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Mechanism of terpene formation in Pogostemon cablin cv. Jing Huoxiang revealed by metabolome and transcriptome analysis

ABSTRACT:

Pogostemon cablin (Blanco) Benth. cv. 'Jing Huoxiang', is a valuable medicinal plant widely studied for its aboveground tissues, which are rich in bioactive compounds such as patchouli alcohol. However, systematic investigations into the biosynthesis of sesquiterpenes in its underground parts (roots) remain limited, with several critical knowledge gaps: (1) the metabolic basis of root-specific accumulation of polycyclic sesquiterpenes is unclear; (2) key terpene synthase (TPS) gene resources remain underexplored; and (3) the regulatory network of terpenoid biosynthesis is poorly understood. Addressing these questions is essential for the rational design and efficient production of terpene synthases.In this study, we integrated metabolomic and transcriptomic approaches to systematically characterize terpenoid profiles across different tissues of P. cablin and elucidate their biosynthetic regulation. Using GC-MS analysis, we identified distinct terpenoid compositions in roots, stems, leaves, flowers, and glandular trichomes. Notably, patchouli alcohol and pogostone accounted for over 60% of the total volatile oil content, while roots specifically accumulated polycyclic sesquiterpenes such as β-caryophyllene and α-humulene. Through transcriptome sequencing and bioinformatic analysis, we comprehensively annotated the TPS gene family, revealing that the TPS-a subfamily (34 genes) was the most abundant in P. cablin, with several members exhibiting root-predominant expression. Co-expression network analysis further identified candidate genes encoding potential high-efficiency polycyclic sesquiterpene synthases and uncovered a β-caryophyllene/α-humulene-regulated tertiary metabolic pathway.Our findings not only fill a critical gap in understanding sesquiterpene biosynthesis in the underground tissues of P. cablin but also provide a foundation for synthetic biology-based optimization of terpenoid production. This research paves the way for the efficient biosynthesis of sesquiterpenes to meet industrial demands in pharmaceuticals, fragrances, and biofuels.

INSTRUMENT(S): Liquid Chromatography MS - - - Gas Chromatography MS - positive

PROVIDER: MTBLS13122 | MetaboLights | 2025-10-12

REPOSITORIES: MetaboLights

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			Action	DRS
	a_MTBLS13122_GC-MS_positive__metabolite_profiling.txt	Txt
	i_Investigation.txt	Txt
	m_MTBLS13122_GC-MS_positive__metabolite_profiling_v2_maf.tsv	Tabular
	s_MTBLS13122.txt	Txt

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Project description:Background: Tobacco leaves are the core raw materials for cigarette production, and their quality has direct and significant impacts on the sensory qualities of cigarette products. Cembrane diterpenoid alcohols are the main precursors of the aroma and fragrance components of tobacco leaves, accounting for approximately 60% of their chemical composition and considerably contributing to their aroma. Initially, we used ⁶⁰Co-γ radiation to induce mutagenesis in the tobacco wildtype variety, GDH64, to obtain the aphid-resistant mutant, MuGHD64. Cembrane diterpenoid alcohol levels were significantly increased in MuGHD64, indicating altered aromatic quality of its tobacco leaves. However, characteristic traits of and molecular mechanisms underlying the increased cembrane diterpenoid alcohol levels in MuGHD64 remain unknown. Therefore, MuGHD64 and GDH64 were investigated in this study. Results: Physiological and biochemical indicator levels were measured in MuGHD64 and GDH64, and transcriptome sequencing was performed to determine the mechanisms responsible for the increased cembrane diterpenoid alcohol levels in MuGHD64. Notably, cembratriene-ol and -diol levels were significantly higher in MuGHD64 than in GDH64. Additionally, MuGHD64 exhibited optimal chemical component levels and improved coordination. Its sensory quality indicators, such as aroma quality and intensity, sweetness, smoothness, and fineness, were superior to those of GDH64. Leaf surface secretory trichome density was significantly higher in MuGHD64 than in GDH64 at all stages. Levels of the cembrane diterpenoid alcohol metabolic precursors, isopentenyl and geranylgeranyl diphosphates, were also significantly higher in MuGHD64 than in GDH64. Transcriptome sequencing revealed that differentially expressed genes were mainly enriched in the metabolic processes related to terpenoid biosynthesis, sesquiterpenoid biosynthesis, isoprenoid biosynthesis, diterpenoid metabolism, diterpenoid biosynthesis, terpenoid skeleton biosynthesis, and plant hormone signal transduction. Cloning and sequencing of the cembratriene-ol synthase gene, NtCYC, revealed that the coding sequence of MuGHD64 harbored nucleotide substitutions at eight sites, resulting in six amino acid alterations. This significantly increased the gene expression levels, enhancing cembratriene-ol synthesis. Consequently, cembratriene-ol and -diol levels were significantly elevated, markedly improving the aroma quality of MuGHD64. Conclusions: Overall, this study revealed new genetic resources for the enhanced synthesis of cembratriene-ol, providing a foundation for breeding aroma-rich high-quality tobacco plants.

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Mechanism of terpene formation in Pogostemon cablin cv. Jing Huoxiang revealed by metabolome and transcriptome analysis

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