Gastrodia elata Blume is an important medicinal orchid, yet its large-scale cultivation is increasingly threatened by fungal diseases. The 4-coumarate–CoA ligase (4CL) gene family directs a key step in phenylpropanoid metabolism and plant defence, but its composition and function in G. elata have not been investigated. We mined the G. elata genome for 4CL homologues, mapped their chromosomal locations, and analysed their gene structures, conserved motifs, phylogenetic relationships, promoter cis-elements and codon usage bias. Publicly available transcriptomes were used to examine tissue-specific expression and responses to fungal infection. Subcellular localisation of selected proteins was verified by transient expression in Arabidopsis protoplasts. Fourteen Ge4CL genes were identified and grouped into three clades. Two members, Ge4CL2 and Ge4CL5, were strongly upregulated in tubers challenged with fungal pathogens. Ge4CL2 localised to the nucleus, whereas Ge4CL5 localised to both the nucleus and the cytoplasm. Codon usage analysis suggested that Escherichia coli and Oryza sativa are suitable heterologous hosts for Ge4CL expression. This study provides the first genome-wide catalogue of 4CL genes in G. elata and suggests that Ge4CL2 and Ge4CL5 may participate in antifungal defence, although functional confirmation is still required. The dataset furnishes a foundation for functional characterisation and the molecular breeding of disease-resistant G. elata cultivars.

Background/Objectives: Blumea balsamifera (L.) DC. (Miao Medicine: Diangd vob bvid), a medicinal plant with a long ethnopharmacological history in Southwest China, is widely used by the Miao, Li, and Zhuang ethnic groups to treat rheumatic diseases. While previous studies indicated that the ethyl acetate fraction of B. balsamifera (BBEA) possesses potent anti-inflammatory properties, its specific bioactive material basis and the underlying molecular mechanisms against rheumatoid arthritis (RA) remain elusive. Methods: In this study, an integrated strategy combining chemical profiling, network pharmacology, and experimental validation was employed. First, UPLC-Q-Exactive-MS/MS analysis was conducted to characterize the chemical constituents of BBEA. Subsequently, network pharmacology and molecular docking were utilized to predict potential active compounds and core signaling pathways. Finally, the therapeutic effects and mechanisms were validated in vivo using a collagen-induced arthritis (CIA) rat model and in vitro using lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Results: A total of 25 active constituents, including Genkwanin and Luteolin, were identified in BBEA via UPLC-Q-Exactive-MS/MS. Network pharmacology analysis predicted that the PI3K/AKT signaling pathway is critical for BBEA’s anti-RA activity, and molecular docking confirmed strong binding affinities between key components (e.g., Genkwanin) and core targets (SRC, AKT1). In vivo experiments demonstrated that BBEA significantly reduced the Arthritis Index (AI) and paw swelling, reversed weight loss, and ameliorated synovial hyperplasia in CIA rats (p < 0.05). Furthermore, BBEA markedly downregulated the levels of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-17) both in serum and synovial tissues. Mechanistically, Western blot analysis verified that BBEA inhibited the phosphorylation of PI3K and AKT in a dose-dependent manner. Conclusions: This study systematically reveals that BBEA alleviates RA symptoms and synovial inflammation primarily by inhibiting the PI3K/AKT signaling pathway. These findings provide a scientific basis for the traditional application of B. balsamifera and suggest BBEA as a promising candidate for RA therapy.

Background: Blumea balsamifera (L.) DC. Is a Miao and Li ethnic medicinal plant traditionally used to activate blood circulation and dispel dampness; however, systematic investigation of its flavonoid-enriched fraction against high-fat diet (HFD)-induced lipid disorder and hepatic steatosis remains limited. Methods: A flavonoid-enriched fraction from B. balsamifera leaves (BBF, total flavonoids 30.96%) was characterised by UPLC-HRMS (83 constituents, 22 high-abundance). Network pharmacology, molecular docking, and serum untargeted metabolomics were integrated, and pivotal pathways were validated in HFD-fed rats (graded BBF doses, 55, 110, 220 mg/kg) via Western blot and RT-qPCR. Results: BBF ameliorated serum lipid abnormalities and hepatic steatosis in a dose-dependent manner; the medium-dose simultaneously improved TC, TG, LDL-C, and HDL-C levels, whereas the high-dose exerted superior triglyceride-lowering and anti-steatotic effects. Integrated two-omics analysis prioritised insulin-signalling and arachidonic acid metabolism; BBF was associated with reduced JNK hyper-phosphorylation, restored AKT activation, and decreased transcript levels of Tnf-α, Ptgs2, Srebf1, and Fasn. Conclusions: BBF alleviates HFD-induced dyslipidaemia and hepatic steatosis. This pharmacological effect is associated with coordinated modulation of inflammatory mediators, JNK/AKT signalling, and hepatic lipogenesis, as suggested by our in vivo assays and multi-omics analysis. These observations generate testable mechanistic hypotheses and provide experimental support for the traditional ethnomedicinal use of this Miao remedy.