Medicinal Plant Research 2025, Vol.15, No.5, 206-213 http://hortherbpublisher.com/index.php/mpr 209 Figure 2 Metabolic Pathways of Active Compounds and Pharmacological Properties of Leonurus japonicus var. Tongzi (Adopted from Lee et al., 2020) 4.2 Applications of proteomics in functional enzyme identification Proteomic methods, alongside enzyme assays, have authenticated the role of catalysts in diterpenoids' and leonurine's biosynthesis pathways. Functional analysis of diterpene synthases, for instance, has identified enzymatic reactions leading to the biosynthesis of characteristic spiro-labdane diterpenoids in L. japonicus (Li et al., 2023). 4.3 Contributions of metabolomics to studying metabolite accumulation Metabolomics, in recent years due to the potency of mass spectrometry, has made large-scale profiling of isomers and signature markers between hundreds of metabolites possible. Studies have identified unique differences in the accumulation of metabolites between plant organs, areas, and states, thus making quality control as well as resource utilization feasible (Garran et al., 2019; Zhang et al., 2025). 4.4 Multi-omics integration and metabolic regulatory network models The integration of genomics, transcriptomics, proteomics, and metabolomics made it possible to create metabolic regulatory networks. Such models indicate how enzyme activity, gene expression, and metabolite accumulation are regulated, permitting system-level understanding of biosynthesis and regulation of bioactive compounds in L. japonicus (Li et al., 2023; Wang et al., 2024). 5 Regulatory Mechanisms of Metabolism 5.1 Roles of transcription factors in metabolic pathway regulations Transcription factors such as MYB, bHLH, and WRKY have been reported to be vital in plant secondary metabolism regulation, including alkaloid, flavonoid, and terpenoid biosynthesis. In L. japonicus, from transcriptomic and multi-omics, biosynthetic gene clusters and regulatory modules for the biosynthesis of some of the major metabolites such as leonurine and stachydrine have been identified. Although direct functional
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