Background: Qianhu is a traditional Chinese medicine. It is thought that Qianhu roots will harden after bolting and not be suitable for medicinal purposes. Bolting Qianhu and unbolting Qianhu are referred to as “Xiong Qianhu” and “Ci Qianhu,” respectively. In this study, the properties, microscopic and chemical characteristics of Ci Qianhu and Xiong Qianhu roots were compared using fluorescence microscopy, laser microdissection coupled with ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry, and high-performance liquid chromatography with diode-array detection. Results: Microscopy results showed that the area of secondary xylem in the root increased after bolting, with the cork and secretory canals showing strong fluorescence intensity. A total of 34 peaks, mostly pyranocoumarins, were identified in the tissues of Ci Qianhu and Xiong Qianhu. The secretory canals contained the highest variability of coumarins, whereas the secondary xylem contained the least coumarins. Moreover, seven coumarins, especially the pyran- coumarin, decreased after bolting. Generally, both before and after bolting, coumarin level was the highest in the bark, followed by the middle part, and the lowest in the inner part. Conclusion: Thus, it was indicated that the area of secondary xylem increased after bolting, however the coumarin variant and content decreased in the secondary xylem of Qianhu. The result shows that the quality of Qianhu decreases after bolting, which supports the viewpoint that Xiong Qianhu is not suitable for medicinal use.

Diterpene alkaloids (DAs) are characteristic compounds in Aconitum, which are classified into four skeletal types: C₁₈, C₁₉, C₂₀, and bisditerpenoid alkaloids. C₂₀-DAs are thought to be the precursor of the other types. Their biosynthetic pathway, however, is largely unclear. Herein, we combine metabolomics and transcriptomics to unveil the methyl jasmonate (MJ) inducible biosynthesis of DAs in the sterile seedling of A. gymnandrum, the only species in the Subgenus Gymnaconitum (Stapf) Rapaics. Target metabolomics based on root and aerial portions identified 51 C₁₉-DAs and 15 C₂₀-DAs, with 40 inducible compounds. The highest content of C₂₀-DA atisine was selected for further network analysis. PacBio Isoform sequencing integrated with RNA sequencing not only provided the full-length transcriptome but also their response to induction, revealing 1994 genes that exhibited up-regulated expression. Further, 38 genes involved in terpenoid biosynthesis were identified, including 7 diterpene synthases. In addition to the expected function of the four diterpene synthases, AgCPS5 was identified to be a new ent-8,13-CPP synthase in Aconitum and could also combine with AgKSL1 to form the C₂₀-DAs precursor ent-atiserene. Combined with multiple network analyses, six CYP450 and seven 2-ODD genes predicted to be involved in the biosynthesis of atisine were also identified. This study not only sheds light on diterpene synthase evolution in Aconitum but also provides a rich dataset of full-length transcriptomes, systemic metabolomes, and gene expression profiles, setting the groundwork for further investigation of the C₂₀-DAs biosynthesis pathway.

Sanghuang (SH), a natural fungal resource used for food and medicinal purposes, has drawn considerable attention due to its pharmacological effects and efficacy. This study focused on Wild Sanghuang (WS) and Sanghuang cultivated using two different methods: Duanmu Sanghuang and Mycelium Sanghuang. Using UPLC-O-TOF-MS, we conducted an in-depth analysis of the secondary metabolites present in SH. The content of 18 amino acids was measured using an automated amino acid analyzer. The results demonstrated major differences in secondary metabolites, including flavonoids, organic acids, amino acids, and their derivatives, between WS and cultivated Sanghuang (CS). The total amino acid content in WS surpassed that of CS, with segmental trunk SH exhibiting a total amino acid content 1.3 times (p < 0.05) greater than that of bag material SH. This variation may be linked to the biosynthetic pathways of valine, leucine, isoleucine, and flavonoids. By comparing the metabolomic and amino acidomic differences between WS and artificially CS, this study aims to provide a scientific basis for understanding the nutritional and medicinal value of various cultivation methods for SH and offer theoretical support for the future development of SH-related products.