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Review
. 2021 May 26;22(11):5671.
doi: 10.3390/ijms22115671.

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Mohsen Hesami  1 , Austin Baiton  1 , Milad Alizadeh  2 , Marco Pepe  1 , Davoud Torkamaneh  3 , Andrew Maxwell Phineas Jones  1
Affiliations
Review

Advances and Perspectives in Tissue Culture and Genetic Engineering of Cannabis

Mohsen Hesami et al. Int J Mol Sci. .

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.

Keywords: gene transformation; genome editing; haploid production; hemp; in vitro culture; marijuana; morphogenic genes; organogenesis; polyploidy; somatic embryogenesis.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Some industrial properties of Cannabis.
Figure 2
Figure 2
The schematic diagram of plant tissue culture procedures.
Figure 3
Figure 3
The schematic diagram of factors affecting in vitro culture procedures.
Figure 4
Figure 4
A schematic view of factors involved in somaclonal variation including genetic mosaicism and mutation as well as epigenetic regulations such as DNA methylation, histone modification, and RNA interference.
Figure 5
Figure 5
Comparing the morphological traits of diploid and tetraploid cannabis (a) Diploid Cannabis leaf, (b) Tetraploid Cannabis leaf, which is noticeably wider than the diploid, (c) Bright light image of diploid Cannabis stomata, (d) Bright light image of tetraploid Cannabis stomata.
Figure 6
Figure 6
The schematic diagram of Agrobacterium-mediated gene transformation.
Figure 7
Figure 7
A schematic view of the morphogenic genes demonstrating their roles in plant growth and development as well as in vitro plant regeneration (AGL15: AGAMOUS-LIKE15; ARR: ARABIDOPSIS RESPONSE REGULATOR; BBM: BABY BOOM; CLV3: CLAVATA3; CUC: CUP-SHAPED COTYLEDON; ESR: ENHANCER OF SHOOT REGENERATION; FUS3: FUSCA3; GA3ox: Gibberellin 3-beta-dioxygenase; IAA30: Indole acetic acid inducible 30; LEC: LEAFY COTYLEDON; PIN1: PIN-FORMED 1; PKL: PICKLE; PLT: PLETHORA; PRC: Polycomb repressive complex; STM: SHOOT MERISTEMLESS; TAA: TRYPTOPHAN AMINOTRANSFERASE ARABIDOPSIS; WOX: WUSCHEL-related homeobox; WUS: WUSCHEL; YUC: YUCCA).
Figure 8
Figure 8
Three types of off-targets induced by CRISPR-mediated genome editing; (a) off-target sites with base mismatch, (b) off-target sites with extra base (DNA bulge or deletion), and (c) off-target site with missing base (RNA bulge or insertion).
Figure 9
Figure 9
A schematic view of Cannabis genome engineering.
See this image and copyright information in PMC

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