Method for tissue culture and quick propagation of brucea javanica
A technology of tissue culture and brucei, applied in the field of biological reproduction, can solve problems such as difficulties in introduction and domestication, endangered wild resources, destruction of ecological environment, etc., and achieves the effects of maintaining excellent characters, increasing reproduction coefficient, and improving safety.
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Embodiment 1
[0041] Step 1, soaking the sterilized explants in a 1% soluble polyvinylpyrrolidone solution for 5 minutes to obtain sterile explants;
[0042] Step 2, utilize the induction medium that comprises 6-BA 1.0mg, 2,4-D 1.0mg, NAA 1.5mg, agar 4.5g, sucrose 25g and distilled water 0.9 liters to induce the described sterile explant and at a temperature of 24 Cultivate for 7 days under dark culture conditions at ℃, then culture for 21 days under the conditions of light intensity of 1500 lux and time of 10 hours / day to obtain callus;
[0043] Step 3. Cultivate the callus with a differentiation medium comprising 6-BA2.0mg, agar 4.5g, sucrose 25g and distilled water 0.9 liters at a temperature of 26°C, a light intensity of 1500 lux, and a light time of 12 hours / day , cultivated for 21 days to obtain the clustered buds.
[0044] Step 4. Cultivate in a proliferation medium comprising 0.3 mg of TDZ, 0.5 mg of 6-BA, 4.5 g of agar, 25 g of sucrose and 0.9 liters of distilled water and culture...
Embodiment 2
[0049] Step 1, soaking the sterilized explant in a soluble polyvinylpyrrolidone solution with a mass concentration of 1% after being sterilized by a filter membrane for 8 minutes to obtain a sterile explant;
[0050] Step 2, utilize the induction medium that comprises 6-BA 1.3mg, 2,4-D 2.0mg, NAA2.0mg, agar 4.5g, sucrose 25g and distilled water 0.92 liters to induce the described sterile explants and at a temperature of 25 Cultivate for 7 days under dark culture conditions at ℃, and then culture for 21 days under the conditions of light intensity of 1500 lux and time of 11 hours / day to obtain callus;
[0051] Step 3. Cultivate the callus with a differentiation medium comprising 1.0 mg of 6-BA, 4.5 g of agar, 25 g of sucrose and 0.92 liters of distilled water and culture the callus at a temperature of 25° C., a light intensity of 1500 lux, and a light time of 11 hours / day Under the condition of culturing for 21 days, the clustered buds were obtained.
[0052] Step 4, using the...
Embodiment 3
[0057] Step 1, soaking the sterilized explant in a 1% soluble polyvinylpyrrolidone solution for 10 minutes to obtain a sterile explant;
[0058] Step 2, utilize the induction medium that comprises 6-BA 1.5mg, 2,4-D 2.5mg, NAA2.5mg, agar 4.5g, sucrose 25g and distilled water 0.95 liters to induce the described sterile explants and at a temperature of 26 Cultivate for 7 days under dark culture conditions at ℃, then culture for 21 days under the conditions of 1500 lux light intensity and 12 hours / day to obtain callus tissue;
[0059] Step 3. Cultivate the callus with a differentiation medium comprising 1.0 mg of 6-BA, 4.5 g of agar, 25 g of sucrose and 0.95 liters of distilled water and culture the callus at a temperature of 25° C., a light intensity of 1500 lux, and a light time of 11 hours / day Under the condition of culturing for 21 days, the clustered buds were obtained.
[0060] Step 4. Cultivate in a proliferation medium containing TDZ 0.5mg, 6-BA 1.5mg, agar 4.5g, sucrose ...
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