Method for cultivating triploid dendrobium huoshanense by fusion breeding method of diploid cells and pollen cells
A technology of diploid cells and pollen cells is applied in the field of biological breeding of medicinal plants of the genus Dendrobium orchid, and can solve the problems that no public report of triploid Dendrobium huoshanii has been reported.
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Embodiment l
[0020] (1) The plants of variety A and variety B are planted and cultivated according to conventional methods;
[0021] (2) The seeds of the above variety A are aseptically sown in a solid medium to obtain the callus of the protocorm. The solid medium is 1 / 2MS medium with halved macronutrients, and 0.2g / L 2,4 -D and 0.2g / L NAA, adjust the sucrose concentration to 5%-8%, add 10-15% coconut milk, cultivate at 25-18 ℃ in the diurnal temperature range, light time 10-12 h, light intensity 2000-3000 Lux, color temperature at 4500-5500 K;
[0022] (3) Aseptically culture the pollen of the above-mentioned variety B in a solid medium to obtain pollen cell populations. The solid medium is H medium, add 0.5g / L 2,4-D and 1.5 g / L NAA, and cultivate The temperature is within the diurnal temperature range of 25-18 ℃, the light time is 10-12 h, the light intensity is 2000-3000 Lux, and the color temperature is 4500-5500 K;
[0023] (4) Prepare the protoplasts of pollen cells and protocorm c...
Embodiment 2
[0027] (1) The plants of variety A and variety B are planted and cultivated according to conventional methods;
[0028] (2) The seeds of the above variety B are aseptically sown in a solid medium to obtain the callus of the protocorm. The solid medium is 1 / 2MS medium with halved macronutrients, and 0.2g / L 2,4 -D and 0.2g / L NAA, adjust the sucrose concentration to 5%-8%, add 10-15% potato juice, cultivate at 25-18 ℃ in the diurnal temperature range, light time 10-12 h, light intensity 2000-3000 Lux, color temperature at 4500-5500 K;
[0029] (3) Aseptically culture the pollen of the above variety A in a solid medium to obtain pollen cell populations, the solid medium is H medium, add 0.5g / L 2,4-D and 1.5 g / L NAA, and culture The temperature is within the diurnal temperature range of 25-18 ℃, the light time is 10-12 h, the light intensity is 2000-3000 Lux, and the color temperature is 4500-5500 K;
[0030] (4) Prepare the protoplasts of pollen cells and protocorm cells, and ca...
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