Method for cultivating tissue cultured gloden line lotus seedling
A culturing method and a technique for tissue culture seedlings, which are applied in the field of tissue culture seedling cultivation of Clematis clematis, can solve problems such as the application of hormone-free culture seedlings of test-tube seedlings, which has not been reported, and are suitable for large-scale production, rapid propagation, and safe consumption. Effect
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Embodiment 1
[0036] Embodiment 1: the cultivation of clematis regenerated seedlings
[0037] Take the stems or tips of wild clematis as materials, soak them in 70% medical ethanol for about 5 minutes, then transfer them to 10% sodium hypochlorite aqueous solution for disinfection, rinse them with sterile water, and inoculate them in high-temperature sterilized MS+ On the solid medium of NAA0.5mg / L+IBA0.5mg / L+ZT0.2mg / L, the culture conditions are: light (1500lx) 12 hours / day, culture at 25°C, and regenerated buds of Aurea clematis can be obtained after 30 days.
Embodiment 2
[0038] Embodiment 2: Rapid propagation of Clematis clematis tissue culture seedling without hormone
[0039] The regenerated shoots obtained in Example 1 were successively placed on the acclimatization medium with hormone concentration from high to low and then to zero for cultivation.
[0040] The regenerated shoots were inoculated on MS+NAA0.2mg / L+IBA0.2mg / L+ZT0.1mg / L medium for cultivation for 30 days at a cultivation temperature of 20-26°C and light (1500lx) for 12 hours / day.
[0041] Take the above tissue cultured seedlings in MS 0 The culture is carried out on the culture medium for 30 days, the culture temperature is 20-26° C., and the light (1500 lx) is 12 hours / day.
[0042] Finally, fast-growing and robust shoots were selected from the hormone-free medium for subculture (once a month).
Embodiment 3
[0043] Embodiment 3: field cultivation of Clematis clematis tissue culture seedling
[0044] After washing away the culture medium of the root of the tissue culture seedling obtained in Example 2 with clear water, soak it in 0.25% carbendazim solution for 10 minutes and then transplant it into the substrate (loose, air-permeable, moist) with a shade shed to keep growing The temperature is 20-25°C, and the relative humidity is 75-85% in a small environment, and attention should be paid to ventilation in the shed.
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