A kind of first-generation tissue culture method of Savannah cypress
A tissue culture and culture medium technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of less branch formation, less ear source, and inability to multiply rapidly and in large quantities, so as to reduce the pollution rate and improve the survival rate. rate effect
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Embodiment 1
[0021] (1) The preparation of medium: select MS medium as basic medium, add sucrose and agar, concentration is respectively 30g / L and 7g / L, then add 6-benzylaminoadenine (6-BA), its concentration is 0.05mg / L, adjust the pH value to 5.8-6.0, and sterilize under high pressure at 121°C for 25 minutes to get it;
[0022] (2) Explant treatment: select explants without buds, first soak them with washing powder aqueous solution, then rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then use 0.1% mercuric chloride Soak for 5 minutes, and finally rinse 5 times with sterile water, and absorb the residual water with sterile filter paper;
[0023] (3) Inoculation and cultivation: the old wound of the explant obtained in step (2) is cut off, inserted into the obtained medium of step (1), and placed in a cultivation room for cultivation, the cultivation conditions are: light 16h / d, light intensity is 2000lx, temperature Control at (25±2)°C.
Embodiment 2
[0025] (1) The preparation of medium: select B5 medium as base medium, add sucrose and agar, concentration is 30g / L and 7g / L respectively, then add 6-benzylaminoadenine, its concentration is 1mg / L, regulates The pH value is 5.8-6.0, and it is sterilized under high pressure at 121°C for 25 minutes, and it is ready;
[0026] (2) Explant treatment: select explants without buds, first soak them with washing powder aqueous solution, then rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then use 0.1% mercuric chloride Soak for 8 minutes, and finally rinse 5 times with sterile water, and absorb the residual water with sterile filter paper;
[0027] (3) Inoculation and cultivation: the old wound of the explant obtained in step (2) is cut off, inserted into the obtained medium of step (1), and placed in a cultivation room for cultivation, the cultivation conditions are: light 16h / d, light intensity is 2000lx, temperature Control at (25±2)°C.
Embodiment 3
[0029] (1) Preparation of medium: select 1 / 2MS medium as the basic medium, add sucrose and agar, the concentration is 30g / L and 7g / L respectively, then add naphthaleneacetic acid, its concentration is 0.05mg / L, adjust the pH The value is 5.8-6.0, and it is sterilized under high pressure at 121°C for 25 minutes to get it;
[0030] (2) Explant treatment: select explants without buds, first soak them with washing powder aqueous solution, then rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then use 0.1% mercuric chloride Soak for 6 minutes, and finally rinse 5 times with sterile water, and absorb the residual water with sterile filter paper;
[0031] (3) Inoculation and cultivation: the old wound of the explant obtained in step (2) is cut off, inserted into the obtained medium of step (1), and placed in a cultivation room for cultivation, the cultivation conditions are: light 16h / d, light intensity is 2000lx, temperature Control at (25±2)°...
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