Method for quickly reproducing hygrophila salicifolia callus tissues and suspension cells
A technology of callus and suspension cells, which is applied in the field of plants, can solve the problems of little research and no research on water raincoats, and achieve the effects of high proliferation rate, low energy consumption and simple operation
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Embodiment 1
[0009] Take the leaves of the water coir raincoat, soak them in bleaching powder for 3 minutes, remove the surface stains with a brush, wash them with running water for 35 minutes, treat them with sodium hypochlorite on the ultra-clean workbench for 14 minutes, rinse them with sterile water 5-7 times, insert the sterilized water coir raincoat leaves The medium formula is improved MS+KT0.5mg / L+2, 4-D2.0mg / L+ sodium thiosulfate 100mg / L medium for callus induction culture, additional sucrose 30g / L, agar 6.5g / L , PH5.8, light intensity 1000lx, temperature 23°C, humidity 70%-80%, the induced callus access medium formula is MS+KT0.2mg / L+2,4-D0.2mg / L+folic acid Culture in the proliferation medium of 0.5mg / L + 3% sucrose + 0.6% agar, pH 5.8, light intensity 1500lx, temperature 23°C, humidity 70%-80%, the callus with good dispersion after proliferation is put into liquid culture Base MS+2,4-D0.2mg / L+KT0.4mg / L+5mg / L phenylalanine for shaking culture, add 30g / L sucrose, inoculate 30ml li...
Embodiment 2
[0011] Take the leaves of the water coir raincoat, soak them in bleaching powder for 3 minutes, remove the surface stains with a brush, wash them with running water for 35 minutes, treat them with sodium hypochlorite on the ultra-clean workbench for 14 minutes, rinse them with sterile water 5-7 times, insert the sterilized water coir raincoat leaves The medium formula is improved MS+KT0.5mg / L+2, 4-D2.0mg / L+ sodium thiosulfate 100mg / L medium for callus induction culture, additional sucrose 30g / L, agar 6.5g / L , PH5.8, light intensity 1000lx, temperature 23°C, humidity 70%-80%, the induced callus access medium formula is MS+KT0.2mg / L+2,4-D0.2mg / L+folic acid Culture in the proliferation medium of 0.5mg / L + 3% sucrose + 0.6% agar, pH 5.8, light intensity 1500lx, temperature 23°C, humidity 70%-80%, the callus with good dispersion after proliferation is put into liquid culture Base MS+2,4-D0.3mg / L+KT0.6mg / L+6mg / L phenylalanine for shaking culture, add 30g / L sucrose, inoculate 30ml li...
Embodiment 3
[0013] Take the leaves of the water coir raincoat, soak them in bleaching powder for 3 minutes, remove the surface stains with a brush, wash them with running water for 35 minutes, treat them with sodium hypochlorite on the ultra-clean workbench for 14 minutes, rinse them with sterile water 5-7 times, insert the sterilized water coir raincoat leaves The medium formula is improved MS+KT0.5mg / L+2,4-D2.0mg / L+thiosulfuric acid 100mg / L medium for callus induction culture, additional sucrose 30g / L, agar 6.5g / L, PH5.8, light intensity 1000lx, temperature 23°C, humidity 70%-80%, the induced callus is inserted into the medium formula as MS+KT0.2mg / L+2, 4-D0.2mg / L+folate 0.5 Cultivate in the proliferation medium of mg / L + 3% sucrose + 0.6% agar, pH 5.8, light intensity 1500lx, temperature 23°C, humidity 70%-80%, callus with good dispersion after proliferation are placed in liquid medium Shaking culture in MS+2,4-D0.3mg / L+KT0.4mg / L+6mg / L phenylalanine, add 30g / L sucrose, inoculate 30ml l...
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