A chromosome-level genome assembly and multi-omics analyses provide insights into the biosynthesis of piperlongumine in Piper sarmentosum

Piper sarmentosum Roxb. is a significant medicinal and edible plant, and its active compound piperlongumine (PL) has garnered attention due to its pharmacological activities, including anticancer and anti-inflammatory effects. However, the key enzymes and regulatory mechanisms of its biosynthetic pathway are not yet fully understood. In this study, we generated a chromosome-level genome assembly, with a contig N50 of 15.36 Mb and a scaffold N50 of 22.52 Mb. The BUSCO assessment indicated high completeness, with a score of 97.4%. Genome annotation revealed 39,154 protein-coding genes and identified three lineage-specific whole-genome duplication (WGD) events that expanded gene families associated with alkaloid biosynthesis. Metabolomic analysis identified 4,456 metabolites, including 238 alkaloids, and demonstrated that flowers and fruits are the primary organs for PL biosynthesis. Molecular docking and the correlation of gene expression with levels of PL suggest that PsHCT1 catalyzes the condensation of sinapoyl-CoA and 5,6-dihydropyridinone, while PsCCoAOMT1 is responsible for the final synthesis of PL. This study provides insights into the mechanism of alkaloid biosynthesis in P. sarmentosum and may help lay the groundwork for enhancing the production of medicinal compounds. A chromosome-level genome assembly for Piper sarmentosum reveals three whole-genome duplication events. PsHCT1 and PsCCoAOMT 1 are identified as potential key enzymes for piperlongumine production, while the loss of Piperine synthases may explain its low piperine content.