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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.
Acknowledgements
The authors acknowledge Dr. Patricio Muñoz for contributing bacterial strains. The MoClo Toolkit (Addgene kit # 1000000044) and plasmid pAGM47523 (Addgene plasmid # 153221; http://n2t.net/addgene:153221; RRID:Addgene_153221) were gifts from Sylvestre Marillonnet. The MoClo Plant Parts Kit (Addgene kit # 1000000047) and plasmid pICSL80036 (Addgene plasmid # 68259; http://n2t.net/addgene:68259; RRID:Addgene_68259) were gifts from Nicola Patron.
Data availability statement
Sequencing data for unique alleles are available in Supplemental Data.
Disclosure statement
The authors confirm that there are no relevant financial or non-financial competing interests to report.