Agropyron mongolicum Keng, a perennial diploid grass with high drought tolerance, belongs to the genus Agropyron, tribe Triticeae. It has made tremendous contributions toward reseeding natural pasture and seeding artificial grassland in China, especially in the arid and semi-arid area of northern China. As a wild relative of wheat, A. mongolicum is also a valuable resource for the genetic improvement of wheat crops. MicroRNAs are small non-coding RNA molecules ubiquitous in plants, which have been involved in responses to a wide variety of stresses including drought, salinity, chilling temperature. To date, little research has been done on drought-responsive miRNAs in A. mongolicum. In this study, two miRNA libraries of A. mongolicum under drought and normal conditions were constructed, and drought-responsive miRNAs were screened via Solexa high throughput sequencing and bioinformatic analysis. A total of 114 new miRNAs were identified in A. mongolicum including 53 conservative and 61 unconservative miRNAs, and 1393 target genes of 98 miRNAs were predicted. Seventeen miRNAs were found to be differentially expressed under drought stress, seven (amo-miR21, amo-miR62, amo-miR82, amo-miR5, amo-miR77, amo-miR44 and amo-miR17) of which were predicted to target on genes involved in drought tolerance. QRT-PCR analysis confirmed the expression changes of the seven drought related miRNAs in A. mongolicum. We then transformed the seven miRNAs into Arabidopsis thaliana plants, and three of them (amo-miR21, amo-miR5 and amo-miR62) were genetically stable. The three miRNAs demonstrated the same expression pattern in A. thaliana as that in A. mongolicum under drought stress. Findings from this study will better our understanding of the molecular mechanism of miRNAs in drought tolerance and promote molecular breeding of forage grass with improved adaption to drought.

Maize stalk lodging causes substantial yield losses worldwide. Although stalk strength is a genetically determined trait, its molecular mechanisms—particularly the dynamic changes during key developmental stages—remain inadequately characterized due to limitations of single-omics approaches. This study employed an integrated transcriptomic and metabolomic analysis strategy to compare stalk tissues from three maize genotypes with contrasting lodging resistance: the highly resistant inbred line PHB1M, the susceptible inbred line Chang 7-2, and their recombinant inbred line 23NWZ561 (abbreviated as P, C, and Z, respectively). Dynamic sampling of all three genotypes was conducted at both grain-filling and maturity stages, with simultaneous measurement of physiological traits related to stalk strength. Phenotypic analysis revealed that the resistant genotype PHB1M exhibited superior rind penetration strength, cell wall composition (cellulose, hemicellulose, and lignin) content, and vascular bundle development. Multi-omics analysis indicated that the molecular basis of lodging resistance is primarily established during the maturity stage. The transcriptomic and metabolomic profiles of the recombinant inbred line Z shifted from clustering with the susceptible parent C at the grain-filling stage to grouping with the resistant parent P at maturity. Key pathways including phenylpropanoid biosynthesis were significantly enriched specifically at maturity, accompanied by upregulation of related genes (PAL, HCT, CCR) and accumulation of metabolites such as lignin precursors in PHB1M. Integrated analysis identified a core co-expression network within the phenylpropanoid pathway comprising three genes and three metabolites. This study systematically demonstrates that lodging resistance in maize is regulated by transcriptional and metabolic reprogramming during late stalk developmental stages, particularly at maturity, where enhanced activation of the phenylpropanoid biosynthesis pathway plays a central role. These findings provide valuable candidate genes and metabolic markers for breeding lodging-resistant maize varieties.

Leaf senescence is essential for the growth and development of deciduous trees in the next season. Larix gmelinii, a deciduous coniferous tree, exhibits its most distinctive feature by turning yellow in the autumn and eventually shedding its leaves, resulting in significant changes in its appearance during the fall. Lysine acetylation plays an important role in diverse cellular processes; however, limited knowledge is available regarding acetylations in the needle senescence of L. gmelinii. In this study, the proteomics and acetylated modification omics of two phenotypic leaves, yellow and green (senescent and non-senescent) needles, were analyzed before autumn defoliation. In total, 5022 proteins and 4469 unique acetylation sites in 2414 lysine acylated proteins were identified, and this resulted in the discovery of 1335 differentially expressed proteins (DEPs) and 605 differentially expressed acetylated proteins (DAPs) in yellow versus green needles. There are significant differences between the proteome and acetylome; only 269 proteins were found to be DEP and DAP, of which 136 proteins were consistently expressed in both the DEP and DAP, 91 proteins were upregulated, and 45 proteins were down-regulated. The DEPs participate in the metabolism of starch and sucrose, while the DAPs are involved in glycolysis and the tricarboxylic acid cycle. Among them, DEPs underwent significant changes in glycolysis and citric acid cycling. Most of the enzymes involved in glycolysis and the citrate cycle were acetylated. DAPs were down-regulated in glycolysis and up-regulated in the citrate cycle. In all, the results of this study reveal the important role of lysine acetylation in the senescence of L. gmelinii needles and provide a new perspective for understanding the molecular mechanism of leaf senescence and tree seasonal growth.

