Methods of regenerating and transforming cannabis
a technology applied in the field of regenerating and transforming cannabis, can solve the problems of difficult to cross, complicating the breeding effort towards improving specific traits, and affecting the quality of cannabis,
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In-Vitro Propagation, Hardening and Rooting of Cannabis Cultivars
[0448]Cannabis has acquired considerable importance as a food, oil, fiber, medicinal and recreational drug source crop all over the world. This extraordinary versatile important plant material naturally calls for development of suitable protocols for production of sufficient number of uniform planting materials from Cannabis varieties through biotechnological intervention. Among the biotechnological approaches, micropropagation is one of the most feasible techniques, allows efficient and rapid clonal propagation of many economically important crops.
[0449]This study describes an efficient in-vitro propagation, hardening and rooting procedures for obtaining plantlets from shoot tips and seedlings of Cannabis sativa L. Ten different cannabis cultivars seedlings and shoot cuttings were sterilized and grown on half-strength ½ MS medium supplemented with 10 g / L sucrose, 5.5 g / L Agar at a pH of 6.8 with different plant hormon...
example 2
Cannabis Regeneration and Transformation Using Tissue Culture
[0478]The development of new Cannabis cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to establish efficient regeneration of Cannabis in tissue culture and to establish a protocol for Agrobacterium-mediated transformation for foreign gene introduction.
[0479]Induction of high-frequency shoot regeneration using nodal segments containing axillary buds from tissue culture plants of Cannabis sativa was achieved on premium treatment media with cuticle nicking enzymes, added to Murashige and Skoog (MS) medium salt mixture, containing 0.05-5.0 μM thidiazuron, supplemented with 100 mg / l myo-inositol, 1 mg / l thiamine-HCl, 2% sucrose (w / v) at pH 5.7) with 5 ml cuticle nicking enzymes reaction mixture (0.25 ml 10% enzyme, fungal mix of pectin and cutinase (BSG HandCraft Liquid Pectic Enzyme; Cutinase—Sigma, Ferdinand Maria Quincy 0.25 ml 200 mM T...
example 3
In Planta Regeneration and Transformation of Cannabis Cultivars
[0504]Alternate methods that avoid / minimize tissue culture would be beneficial for the development of new transgenic Cannabis cultivars. Transgenic Cannabis plants have been produced by a tissue-culture independent Agrobacterium tumefaciens-mediated transformation procedure. One of the two cotyledons of germinated Cannabis seedlings was excised. Unique regeneration method using plant hormones in a nano-encapsulation paste were introduced to the excised apical meristem of the germinating seedling. Regeneration of more than 80% of the germinating seedlings was obtained. A similar regeneration ratio was achieved with adult Cannabis plants of three different cultivars, suggesting that the nano encapsulation paste induces efficient regeneration. Agrobacterium strain EHA 105 harboring the binary vector pME504 that carries the genes for β-glucuronidase (GUS) and neomycin phosphotransferase (npt II), or the plasmid carrying the ...
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