A method for rapid evoking adventive bud panax ginseng stem segment
A ginseng stem and rapid technology is applied in the field of rapidly obtaining adventitious buds of ginseng, which can solve the problems of long time consumption and complicated pathway steps, and achieves the effects of high speed, small variation of somatic clones, and maintaining genetic stability.
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Embodiment 1
[0017] a. Select 10 ginseng seeds, soak them with 75% ethanol for 30s on the ultra-clean workbench, wash them with sterile water for 4 times, and use 0.1% HgCl 2 Soak in the solution for 8 minutes, then rinse with sterile water for 4 times, inoculate on 1 / 2 MS medium and cultivate for 35 days to germinate the seeds and form ginseng aseptic seedlings;
[0018] b. Remove the leaves and roots of ginseng aseptic seedlings, cut the stem into 1cm segments, insert them into the adventitious bud induction medium to induce adventitious buds, and the adventitious bud induction medium is MS medium + 3% sucrose + 2,4- D 0.2mg / L+NAA 0.5mg / L+6-BA 1.0mg / L+agar 0.7%, the pH value is 5.9, the culture temperature is 20°C, the humidity is 40%, the light intensity is 2000lux, and the light time is 14 hours / After 15 days of culturing, the stem segment began to appear bud point protrusions, and adventitious buds grew after 25 days, and each stem segment explant could induce 1-3 adventitious buds; ...
Embodiment 2
[0021] a. Select 10 ginseng seeds, soak them with 75% ethanol for 25s on the ultra-clean workbench, wash them with sterile water for 3 times, and use 0.2% HgCl 2 Soak in the solution for 6 minutes, then rinse with sterile water for 5 times, inoculate on 1 / 2 MS medium and cultivate for 30 days to germinate the seeds and form ginseng aseptic seedlings;
[0022] b. Remove the leaves and roots of the ginseng sterile seedlings, cut the stems into 1.5cm segments, insert them into the adventitious bud induction medium to induce adventitious buds, and the adventitious bud induction medium is MS medium + 3% sucrose + 2,4 -D 0.1mg / L+NAA 1.0mg / L+6-BA 0.5mg / L+agar 0.8%, the pH value is 5.8, the culture temperature is 22°C, the humidity is 50%, the light intensity is 2300lux, and the light time is 14 hours / day, after cultivating for 20 days, the stem section begins to appear bud point protrusions, and after 30 days, adventitious buds grow, and each stem section explant can induce 1-3 adv...
Embodiment 3
[0025] a. Select 10 ginseng seeds, soak them with 75% ethanol for 35s on the ultra-clean workbench, wash them with sterile water for 5 times, and use 0.1% HgCl 2 Soak in the solution for 10 minutes, then rinse with sterile water for 3 times, inoculate on 1 / 2 MS medium and cultivate for 45 days to germinate the seeds and form aseptic ginseng seedlings;
[0026] b. Remove the leaves and roots of ginseng aseptic seedlings, cut the stem into 2cm segments, and insert them into the adventitious bud induction medium to induce adventitious buds. The adventitious bud induction medium is MS medium+sucrose 3%+2,4- D 0.3mg / L+NAA 0.1mg / L+6-BA 2.0mg / L+agar 0.6%, the pH value is 6.0, the culture temperature is 25°C, the humidity is 60%, the light intensity is 2500lux, and the light time is 14 hours / After 15 days of cultivation, the stem segments began to protrude from bud points, and adventitious buds grew after 25 days, and each stem segment explant could induce 1-3 adventitious buds;
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