A method for screening dendrobium cultures with high content of functional components
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A technology of differentiation medium and dendrobium, applied in the biological field, can solve problems such as unstable genetics, and achieve the effect of strong metabolic synthesis ability
Active Publication Date: 2016-05-04
SHANGHAI ACAD OF AGRI SCI
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However, the synthesis of these functional components is derived from the products of inducing stimuli and cannot be genetically stable.
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Embodiment 1
[0079] Embodiment 1: seed pretreatment
[0080] Take wild Dendrobium officinale capsules (uncracked), rinse with tap water to remove dust, treat with 75% ethanol for 1 minute, then sterilize the surface with 1% sodium hypochlorite or chlorfenapyr for 15-20 minutes, rinse with sterile water for 5 times , cut out the seed embryo, and make a seed suspension with an appropriate amount of sterile water. Add final concentrations of 0.2-0.3% NaCl and / or 0.1-0.3mol / L mannitol to the seed suspension, and place them in low temperature (4-10°C) for stress treatment for 3-5 days.
Embodiment 2
[0081] Embodiment 2: Seed induced germination
[0082] After the treatment of the seed suspension obtained in Example 1 is completed, use a sterile filter to filter out the treatment solution, rinse it with sterile water for 5 times, and carefully transfer the seeds to the medium containing the induction medium with a medicine spoon (cooled by flame sterilization). On a plate, spread evenly. The components and content of the induction medium are as mentioned above, 2 / 3MS, supplemented with 500mg / LCH, naphthaleneacetic acid (NAA), indoleacetic acid (IAA), 2-4% sucrose, 0.6% agar, and containing 0.2- 0.3% NaCl and / or 0.1-0.3mol / L mannitol.
[0083] The culture conditions are light of 2000Lx, temperature of 25±2°C, and culture time of 45-60 days.
Embodiment 3
[0084] Example 3: Induced Proliferation of Seed Embryo Callus
[0085] The germinated seed embryos obtained in Example 2 were carefully picked into the liquid callus proliferation medium (2 / 3N6 (the N element content was reduced to 1 / 10 of the normal amount), 500mg / LCH, glutamine 500mg / L, 2, 4-Dichlorophenoxyacetic acid (2,4-D) 0.5mg / L, NAA0.1mg / L, 2-3% sucrose), cultured in the dark on a shaker (120rpm), subcultured once every 3 days at first, and cultured After 2 weeks, subculture once a week instead.
[0086] The culture condition is dark culture, the temperature is 25±2°C, and the culture time is 20-30 days.
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Abstract
The invention provides a method for screening a high-content functional component dendrobe culture, a method for improving polysaccharide and / or dendrobine in a dendrobe tissue culture seedling and a kit for implementing the methods. The method disclosed by the invention comprises: a seed pretreatment step: processing sterile dendrobe seeds by using NaCl and / or mannitol; a seed induced germination step: cultivating the seeds in an induced culture medium containing NaCl and / or mannitol, so as to form a protocorm after the seeds germinate; an induced proliferation step of embryo callus: cultivating the protocorm by using callus proliferation medium, and forming a cell mass or callus, so as to obtain the dendrobe tissue culture. The kit disclosed by the invention contains various culture mediums for implementing the methods disclosed by the invention. By adopting the methods disclosed by the invention and the kit, tissue culture seedlings with healthy growth and vigorous metabolism can be obtained; the alkali content of main functional components, namely polysaccharide and dendrobe is higher than that of a common tissue culture seedling; the dendrobe culture can be stably inherited, and can serve as an excellent seedling breeding source.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a screening method and application of a dendrobium high-content culture. Background technique [0002] Dendrobium is a precious Chinese medicinal material in my country, ranking first among the "Nine Great Immortal Grasses of China". It has good functions such as anti-aging, anti-tumor, lowering blood sugar and improving immunity. However, the growth of Dendrobium itself is slow, the habitat requirements are harsh, and the wild resources are on the verge of extinction. In 1987, it was listed as a rare and endangered medicinal plant under special protection by the state. Carrying out and expanding artificial planting can not only protect resources, but also realize rational utilization, which is the road to sustainable development that kills multiple birds with one stone. At present, the tissue culture technology of Dendrobium is relatively mature, and the large-scale arti...
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