Sterile seedling source plant cell and preparation method thereof
A technology of plant cells and sterile seedlings, applied in plant cells, cell culture medium, etc., can solve the problems of successful cultivation of ginseng root tip meristem without visible main root, material contamination, inability to completely eliminate plant endophytes, etc.
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Embodiment 1
[0134] This embodiment provides a preparation method of sterile seedling-derived plant stem cells, the specific steps are as follows:
[0135] 1.1 Seed disinfection
[0136] Select ginseng seeds after ripening ( figure 1 Shown in A), remove seed shell, will remove the ginseng seed after seed shell ( figure 1 Shown in B) rinse with running water for 3 hours, soak in 70% ethanol for 5 minutes, rinse with sterile water for 3 times, soak in 7.5% sodium hypochlorite for 1 minute, soak in 1% sodium hypochlorite for 15 minutes, and rinse with sterile water for 5 times.
[0137] 1.2 Cultivation of ginseng sterile seedlings
[0138] MS medium was selected, and the sterilized ginseng seeds were inoculated on the medium for light culture ( figure 1 C shown). The MS medium includes: 20 g / l sucrose, 7 g / l agar powder, 100 mg / l L-ascorbic acid, 150 mg / l citric acid, 2 mg / l IAA, and 2 mg / l IBA. The culture condition is: culture temperature 22 ℃, humidity 30% rH, illuminance 60%, 16h lig...
Embodiment 2
[0159] This embodiment provides a preparation method of sterile seedling-derived plant stem cells, the specific steps are as follows:
[0160] 2.1 The aseptic seedling-derived plant stem cells prepared by the same method as described in 1.1-1.3 in Example 1 were placed in the cell chamber for culture, and B5 liquid medium was added, and the liquid level of the medium was slightly higher than the filter screen of the chamber, but not higher than The CMCs tissue was then covered with a sterile sealing film, cultured with shaking, supplemented with liquid medium every 2 days, the rotation speed was 100r / min, the culture temperature was 24.6°C, and cultured in the dark for 14 days.
[0161] 2.2 Aspirate the CMCs tissue in the cell chamber in step 1, place it in a well plate for culture, add B5 liquid medium, cover the plate with a sterile sealing film, shake the culture, replenish the liquid medium every 2 days, rotate at 100r / min, and cultivate at temperature Culture at 24.6°C in...
Embodiment 3
[0165] This embodiment provides a preparation method of sterile seedling-derived plant stem cells, the specific steps are as follows:
[0166] 3.1 The aseptic seedling-derived plant stem cells prepared by the same method as described in 1.1-1.3 of Example 1 were cultured in a well plate containing B5 solid medium, and then covered with a sterile sealing film, cultured at 24.6°C in the dark , cultivated for 14d.
[0167] 3.2 Take the CMCs tissue cultured in the 12-well plate after the above step 3.1, then add a certain amount of B5 liquid medium, cover the plate with a sterile sealing film, shake the culture, replenish the liquid medium every 2 days, and rotate at 100r / min. The culture temperature was 24.6°C, cultured in the dark, and cultured for 14 days.
[0168] 3.3 Take the culture solution obtained in the above step 3.2, place it in a 150ml Erlenmeyer flask (with a gas-ventilating cap) for culture, add B5 liquid medium, shake the culture, replenish the liquid medium every...
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