Euphorbia lathyris callus cell suspension culture method capable of extracting and obtaining ingenol
A callus, cell suspension technology, applied in tissue culture, cell culture medium, plant cells, etc., can solve the problems of low yield, high cost, and unsatisfactory control effect of secondary metabolite synthesis.
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Embodiment 1
[0014] In the ultra-clean workbench, take out the aseptic seedlings of caperia seedlings from the culture bottle for aseptic sowing and culture for 30 days, cut the roots with sterilized knife tweezers, cut them into small sections with a length of 1 cm, and insert them into callus induction culture cultured in the base. The formula of the callus induction medium is: MS medium+1.5mg / L 2,4-D+0.5mg / L 6-BA+30g / L sucrose+7g / L agar, the pH value is 5.8. The culture temperature is 24°C, the light intensity is 2000lx, and the light time is 12h / d. After 20 days, it was observed that white and light yellow callus grew from both ends of the root segment.
[0015] Select light yellow, loose callus and cut it into 0.5cm 2 Insert the small pieces into the subculture medium for subculture of callus. The subculture medium is a solid medium, and its formula is: MS medium + 1.0mg / L 2, 4-D + 0.3mg / L 6 -BA+40g / L sucrose+8g / L agar, the pH value is 5.8. The subculture conditions of callus were...
Embodiment 2
[0020] In the ultra-clean workbench, take out the aseptic seedlings of Caperia spp. seeds from the culture bottle for aseptic sowing and culture for 30 days, cut the roots with sterilized knife tweezers, cut them into small sections with a length of 1.5 cm, and insert them into callus induction cultured in culture medium. The formula of the callus induction medium is: MS medium+1.5mg / L 2,4-D+0.5mg / L 6-BA+30g / L sucrose+7g / L agar, the pH value is 5.8. The culture temperature is 26°C, the light intensity is 1800lx, and the light time is 12h / d. After 25 days, it was observed that white and light yellow callus grew from both ends of the root segment.
[0021] Select light yellow, loose callus and cut it into 0.5cm 2 Insert the small pieces into the subculture medium for subculture of callus. The subculture medium is a solid medium, and its formula is: MS medium + 1.0mg / L 2, 4-D + 0.3mg / L 6 -BA+40g / L sucrose+8g / L agar, the pH value is 5.8. The subculture conditions of callus wer...
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