Tissue culture method of corydalis pygmaea (endangered Tibetan medicine)
A tissue culture, Viola dwarfs technology, applied in horticultural methods, botanical equipment and methods, horticulture and other directions, can solve the problems of destruction of the native environment, blank artificial breeding technology of Viola dwarfs, etc. low temperature effect
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Embodiment 1
[0029] This embodiment provides a tissue culture method of the endangered Tibetan medicinal material Corydalis dwarfia, the steps comprising:
[0030] (1) Select robust plant buds, wash them quickly with ethanol with a volume concentration of 60%, and then use a concentration of 0.05% HgCl 2 The solution was disinfected for 1 min;
[0031] (2) Rinse the buds treated in step (1) three times quickly with sterile water, then blot the water on the surface of the buds to obtain sterile explants for use;
[0032] (3) Under sterile conditions, inoculate the sterile explants described in step (2) onto the Ms medium, and after cultivating in the dark at 15°C for 10 days, transfer to Ms+6-BA1mg / l+NAA0.01mg On the / l medium, cultivate and induce cluster buds for 30 days under light at 15°C to form cluster buds; the time of the light per day is 18h;
[0033] (4) Transfer the cluster buds into 1 / 2Ms+6-BA0.2mg / l+NAA0.5mg / l, cultivate and take root at 15°C, 1500LX intensity light+natural l...
Embodiment 2
[0036] This embodiment provides a tissue culture method of the endangered Tibetan medicinal material Corydalis dwarfia, the steps comprising:
[0037] (1) Select robust plant buds, first rinse quickly with ethanol with a volume concentration of 80%, and then use a concentration of 0.2% HgCl 2 The solution was disinfected for 3 minutes;
[0038] (2) Rinse the sprouts treated in step (1) rapidly with sterile water 5 times, then blot the water on the surface of the sprouts to obtain sterile explants for use;
[0039] (3) Under sterile conditions, inoculate the sterile explants described in step (2) onto the Ms medium, and after cultivating in the dark at 20°C for 7 days, transfer to Ms+6-BA1mg / l+NAA0.01mg On the / l medium, cultivate and induce cluster buds for 35 days under light at 25°C to form cluster buds; the time of the light per day is 24h;
[0040] (4) Transfer the cluster buds into 1 / 2Ms+6-BA0.2mg / l+NAA0.7mg / l, cultivate and take root at 25°C, 2000LX intensity light+nat...
Embodiment 3
[0043] This embodiment provides a tissue culture method of the endangered Tibetan medicinal material Corydalis dwarfia, the steps comprising:
[0044] (1) Select robust plant buds, wash them quickly with ethanol with a volume concentration of 70%, and then use a concentration of 0.1% HgCl 2 The solution was disinfected for 2 minutes;
[0045] (2) The sprouts treated in step (1) are quickly rinsed 4 times with sterile water, and then the water on the surface of the sprouts is blotted dry to obtain sterile explants for use;
[0046] (3) Under sterile conditions, inoculate the sterile explants described in step (2) onto the Ms medium, and after cultivating in the dark at 18°C for 8 days, transfer to Ms+6-BA1mg / l+NAA0.01mg / l culture medium, at 22 DEG C, cultivate and induce cluster buds under light for 32 days to form cluster buds; the time of light per day is 21h;
[0047] (4) Transfer the cluster buds into 1 / 2Ms+6-BA0.2mg / l+IAA0.5mg / l, cultivate and root at 22°C, 1800LX int...
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