Ultra-low temperature preserving method for nutrient breeding flower and short-tube lycoris
A technology of ultra-low temperature preservation and vegetative reproduction, which is applied in the field of ultra-low temperature preservation of vegetatively propagated flowers of Lycoris chinensis, can solve the problem that the ultra-low temperature preservation technology of Lycoris is not yet established, and achieves the effect that the ultra-low temperature preservation method is simple and easy to implement and restores a good growth condition.
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Embodiment 1
[0023] Take 3-4 year old bulbs of Lycoris radiata Herb, disinfect with 70% alcohol for 1 min, 0.1% mercuric chloride for 15 min, rinse with sterile water for several times and remove 3-4 layers of scales and part of the stem disk. Bulb cuts (about 1cm in size) were inoculated with 3mg·L -1 BA and 0.5 mg·L -1 Induction of bulblets on MS medium with NAA. The culture temperature is 25±2℃, and the light is 12h·d -1, light intensity 36μmol m -2 the s -1 .
Embodiment 2
[0024] Example 2 Lycoris radiata Herb (Lycoris radiata Herb) pre-cultivation
[0025] Get the bulblet of embodiment 1 gained, cut out the shoot tip that has root primordia and bud primordia simultaneously of 2.5mm size, inoculate to contain 0.4mol L -1 In sucrose MS medium, culture in dark at 25°C for 5 days.
Embodiment 3
[0027] Put the pre-cultured shoot tips in the pretreatment solution (2mol L -1 glycerol and 0.4mol L -1 Sucrose MS liquid medium) was treated at room temperature for 20min, then transferred to vitrification agent (3.26mol·L -1 Glycerin+2.42mol·L -1 Ethylene glycol+1.9mol·L -1 DMSO+0.4mol L -1 sucrose) in ice bath for 80min, and then transfer the shoot tips to 2mL cryovials filled with pre-cooled fresh vitrification agent (10 shoot tips·tube -1 ), quickly put into liquid nitrogen for storage.
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