A kind of genetic transformation method of grape summer black
A gene transformation and grape technology, applied in the field of genetic engineering, can solve the problems of Agrobacterium being unable to inhibit, cumbersome, and unsuccessful transgenesis, and achieve the effects of improving transgenic efficiency, simplifying the operation process, and avoiding blindness
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[0027] Example: First, prepare the main solution and the preparation of the culture medium:
[0028] Grape subculture medium (S1): MS 2.2g / L, sucrose 30g / L, agar 5.5g / L, IBA 0.4mg / L, pH 5.8, autoclaved, set aside.
[0029] Grape co-cultivation medium (S2): MS2.2g / L, sucrose 30g / L, agar 5.5g / L, TDZ1mg / L, ZT1mg / L, IBA 0.4mg / L, pH5.4, autoclaved, cooled to At about 60°C, divide into flat plates for later use.
[0030] Grape screening medium (S3): MS2.2g / L, sucrose 30g / L, agar 5.5g / L, TDZ1mg / L, ZT 1mg / L, IBA 0.4mg / L, pH5.4, autoclaved, cooled to Add 20 mg / L kanamycin and 400 mg / L carbenicillin at 60°C, and fill in culture bottles for later use.
[0031] Grape rooting screening medium (S4): MS2.2g / L, sucrose 30g / L, agar 5.5g / L, IBA 0.4mg / L, pH5.8, autoclaved, cooled to 60°C, added kanamycin 20mg / L, carbenicillin 300mg / L subpackage culture bottle for standby.
[0032] Secondly, the activation of the bacterial species in the bacterial liquid is carried out:
[0033] ① Streak th...
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