Method for regenerating Onosma paniculatum plants
A technology for comfrey and plant is applied in the field of plant regeneration of comfrey, which can solve the problems of high cell culture cost and technical content, difficult to popularize and apply on a large scale, and plant regeneration of comfrey has not yet been seen, and achieves a good growth state. , the effect of high survival rate
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Embodiment 1
[0015] 1) Take the tender leaves of the stem tip of Comfrey yunnanensis, rinse the surface of the leaves with distilled water, then soak them in 75% ethanol solution for 20s, remove the ethanol solution quickly, rinse the leaves twice with sterilized distilled water, and then transfer to 2% ethanol solution. Soak in sodium hypochlorite solution for 2 minutes, take out the leaves, and rinse 3 times with sterilized distilled water. Put the sterilized leaves into a sterilized petri dish and cut them into about 0.5cm in size with a blade 2 and transferred to callus induction medium to induce callus. Induction medium and culture conditions are: MS basal medium + NAA0.3mg / L + 2,4-D 0.8mg / L, cultured on solid agar, pH 5.6, temperature 25°C, cultured in the dark for 30 days to obtain embryogenic healing damage tissue.
[0016] 2) The induced embryogenic callus was subcultured twice and then used for organ differentiation to obtain plants. The subculture medium and culture condition...
Embodiment 2
[0023] 1) Take the tender leaves of the stem tip of Comfrey yunnanensis, rinse the surface of the leaves with distilled water, then soak them in 75% ethanol solution for 20s, remove the ethanol solution quickly, rinse the leaves twice with sterilized distilled water, and then transfer to 2% ethanol solution. Soak in sodium hypochlorite solution for 2 minutes, take out the leaves, and rinse 3 times with sterilized distilled water. Put the sterilized leaves into a sterilized petri dish and cut them into about 0.5cm in size with a blade 2 and transferred to callus induction medium to induce callus. Induction medium and culture conditions are: MS basal medium + NAA 0.1mg / L + 2,4-D 1.5mg / L, cultured on solid agar, pH 5.6, temperature 25°C, cultured in the dark for 30 days to obtain embryogenic healing damage tissue.
[0024] 2) The induced embryogenic callus was subcultured twice and then used for organ differentiation to obtain plants. The subculture medium and culture conditio...
Embodiment 3
[0031]1) Take the tender leaves of the stem tip of Comfrey yunnanensis, rinse the surface of the leaves with distilled water, then soak them in 75% ethanol solution for 20s, remove the ethanol solution quickly, rinse the leaves twice with sterilized distilled water, and then transfer to 2% ethanol solution. Soak in sodium hypochlorite solution for 2 minutes, take out the leaves, and rinse 3 times with sterilized distilled water. Put the sterilized leaves into a sterilized petri dish and cut them into about 0.5cm in size with a blade 2 and transferred to callus induction medium to induce callus. Induction medium and culture conditions are: MS basal medium + NAA 0.2mg / L + 2,4-D 0.8mg / L, cultured on solid agar, pH 5.6, temperature 25°C, cultured in the dark for 30 days to obtain embryogenic healing damage tissue.
[0032] 2) The induced embryogenic callus was subcultured twice and then used for organ differentiation to obtain plants. The subculture medium and culture condition...
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