Cucumber Flamingo-bill explant transgenic method
An explant and transgene technology, applied in the field of transgene, achieves the effect of short regeneration time, high survival rate and high transformation frequency
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Embodiment 1
[0082] Follow the steps below
[0083] a. The cucumber inbred line 'IL 69' was used as the material. The inbred line 'IL 69' was isolated from the offspring of Jinza 2. It has the characteristics of strong growth vigor, high resistance to downy mildew, long fruit shape, and ribs. . Disinfect the surface of 50 inbred seeds, inoculate them into MS medium, and cultivate them to form seedlings. The culture conditions are light intensity 54 μmol·m -2 ·s -1 , photoperiod 14h / d, temperature 25°C. A cotyledon and growth point were excised from sterile seedlings with a seedling age of 2 days. The depth of the incision was 1 / 2 of the diameter of the hypocotyl, and the length of the slope of the incision was 0.4 cm to obtain 50 Flamingo-bill explants. And the Flamingo-bill explants were placed in MS liquid medium containing 0.1 mg / L 6-benzyl adenine and 30 g / L sucrose for pre-culture for 2 days.
[0084] b. Pick the colony of Agrobacterium LBA4404 containing the target gene, add it t...
Embodiment 2
[0090] Follow the steps below
[0091] a. The cucumber inbred line 'IL 103' was used as the material. The inbred line 'IL 103' was isolated from the offspring of Jinchun 4. It has strong growth vigor, high resistance to downy mildew, medium fruit shape, no ribs, Prickly and other characteristics. Disinfect the surface of 50 inbred seeds, inoculate them into MS medium, and cultivate them to form seedlings. The culture conditions are light intensity 56 μmol·m -2 ·s -1 , photoperiod 16h / d, temperature 26°C. A cotyledon and growth point were excised from sterile seedlings with a seedling age of 3 days. The depth of the incision was 1 / 2 of the diameter of the hypocotyl, and the length of the slope of the incision was 0.5 cm to obtain 49 Flamingo-bill explants. And the Flamingo-bill explants were placed in MS liquid medium containing 0.3mg / L 6-benzyl adenine and 32g / L sucrose for pre-culture for 3 days.
[0092] b. Pick the Agrobacterium LBA4404 colony containing the target gene...
Embodiment 3
[0098] Follow the steps below
[0099] a. The cucumber inbred line 'IL 67' was used as the material. The inbred line 'IL 67' was isolated from the offspring of Yuanfengyuan No. 6. It has strong growth potential, long growth period, high resistance to downy mildew, and short fruit shape. , No edges and other features. Disinfect the surface of 52 inbred seeds, inoculate them into MS medium, and cultivate them to form seedlings. The culture conditions are light intensity 55 μmol·m -2 ·s -1 , photoperiod 15h / d, temperature 28°C. A cotyledon and growth point were excised from sterile seedlings with a seedling age of 2 days. The depth of the incision was 1 / 2 of the diameter of the hypocotyl, and the length of the slope of the incision was 0.5 cm to obtain 50 Flamingo-bill explants. And the Flamingo-bill explants were placed in MS liquid medium containing 0.5mg / L 6-benzyl adenine and 35g / L sucrose for pre-culture for 2 days.
[0100] b. Pick the colony of Agrobacterium LBA4404 co...
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