Agrobacterium rhizogenes mediated aconitum coreanum and its obtaining method
A technology of Aconitum chrysalis and hair, which is applied in the field of Aconitum chrysantha root and its acquisition, can solve the problems affecting the output and quality of Aconitum chrysalis, not conforming to land resources, long growth cycle, etc., and achieve broad application prospects and utilization value, To overcome the effects of slow reproduction speed and short production cycle
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Embodiment 1
[0015] Embodiment 1, the cultivation and identification of the root of Aconitum japonica
[0016] 1. Cultivation of the roots of Aconitum japonica
[0017] 1. Materials
[0018] Fresh, healthy leaves of Aconitum japonica in pot, Agrobacterium rhizogenes A 4 .
[0019] 2. Conversion
[0020] 1) Obtaining of explants: Take fresh and healthy leaves of potted Aconitum chrysanthemum, wash them; sterilize them with 0.1% mercuric chloride for 10 minutes, rinse them with sterile water for 4 times, each time for no less than 1 minute, and then blot the surface dry Water; then cut the leaves into 0.5cm×1.0cm size on the ultra-clean bench, transfer to sterilized 1 / 2MS solid medium (5 pieces per bottle); pre-cultivate in the dark for 3 days.
[0021] 2) Cultivation of Agrobacterium rhizogenes: Pick a single colony of Agrobacterium rhizogenes, inoculate it in 5mL liquid YEB, culture it with shaking at 28°C in the dark overnight, inoculate it into 50ml liquid YEB for secondary culture f...
Embodiment 2
[0040] Embodiment 2, the cultivation of Aconitum japonica root
[0041] Take fresh and healthy stems of potted Aconitum chrysanthemum, wash them; sterilize them with 0.1% mercuric chloride for 10 minutes, rinse them with sterile water for 4 times, each time for no less than 1 minute, then blot the surface moisture; then put them on the ultra-clean bench The leaves were cut into 0.5cm×1.0cm size, transferred to sterilized 1 / 2MS solid medium (5 pieces per bottle); pre-cultivated in the dark for 6 days.
[0042] Pick a single colony of Agrobacterium rhizogenes 15834, inoculate it in 5 mL of liquid YEB, culture it overnight with shaking at 28°C in the dark, and inoculate it into 50 mL of liquid YEB for secondary culture for 4-6 hours the next day before use.
[0043] Immerse the pre-cultured Aconitum chrysogenum stem in the cultured Agrobacterium rhizogenes bacterial liquid, infect it for 20min, then use sterile filter paper to suck the bacterial liquid, and then inoculate it on 1...
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