A 2OGD multi-gene cluster encompasses functional and tissue specificity that direct furanocoumarin and pyranocoumarin biosynthesis in citrus

Abstract

• Furanocoumarins (FCs) are plant defence compounds derived from the phenylpropanoid pathway via the coumarin umbelliferone that harbour some therapeutic benefits yet are the underlying cause of ‘grapefruit–drug interactions’ in humans. Most of the pathway genes have not been identified in citrus.
• We employed a genetic/Omics approach on citrus ancestral species and F1 populations of mandarin × grapefruit and mandarin × pummelo. Enzyme specificity was characterized by In vivo 2-oxoglutarate-dependent dioxygenase family (2OGD) activity assays.
• We identified a 2OGD multi-gene cluster involved in coumarin/FC/pyranocoumarin biosynthesis; Species lacking FCs in leaves/fruit were homozygous for a 655-base solo-LTR frame-disrupting insertion within one dual specificity C2′H/F6′H encoding 2OGD gene, demonstrating that integrity of this gene is fully correlated with the capacity to biosynthesize metabolites of the extended FC pathway in leaves/fruit. A second 2OGD is the prominent gene expressed in citrus roots, which contain a unique pattern of extended FC pathway metabolites, including the predominant pyranocoumarins. A third 2OGD gene encodes a single activity F6′H, which appears to be induced at the transcript level by citrus pathogens.
• The results provide insights into the genetic basis underlying the difference between citrus fruit FC producers (grapefruit and pummelo) and nonproducers (mandarin and orange) and provide a gene target to breed for FC-free varieties by marker-assisted breeding or genome editing.