Pittosporum tobira somatic embryogenesis and plant regeneration method
A technology of embryogenesis and somatic cells, which is applied in the field of plant propagation, can solve the problems of difficult germination of Pittosporum seeds and low seedling rate of seeds, and achieve the effects of shortening the cultivation period, uniform emergence of seedlings, and easy management
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Embodiment 1
[0015] Pittosporum somatic embryogenesis and plant regeneration method of the present invention, comprises the following steps:
[0016] (1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.
[0017] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the M...
Embodiment 2
[0023] (1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.
[0024] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the MS medium, and the pH value of the medium was 5.8.
[0025] (3) Induction of embryogenic callus: the cotyledon of the aseptic test-...
Embodiment 3
[0030](1) Selection and disinfection of explants: Take Pittosporum fresh seeds as explants, soak them in 2v / v% detergent solution for 5min, rinse them with tap water for 15-30min, add 2-3 drops of Tween -20 0.1v / v% mercuric chloride 100mL was sterilized for 8-10min, rinsed with sterile water for 3-5 times, and finally removed the surface moisture with sterile filter paper to obtain explants. Wherein, sterile water is distilled water sterilized by high temperature and high pressure.
[0031] (2) Induction of explants to obtain sterile test-tube plantlets: place the explants obtained in step (1) in an ultra-clean workbench, inoculate them into MS medium, and culture them at a temperature of 23-27°C under dark conditions Cultivate for 20 days to obtain sterile test-tube plantlets. Among them, GA30.1mg / L, sucrose30g / L and agar5g / L were added to the MS medium, and the pH value of the medium was 5.8.
[0032] (3) Induction of embryogenic callus: the cotyledon of the aseptic test-t...
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