Method for ultralow-temperature preservation of winged bean seeds
A technology for ultra-low temperature preservation and quadrangular bean, which is applied in the preservation of plants, botanical equipment and methods, applications, etc., can solve the problems of non-permanent preservation, reduction, and vitality reduction, and achieve safe preservation, simple process, and stability. reliable results
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Embodiment 1
[0023] Take the Xiangleng bean No. 1 harvested in a certain place in Hunan, and dry it naturally, and use the B5 liquid culture solution containing 2.2mol / L glycerol and 1.3mol / L dimethyl sulfoxide to the four-lens bean seeds with a water content of about 11%. Treatment, after 20 minutes of treatment, take out the treated square bean seeds, and put them into the cryopreservation tube of B5 liquid culture medium containing 3.2mol / L glycerol, 1.7mol / L dimethyl sulfoxide and 0.6mol / L sucrose , incubate at room temperature for 30 minutes, and then put it into the gradient cooling box; put the gradient cooling box into a -70°C low-temperature refrigerator for gradient cooling, so that the temperature of the cooling box is at -45°C, and the cooling rate is -1°C / min , then take out the cooling box, put the cooling box into liquid nitrogen immediately, and store it at a low temperature in liquid nitrogen for 5 years, then take the cryopreservation tube out of the liquid nitrogen, place...
Embodiment 1
[0025] Take Xiangleng bean No. 1 harvested in a certain place in Hunan that year, and dry it naturally. Treat the four-lens bean seeds with a water content of about 11% with B5 liquid culture solution containing 2.2mol / L glycerol. After 20 minutes of treatment, take out the treated Lens bean seeds, then put into the cryopreservation tube of the B5 liquid culture medium containing 3.2mol / L glycerol, 1.7mol / L dimethyl sulfoxide and 0.6mol / L sucrose, and then put it into the gradient cooling box ;Put the gradient cooling box into a -70°C low-temperature refrigerator for gradient cooling, so that the temperature of the cooling box is at -45°C, and the cooling rate is -1°C / min, then take out the cooling box, and immediately put the cooling box into liquid nitrogen , after 5 years of cryopreservation in liquid nitrogen, the cryopreservation tube was taken out of the liquid nitrogen, placed at a room temperature of about 23°C, and thawed naturally at room temperature. The tap water o...
Embodiment 2
[0027] Take the Xiangleng bean No. 1 harvested in a certain place in Hunan, and dry it naturally, and use the B5 liquid culture solution containing 2.2mol / L glycerol and 1.3mol / L dimethyl sulfoxide to the four-lens bean seeds with a water content of about 11%. Treatment, after 20 minutes of treatment, take out the treated square bean seeds, and put them into the cryopreservation tube of B5 liquid culture medium containing 3.2mol / L glycerol, 1.7mol / L dimethyl sulfoxide and 0.6mol / L sucrose , incubate at room temperature for 30 minutes, and then store it in a low-temperature refrigerator at -5°C for 5 years. Take the cryopreservation tube out of the liquid nitrogen, place it at a room temperature of about 23°C, and thaw it naturally at room temperature. After thawing, The four-lens bean seeds in the frozen storage tube were taken out, and then washed four times with tap water or distilled water. After washing, the four-lens bean seeds were directly used for germination experiment...
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