Method for screening high-content functional component dendrobe culture
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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: 2014-03-19
SHANGHAI ACAD OF AGRI SCI
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[0079] Example 1: Seed pretreatment
[0080] Take wild Dendrobium officinale capsules (uncracked), rinse with tap water to show dust, after treatment with 75% ethanol for 1 minute, then use 1% sodium hypochlorite or sterile to sterilize the surface for 15-20 minutes, rinse with sterile water 5 times , Cut out the seed embryos, use sterile water to make seed suspension. The seed suspension was added with a final concentration of 0.2-0.3% NaCl and / or 0.1-0.3mol / L mannitol, and placed in a low temperature (4-10°C) stress treatment for 3-5 days.
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[0081] Example 2: Induced germination of seeds
[0082] After the treatment of the seed suspension obtained in Example 1, the treatment solution was filtered off with a sterile filter, and washed with sterile water for 5 times, and the seeds were carefully transferred to the induction medium with a medicine spoon (cooled by flame sterilization) Spread evenly on the plate. The components and content of the induction medium are as described above, 2 / 3MS, added with 500mg / L CH, naphthaleneacetic acid (NAA), indole acetic 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 2000Lx light, 25±2°C, and culture time 45-60 days.
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[0084] Example 3: Induced propagation of seed embryo callus
[0085] The germinated seed embryos obtained in Example 2 were carefully picked into the liquid callus proliferation medium (2 / 3N6 (N element content reduced to 1 / 10 of the normal amount), 500mg / L CH, 500mg / L glutamine, 2 ,4-Dichlorophenoxyacetic acid (2,4-D) 0.5mg / L, NAA0.1mg / L, 2-3% sucrose), culture in the dark on a shaker (120rpm), and subculture once every 3 days at first. After 2 weeks of cultivation, it was changed to subculture once a week.
[0086] The culture condition is dark culture, the temperature is 25±2℃, 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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