Method for obtaining eustoma regeneration plant by anther culture
A technology for regenerating plants and eustoma, applied in the biological field, can solve the problems of difficulty in producing eustoma cut flowers, and achieve the effect of reliable technical support and solving the difficulties of cut flowers
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Embodiment 1
[0016] A. The material of eustoma used is 'Ceremony Orange'. Take flower buds with calyx and corolla equal length; put the flower buds in the refrigerator for 4 days at 3°C; take out the flower buds and disinfect them with 75% alcohol for 30 seconds, and then use mass The concentration is 0.2% HgCl2 Sterilize for 10 minutes, and finally rinse with sterile water 3 times, dry the water on the flower buds with high-pressure sterilized filter paper; cut off the corolla of the flower buds, peel off the anthers, remove the filaments, and inoculate in : MS+0.5mg / L KT+0.5mg / L 2,4-D, pH 5.5 induction medium, light intensity: 1500lx, light time: 12 hours / day, temperature 20°C, Cultivate for 40 days, take it out, put it into a new induction medium same as above until the anther grows a callus, cut off the callus, and continue to use the same induction medium for the anther that has not grown a callus. Subculture under the conditions until the callus grows;
[0017] B. Place the subcultu...
Embodiment 2
[0021] A. Choose the Eustoma material 'Art Peach', take the flower buds whose calyx is slightly longer than the corolla; put the flower buds in the refrigerator at 6°C for 2 days; take out the flower buds and disinfect them with alcohol with a volume concentration of 75% for 60 seconds, and then use a mass concentration Sterilize the species with 0.05% HgCl2 for 20 minutes, rinse twice with sterile water, and dry the water on the flower buds with filter paper that has been sterilized by autoclaving; cut off the corolla of the flower buds, peel off the anthers, remove the filaments, and inoculate the anthers on the : MS+3mg / L KT+3mg / L 2,4-D, pH 6.0 induction medium, light intensity: 2000lx, light time: 8 hours / day, temperature 28°C, culture 20 day, take it out, put it on the above-mentioned induction medium until the anther grows callus tissue, cut off the callus tissue, and continue to subculture the anther without callus tissue in the same induction medium and under the same c...
Embodiment 3
[0026] A. Choose the Eustoma material 'Cessna White', take the flower buds whose calyx is slightly longer than the corolla; put the flower buds into the refrigerator at 4°C for 3 days; disinfect the flower buds with 75% alcohol for 40 seconds, and then use 0.1% HgCl2 Sterilize for 15 minutes, rinse 4 times with sterile water at last, dry the water on the flower buds with filter paper that has been sterilized by autoclaving; cut off the corolla of the flower buds, peel off the anthers, remove the filaments, and inoculate the anthers on: MS+1mg / L KT+1mg / L 2,4-D, pH 5.8 induction medium, light intensity: 1800lx, light time: 10 hours / day, temperature 25°C, culture for 30 days , take it out, put it on the above-mentioned induction medium until the anther grows callus, cut off the callus, and continue to subculture the anther without callus in the same induction medium and under the same culture conditions until the anther grows callus. Callus out;
[0027] B. Place the subcultured...
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