As one of the microelements, nitrogen play essential roles in cereal production. Although the use of chemical fertilizers has significantly improved the yield of wheat, it has also caused increasingly adverse environmental pollution. Revealing the molecular mechanism manipulating wheat nitrogen use efficiency (NUE), and cultivating wheat germplasms with high nitrogen use efficiency has become important goals for wheat researchers. In this study, we investigated the physiological and transcriptional differences of three wheat cultivars with different NUE under low nitrogen stress. The results showed that, under low nitrogen conditions, the activities of nitrogen metabolism-related enzymes (GS, NR, GDH), antioxidant enzymes (SOD, POD, CAT) and soluble protein contents of ZM366 (high NUE cultivar) were higher than those of JD8 (low NUE cultivar). The hybrid cultivar of ZM366 and JD8 showed mid-parent or over-parent heterosis. Transcriptome analysis revealed that ‘alanine, aspartate and glutamate metabolism’, ‘terpenoid backbone biosynthesis’ and ‘vitamin B6 metabolism’ pathways play key roles in nitrogen use efficiency in wheat. The significant enhancement of the ‘Calvin cycle’ and ‘photorespiration’ in ZM366 contributed to its higher level of carbon metabolism under low nitrogen stress, which is an important attribute differs from the other two varieties. In addition, the activation of ABA signal transduction and biosynthesis pathways also helps to maintain NUE under low- nitrogen conditions. Moreover, bHLH transcription factors were also found to play a positive role in wheat NUE. In conclusion, these results enriched our knowledge of the mechanism of wheat NUE, and provided a theoretical basis for improving wheat NUE and breeding new cultivars.

Sucrose transporters (SUTs) play vital roles in phloem sucrose unloading and transportation in wheat grains. However, the genomic information regarding the SUT gene family and their expression patterns in response to heat stress in grains of male-sterile wheat (Triticum aestivum L.) lines has not been systematically studied. In this study, a thorough examination of the wheat SUT gene family was conducted, focusing on their expression patterns in male-sterile lines under heat stress conditions in grain tissues. A total of 19 SUT genes were identified, with phylogenetic analysis indicating their classification into five distinct groups. Polyploidization was identified as a substantial factor in the expansion of SUT genes, with segmental duplication being the predominant mechanism driving the evolutionary expansion of the SUT gene family in wheat. Transcriptome data indicate that the expression levels of TaSUT1 and TaSUT2 were higher than other SUT genes in grains of male-sterile lines. The TaSUT1 expression showed a gradual decreasing trend, while TaSUT2 showed a reverse trend with the process of grain filling. After heat stress, the TaSUT1 expression in grains of male-sterile lines was first significantly increased and then significantly decreased with the filling stage extension, aligning with the observed trend of sucrose levels, indicating that heat stress may decrease the grain weight by reducing sucrose unloading and transportation process in grains. These results provide a systematic analysis of the SUT gene family and lay a theoretical foundation for us to study the grain filling of male-sterile lines in response to abiotic stress.

High-temperature (HT) stress frequently affects the early and middle stages of grain filling in hybrid seed production regions. Photo-thermo-sensitive male-sterile (PTMS) wheat lines, which play a critical role as female parents in hybrid seed production, face challenges under HT conditions. However, the mechanisms governing grain filling in PTMS lines under HT stress remain poorly understood. This study used the BS253 line to investigate the effects of HT on grain filling, primarily focusing on the transition from sucrose unloading to starch synthesis. The findings indicated that HT significantly reduced the grain starch content and weight by 7.65% and 36.35% at maturity, respectively. Further analysis revealed that the expression levels of TaSUT1 and TaSWEETs in grains initially increased after HT stress, paralleling the rise in sucrose content during the same period. The activities of ADP-glucose pyrophosphorylase, UDP-glucose pyrophosphorylase, granule-bound starch synthase, and soluble starch synthase were markedly decreased, indicating that impaired starch synthesis was a key factor limiting grain filling immediately after HT exposure. A total of 41 key regulatory genes involved in sucrose-to-starch metabolism were identified, with HT significantly reducing the expression of genes associated with pathways from sucrose unloading to starch synthesis during the middle and late stages post-HT. Based on the observed ultrastructural changes in the abdominal phloem and sucrose transporter expression levels under HT, we concluded that limited sucrose supply, degradation, and inhibition of starch synthesis collectively constrained grain filling during these stages. Additionally, 11 heat shock proteins and two catalase genes were identified and significantly upregulated during the initial phase post-HT, suggesting their potential role in enhancing sucrose supply at this critical time. More importantly, seven key genes involved in the sucrose-to-starch pathway were identified by weighted gene co-expression network analysis (WGCNA), which provides target genes for their functional research for starch synthase. These findings provide a comprehensive understanding of how HT limits grain filling, identify several genes involved in the sucrose-to-starch pathway, and offer a novel perspective for future research on HT-restricted grain filling across the entire process from sucrose unloading to starch synthesis in developing grains.

Drought severely limits global wheat production, and plants can mitigate this stress by recruiting beneficial rhizosphere microbiomes. However, the role of host genotype in shaping this recruitment and influencing microbial inoculant efficacy is poorly understood. Here, we aimed to elucidate the interplay among host genotype, drought stress, and inoculation with Trichoderma citrinoviride. A factorial pot experiment investigated the responses of two wheat genotypes (JN 14-95, constitutively drought-tolerant; JN 72, drought-responsive) to water regimes and inoculation with T. citrinoviride. Metagenomic analysis was performed to characterize rhizosphere bacterial community structure and functional potential. Host genotype was the primary driver of bacterial community structure (42.3% of variation), exceeding water stress (23.8%) and inoculation (11.5%). JN 14-95 adopted a “physiological autonomy” strategy with constitutive high root-to-shoot ratio and inferred enrichment of auxin and SOD biosynthesis genes. JN 72 employed a “microbial outsourcing” strategy, enriching beneficial bacteria (Pseudomonas, Bacillus, Streptomyces) and showing inferred enrichment of central carbon metabolism genes. T. citrinoviride amplified these genotype-specific responses, increasing the total dry matter by 21.3% in JN 14-95 and 30.3% in JN 72 under drought. Our findings suggest a need to move from uniform inoculation practices toward genotype-informed microbiome management. The two strategies provide a framework for leveraging host genetics in sustainable agriculture, highlighting that breeding and microbiome management should be integrated.