MicroRNA (miRNA) is an essential endogenous post-transcriptional regulatory factor, and foxtail millet (Setaria italica L.) is an ideal C4 model cereal that is a highly valuable crop in semiarid and arid areas. The Research on comprehensive and high confidence identification and annotation of foxtail millet miRNAs needs to be strengthened, and to our knowledge, there is no information on the regulatory network of foxtail millet miRNA. In this study, 136 high confidence miRNAs were identified through high-throughput sequencing of the small RNAs in seven tissues at the shooting and grain filling stages of foxtail millet. A total of 2,417 target genes were obtained by combining computational biology software and degradome sequencing methods. Furthermore, an analysis using transcriptome sequencing revealed the relationships between miRNAs and their target genes and simultaneously explored key regulatory modules in panicles during the grain filling stage. An miRNA regulatory network was constructed to explore the functions of miRNA in more detail. This network, centered on miRNAs and combining upstream transcriptional factors and downstream target genes, is primarily composed of feed forward loop motifs, which greatly enhances our knowledge of the potential functions of miRNAs and uncovers numerous previously unknown regulatory links. This study provides a solid foundation for research on the function and regulatory network of miRNAs in foxtail millet.

Foxtail millet (Setaria italica), a significant C4 model crop known for its exceptional photosynthetic efficiency and robust environmental adaptability, serves as an excellent model for investigating C4 photosynthesis and crop stress resilience. When subjected to abiotic stress, foxtail millet employs a sophisticated signal transduction network to regulate its physiological processes, ensuring sustained high photosynthetic efficiency and normal growth. The mitogen-activated protein kinase (MAPK) family plays a key role in plant growth, development, and stress response. Here, we identified and named a MAPK in S. italica as SiMPK6. Fluorescence quantitative PCR analysis revealed that SiMPK6 is mainly expressed in the leaves during the early shooting stage, with induction under various abiotic stresses such as low temperature, high osmotic pressure, high salt, high temperature, and high light. Overexpressing the SiMPK6 in Arabidopsis thaliana mitigated damage to photosystem II induced by stress, underscoring the gene’s crucial role in foxtail millet’s stress signal transduction and maintenance of high photosynthetic efficiency.

Salix psammophila, commonly known as the sandlive willow, is a vital shrub species within the Salicaceae family, particularly significant for its ecological role in regions susceptible to desertification and sandy soils. In this study, we assembled the complete S. psammophila mitochondrial genome using Pacbio HiFi third-generation sequencing data. The genome was found to be a typical single circular structure, with a total length of 715,555 bp and a GC content of 44.89%. We annotated 33 unique protein-coding genes (PCGs), which included 24 core mitochondrial genes and 9 variable genes, as well as 18 tRNA genes (5 of which were multicopy genes) and 3 rRNA genes. Comparative analysis of the PCGs from the mitochondrial genomes of S. psammophila, Populus deltoides, Populus simonii, Salix wilsonii, and Salix suchowensis revealed that these genes are relatively conserved within the Salicaceae family, with variability primarily occurring in the ribosomal protein genes. The absence of the rps14, which encodes a ribosomal protein, may have played a role in the evolution of stress tolerance in Salicaceae plants. Additionally, we identified 232 SSRs, 19 tandem repeat sequences, and 236 dispersed repeat sequences in the S. psammophila mitochondrial genome, with palindromic and forward repeats being the most abundant. The longest palindromic repeat measured 260 bp, while the longest forward repeat was 86,068 bp. Furthermore, 324 potential RNA editing sites were discovered, all involving C-to-U edits, with the nad4 having the highest number of edits. These findings provide valuable insights into the phylogenetic and genetic research of Salicaceae plants.

