Method for industrial large-scale production of flavones-enriched Tianshan Tianshan saussurea involucrate culture
A kind of snow lotus culture and large-scale technology, applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of harsh growth conditions, low reproduction rate, and insufficient resources of Tianshan snow lotus, and achieve adaptation to normal temperature Effects of growth conditions, short production cycle, and stable traits
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Embodiment 1
[0020] (1) Use Tianshan snow lotus seeds to induce aseptic seedlings after sterilization; the sterilization steps include soaking in 75% ethanol for 30 seconds, washing with sterile water twice, soaking in 0.1% mercury chloride solution for 15 minutes, and washing with sterile water for 3 Inoculate on MS solid medium, light 2000Lux, 12h / d.
[0021] (2) Transfer the sterile seedlings (leaves, roots, stems) to MS-added phytohormone medium to induce callus; the induction medium is 0.5mg / L 6-BA+0.5mg / LNAA+MS+ 200g / sucrose+5.6g / L agar, the culture condition is light 2000Lux, 12h / d.
[0022] (3) Select the callus with the darkest color, close to purple red, and grow well, and subculture it on the screening medium to obtain purple, fast-proliferating, and stable Saussurea culture A; the screening medium is 0.5mg / L 6-BA+0.8mg / LNAA+MS+200g / sucrose+5.6g / agar, the culture condition is light 2000Lux, 12h / d.
[0023] (4) Put Saussurea culture A at 10°C for 20 days, subculture on the scre...
Embodiment 2
[0026] (1) Use Tianshan snow lotus seeds to induce sterile seedlings after sterilization; the sterilization steps include soaking in 75% ethanol for 30 seconds, washing with sterile water three times, soaking in 0.1% mercuric chloride solution for 15 minutes, and washing with sterile water for 5 minutes. Inoculate on MS solid medium, light 3000Lux, 10h / d.
[0027] (2) Transfer the sterile seedlings (leaves, roots, stems) to MS-added phytohormone medium to induce callus; the induction medium is 2.5mg / L6-BA+2.5mg / LNAA+MS+250g / L sucrose+6g / L agar, the culture condition is light 3000Lux, 10h / d.
[0028] (3) Select the callus with the darkest color, close to purple red, and grow well, and subculture it on the screening medium to obtain the purple, fast-proliferating, and stable snow lotus culture A; the screening medium is 2mg / L 6 -BA+2.5mg / LNAA+MS+250g / L sucrose+6g / L agar, the culture condition is light 3000Lux, 10h / d.
[0029] (4) Put Saussurea culture A at 15°C for 15 days, s...
Embodiment 3
[0032] (1) Use Tianshan snow lotus seeds to induce sterile seedlings after sterilization; the sterilization steps include soaking in 75% ethanol for 30 seconds, washing with sterile water twice, soaking in 0.1% mercury chloride solution for 15 minutes, and washing with sterile water for 4 minutes. Inoculate on MS solid medium, light 3000Lux, 11h / d.
[0033] (2) Transfer the sterile seedlings (leaves, roots, stems) to MS medium added with phytohormones to induce callus; the induction medium is 1mg / L 6-BA+2mg / LNAA+MS+250g / L sucrose + 5.8g / L agar, the culture condition is light 2800Lux, 11h / d.
[0034] (3) Select the callus with the darkest color, close to purple red, and grow well, and subculture it on the screening medium to obtain the purple, fast-proliferating, and stable snow lotus culture A; the screening medium is 2mg / L 6 -BA+2mg / L NAA+MS+230g / L sucrose+5.8g / L agar, the culture condition is light 2500Lux, 11h / d.
[0035] (4) Cultivate Saussurea culture A at 13°C for 18 ...
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