Histone acetylation and methylation, governed by various histone modification (HM) gene families, are vital for plant biological processes. However, there are limited studies that have explored HMs in ornamental horticultural trees, including sweet osmanthus (Osmanthus fragrans). We performed genome-wide search and identified 208 OfHMs, encompassing 81 histone methyltransferases (OfHMTs), 51 histone demethylases (OfHDMs), 49 histone acetyltransferases (OfHATs) and 27 histone deacetylases (HDACs). Our comprehensive analysis covered chromosome locations, gene structures, conserved domains, cis-acting elements, phylogenetic comparisons, protein interaction networks and functional enrichment pathways for these gene families. Additionally, tandem and fragment replications were unveiled as contributors to the expansion of OfHMs, with some genes exhibiting positive selection. Furthermore, we examined OfHM expression profiles across various tissues and flowering stages, and under 5′-azacytidine (Aza) and ethylene treatments. Most OfHMs displayed heightened expression in leaves, and were downregulated during the flower opening and senescence stages, including OfPRMTs, OfHDTs, OfHDAs, OfSRTs, OfJMJs and OfHAGs; 75.86% and 80.77% of the differentially expressed OfHMs were upregulated after Aza and ethylene treatments, including OfHAGs, OfHDAs and OfSDGs. This study offers a comprehensive analysis of the OfHM gene family, which indicated their potential involvement in ethylene and Aza responses, and in the flowering process. These findings provide valuable insights into the role of OfHMs in flowering and stress responses.

Ethylene-insensitive 3/ethylene-insensitive 3-like (EIN3/EIL) transcription factors are central regulators of ethylene signaling and stress adaptation in plants. However, their roles in Osmanthus fragrans, a globally cherished ornamental and aromatic plant with significant economic value, remain poorly characterized. Here, we identified nine OfEIL genes across eight chromosomes in the O. fragrans “Liuye Jingui” genome. Conserved motif analysis revealed core domains (Motif1/2/4/7), and promoter cis-elements highlighting hormone-related, stress-related, and growth-related regulatory potential. During late flowering stages, six OfEILs (3/4/5/6/7/9) were significantly upregulated. Under 5-azacytidine (AZA, a DNA demethylation agent), OfEIL2 and OfEIL7 were downregulated, whereas the ETH treatment activated OfEIL3/7/8/9. Strikingly, OfEIL7 exhibited dual regulatory roles, correlating strongly with natural flowering progression, AZA-induced demethylation, and ETH responses. Functional divergence was observed in petal senescence, with OfEIL2–5 and OfEIL7–9 showing stage-specific and tissue-specific expression patterns. These results position OfEIL7 as a key hub integrating epigenetic and hormonal signals to modulate floral longevity and stress adaptation. Our study provides the first genome-wide characterization of the EIL family in O. fragrans, offering critical insights for molecular breeding aimed at enhancing ornamental traits and environmental resilience in this economically significant species.

The floral color phenotypes of Osmanthus fragrans cultivars range from light yellow to orange yellow, with ‘Yanzhi Hong’ being the only reported cultivar with a red color. However, the underlying reason for this unique floral coloration remains unclear. The study conducted targeted metabolomics and transcriptomics analyses on the petals of ‘Yanzhi Hong’ at both initial and peak flowering stages. Candidate gene expression was validated, and expression levels of the petals of three cultivars were compared using RT-qPCR. The results revealed the presence of 27 components in the petals of ‘Yanzhi Hong’, including 5 carotenoids, 8 xanthophylls, and 14 xanthophyll esters. Notably, lycopene was detected in abundance for the first time in O. fragrans cultivars. Carotenes accounted for 78.82 ± 3.17% and 91.19 ± 1.69% of the total carotenoid content in petals during the initial and peak flowering stages, respectively, with all carotene contents increasing during the peak flowering period. β-carotene, lycopene, and γ-carotene were identified as the top three carotene components in petals during both initial and full flowering stages. The unique blush red color of ‘Yanzhi Hong’ petals could be attributed to the low content of α-carotene and the rich accumulation of lycopene. Furthermore, a total of 1550 differentially expressed genes (DEGs) were identified in petals at the peak flowering stage relative to the initial flowering stage, with 1003 genes being downregulated and 547 genes being upregulated during the full flowering stage. There are 926 differentially expressed genes (DEGs) annotated in the Gene Ontology (GO) database. Among these DEGs, those that were downregulated and upregulated during the peak flowering period showed significant enrichment in carbohydrate metabolism and oxidation–reduction processes, respectively. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis identified 14 structural genes associated with phenylpropanoid biosynthesis and 7 structural genes linked to carotenoid biosynthesis. Expression levels of candidate genes involved in carotenoid biosynthesis were examined in the petals of three cultivars (‘Yanzhi Hong’, ‘Liuye Jingui’, and ‘Gecheng Dangui’) at both the initial and peak flowering stages. The results indicated that the decreased expression of LYG009054 (LYCE) and LYG018651 (LYCB) in ‘Yanzhi Hong’ resulted in higher lycopene accumulation and lower α-carotene content in the petals. This study offers valuable insights into the mechanisms underlying the unique flower color phenotype of O. fragrans, proving a basis for further research on carotenoid metabolism pathways and the breeding of new cultivars with a variety of flower colors in O. fragrans.

