Spiraea ultralow-temperature storage and culture method
A technology of cryopreservation and cultivation method, applied in the field of germplasm resource preservation, to achieve the effects of genetic stability, high recovery rate and regeneration rate, and good activity ability
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Embodiment 1
[0028] The invention provides a method for ultra-low temperature preservation and cultivation of spirea, comprising: placing the spirea tip of 1 mm in length cultured in the pre-culture solution in a shaker shaker at room temperature and cultivating in the dark for 1 day, and then using a loading After 30 minutes of liquid treatment, the stem tip of Spiraea was taken out and transferred to a vitrification solution at 0°C for dehydration for 20 minutes, then put into a cryopreservation tube, and then the cryopreservation tube was stored in liquid nitrogen for ultra-low temperature preservation.
[0029] Wherein, the preculture solution is: 1 / 2MS+0.8mol / L sucrose+7g / L agar;
[0030] The loading solution is: 1 / 2MS+2mol / L glycerol+0.5mol / L sucrose
[0031] The vitrification solution is: PVS1
Embodiment 2
[0033] The invention provides a method for ultra-low temperature preservation and cultivation of Spiraea, comprising: placing the stem tip of Spiraea with a length of 2 mm in the pre-culture solution in a shaker shaker for 2 days in the dark for 2 days, and then using a loading After 40 minutes of liquid treatment, the stem tip of Spiraea was taken out and transferred to a vitrification solution at 0°C for dehydration for 40 minutes, then put into a cryopreservation tube, and then the cryopreservation tube was stored in liquid nitrogen for ultra-low temperature preservation.
[0034]Among them, the pre-culture solution is: MS+1.2mol / L glucose+0.2mg / mL NAA;
[0035] The loading solution is: 1 / 2MS+2mol / L glycerol+0.7mol / L sucrose
[0036] The vitrification solution is: PVS3
Embodiment 3
[0038] The invention provides a method for ultra-low temperature preservation and cultivation of Spiraea, which comprises: placing the stem tip of Spiraea with a length of 2 mm in the pre-culture solution in a shaker shaker for 3 days in the dark for 3 days, and then using a loading After 50 minutes of liquid treatment, the stem tip of Spiraea was taken out and transferred to a vitrification solution at 0°C for dehydration for 60 minutes, then put into a cryopreservation tube, and then the cryopreservation tube was stored in liquid nitrogen for ultra-low temperature preservation.
[0039] Among them, the pre-culture solution is: MS+1.2mol / L glucose+5%DMSO+1.0mg / mL6-BA
[0040] The loading solution is: 1 / 2MS+2mol / L glycerol+0.7mol / L sucrose
[0041] The vitrification solution is: PVS1
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