Coffee senna tissue culture regeneration system construction method
A construction method and tissue culture technology, applied in culture medium, horticultural methods, plant regeneration, etc., can solve the problems of browning of explants, limitation of application of tissue culture technology, difficulty in rooting, etc., and achieve the effect of solving the shortage
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Embodiment 1
[0013] (1) Seed disinfection: select immature fruits, remove leaves, soak in washing powder water for 15 minutes, rinse with tap water for 2 hours after brushing, take out immature milky white or slightly brown seeds, and peel off the outer testa in an ultra-clean workbench , first sterilized with 75% ethanol for 6 seconds, then washed 6 times with sterile water, then sterilized with 0.1% mercuric chloride solution for 12 minutes, rinsed with sterile water 5 times and then set aside. The statistical pollution rate can be as low as 3% one week after inoculation.
[0014] (2) Embryo callus induction: the immature embryos treated in step (1) were cut into 0.5cm×0.5cm×0.5cm and inoculated on the induction medium, and cultured at 28°C in total darkness for 30 days. Then placed in light for 12 hours a day, the light intensity was 1400x, until embryogenic callus was induced, and the callus induction rate was 67%. The induction medium is ER+6-BA1.0mg / L+NAA0.5mg / L+GA 3 5 mg / L+sucrose ...
Embodiment 2
[0019] (1) Seed disinfection: select immature fruits, soak in washing powder water for 16 minutes after removing leaves, rinse with tap water for 3 hours after brushing, take out immature milky white or slightly brown seeds, and peel off the outer testa in an ultra-clean workbench , first sterilized with 75% ethanol for 8 seconds, then washed 9 times with sterile water, then sterilized with 0.1% mercuric acid solution for 11 minutes, rinsed with sterile water 9 times and then used for later use. The statistical pollution rate can be as low as 6% one week after inoculation.
[0020] (2) Embryo callus induction: the immature embryos treated in step (1) were cut into 0.5cm×0.5cm×0.5cm and inoculated on the induction medium, and cultured at 28°C in total darkness for 31 days. Then placed in light for 13 hours a day, the light intensity was 1400x, until embryogenic callus was induced, and the callus induction rate was 63%. The induction medium is ER+6-BA1.0mg / L+NAA0.2mg / L+GA 3 3 m...
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