Multi-omics analysis Taiwanofungus gaoligongensis fermentation with traditional Chinese medicine

This study employed a multi-omics approach to investigate the accumulation of bioactive terpenoids and flavonoids in Taiwanofungus gaoligongensis during liquid culture supplemented with traditional Chinese medicine materials, including Coix lacryma-jobi powder (YM), Astragalus membranaceus powder (HQ), and Pleuropterus multiflorus powder (HSW). Compared with pea flour (WDF), total terpenoid content increased significantly in the HQ and HSW groups, reaching 2.86-fold and 4.23-fold, respectively. Sesquiterpenoids were the predominant class, among which (+ /-)-α-bisabolol and nootkatone in HQ were 8.81-fold and 9.07-fold higher than those in WDF, respectively, whereas drechslerene E in HSW was elevated by 11.17-fold. Flavonoid accumulation was highest in the HQ group, reaching 16.05 times that of WDF, and was primarily composed of isoflavones. Specifically, the levels of maximaisoflavone J, glycitein 1, and formononetin in HQ were 56.59-, 460.39-, and 4570.37-fold higher than those in WDF, respectively. Moreover, the characteristic active component astragaloside IV from A. membranaceus and the flavonoids quercetin and rutin from P. multiflorus were detected in the HQ and HSW fermentation products, respectively. In contrast, potentially hepatotoxic constituents of P. multiflorus, including emodin and anthraquinones, were not detected in the HSW fermentation products, suggesting that co-fermentation with T. gaoligongensis may attenuate the toxicity of P. multiflorus. Transcriptomic analysis revealed that the relative expression levels of multiple genes involved in terpenoid biosynthesis were significantly upregulated in the HQ and HSW groups. In HSW, the expression levels of TgHMGS, TgHMGR, TgMVD, and TgOSC were 2.68-, 2.62-, 2.30-, and 1.86-fold higher than those in WDF, respectively, whereas TgAACT, TgIDI, TgSES, and TgErg25 were upregulated by 2.11-, 1.82-, 2.68-, and 2.25-fold in HQ. These genes may represent key regulatory nodes in the terpenoid biosynthetic pathway of T. gaoligongensis. Furthermore, co-expression trend analysis combined with promoter-binding site prediction suggested that TgZnF10, TgHMG11, and TgZnF19 may regulate the transcriptional activity of TgHMGR, TgPMK, and TgFPPS_2, respectively, while TgErg26, TgErg62, TgErg4, and TgErg5 may be regulated by TgBHLH7, TgBZIP4, TgMYB3, and TgHOX2, respectively. Collectively, these findings provide a theoretical basis for the comprehensive utilization and industrial development of T. gaoligongensis through bidirectional fermentation with Chinese medicinal materials.