Salix psammophila C. Wang & C. Y. Yang is an important windbreak and sand-fixing shrub species in Northwest China, with excellent characteristics such as resistance to drought, wind, and sand. S. psammophila needs to be stubbed flat after several years of growth to continue to grow, otherwise, its growth rate will slow down and even begin to die. To understand the genetic regulatory mechanism of secondary growth in S. psammophila, cell structure and transcriptome analysis were performed on the secondary xylem and secondary phloem of stems. The results showed that the secondary xylem and the secondary phloem of S. psammophila were well developed at 1, 2, and 3-year-old stages, and the secondary growth changes mainly occurred in the secondary xylem at the 2 to 3-year-old stage, with a faster growth rate. The CSE2 and CSE1 genes that regulate CSE (caffeoyl shikimate esterase) have high sequence similarity (92% and 93%) with the CSE2 and CSE1 genes of the genus Populus, respectively, and regulate lignin biosynthesis. Notably, the expression levels of these two genes decreased in the secondary xylem of 3-year-old S. psammophila, indicating that the rapid growth of S. psammophila may be related to lignin biosynthesis. Weighted gene co-expression network analysis (WGCNA) was utilized to screen candidate TFs and genes involved in the secondary growth processes of S. psammophila, which were categorized into six co-expression modules. A total of 79 genes were selected from these co-expression modules, and co-expression network maps of the genes were constructed. The results indicate that the secondary growth of S. psammophila was regulated by a TF regulatory network. Interestingly, PLATZ TFs were involved in the rapid secondary growth and stress tolerance in S. psammophila. This hints that S. psammophila may promote secondary growth by increasing stress tolerance.

Foxtail millet ( Setaria italica ), a typical NADP-ME-type C4 crop, demonstrates superior light-use and water-use efficiencies compared to C3 species. Beyond its photosynthetic edge, it employs a range of stress-resilience mechanisms that optimize growth-defense trade-offs under drought, salinity, or nitrogen scarcity. The combination of high C4 photosynthetic efficiency with extensive stress tolerance is rare among cultivated species, positioning foxtail millet as an ideal model for studying the integration of yield and stress resilience. Phosphoinositide-specific Phospholipase C (PI-PLC) plays a crucial role in lipid- and Ca ²⁺ -dependent signaling pathways. In plants, it modulates responses to biotic and abiotic stresses, though the mechanisms remain partially understood. In this study, we identified five PI-PLC -encoding genes in foxtail millet, named SiPLC1-SiPLC5 , and analyzed their systematic phylogeny, gene structure, protein characteristics, distribution of the chromosome, collinearity relationship, and cis- acting elements prediction at the promoter region. Phylogenetic analysis revealed that the members of the SiPLCs gene family were divided into three subgroups. Structural analysis that all of them have four conserved motifs and these motifs were evenly distributed. Notably, SiPLC1 harbors an exceptionally large first intron and falls within subgroup II; its protein sequence is highly homologous to AtPLC1 and AtPLC3 of Arabidopsis thaliana (L.) Heynh. and to OsPLC4 of Oryza sativa L. RT-qPCR indicated that SiPLC1 is predominantly expressed in roots during early stem elongation and is significantly upregulated under salt stress. Overexpression of SiPLC1 in Arabidopsis mitigated salt-induced damage, highlighting its critical role in salt-stress signal transduction in foxtail millet.

Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein family unique to eukaryotes, yet its functional characterization in woody plants remains limited. In this study, we identified four eIF5A genes (PtoeIF5A1–PtoeIF5A4) from the genome of Populus tomentosa, a fast-growing tree species indigenous to China, and characterized their expression patterns and functional roles through bioinformatics analysis, quantitative real-time PCR, stable overexpression in Arabidopsis thaliana, and transient expression in Nicotiana benthamiana leaves. Our results demonstrated that all PtoeIF5A proteins contain a conserved OB-fold domain and multiple phosphorylation sites, with PtoeIF5A1 showing predominant expression in roots and secondary xylem. Functional assays revealed that PtoeIF5A1 overexpression accelerated inflorescence stem elongation and early flowering in Arabidopsis, induced visible chlorosis and programmed cell death (PCD) in tobacco leaves, and significantly enhanced salt tolerance under NaCl treatment. Collectively, these findings establish PtoeIF5A1 in poplar as a pleiotropic regulator integrating developmental cues, programmed cell death, and stress responses; and as a valuable genetic resource for breeding stress-resilient woody plants.