In northern China’s arid and semi-arid regions, evergreen trees demonstrate significant cold tolerance to natural low-temperature stress during winter. However, the metabolic strategies and their associated properties underlying their overwintering adaptation remain incompletely elucidated. This study aims to reveal the metabolic properties of natural low-temperature adaptation strategies in five evergreen trees through metabolomic analysis and to identify key metabolites and their dynamic variation patterns. The GC-TOF-MS platform was used to investigate seasonal differential metabolites in five evergreen trees across January, April, July, and October and further explore core differentially expressed metabolites responsive to low-temperature stress. The results demonstrated that the seasonal changes in the chlorophyll content of five evergreens exhibited distinct patterns, that significant differences were observed between Juniperus sabina L. and Picea meyeri R., Ammopiptanthus mongolicus M., Buxus sinica var. parvifolia M.Cheng, and Pinus tabuliformis C., and that no significant differences were found among the other tree species. A total of 427 metabolites were detected in the metabolome; when assessing seasonal dynamics, it was found that the types of differentially expressed metabolites in the five evergreens underwent significant changes. In spring, the differentially expressed metabolites included some carbohydrates, alcohols, organic acids, and lipids. During summer and autumn, the largest number of differentially expressed metabolites accumulated, mainly including carbohydrates, organic acids, and amino acid compounds. In winter, while Picea meyeri primarily accumulated carbohydrates, the remaining four species mainly accumulated organic acids, along with a small number of alcohols, phenylpropanoids, and polyketides. Three shared carbohydrate metabolites, L-threose, galactinol, and gluconic lactone, were commonly downregulated across all species. Additionally, coniferous trees collectively accumulated 3,6-anhydro-D-galactose, showing downregulation. The KEGG enrichment analysis of winter-accumulated metabolites revealed significant associations with the pentose phosphate pathway, amino acid metabolism, phenylpropanoid biosynthesis, the tricarboxylic acid cycle, and ascorbate–aldarate metabolism pathways. Through comparative analysis with the summer growth season, we ultimately identified the core differentially expressed metabolites of the five evergreens, providing potential metabolic markers for the breeding of cold-tolerant species. In summary, these findings provide critical metabolomic insights into how plants adapt to low temperatures, significantly enhancing our understanding of the metabolic foundations of cold tolerance in evergreen species.

In mitigating agricultural nitrous oxide (N₂O) emissions, partially substituting chemical nitrogen (N) fertilizers with organic amendments is widely advocated for; however, the effectiveness of this approach varies, and the optimal substitution ratio remains unclear. This study employed a long-term field experiment in a maize cropping system, integrating in situ N₂O flux monitoring, soil analysis, and metagenomic sequencing to investigate the effects of substituting synthetic N fertilizer with sheep manure at different ratios (one-sixth, one-third, and full substitution) on cumulative N₂O emissions and underlying microbial mechanisms. The results showed that substituting one-third of chemical N with sheep manure (OF2) most effectively reduced cumulative N₂O emissions, by 18.64% (2021) and 47.00% (2022), compared to conventional chemical fertilization (NPK). This treatment optimized key soil properties, significantly lowering soil nitrate (NO₃⁻-N) and ammonium (NH₄⁺-N) concentrations while increasing soil organic matter. Metagenomic analysis revealed that the mitigation was primarily driven by a coordinated shift in the nitrogen-cycling microbial functional gene network: OF2 significantly enhanced the abundance of N₂O reductase genes (nosZ clades I and II) and increased the nosZ/(nirK + nirS) gene ratio, thereby strengthening the genetic potential for complete denitrification (N₂O → N₂). Partial least squares path modeling confirmed that soil properties indirectly reduced N₂O emissions, primarily by regulating the microbial community and functional gene abundances. This study provides mechanistic evidence that partial organic substitution (with one-third sheep manure) mitigates N₂O emissions by optimizing the soil habitat and reprogramming the functional gene network of nitrogen-cycling microbes, with a key emphasis on enhancing the N₂O reduction pathway.