Tobacco explant anti-browning method based on active carbon and citric acid
A technology of explants and activated carbon, applied in the field of plant tissue culture, can solve the problems of insignificant effect, achieve good effect, improve differentiation state, and shorten the days of rooting
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Embodiment 1
[0028] The tobacco explant anti-browning method based on gac and citric acid of the present embodiment may further comprise the steps:
[0029] Step (1), disinfection of seeds: take plump tobacco seeds, then move them to an ultra-clean workbench, soak the seeds with 75% alcohol for 30s, rinse them twice with sterile water, and then use 1% AgNO 3 Disinfect for 10 minutes, soak and wash in sterile water for 5 times, dry the surface moisture of seeds with sterile filter paper and then inoculate, wherein the sterile water is ultrapure water sterilized by autoclaving.
[0030] Step (2), seed cultivation: inoculate the seeds obtained in step (1) into MS medium, under the conditions of cultivation temperature of 25±1°C, light intensity of 30 μmol / (m2 s), and light time of 16 h / d Under culture for 60 days, the seeds germinate and grow into explant seedlings, and then coat the surface with water soaked in raw potatoes. The water soaked in raw potatoes contains substances such as starch...
Embodiment 2
[0037] The tobacco explant anti-browning method based on gac and citric acid of the present embodiment, wherein:
[0038] Steps (1)-(2) are the same as in Example 1;
[0039] Step (3), pre-cultivation of tobacco explants: transfer and inoculate the explant seedlings obtained in step (2) into a medium containing MS basic medium, 4g / L agar, 30g / L walnut kernels added with 10g / L L sucrose MS pre-cultured on solid medium. The control temperature is 24°C, the light intensity is 1500lux, the light time is 10 hours / day, and the culture time is 3 days. Finally, the pre-cultured explants are repeatedly inoculated in MS medium 4-5 times to make the tissue Partial infiltration of phenolic substances in the medium can promote the rapid germination of explants. Walnut grains are rich in melatonin, which can scavenge free radicals and prevent cell oxidative damage from browning.
[0040] Step (4) is identical with embodiment 1;
[0041] Step (5), explant anti-browning culture: place the e...
Embodiment 3
[0045] The tobacco explant anti-browning method based on gac and citric acid of the present embodiment, wherein:
[0046] Steps (1)-(2) are the same as in Example 1;
[0047] Step (3), pre-cultivation of tobacco explants: transfer and inoculate the explant seedlings obtained in step (2) into a medium containing MS basic medium, 4g / L agar, 30g / L walnut kernels added with 15g / L L sucrose MS pre-cultured on solid medium. The control temperature is 24°C, the light intensity is 1500lux, the light time is 10 hours / day, and the culture time is 3 days. Finally, the pre-cultured explants are repeatedly inoculated in MS medium 4-5 times to make the tissue Partial infiltration of phenolic substances in the medium can promote the rapid germination of explants. Walnut grains are rich in melatonin, which can scavenge free radicals and prevent cell oxidative damage from browning.
[0048] Step (4) is identical with embodiment 1;
[0049] Step (5), explant anti-browning culture: place the...
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