N⁶-methyladenosine (m⁶A) is the most abundant modification of eukaryotic mRNA. Although m⁶A has been demonstrated to affect almost all aspects of RNA metabolism, its global contribution to the post-transcriptional balancing of translational efficiency remains elusive in plants. In this study, we performed a parallel analysis of the transcriptome-wide mRNA m⁶A distribution and polysome profiling in two maize (Zea mays) inbred lines to assess the global correlation of m⁶A modification with translational status. m⁶A sites are widely distributed in thousands of protein-coding genes, confined to a consensus motif and primarily enriched in the 3′ untranslated regions, and highly coordinated with alternative polyadenylation usage, suggesting a role of m⁶A modification in regulating alternative polyadenylation site choice. More importantly, we identified that the m⁶A modification shows multifaceted correlations with the translational status depending on its strength and genic location. Moreover, we observed a substantial intraspecies variation in m⁶A modification, and this natural variation was shown to be partly driven by gene-specific expression and alternative splicing. Together, these findings provide an invaluable resource for ascertaining transcripts that are subject to m⁶A modification in maize and pave the way to a better understanding of natural m⁶A variation in mediating gene expression regulation.

Background: MicroRNA (miRNA) plays vital roles in the regulation of both plant architecture and stress resistance through cleavage or translation inhibition of the target messenger RNAs (mRNAs). However, miRNA-induced gene silencing remains a major challenge in vivo due to the low delivery efficiency and instability of miRNA, thus an efficient and simple method is urgently needed for miRNA transformation. Previous researches have constructed a star polycation (SPc)-mediated transdermal double-stranded RNA (dsRNA) delivery system, achieving efficient dsRNA delivery and gene silencing in insect pests. Results: Here, we tested SPc-based platform for direct delivery of double-stranded precursor miRNA (ds-MIRNA) into protoplasts and plants. The results showed that SPc could assemble with ds-MIRNA through electrostatic interaction to form nano-sized ds-MIRNA/SPc complex. The complex could penetrate the root cortex and be systematically transported through the vascular tissue in seedlings of Arabidopsis and maize. Meanwhile, the complex could up-regulate the expression of endocytosis-related genes in both protoplasts and plants to promote the cellular uptake. Furthermore, the SPc-delivered ds-MIRNA could efficiently increase mature miRNA amount to suppress the target gene expression, and the similar phenotypes of Arabidopsis and maize were observed compared to the transgenic plants overexpressing miRNA. Conclusion: To our knowledge, we report the first construction and application of star polycation nanocarrier-based platform for miRNA delivery in plants, which explores a new enable approach of plant biotechnology with efficient transformation for agricultural application. Graphical

Drought is a major abiotic stress that restricts plants growth, development, and yield. Coronatine (COR), a mimic of JA-Ile, functions in plant tolerance to multiple stresses. In our study, we examined the effects of COR in tobacco under polyethylene glycol (PEG) stress. COR treatment improved plant growth under stress as measured by fresh weight (FW) and dry weight (DW). The enzyme activity assay indicated that, under osmotic stress conditions, the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) were enhanced by COR treatment. Histochemical analyses via nitrotetrazolium blue chloride (NBT) and 3,3′-diaminobenzidine (DAB) staining showed that COR reduced reactive oxygen species (ROS) accumulation during osmotic stress. Metabolite profiles revealed that COR triggered significant metabolic changes in tobacco leaves under osmotic stress, and many essential metabolites, such as sugar and sugar derivatives, organic acids, and nitrogen-containing compounds, which might play active roles in osmotic-stressed tobacco plants, were markedly accumulated in the COR-treated tobacco. The work presented here provides a comprehensive understanding of the COR-mediated physiological, biochemical, and metabolic adjustments that minimize the adverse impact of osmotic stress on tobacco.

Summary: Stomatal density (SD) is closely related to crop drought resistance. Understanding the genetic basis for natural variation in SD may facilitate efforts to improve water‐use efficiency. Here, we report a genome‐wide association study for SD in maize seedlings, which identified 18 genetic variants that could be resolved to seven candidate genes. A 3‐bp insertion variant (InDel1089) in the last exon of Zea mays (Zm) IRX15A (Irregular xylem 15A) had the most significant association with SD and modulated the translation of ZmIRX15A mRNA by affecting its secondary structure. Dysfunction of ZmIRX15A increased SD, leading to an increase in the transpiration rate and CO₂ assimilation efficiency. ZmIRX15A encodes a xylan deposition enzyme and its disruption significantly decreased xylan abundance in secondary cell wall composition. Transcriptome analysis revealed a substantial alteration of the expression of genes involved in stomatal complex morphogenesis and drought response in the loss‐of‐function of ZmIRX15A mutant. Overall, our study provides important genetic insights into the natural variation of leaf SD in maize, and the identified loci or genes can serve as direct targets for enhancing drought resistance in molecular‐assisted maize breeding.