CRISPR/Cas9-mediated editing of the phytoene desaturase gene in Vanilla planifolia enabling targeted domestication

Abstract

ABSTRACT Vanilla is an undomesticated species and the botanical source of vanilla extract. There has been increasing demand to expand vanilla cultivation in the Americas to strengthen the vanilla supply chain. Recent development in vanilla genomics resources has opened opportunities for interrogation of vanilla diversity and discovery of candidate genes responsible for key traits. The ability to precisely change genomic sequences could be a valuable tool for domesticating vanilla while enabling functional genomics. Here we report the successful biolistic-mediated transformation of seed-derived callus and CRISPR/Cas9 gene editing of the phytoene desaturase (PDS) gene in vanilla. Sequencing of leaves from regenerated plantlets displaying the expected albino phenotype showed short deletions from 1 to 163 base pairs in the phytoene desaturase gene at each of three target loci. The ability to apply gene editing to vanilla can now be used in combination with the vanilla genome to create targeted knockouts to support the domestication of vanilla.