominent enrichment, with TaEREBP1-L (TraesCS5A02G215900) identified as a central regulator. Dual-luciferase reporter assays confirmed that TaEREBP1-L directly activated the promoters of ABA- and JA-pathway genes (AAO3, AOC2). Co-expression analysis further revealed its coordination with membrane transporters (e.g., ABC and MATE families), suggesting an integrative regulatory function under HD conditions. Transgenic Arabidopsis overexpressing TaEREBP1-L displayed significantly enhanced drought and heat tolerance, evidenced by increased proline content, antioxidant enzyme activity (SOD, CAT), and elevated ABA/JA levels. These results demonstrate that TaEREBP1-L confers broad-spectrum stress tolerance by orchestrating hormonal and physiological pathways. Our findings provide valuable insights into combinatorial stress adaptation and identify TaEREBP1-L as a promising target for improving wheat resilience under climate-related challenges.

Introduction: Wheat is a food crop with a large global cultivation area, and the content and quality of wheat glutenin accumulation are important indicators of the quality of wheat flour.Methods: To elucidate the gene expression regulation and metabolic characteristics related to the gluten content during wheat grain formation, transcriptomic and metabolomic analyses were performed for the high gluten content of the Xinchun 26 cultivar and the low proteins content of the Xinchun 34 cultivar at three periods (7 d, 14 d and 21 d) after flowering.Results: Transcriptomic analysis revealed that 5573 unique differentially expressed genes (DEGs) were divided into two categories according to their expression patterns during the three periods. The metabolites detected were mainly divided into 12 classes. Lipid and lipid-like molecule levels and phenylpropanoid and polyketide levels were the highest, and the difference analysis revealed a total of 10 differentially regulated metabolites (DRMs) over the three periods. Joint analysis revealed that the DEGs and DRMs were significantly enriched in starch and sucrose metabolism; the citrate cycle; carbon fixation in photosynthetic organisms; and alanine, aspartate and glutamate metabolism pathways. The genes and contents of the sucrose and gluten synthesis pathways were analysed, and the correlation between gluten content and its related genes was calculated. Based on weighted correlation network analysis (WGCNA), by constructing a coexpression network, a total of 5 specific modules and 8 candidate genes that were strongly correlated with the three developmental stages of wheat grain were identified.Discussion: This study provides new insights into the role of glutenin content in wheat grain formation and reveals potential regulatory pathways and candidate genes involved in this developmental process.

Wheat ( Triticum aestivum L.), a globally significant cereal crop, plays a crucial role in human nutrition and agricultural systems. Recent studies have extensively examined the role of transcription factors across various plant genomes. However, comprehensive comparative genomic analyses of the WRKY gene family across various diploid, tetraploid, and hexaploid wheat genomes remain limited. Here, we conducted a detailed analysis of the WRKY gene family in one diploid, two tetraploid, and five hexaploid wheat genomes, identifying a total of 1912 WRKY genes. Consistent with previous reports, the genes were classified into three groups according to their conserved domain and motif architectures. Phylogenetic reconstruction resolved clear clades, with Group II forming the largest and most diverse branch. Tandem duplication events were found to significantly impact the expansion of the WRKY gene family, particularly in the T. aestivum acc. Chinese Spring genome, which exhibited the highest number of tandemly duplicated genes. Analysis of A subgenomes in polyploid wheat genomes suggested potential gene loss following polyploid formation. Expression profiling of WRKY genes in bread wheat under salt stress and autophagy inhibition conditions uncovered differential expression patterns and significant divergence among tandemly duplicated genes. This study provides valuable insights into the distribution, phylogenetic relationships, and expression patterns of WRKY genes in wheat. Our findings highlight the role of tandem duplication in driving functional divergence, offering important genetic resources for wheat gene editing and genome‐assisted breeding.