Amaryllidaceae alkaloids (AAs) are a structurally diverse group of plant specialized metabolites with powerful biological activities. The medicinal properties of many AAs have been identified including the antitumor agent narciclasine and galanthamine, used for Alzheimer’s disease. Tracer studies have led to proposed pathways but AA biosynthesis remains molecularly uncharacterized. The use of systems biology-based approaches could lead to the unraveling of AA metabolic pathways. The elucidation of AA biosynthesis will provide necessary tools required to enhance AA production in plants as well as the development of microbial production platforms as an alternative to plants as a commercial source of valuable AAs.