Tissue culture and Agrobacterium-mediated transformation of hemp (Cannabis sativa L.)

Feeney, Mistianne; Punja, Zamir K.

doi:10.1079/ivp2003454

Cited by 88 publications

(101 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Moreover hairy roots induced by AR10GUS strain, harboring the binary p35SGUS intron plasmid, showed normal pattern of GUS positive staining (Figure 2f), demonstrating cotransfer of the binary vector and the expression of the foreign GUS gene. All the above provides unequivocal proof that hemp is transformable by Agrobacterium, supporting and extending results of Feeney and Punja (2003), who reported the transformation of cell suspensions with A. tumefaciens. As previously indicated, two morphological phenotypes were distinguished within hairy root lines, referred as thin and thick morphology (Figure 2g and 2h, respectively), which were stable for more than two years.…”

Section: Discussionsupporting

confidence: 86%

“…These genes, located in the TL-DNA of A. rhizogenes, are responsible for the root induction process (Palazon et al 1998, Tiwari et al 2007). At present, there is only one report of Agrobacterium infection on hemp, which describes the transformation of cell suspension cultures with A. tumefaciens strain EHA101 carrying the binary vector pNOV3635 with a gene encoding phosphomannose isomerase (Feeney & Punja 2003).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Agrobacteriuminfection of hemp (Cannabis sativaL.): establishment of hairy root cultures

Wahby

Caba

Ligero

2013

Journal of Plant Interactions

View full text Add to dashboard Cite

Experimental conditions were optimized for hemp, a difficult to transform plant, to be effectively infected with either Ri or Ti plasmid-bearing agrobacteria and to establish stably transformed tissues. Hypocotyl of intact seedlings was the most responsive material and the response depended on both bacterial strain and plant variety. Transformed tissues, hairy roots and tumors, were cultured and stabilized in vitro and showed the characteristic traits of fast and phytohormone-independent growth as well as high incidence of lateral branching and abundance of root hairs in the case of roots. They all contained T-DNA of the corresponding Ri or Ti plasmid as revealed by PCR analysis with specific primers and further hairy roots induced by AR10GUS strain showed normal pattern of b-glucuronidase positive staining. To our knowledge, this represents the first reported protocol for the establishment of Cannabis sativa hairy root cultures.

show abstract

Section: Discussionsupporting

confidence: 86%

Section: Introductionmentioning

confidence: 99%

Agrobacteriuminfection of hemp (Cannabis sativaL.): establishment of hairy root cultures

Wahby

Caba

Ligero

2013

Journal of Plant Interactions

View full text Add to dashboard Cite

show abstract

“…Cannabis cells were cultured in Gamborg B5 medium supplemented with 2.5 M 2,4-dichlorophenoxyacetic acid at 25°C (24). Cannabis cells used here were confirmed to produce quite low levels of CBCA and THCA (lower than 0.1 g/g fresh cells in each cannabinoid).…”

Section: Methodsmentioning

confidence: 74%

Identification and Characterization of Cannabinoids That Induce Cell Death through Mitochondrial Permeability Transition in Cannabis Leaf Cells

Morimoto

Tanaka

Sasaki

et al. 2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

Cannabinoids are secondary metabolites stored in capitatesessile glands on leaves of Cannabis sativa. We discovered that cell death is induced in the leaf tissues exposed to cannabinoid resin secreted from the glands, and identified cannabichromenic acid (CBCA) and ⌬ 1 -tetrahydrocannabinolic acid (THCA) as unique cell death mediators from the resin. These cannabinoids effectively induced cell death in the leaf cells or suspension-cultured cells of C. sativa, whereas pretreatment with the mitochondrial permeability transition (MPT) inhibitor cyclosporin A suppressed this cell death response. Examinations using isolated mitochondria demonstrated that CBCA and THCA mediate opening of MPT pores without requiring Ca 2؉ and other cytosolic factors, resulting in high amplitude mitochondrial swelling, release of mitochondrial proteins (cytochrome c and nuclease), and irreversible loss of mitochondrial membrane potential. Therefore, CBCA and THCA are considered to cause serious damage to mitochondria through MPT. The mitochondrial damage was also confirmed by a marked decrease of ATP level in cannabinoid-treated suspension cells. These features are in good accord with those of necrotic cell death, whereas DNA degradation was also observed in cannabinoid-mediated cell death. However, the DNA degradation was catalyzed by nuclease(s) released from mitochondria during MPT, indicating that this reaction was not induced via a caspasedependent apoptotic pathway. Furthermore, the inhibition of the DNA degradation only slightly blocked the cell death induced by cannabinoids. Based on these results, we conclude that CBCA and THCA have the ability to induce necrotic cell death via mitochondrial dysfunction in the leaf cells of C. sativa.Multicellular organisms such as plants and animals possess cell death-inducing systems to eliminate damaged, superfluous, and ectopic cells. In higher plants, these systems participate in a variety of physiologically important events (e.g. root cap elimination, somatic embryogenesis, xylogenesis, leaf senescence, and defense against microbial pathogens) (1, 2), and therefore plant cell death has attracted a great deal of attention. To reveal the regulatory mechanism of cell death induction, biochemical and molecular examinations have been extensively carried out to date and have demonstrated that, in many cases, plant cell death can be classified into apoptosis or necrosis and that mitochondria control both types of cell death in plants as well as animals (3-5). For example, plant apoptosis, like animal apoptosis, is caused by release of apoptotic proteins (cytochrome c and others) from mitochondria, followed by activation of cellexecuting enzymes (caspases and nucleases) (3, 4). On the other hand, a decrease of the ATP level due to serious mitochondrial dysfunction has been shown to induce necrosis in plants and animals (5, 6).Apoptotic-protein release and mitochondrial dysfunction are often reported to be controlled by mitochondrial membrane permeability, and several systems that regulate this pe...

show abstract

“…In the literature several reports can be found on the growth of callus and cell suspension cultures [60][61][62]. Most of them document that no cannabinoids can be found within these cultures.…”

Section: Cell Culturesmentioning

confidence: 99%

Chemistry and Biological Activity of Tetrahydrocannabinol and its Derivatives

Flemming

Muntendam

Steup³

et al. 2007

Topics in Heterocyclic Chemistry

View full text Add to dashboard Cite

Tissue culture and Agrobacterium-mediated transformation of hemp (Cannabis sativa L.)

Cited by 88 publications

References 21 publications

Agrobacteriuminfection of hemp (Cannabis sativaL.): establishment of hairy root cultures

Agrobacteriuminfection of hemp (Cannabis sativaL.): establishment of hairy root cultures

Identification and Characterization of Cannabinoids That Induce Cell Death through Mitochondrial Permeability Transition in Cannabis Leaf Cells

Chemistry and Biological Activity of Tetrahydrocannabinol and its Derivatives

Contact Info

Product

Resources

About