Chamaecyparis pisifera primary tissue culture method
A technology of tissue culture and culture medium, applied in horticultural methods, botany equipment and methods, horticulture, etc., can solve the problems of less branch formation, less ear sources, and inability to reproduce rapidly and in large quantities, so as to reduce pollution rate and improve survival rate effect
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Embodiment 1
[0021] (1) Preparation of the medium: MS medium was selected as the basic medium, sucrose and agar were added to the concentration of 30g / L and 7g / L, and 6-benzylamino adenine (6-BA) was added to the concentration of 0.05mg / L, adjust the pH to 5.8~6.0, autoclave at 121℃ for 25min, it is obtained;
[0022] (2) Treatment of explants: Choose explants without buds, soak them with detergent, rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then soak them with 0.1% mercury. 5min, finally rinse with sterile water for 5 times, and absorb the residual water with sterile filter paper;
[0023] (3) Inoculation culture: Cut off the old wound of the explant obtained in step (2), insert it into the culture medium obtained in step (1), and place it in a culture room for culture. The culture conditions are: light 16h / d, light intensity of 2000lx, temperature Control at (25±2)℃.
Embodiment 2
[0025] (1) Preparation of medium: choose B5 medium as the basic medium, add sucrose and agar, the concentrations are 30g / L and 7g / L, and then add 6-benzylamino adenine, the concentration is 1mg / L, adjust The pH value is 5.8~6.0, and it is sterilized under high pressure at 121℃ for 25min, and it is obtained
[0026] (2) Treatment of explants: Choose explants without buds, soak them with detergent, rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then soak them with 0.1% mercury. 8min, finally rinse with sterile water for 5 times, and absorb the residual water with sterile filter paper;
[0027] (3) Inoculation culture: Cut off the old wound of the explant obtained in step (2), insert it into the culture medium obtained in step (1), and place it in a culture room for culture. The culture conditions are: light 16h / d, light intensity of 2000lx, temperature Control at (25±2)℃.
Embodiment 3
[0029] (1) Preparation of the medium: Choose 1 / 2MS medium as the basic medium, add sucrose and agar, the concentrations are 30g / L and 7g / L, and then add naphthalene acetic acid, the concentration is 0.05mg / L, adjust the pH The value is 5.8~6.0, sterilized under high pressure at 121℃ for 25min, it is obtained;
[0030] (2) Treatment of explants: Choose explants without buds, soak them with detergent, rinse with pure water for 2 hours, then treat them with 75% alcohol for 30 seconds, and then soak them with 0.1% mercury. 6min, finally rinse with sterile water 5 times, and absorb the residual water with sterile filter paper;
[0031] (3) Inoculation culture: Cut off the old wound of the explant obtained in step (2), insert it into the culture medium obtained in step (1), and place it in a culture room for culture. The culture conditions are: light 16h / d, light intensity of 2000lx, temperature Control at (25±2)℃.
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