Method for inducing litsea cubeta cluster buds
A technology of Radix chinensis and explants is applied in the field of tissue culture and rapid propagation of inducing the clustered buds of the woody aromatic plant Radix vulgaris Low seedling rate and other problems, to achieve the effects of inhibiting the browning of the medium and explants, improving the disinfection effect, and stabilizing the genetic traits
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Embodiment 1
[0020] Get the young and tender branches of Litsea Cubeba dry base, remove the leaves, put in 3% H2O after washing with running water. 2 o 2 Soak in medium for 15 minutes, take it out and immediately transfer to 0.1% HgCl containing 5% Tween-80 2 Soak in the solution for 20 minutes, constantly brush the branches with a soft brush, then take them out and rinse them with sterile water for 5 to 6 times and dry the water with sterile absorbent paper, then cut out the stems containing nodes as explants, and quickly inoculate them on the The explants were cultured in the dark on MS medium, and the explants were transferred to new MS medium the next day.
[0021] Transfer the explants that are no longer browned on MS medium to MS+6-BA1.0mg / L+IBA0.5mg / L+GA2.0mg / L+agar 7.0g / L+sucrose 30g / L, pH6.0 on the induction medium, placed at a temperature of 25±2°C, with a photoperiod of 12h.d -1 , Induced culture was carried out in an incubator with a light intensity of 1200 lux. After 21 da...
Embodiment 2
[0025] Get the young and tender branches of Litsea Cubeba dry base, remove the leaves, put in 3% H2O after washing with running water. 2 o 2 Soak in medium for 20 minutes, take it out and immediately transfer to 0.1% HgCl containing 5% Tween-80 2 Soak in the solution for 10 minutes, constantly brush the branches with a soft brush, then take them out and rinse them with sterile water for 5 to 6 times and dry the water with sterile absorbent paper, then cut out the stems containing nodes and inoculate them on MS medium, put Culture in the dark, and transfer to new MS medium the next day.
[0026] Transfer the explants that are no longer browned on MS medium to MS+6-BA0.5mg / L+IBA0.2mg / L+GA1.0mg / L+agar 7.0g / L+sucrose 30g / L, pH6.0 On the culture medium, put it at a temperature of 25±2°C and a photoperiod of 12h.d -1 , Induced culture was carried out in an incubator with a light intensity of 1200 lux. After 21 days, induced shoots were obtained.
[0027] After 30 days, cut out ...
Embodiment 3
[0030] According to the method of Example 1, only the hormone concentration in the subculture stage was reduced. Specific method: After 30 days of multiplication and culture, cut out the section-containing stem section of the bud and inoculate it into MS+6-BA1.0mg / L+IBA0.3mg / L+GA1.0mg / L agar 7.0g / L+sucrose 30g / L, pH6. 0 medium; after 30 days, transfer the buds to MS+6-BA2.0mg / L+IBA0.3mg / L+agar 7.0g / L+sucrose 30g / L pH 6.0 medium in the same way for subculture. The temperature is 25±2°C, the photoperiod is 14h.d-1, and the light intensity is 2000lux; such alternate culture, the multiplication coefficient is more than 4 times, and the sprouts obtained are strong and neat.
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