Multi-omics combined with MALDI mass spectroscopy imaging reveals the mechanisms of biosynthesis of characteristic compounds in Tetrastigma hemsleyanum Diels et Gilg

Yan Lin; Xuechun Jiang; Sheng Zhu; Junling Dun; Jinbao Pu; Weiqing Liang

doi:10.3389/fpls.2023.1294804

Multi-omics combined with MALDI mass spectroscopy imaging reveals the mechanisms of biosynthesis of characteristic compounds in Tetrastigma hemsleyanum Diels et Gilg

Front Plant Sci. 2024 Jan 9:14:1294804. doi: 10.3389/fpls.2023.1294804. eCollection 2023.

Authors

Yan Lin¹, Xuechun Jiang², Sheng Zhu³, Junling Dun⁴, Jinbao Pu², Weiqing Liang²

Affiliations

¹ Department of Pharmacy, Tongde Hospital of Zhejiang Province, Hangzhou, China.
² Key Laboratory of Research and Development of Chinese Medicine of Zhejiang Province, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, China.
³ Zhejiang Guangsheng Pharmaceutical Co., Ltd., Quzhou, China.
⁴ Analytical Applications Center, Shimadzu (China) Co., Ltd., Shanghai, China.

Abstract

Tetrastigma hemsleyanum Diels et Gilg is recognized as a source of extracts with various desirable bioactivities. However, current knowledge regarding the mechanisms of biosynthesis of flavonoids, phenolic compounds, and other bioactive chemicals is limited. We conducted comprehensive tissue distribution studies and biosynthetic analyses of the 26 main bioactive compounds of this plant. The majority of flavonoids exhibited higher concentrations in the cortex (CT) compared to the vascular cylinder (VC). The expression levels of genes and proteins in CT and VC were quantified using mRNA sequencing and isobaric tags for relative and absolute quantification (iTRAQ). A total of 31,700 genes were identified, among which 4921 exhibited differential expression between CT and VC. A total of 13,996 proteins were identified in the proteomes of CT and VC, with 927 showing differential expression. Co-expression network analyses of DEGs and DEPs from multiple sites demonstrated substantial pathway variations linked to flavonoid biosynthesis. Through differential enrichment analysis, a total of 32 genes involved in the flavone biosynthesis pathway were identified, with iTRAQ specifically detecting C3'H, F3H and FLS. Pearson correlation analysis revealed a strong association between the expression levels of C3'H, F3H, and FLS and the concentrations of flavonoids. The validation of multiple genes encoding pivotal enzymes was conducted using real-time fluorescence quantitative PCR (RT-qPCR). The findings provide a foundation for future investigations into the molecular mechanisms and functional characterization of T. hemsleyanum candidate genes associated with characteristic compounds.

Keywords: MALDI mass spectrometry imaging; Tetrastigma hemsleyanum; biosynthesis mechanism; characteristic compound; proteomics; transcriptomics.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the Zhejiang Traditional Chinese Medicine Science and Technology Planning Project (2020ZX004) and Chinese Medicine Health Services Research Project (2024ZF055) from Administration of Traditional Chinese Medicine of Zhejiang Province, and the program of Zhejiang Provincial Department of Science and Technology (LGF20H280003).