Tissue culture propagation method for obtaining cibotium barometz green globular body
A technology of green spheroids and golden retriever dogs, applied in the field of plant tissue culture, to achieve the effects of inhibiting differentiation, high proliferation rate, and high reproduction efficiency
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Embodiment 1
[0039] Embodiment 1 The inventive method
[0040] (1) Obtaining sterile explants of golden retriever dogs
[0041] Inoculate the sterile spores of Golden Retriever on the medium of 1 / 4MS+sucrose 30.0g / L+agar 7.0g / L, pH 5.80. 2000lx, to obtain young tender aseptic seedlings. In the ultra-clean workbench, the obtained sterile seedlings were peeled off the shoot tips with tweezers under a dissecting microscope, and the shoot tips were used as sterile explants induced by golden retriever green spheroids.
[0042] The method for obtaining aseptic spores of golden retriever dogs is as follows: put 10 mg of spores in a 1.5 ml centrifuge tube, soak in sterile water for 1-2 hours, centrifuge at 6000 r / min for 2 minutes to obtain spore precipitate, and add a volume fraction of 5% NaClO solution Sterilize for 4 minutes, wash with sterile water for 3-5 times to obtain aseptic spores of Golden Retriever.
[0043] (2) Induction of golden retriever green spheroids
[0044] ①The first induc...
Embodiment 2
[0061] Embodiment 2 and embodiment 3 are the method of the present invention
[0062] Example 2 and Example 3 except that the concentration of each component in each culture medium listed in Table 1 is different, other measures are the same as Example 1, and will not be repeated.
[0063] The difference between the steps of Table 1 Embodiment 2-Embodiment 3 and Embodiment 1
[0064]
[0065]
[0066] The propagation result of embodiment 1: the induction rate of green spheroid is 94%, the differentiation rate is 100%, and the rooting rate is 100%, that is, the formation rate of tissue cultured shoots is 100%, rooted and cultivated for 28 days, and the average height of tissue cultured plants is 3.15cm . The green spheroid with a diameter of 2mm has a fresh weight of 8 mg, and the average fresh weight of the green spheroid increases by 76 mg every 18 days, and the average multiplication factor is 9.5 times. The obtained green spheroids are intermediate propagules, which ...
Embodiment 3
[0068] The propagation result of embodiment 3: the induction rate of green spheroid is 92%, the differentiation rate is 100%, the rooting rate is 100%, that is, the formation rate of tissue cultured seedlings is 100%, rooting is cultivated for 28 days, and the average height of tissue cultured seedlings is 3.30cm . The green spheroids with a diameter of 2mm have a fresh weight of 8 mg, and the average fresh weight of the green spheroids increases by 77.6 mg every 21 days, and the average multiplication factor is 9.7 times. The obtained green spheroids are intermediate propagules, which are cultured for 42 days by two steps, differentiated for 28 days, rooted and cultured to form tissue cultured seedlings for 28 days, and propagated for 98 days.
[0069] A 1-3 mm green spheroid can be differentiated once to obtain 20-25 tissue cultured seedlings.
[0070] Above-mentioned embodiment 1 to embodiment 3 show: the induction rate of green spheroid of the present invention is 90~94%,...
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