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Hey guys, I have been researching cannabis tissue culture lately and was wondering if anyone around here is doing any current tissue cultures, and if they could suggest a good agar-based medium. I have a few recipes to produce some TC medium, but it seems very tedious to prepare myself, and I won't have access to my school's plant physiology lab until Thursday. I would love to pick up some sterile pre-mixed agar. Here is one thorough recipe if anyone is interested:
Hemp (Cannabis sativa L.) is cultivated in many parts of the world for ils fiber, oil, and seed. The development of new hemp cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to investigate the propagation of hemp in tissue culture and to establish a protocol for Agrobacterium-mediated transformation for foreign gene introduction. Stem and leaf segments from seedlings of four hemp varieties were placed on Murashige and Skoog medium with Gamborg B5 vitamins (MB) supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 μM kinetin, 3% sucrose, and 8 gl−1 agar. Large masses of callus were produced within 4 wk for all cultivars. Suspension cultures were established in MB medium containing 2.5 μM 2,4-D. To promote embryogenesis or organogenesis, explants, callus, and suspension cultures derived from a range of explant sources and seedling ages were exposed to variations in the culture medium and changes to the culture environment. None of the treatments tested were successful in promoting plantlet regeneration. Suspension cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary vector pNOV3635 with a gene encoding phosphomannose isomerase (PMI). Transformed callus was selected on medium containing 1–2% mannose. A chlorophenol red assay was used to confirm that the PMI gene was expressed. Polymerase chain reaction and Southern hybridization detected the presence of the PMI gene. Copy number in different lines ranged from one to four. (http://www.springerlink.com/content/a373754v5658q576/)
I think the idea of keeping 50+ mother plants in a small space is very intriguing and I know it can be done with proper tissue culture techniques. There have also been reports (cited below) of tissue culture ridding clones of vascular tissue infections carried by the mother plant, which is one of the main reasons regular clones lose vigor. If anyone is interested, there is a very detailed explanation of the process of cannabis tissue culture that starts on page 18 of the article (cited below). The person who wrote the article is trying to produce a transgenic cannabis plant! That means that he could use that transgenic cannabis plant and manipulate and separate out the THC producing gene (once identified), and insert that gene into other plants! With this kind of research going on, eventually, we could be eating corn that is naturally producing THC lol. I suggest a full read of the article, very intriguing.
The Biotechnology of Cannabis Sativa
http://www.scribd.com/doc/14571756/The-Biotechnology-of-Cannabis-Sativa
Hemp (Cannabis sativa L.) is cultivated in many parts of the world for ils fiber, oil, and seed. The development of new hemp cultivars with improved traits could be facilitated through the application of biotechnological strategies. The purpose of this study was to investigate the propagation of hemp in tissue culture and to establish a protocol for Agrobacterium-mediated transformation for foreign gene introduction. Stem and leaf segments from seedlings of four hemp varieties were placed on Murashige and Skoog medium with Gamborg B5 vitamins (MB) supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 μM kinetin, 3% sucrose, and 8 gl−1 agar. Large masses of callus were produced within 4 wk for all cultivars. Suspension cultures were established in MB medium containing 2.5 μM 2,4-D. To promote embryogenesis or organogenesis, explants, callus, and suspension cultures derived from a range of explant sources and seedling ages were exposed to variations in the culture medium and changes to the culture environment. None of the treatments tested were successful in promoting plantlet regeneration. Suspension cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary vector pNOV3635 with a gene encoding phosphomannose isomerase (PMI). Transformed callus was selected on medium containing 1–2% mannose. A chlorophenol red assay was used to confirm that the PMI gene was expressed. Polymerase chain reaction and Southern hybridization detected the presence of the PMI gene. Copy number in different lines ranged from one to four. (http://www.springerlink.com/content/a373754v5658q576/)
I think the idea of keeping 50+ mother plants in a small space is very intriguing and I know it can be done with proper tissue culture techniques. There have also been reports (cited below) of tissue culture ridding clones of vascular tissue infections carried by the mother plant, which is one of the main reasons regular clones lose vigor. If anyone is interested, there is a very detailed explanation of the process of cannabis tissue culture that starts on page 18 of the article (cited below). The person who wrote the article is trying to produce a transgenic cannabis plant! That means that he could use that transgenic cannabis plant and manipulate and separate out the THC producing gene (once identified), and insert that gene into other plants! With this kind of research going on, eventually, we could be eating corn that is naturally producing THC lol. I suggest a full read of the article, very intriguing.
The Biotechnology of Cannabis Sativa
http://www.scribd.com/doc/14571756/The-Biotechnology-of-Cannabis-Sativa
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