Multiomics analysis reveals flavonoid accumulation and biosynthesis across different cultivation years and localities of Gongronemopsis tenacissima (Dai-Bai-Jie).

BACKGROUND

The dried root of Gongronemopsis tenacissima (Roxb.) S. Reuss, Liede & Meve, traditionally known as Dai-Bai-Jie, is a medicinal plant used by the Dai ethnic group, primarily for detoxification. Due to extensive use, wild resources have become increasingly scarce, prompting domestication efforts in China. However, the accumulation patterns of secondary metabolites-particularly flavonoids, the main detoxifying components-and their biosynthetic mechanisms remain unclear.

METHODS

This study investigated differences in flavonoid accumulation and transcriptional regulation in Dai-Bai-Jie cultivated for one, two, and three years at high altitudes, and three years at low altitudes. Transcriptome and widely targeted metabolome analyses were conducted. A total of 1,495 metabolites were identified using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS), with 943 showing differential accumulation across the four groups. All flavonoids were classified into six clusters using k-means clustering. Flavonoid levels were generally higher in plants cultivated for two and three years, with total metabolite content also more abundant in these groups. Two-year cultivation was recommended as the optimal harvesting strategy.

RESULTS

A regulatory relationship was observed between genes such as phenylalanine ammonia-lyase, CYP73A, 4-coumarate: coenzyme A ligase, and flavonol synthase, and the flavonoid components in Dai-Bai-Jie. No significant differences were found in Shannon, Chao1, or abundance coverage estimator (ACE) indices of rhizosphere microorganisms across different cultivation years and locations.

CONCLUSIONS

This study elucidates the mechanisms of flavonoid accumulation and supports the scientific rationale for optimal harvesting years of Dai-Bai-Jie. The findings provide a valuable foundation for guiding large-scale cultivation and reducing reliance on wild resources.