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Rheum palmatum  L. is a perennial herb of the Rheum genus Polygonaceae. It is a traditional Chinese herbal medicine which has extremely high medicinal value. Rhubarb has scarce seeds in subtropical regions, and natural planting conditions have limitations. In recent years, the demand of rhubarb has increased, and the means of sowing and raising seedlings can no longer meet the needs of the market. Traditional sowing and propagation methods have limitations such as long cycle and low reproduction coefficient. Tissue culture technology can shorten the reproduction cycle that increase the reproduction coefficient and maintain the excellent characteristics of the parents, which is an effective way to promote the large-scale and commercial production of fine varieties of rhubarb. The results of the study showed that: (1) Analysis of the characteristics of Qinghai Rhubarb seeds. (2) Seed sterilization treatment. (3) Primary culture medium selection. (4) Callus induction. (5) Callus proliferation culture. (6) Hypocotyl callus bud differentiation.(7)Rooting culture. (8) Qinghai Rhubarb tissue culture seedling transplantation. Using the method of the present invention to establish a Qinghai Rhubarb in vitro culture aseptic propagation system, high-quality Qinghai Rhubarb seeds as explants and different hormone medium ratios, Qinghai Rhubarb hypocotyl induction rate reached 90%, and the proliferation coefficient reached 4.3. The survival rate of transplanting tissue cultured seedlings reaches 95% which can improve the reproduction efficiency of Qinghai Rhubarb, and provide theoretical basis and technical support for subsequent research on transgenic and genetic improvement. It can also provide a basis for medicinal value for the systematic research and commercial production of Qinghai Rhubarb.

Research on key techniques of tissue culture of Qinghai Rhubarb