Rapid propagation method of polyploidy mutation of cissampelos pareira
A polyploid, rapid technology, applied in the field of plants, to achieve the effect of high utilization rate, large-scale planting and high survival rate
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Embodiment 1
[0009] Take the pollen of vine vine, disinfect it with 70% ethanol for 0.5min, then sterilize it with 1% hydrogen peroxide solution for 5 minutes, wash it with sterile water 5 times, and inoculate the sterilized vine pollen into MS+NAA0. 4mg / L+KT6mg / L+colchicine 50mg / L medium for callus induction, add sucrose 30g / L, agar 6.5g / L, pH5.8, dark light, temperature 22±2℃, induction The dispersed and stable callus was inoculated into the medium of VR+TDZ2mg / L+NAA0.1mg / L+tryptophan 24μmol / L for callus differentiation, adding sucrose 30g / L, agar 6.5g / L, pH 5.8, light 2500lx, light period 12h / d, temperature 23±2°C, differentiated embryos were placed in MS medium for strong seedling growth, supplemented with sucrose 60g / L, agar 7g / L, pH 5.8, light 20000lx, photoperiod 14h / d, temperature 28℃, survival rate 80%.
Embodiment 2
[0011] Take the pollen of vine vine, disinfect it with 70% ethanol for 0.5min, then sterilize it with 1% hydrogen peroxide solution for 5 minutes, wash it with sterile water 5 times, and inoculate the sterilized vine pollen into MS+NAA0. 4mg / L+KT6mg / L+colchicine 100mg / L medium for callus induction, add sucrose 30g / L, agar 6.5g / L, pH5.8, dark light, temperature 22±2℃, induction The dispersed and stable callus was inoculated into the medium of VR+TDZ2mg / L+NAA0.1mg / L+tryptophan 24μmol / L for callus differentiation, adding sucrose 30g / L, agar 6.5g / L, pH 5.8, light 2500lx, light period 12h / d, temperature 23±2°C, differentiated embryos were placed in MS medium for strong seedling growth, supplemented with sucrose 60g / L, agar 7g / L, pH 5.8, light 20000lx, photoperiod 14h / d, temperature 28℃, survival rate 82%.
Embodiment 3
[0013] Take the pollen of vine vine, disinfect it with 70% ethanol for 0.5min, then sterilize it with 1% hydrogen peroxide solution for 5 minutes, wash it with sterile water 5 times, and inoculate the sterilized vine pollen into MS+NAA0. 4mg / L+KT6mg / L+colchicine 150mg / L medium for callus induction, add sucrose 30g / L, agar 6.5g / L, pH5.8, dark light, temperature 22±2℃, induction The dispersed and stable callus was inoculated into the medium of VR+TDZ2mg / L+NAA0.1mg / L+tryptophan 24μmol / L for callus differentiation, adding sucrose 30g / L, agar 6.5g / L, pH 5.8, light 2500lx, light period 12h / d, temperature 23±2°C, differentiated embryos were placed in MS medium for strong seedling growth, supplemented with sucrose 60g / L, agar 7g / L, pH 5.8, light 20000lx, photoperiod 14h / d, temperature 28℃, survival rate 83%.
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