Species

Strain

Vector

Techniques

Genes of interest

Objectives

Outcomes

Ref.

Cannabis sativa L.

Agrobacterium tumefaciens strain LBA4404

binary plasmid pBIN19

Agrobacterium-mediated genetic transformation

β-glucuronidase (uidA) reporter gene and the kanamycin resistance neomycin phosphotransferase (nptII) genes

Regenerating Cannabis sativa plants may effectively and quickly         undergo genetic change, resulting       in a  particular metabolic profile or enhanced resilience to stress.

Created an innovative and effective procedure for cultivating Cannabis sativa plants, leading to improved rates of regeneration and transformation compared to current procedures.

[76]

Cannabis sativa L.

Agrobacterium tumefaciens strain AGL1 .

pG41sg

Agrobacterium-mediated transformation and CRISPR/Cas9-mediated targeted mutagenesis

Phytoene desaturase gene and candidate genes homologous to ZmWUS2, NbSTM, NbIPT, OsGRF4, and AtGIF1

Provide a reliable Cannabis sativa transformation system using Agrobacterium tumefaciens and produce plants with altered DNA.

Development of a stable Agrobacterium-mediated transformation system for Cannabis sativa, enabling successful gene editing and stable integration of T-DNA in the Cannabis genome.

[77]

Cannabis sativa L.

p326GFP3G vector

Protoplast isolation and transformation.

CsCBCAS, CsCBDAS, and CsTHCAS

Effectively Isolating Cannabis Protoplasts and Characterizing Enzymes Involved in Cannabinoid Biosynthesis

Confirmed the practical separation of Cannabis protoplasts, the subcellular localisation of critical enzymes, and the improvement of mesophyll protoplast separation.

[66]

Cannabis sativa L.

Vector pDONR221-GFP

Construct a resilient transient expression screening system utilising Cannabis protoplast.

Cannabis cannabinoid production genes and regulators

Create a reliable screening system for transient expression in Cannabis protoplasts, optimise protoplast isolation and GFP expression

High protoplast yields from Cannabis leaf mesophyll protoplasts in an efficient protoplast separation and transient expression method.

[78]

Cannabis sativa var. Cherry x Otto II: Sweetened

The p35S: GFP expression cassette and the DR5::GFP auxin-sensitive reporter gene.

Separating protoplasts, transforming them temporarily, and analysing reporter genes using flow cytometry

DR5::GFP auxin-sensitive reporter gene

Develop hormone-free in vitro micropropagation and protoplast transformation for transient gene expression and verify Cannabis research methods.

Effective isolation, transformation, and analysis of Cannabis leaf protoplasts with high yields, viability, and efficiency.

[79]

Cannabis sativa L.

Agrobacterium tumefaciens strain EHA105

Binary vector pCAMBIA1301 with uidA gene

Agrobacterium tumefaciens-mediated transformation

uidA gene, which encodes for β-glucuronidase

Create a fast and effective transient expression technique for Cannabis sativa seedlings to exhibit gene function research and genetic enhancement potential.

Agrobacterium tumefaciens-mediated transient expression in Cannabis sativa seedlings is fast and effective and might be helpful for gene-function investigations and genetic improvement.

[80]