Atrac tylodes macrocephala koidz conservation method and re-cultivating method
A preservation method, Atractylodes macrocephala technology, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of genetic variation, labor and material resources, etc., achieve low cost, easy operation, and avoid germplasm mutation or destruction. Effect
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Embodiment 1
[0040] Embodiment 1, the cryopreservation of Atractylodes Rhizome
[0041] 1. Reagent formulations
[0042] Pre-culture medium: MS liquid medium containing 0.3mol / L sucrose.
[0043] Pretreatment solution: MS liquid medium containing 2.0mol / L glycerol and 0.4mol / L sucrose.
[0044]Vitrification protectant: 3.26mol glycerol, 2.42mol ethylene glycol, 1.9mol dimethyl sulfoxide and 0.4mol sucrose were thoroughly mixed with MS liquid medium to 1L.
[0045] 2. Ultra-low temperature preservation of Atractylodes macrocephala
[0046] Take 2-month-old Atractylodes macrocephala tissue-cultured seedlings, peel off the outer leaves layer by layer until the remaining 1-2 leaf primordium surrounds the shoot apex meristem, and cut out 1.5-2mm shoot tips (a total of 100 shoot tips were obtained), Soak in the pre-culture solution for 3 days in the dark at 25°C, then soak in the pretreatment solution for 30 minutes with shaking at 25°C (normal light), then soak in the vitrification protectan...
Embodiment 2
[0055] Embodiment 2, the cryopreservation of Atractylodes macrocephala
[0056] 1. Reagent formulations
[0057] All the same as Step 1 of Example 1.
[0058] 2. Ultra-low temperature preservation of Atractylodes macrocephala
[0059] The time of dark cultivation at 25° C. in the pre-culture solution is 1 day, 2 days, 3 days, 4 days or 5 days respectively, and the others are the same as step 2 of Example 1.
[0060] 3. Statistics on recultivation, survival rate and regeneration rate
[0061] Same as Step 3 of Example 1.
[0062] The experiments were repeated three times, and the results were averaged.
[0063] When cultured in the dark at 25°C for 1 day in the pre-culture solution, the average plant survival rate of the three experiments was 69.8%. The average plant regeneration rate of the three experiments was 54.9%.
[0064] When cultured in the dark at 25°C for 2 days in the pre-culture solution, the average plant survival rate of the three experiments was 66.0%. Th...
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