Development of a Comprehensive Tissue Culture System for Genome Editing in Cannabis

Matchett-Oates, L.1,2, Mohamaden, E.1, Spangenberg, G.C.1,2 and Cogan, N.O.I.1,2

1Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Bundoora, Victoria 3083, Australia

2School of Applied Systems Biology, La Trobe University, Bundoora, Victoria 3086, Australia

Cannabis is dioecious in nature, limiting the ability to maintain elite cultivars with selected traits through seed multiplication. Standard practice for the maintenance of key production strains is via vegetative mother plants, this is a high risk to the industry through inadvertent pest and disease outbreaks eliminating product strains. The development of biotechnological approaches, for genetic modification as well as tissue culture systems for the conservation, storage and rapid multiplication of elite strains is crucial. Currently, there is limited literature on successful tissue culture methods in Cannabis, with biotechnologically important protocols yet to be developed. Micropropagation of selected Cannabis genotypes have been developed, however considerable response variation has been reported. Similarly, cell suspension cultures and callus induction from Cannabis explants show high levels of response variation between genotypes from plant growth regulators and media compositions. A broad range of tissue culture protocols are also critical for the establishment of capability in genome editing to enable designer strains with novel chemotypes to be developed. A broad range of tissue culture protocols have been developed for Cannabis and are shown to be robust across genotypes with the first reported details of protoplast isolation and transformation, along with whole plant regeneration from callus. These protocols will allow for the conservation and rapid multiplication of elite strains to be used for genetic improvement through genome editing.