Mutagenesis method of tender shoots of dianthus flowers and application of mutagenesis method
A technology of Dianthus and young shoots, applied in the biological field, can solve the problems of certain risks to the health of breeding operators, environmental pollution, improper protection, etc., and achieve the effects of overcoming obstacles to variety improvement, not polluting the environment, and simple operation.
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Embodiment 1
[0035] Take 10 μL of GV3101 Agrobacterium in an ultra-low temperature refrigerator and put it into 20ml of YEB liquid medium, add rifampicin with a final concentration of 60mg / L, shake and culture at 28°C and 200rpm for 20h, and obtain activated Agrobacterium liquid, Agrobacterium liquid OD 600 The value is 0.3.
[0036] Get 50 shoots of 4 cm in carnation variety 'Lan Guiren', peel off the young leaves to expose the stem tip, prick the wound around the shoot tip with a dissecting needle, then soak the shoots in the bacterium solution of activated GV3101 Agrobacterium, in Shake culture at 26°C and 30rpm in the dark for 16 hours; take out the shoots and wash them with water for 3 times, then immerse them in 400mg / L cefotaxime sodium solution for 4 hours; take out the shoots and wash them with water for 3 times, put in the rooting powder In the substrate, the substrate is made of humus, perlite and coconut shell in a mass ratio of 1:1:1; after 20 days of rooting, the rooted seedl...
Embodiment 2
[0038] Take 20 μL of EHA105 Agrobacterium in an ultra-low temperature refrigerator and put it into 30ml of YEB liquid medium, add rifampicin with a final concentration of 70mg / L, shake and culture at 28°C and 200rpm for 22h, and obtain activated Agrobacterium liquid, Agrobacterium liquid OD 600 The value is 0.32.
[0039]Get 50 shoots of 3 cm in carnation variety 'Fen Lian', peel off the young leaves to expose the stem tip, prick the wound with a dissecting needle around the shoot tip, then soak the shoots in the bacterium solution of activated EHA105 Agrobacterium. Shake culture at 27°C and 40rpm in the dark for 24 hours; take out the shoots and wash them with water for 3 times, then immerse them in 450 mg / L cefotaxime sodium solution for 6 hours; take out the shoots and wash them with water for 3 times, put in the rooting powder In the matrix, the matrix is made of humus, perlite and coconut shell in a mass ratio of 2:2:1. Put the rooted seedlings into the greenhouse for c...
Embodiment 3
[0041] Take 30 μL of LBA4404 Agrobacterium in an ultra-low temperature refrigerator and put it into 40ml of YEB liquid medium, add rifampicin with a final concentration of 65mg / L, shake and culture at 28°C and 200rpm for 24 hours, and obtain activated Agrobacterium liquid, Agrobacterium liquid OD 600 The value is 0.4.
[0042] Take 50 shoots of carnation variety 'Master' 3cm, peel off the young leaves to expose the stem tip, prick the wound with a dissecting needle around the shoot tip, then soak the shoots in the bacterium liquid of activated LBA4404 Agrobacterium, Shake culture at 27°C and 40rpm in the dark for 24 hours; take out the shoots and wash them with water for 3 times, then immerse them in 500 mg / L cefotaxime sodium solution for 8 hours; take out the shoots and wash them with water for 3 times, and add root-growing powder Insert it into the substrate, which is made of humus, perlite and coconut shell in a mass ratio of 1:3:1; 30 days after rooting, put the rooted se...
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