Abstract
For a long time, Cannabis sativa has been used for therapeutic and industrial purposes. Due to its increasing demand in medicine, recreation, and industry, there is a dire need to apply new biotechnological tools to introduce new genotypes with desirable traits and enhanced secondary metabolite production. Micropropagation, conservation, cell suspension culture, hairy root culture, polyploidy manipulation, and Agrobacterium-mediated gene transformation have been studied and used in cannabis. However, some obstacles such as the low rate of transgenic plant regeneration and low efficiency of secondary metabolite production in hairy root culture and cell suspension culture have restricted the application of these approaches in cannabis. In the current review, in vitro culture and genetic engineering methods in cannabis along with other promising techniques such as morphogenic genes, new computational approaches, clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR/Cas9-equipped Agrobacterium-mediated genome editing, and hairy root culture, that can help improve gene transformation and plant regeneration, as well as enhance secondary metabolite production, have been highlighted and discussed.
Highlights
Cannabis sativa L. is a high-demand plant with a long history of medicinal, industrial, recreational, and agricultural uses [1,2]
FloresSanchez et al [54] used various biotic (Pythium aphanidermatum and Botrytis cinerea) and abiotic elicitors in cannabis cell suspension cultures; improved cannabinoid production was not obtained. These results suggest that the biosynthesis of cannabinoids is completely linked to tissue and organ-specific development and complex gene regulatory networks that can only be efficiently produced by trichomes, which are most abundant in differentiated floral tissues
Efficient and reliable in vitro culture procedures can be considered as an important prerequisite for successful gene transformation, genome editing, micropropagation, and conservation of cannabis
Summary
Cannabis sativa L. is a high-demand plant with a long history of medicinal, industrial, recreational, and agricultural uses [1,2]. Cannabis can be categorized based on taxonomic relationships or chemotype but is often divided into two main groups and regulated based on the level of psychoactive cannabinoids that are produced. In most countries, anything below 0.3% ∆9-tetrahydrocannabinol (THC) is classified as hemp and plants that produce 0.3% or greater are categorized as marijuana [3]. More than 115 cannabinoids, isoprenylated polyketides, have been identified in cannabis, which are mainly produced in glandular trichomes of female flowers. Cannabidiol (CBD), THC, and cannabichromene (CBC) can be considered as the major cannabinoids in the crop, but new genetics that express other cannabinoids such as cannabigerol (CBG) are emerging [4]. As the demand for these products increases, there is a pressing need to develop improved genetics and cultivation techniques [10,11]
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