SWATH-MS-Based Proteomics Reveals the Regulatory Metabolism of Amaryllidaceae Alkaloids in Three Lycoris Species

Alkaloids are a class of nitrogen-containing alkaline organic compounds found in nature, with significant biological activity, and are also important active ingredients in Chinese herbal medicine. Amaryllidaceae plants are rich in alkaloids, among which galanthamine, lycorine, and lycoramine are representative. Since the difficulty and high cost of synthesizing alkaloids have been the major obstacles in industrial production, particularly the molecular mechanism underlying alkaloid biosynthesis is largely unknown. Here, we determined the alkaloid content in Lycoris longituba, Lycoris incarnata, and Lycoris sprengeri, and performed a SWATH-MS (sequential window acquisition of all theoretical mass spectra)-based quantitative approach to detect proteome changes in the three Lycoris. A total of 2193 proteins were quantified, of which 720 proteins showed a difference in abundance between Ll and Ls, and 463 proteins showed a difference in abundance between Li and Ls. KEGG enrichment analysis revealed that differentially expressed proteins are distributed in specific biological processes including amino acid metabolism, starch, and sucrose metabolism, implicating a supportive role for Amaryllidaceae alkaloids metabolism in Lycoris. Furthermore, several key genes collectively known as OMT and NMT were identified, which are probably responsible for galanthamine biosynthesis. Interestingly, RNA processing-related proteins were also abundantly detected in alkaloid-rich Ll, suggesting that posttranscriptional regulation such as alternative splicing may contribute to the biosynthesis of Amaryllidaceae alkaloids. Taken together, our SWATH-MS-based proteomic investigation may reveal the differences in alkaloid contents at the protein levels, providing a comprehensive proteome reference for the regulatory metabolism of Amaryllidaceae alkaloids.

Automated summary

Alkaloids are nitrogen-containing alkaline organic compounds with important biological activity and are essential active ingredients in Chinese herbal medicine. Amaryllidaceae plants are rich in alkaloids, including galanthamine, lycorine, and lycoramine. This study analyzed the alkaloid content in Lycoris longituba, Lycoris incarnata, and Lycoris sprengeri, and used a SWATH-MS-based quantitative approach to detect proteome changes. The results revealed differentially expressed proteins involved in specific biological processes, suggesting a supportive role for Amaryllidaceae alkaloid metabolism. Key genes responsible for galanthamine biosynthesis were identified, and RNA processing-related proteins were found abundantly in alkaloid-rich Lycoris, indicating a potential role of posttranscriptional regulation in alkaloid biosynthesis. Overall, this proteomic investigation provides a comprehensive reference for understanding the regulatory metabolism of Amaryllidaceae alkaloids.