Tissue culture method for directly growing seedling from barley embryo and culture medium used in method
A tissue culture medium and culture medium technology, applied in the field of plant tissue culture, can solve the problems of easy variation, long callus-differentiation-seedling process cycle, and high pollution rate
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Embodiment 1
[0049] Embodiment 1, barley immature embryo culture medium in vitro, is made up of these 2 types of culture medium of seedling medium and rooting strong seedling medium:
[0050] 1), seedling growth medium:
[0051] First prepare the mother liquor of the seedling medium:
[0052] The mother liquor of seedling growth medium is composed of the following components:
[0053] KNO 3 1900mg / L, NH 4 NO 3 165mg / L, KH 2 PO 4 170mg / L, CaCl 2 2H 2 O165mg / L, MgSO 4 ·7H 2 O185mg / L, Na 2 -EDTA·2H 2 O37.3mg / L, FeSO 4 ·7H 2 O27.8mg / L, H 3 BO 3 6.2mg / L, MnSO 4 4H 2 O22.3mg / L, ZnSO 4 ·7H 2 O8.6mg / L, Na 2 MoO 4 2H 2 O0.25mg / L, KI0.83mg / L, CuSO 4 ·5H 2 O0.025mg / L, CoCl 2 ·6H 2 O0.025mg / L, proline 600mg / L, glutamine 250mg / L, hydrolyzed casein 400mg / L, inositol 100mg / L and maltose 30g / L, the rest is distilled water;
[0054] The preparation method of the mother liquor of seedling growth medium is as follows:
[0055] ①. Weigh KNO separately 3 19.0g, NH 4 NO 3 1.65g, K...
Embodiment 2
[0068] Embodiment 2, a method for in vitro culture of barley immature embryos, using the corresponding medium in Example 1, with "Zhexiu 12 / Subei No. 3" F 1 Barley was used as the experimental material in the following steps:
[0069] 1) Select "Zhexiu 12 / Su Beer No. 3" F 1 The immature seeds of barley 12-15 days after pollination, peel off the shell to expose the embryo, then soak the above-mentioned treated seeds in 50% (mass concentration) sodium hypochlorite solution for 30 minutes, discard the sodium hypochlorite solution and add 70% (v / v) Soak in alcohol for 5 minutes, then discard 70% alcohol and add 3% (v / v) hydrogen peroxide to soak for 5 minutes, and finally wash 5 times with sterile water.
[0070] On the aseptic operating table, use a sterilized scalpel to gently puncture the sterilized immature seeds from the side of the embryo, then slowly take out the embryo from the front end of the embryo and inoculate it into the on seedling medium.
[0071] 2) Place the ...
Embodiment 3
[0077] Example 3. According to the method of Example 2, the immature embryos of the barley variety "Zhepi 33" were cultured in vitro. Three repetitions were set up, and 60 embryos were inoculated each time. The seedling rate (average value) was 90.6%.
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