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. 2022 Jul 13:13:933849.
doi: 10.3389/fpls.2022.933849. eCollection 2022.

Metabolomics Analysis Reveals the Differences Between Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd

Xuejie Qu  1 , Shanqun Hu  1 , Tong Li  1 , Jiaqi Zhang  1 , Baoshun Wang  1 , Changli Liu  1
Affiliations

Metabolomics Analysis Reveals the Differences Between Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd

Xuejie Qu et al. Front Plant Sci. .

Abstract

Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. are two varieties of Bupleuri Radix in Chinese Pharmacopoeia 2020. The clinical efficacy of the two bupleurum species is different. The difference in clinical efficacy is closely related to the composition of plant metabolites. In order to analyze the difference in metabolites, we used liquid chromatography coupled with mass spectrometry (LC-MS) for untargeted metabolome and gas chromatography coupled with mass spectrometry (GC-MS) for widely targeted metabolome to detect the roots (R), stems (S), leaves (L), and flowers (F) of two varieties, and detected 1,818 metabolites in 25 classes. We performed a statistical analysis of metabolites. Differential metabolites were screened by fold-change and variable importance in the projection values of the OPLS-DA model, and significant differences were found among different groups. The content of active components (triterpenoid saponins) was found to be high in the BcR group than in the BsR group. Other pharmacological metabolites were significantly different. By Kyoto Encyclopedia of Genes and Genomes annotation and enrichment analysis, we found that differential metabolites of the aboveground parts mainly concentrated in monoterpenoid biosynthesis, while the differential metabolites of the root mainly concentrated in sesquiterpenoid and triterpenoid biosynthesis. Differences in metabolic networks may indirectly affect the metabolic profile of Bc and Bs, leading to differences in clinical efficacy. Our study provides a scientific basis for subsequent biosynthesis pathway and related bioactivity research, and provides a reference for developing non-medicinal parts and guiding the clinical application of Bupleuri Radix.

Keywords: Bupleurum chinense DC.; Bupleurum scorzonerifolium Willd.; differential metabolites; saikosaponins; statistical analysis.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
The specimens of mature Bupleurum chinense DC. (A) and Bupleurum scorzonerifolium Willd. (B).
FIGURE 2
FIGURE 2
Quality control (QC) sample with the coefficient of variation (CV) of LC-MS (A) and GC-MS (B) less than 0.3 accounted for more than 75% of the samples, indicating that the experimental data were very stable. Overview of annotated metabolites (C). Venn diagram of the detected metabolites in four tissues of Bc (D) and Bs (E).
FIGURE 3
FIGURE 3
Heat map of HCA and PCA of the relative differences in the metabolite profiles between different varieties and parts. The metabolites are divided into four clusters (A), 3D PCA plot of LC-MS (B), GC-MS (C), and incorporated PCA plot (D).
FIGURE 4
FIGURE 4
The groups of BcR and BsR are shown, including the score OPLS-DA plot (A), OPLS-DA model simulation verification diagram (B), and volcano plots of differential metabolites (C). The statistical analysis of the number of differential metabolites for all different groups is presented (D), with the red color indicating upregulated metabolites and the blue color indicating downregulated metabolites.
FIGURE 5
FIGURE 5
The top 10 downregulated and upregulated metabolites in the BcR and BsR groups (A). Venn diagram between different tissues showed the overlapping and accession-specific differential metabolites (B).
FIGURE 6
FIGURE 6
Differential metabolite pathway bubble map. (A) BcR and BsR. (B) BcS and BsS. (C) BcL and BsL. (D) BcF and BsF. (E) BcR and BcF. (F) BsR and BsF.
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