Daphne genkwa stem explant primary tissue culture method
A tissue culture and explant technology, which is applied in the directions of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems such as no report on the tissue culture and rapid propagation technology of stem explants, and achieves convenient and easy experimental operation. The effect of strong tolerance and high success rate
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Embodiment 1
[0036] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.0mg·L -1 ZT, add 30g / L sucrose and 7g / L agar to adjust the pH to pH=5.8.
[0037] Training method:
[0038] (1) Selection of Daphne genkwa explants: collection of robust growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 10cm stems with axillary buds as explants;
[0039] (2) Treatment of Daphne genkwa explants: soak the explants in a 0.5% washing powder aqueous solution for 20 minutes with a mass-volume ratio, then rinse them with running water for 12 hours, and place the explants under the aseptic environment of the ultra-clean workbench in the inoculation room. The body was cut into 4cm stem segments, sterilized with 75% ethanol aqueous solution for 30s, and then 1g·L -1Soak in a mixed solution of mercuric chloride and 5 drops of Tween 80 for 12 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a l...
Embodiment 2
[0042] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.5mg·L -1 ZT, add 30g / L sucrose and 7g / L agar to adjust the pH to pH=5.8.
[0043] Training method:
[0044] (1) Selection of Daphne genkwa explants: collection of robust growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 10cm stems with axillary buds as explants;
[0045] (2) Treatment of Daphne genkwa explants: soak the explants in 0.5% washing powder aqueous solution for 20 minutes, then rinse them with running water for 12 hours, and place the explants in the aseptic environment of the ultra-clean workbench in the inoculation room. Cut into 4cm stem segments, sterilize with 75% ethanol aqueous solution for 30s, and then use 1g·L -1 Soak in a mixed solution of mercuric chloride and 5 drops of Tween 80 for 12 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a length of 2 cm, the upper cut of the ste...
Embodiment 3
[0048] Primary tissue culture medium: MS+0.1mg·L -1 NAA+1.5mg·L -1 ZT, add 25g / L sucrose and 6.5g / L agar to adjust the pH to pH=5.7.
[0049] Training method:
[0050] (1) Selection of explants of Daphne genkwa: collection of strong growth, full axillary buds, no pests and diseases of Daphne genkwa explants in the same year, remove all leaves, cut into 8cm stems with axillary buds as explants;
[0051] (2) Treatment of Daphne genkwa explants: Soak the explants in a 0.2% washing powder aqueous solution for 20 minutes, then rinse them with running water for 12 hours, and inoculate the explants under the aseptic environment of the ultra-clean workbench in the inoculation room. The body was cut into 5cm stem segments, sterilized with 70% ethanol aqueous solution for 45s, and then 1g·L -1 Soak in a mixed solution of mercuric chloride and 3 drops of Tween 80 for 10 minutes for disinfection; rinse with sterile water after disinfection, cut the stem section into a length of 2.5 cm,...
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