The MADS-box transcription factors play essential roles in various processes of plant growth and development. Here, we identified 107 MADS-box genes in Osmanthus fragrans Lour. genome (OfMADS), encoding proteins ranging from 61 to 608 amino acids. Phylogenetic analysis classified these genes into five subfamilies: MIKC*, MIKCC, Mα, Mβ, and Mγ, with conserved motif architectures within subfamilies. Tandem and whole-genome duplications were identified as key drivers of OfMADS expansion. Cis-regulatory element analysis revealed enrichment for hormone response and developmental regulatory motifs, implicating roles in growth and flowering processes. Transcriptome dynamics across six floral developmental stages (bolting to petal shedding) uncovered 78 differentially expressed OfMADS genes, including 16 exhibiting flower-specific expressions. Integrated metabolome profiling demonstrated robust correlations between critical OfMADS regulators and scent metabolites. This nexus suggests a potential role of these OfMADS in regulating specialized metabolite biosynthesis pathways. Our multi-omics study provides insights into the regulatory hierarchy of OfMADS in coordinating floral morphogenesis and the accumulation of economically significant metabolites in O. fragrans. These findings establish a foundation for subsequent functional validation and molecular breeding of horticultural traits.

The glycine-aspartic acid-serine-leucine (GDSL) esterase/lipase family is a functionally diverse group of hydrolytic enzymes involved in multiple plant biological processes, including stress adaptation and development. However, its evolutionary patterns, functional conservation, and stress-responsive mechanisms in grasses remain not fully elucidated. In this study, a comprehensive comparative genomic analysis was performed on the GDSL gene family across nine representative grass species and Arabidopsis thaliana. Genome-wide identification, phylogenetic analysis, duplication pattern detection, synteny analysis, cis-regulatory element prediction, protein–protein interaction (PPI) network construction, and RNA-seq-based expression profiling were employed. A total of 1707 GDSL genes were identified, with substantial expansion in grasses, especially hexaploid wheat. Whole-genome and segmental duplications were the major drivers of family expansion, with most duplicated genes under strong purifying selection. A grass-specific clade (C3-2) was identified, and extensive syntenic conservation was observed among closely related grasses. Promoter analysis revealed enrichment of stress- and hormone-responsive cis-elements, and RNA-seq showed dynamic GDSL expression under low-temperature stress in rice and wheat. These findings demonstrate that the expansion of the GDSL gene family in grasses is driven by polyploidization and lineage-specific duplication, accompanied by the emergence of a grass-specific clade (C3-2) and regulatory diversification, collectively shaping stress-responsive evolutionary innovation in Poaceae.

Osmanthus fragrans, a fragrant plant native to China, is extensively utilized in the food and cosmetics industries. However, its optimal harvest period spans only 2–3 days, restricting industrial utilization. Long non-coding RNAs (lncRNAs) have emerged as key regulators of plant growth, yet their roles in O. fragrans petal opening and senescence remain unexplored. Here, we performed high-throughput sequencing of O. fragrans petals across six developmental stages, generating a valuable transcriptomic resource. We identified 57,860 mRNAs and 2414 lncRNAs, including 6499 differentially expressed mRNAs (DEMs) and 494 differentially expressed lncRNAs (DELs). Co-expression and WGCNA analyses revealed key modules associated with petal opening (Turquoise module) and senescence (Blue module). The Blue module, significantly correlated with the S6 senescence stage, contained hub lncRNAs novel_00098659 and novel_00077227 co-expressed with multiple transcription factors and hormone signaling components, including 10 ERFs, OfEIN3, OfJAZ, and OfMYC2. Transient overexpression of novel_00077227 in tobacco accelerated leaf senescence in a hormone-dependent manner, with significant increases in electrolyte leakage and MDA content, and reductions in chlorophyll content and antioxidant enzyme activities. qRT-PCR analysis confirmed that novel_00077227 alters the expression of tobacco homologs of predicted target genes involved in ethylene, auxin, jasmonate, and ABA signaling pathways. This study provides a valuable transcriptomic resource of senescence-associated lncRNAs in O. fragrans and offers evidence for the potential involvement of a key candidate, novel_00077227, in hormone-associated senescence regulation. These findings provide a foundation for future functional studies and potential applications in extending flower longevity.