Method for preparing paper mulberry somatic embryo and application thereof to paper mulberry expanding propagation
A somatic embryo and paper mulberry technology, applied in the biological field, can solve the problems of low efficiency, high labor cost and high cost, and achieve the effect of solving the complicated preparation and solving the market shortage.
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Embodiment 1
[0049] Embodiment 1, preparation somatic embryo
[0050] 1. Preparation of embryogenic callus
[0051] (1) Get the leaves of the aseptic seedlings of the tree, cut into 0.5 × 0.5cm 2, inoculated on Medium A, and cultivated in the dark at 23±2°C for 25 days. At this time, it can be observed that light yellow callus grows from the leaf cut, that is, embryogenic callus.
[0052] (2) The embryogenic callus tissue obtained in step (1) was taken, inoculated on medium A, and cultured in the dark at 23±2° C. for 25 days.
[0053] (3) The embryogenic callus tissue obtained in step (2) was taken, inoculated on medium A, and cultured in the dark at 23±2° C. for 25 days.
[0054] In actual operation, the subculture can be continuously carried out according to the method of step (2).
[0055] Medium A (solid): MS medium containing 0.5mg / L 2,4-dichlorophenoxyacetic acid, 30g / L sucrose and 5g / L agar powder.
[0056] 2. Liquid suspension culture
[0057] (1) Add 1 g of the embryogenic c...
Embodiment 2
[0067] Embodiment 2, somatic embryo plant regeneration
[0068] Dissolve sodium alginate in 1 / 2 MS medium to obtain sodium alginate solution.
[0069] 1. Put the somatic embryo obtained in Example 1 into 3.0g / 100mL sodium alginate solution and soak at room temperature for 3-4min, then transfer to 0.5g / 100mL CaCl 2 Soak in an aqueous solution for 3 minutes at room temperature to form artificial seeds with a certain hardness, then rinse with sterile deionized water for 3-4 times to terminate the reaction, and place them on sterile paper to absorb the surface moisture of the artificial seeds.
[0070] 2. Place the artificial seeds on the culture medium, light at 24°C for 12 hours (the light intensity in this example is 1500 lx; in actual operation, the light intensity is 1500-2000 lx) / 12 hours dark, static culture 30d.
[0071] Medium D (solid): containing 0.5mg / L NAA, 0.5mg / L 6-BA, 1mg / L GA 3 (Gibberellin), 1 / 2MS medium of 30g / L sucrose and 7g / L agar.
[0072] Experiments ...
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