Transcriptome and Metabolome Analysis of Atractylodes lancea Across Different Developmental Stages

Guanyu Zhang¹Zhiqiang Zhao²Yaqian Li²Leheng Zhang¹Jimei Lu²Kangru Qi²Hua Liang²Liangping Zha^2*Jin Xie^1*

• ¹Anhui Medical University, Hefei, China
• ²Anhui University of Chinese Medicine, Hefei, China

Atractylodes lancea (Thunb.) DC is a medicinal plant known for its rhizome's 20 production of valuable sesquiterpenoids, although the molecular mechanisms 21 underlying their biosynthesis are not well understood. This study utilized integrated 22 metabolomic and transcriptomic analyses to examine terpenoid dynamics across four 23 developmental stages (June, July, September, November) in A. lancea. Metabolite 24 profiling indicated distinct accumulation patterns: monoterpenoids reached their peak 25 in July, while sesquiterpenoids were most abundant in September. Transcriptome 26 analysis revealed the presence of 36 structural genes linked to the mevalonate (MVA) 27 and methylerythritol phosphate (MEP) pathways, alongside 55 terpene synthase (TPS) 28 genes. Subsequent phylogenetic analysis categorized the TPS genes into distinct 29 subfamilies. Within the TPS-a subfamily, a comprehensive screening process, which 30 considered significant correlations with terpenoid metabolites and the preservation of 31 key conserved motifs, identified eight candidate genes, including AlTPS21 and 32 AlTPS42. Functional characterization demonstrated that the AlTPS21 protein 33 catalyzes the conversion of farnesyl diphosphate to δ-cadinene and α-cadinol, while 34 the AlTPS42 protein catalyzes the conversion of farnesyl diphosphate to δ-cadinene 35 and α-copaene. Subcellular localization studies showed that both enzymes are 36 localized to the nucleus and cell membrane. These findings enhance the understanding 37 of the temporal regulation of terpenoid biosynthesis in A. lancea and provide crucial 38 genetic insights for future metabolic engineering efforts.