Method for increasing content of alkaloid in lycoris radiata through exogenous substance
A technology of alkaloids and exogenous substances, applied in botany equipment and methods, plant regeneration, horticultural methods, etc., can solve problems such as shortage
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Embodiment 1
[0038]Select 3-month-old tissue-cultured seedlings of Lycoris in uniform growth state, inoculate them on the liquid medium of MS+6-BA 3.0mg / L+NAA 1mg / L+30g / L sucrose, and fill each bottle with 20ml of medium. Add 0.22 μm filter-sterilized 4'-O-methylnoramaryllin (MN) solution to the medium, and set the final concentrations to 0.1 g / L MN (T1) and 0.2 g / L MN (T2), respectively. Each bottle of liquid medium was inoculated with 2g of Amaryllis lycoris tissue culture seedlings, and each liquid medium with different treatments needed to inoculate 3 bottles, with 3 replicates, at 23°C, 50μmol / m 2 / s light intensity for 12 hours, the oscillation frequency is 100r / min. They were taken out after 7d, 14d and 30d of culture in liquid medium.
Embodiment 2
[0040] Select Lycoris annual tissue culture seedlings with uniform growth state and inoculate them on the liquid medium of MS+6-BA 3.0mg / L+NAA 1mg / L+30g / L sucrose, and fill each bottle with 20ml of medium. Add 0.22 μm filter-sterilized 4'-O-methylnoramaryllin (MN) solution to the medium, and set up 3 treatments so that the final concentrations are 0g / L (T0), 0.1g / L MN (T1), 0.2g / L MN(T2), each bottle of liquid medium was inoculated with 2g long-tube Lycoris tissue culture seedlings, each liquid medium with different treatments needed to inoculate 3 bottles, set 3 replicates, at 25°C, 50μmol / m 2 Under the light intensity of / s light for 12 hours, the oscillation frequency is 110r / min. After being cultured in liquid medium for 7d, 14d and 30d, they were taken out, and the contents of three alkaloids, galantamine, ricolamine and lycorine, were determined by a chromatograph connected in series with a secondary mass spectrometer.
[0041] The measurement results are shown in Tab...
Embodiment 3
[0049] Select 1-year-old tissue culture seedlings of Amaryllis lycoris with uniform growth state, inoculate them on the liquid medium of MS+6-BA 3.0mg / L+NAA 1mg / L+30g / L sucrose, and fill each bottle with 20ml of medium. Add 0.22 μm filter-sterilized methyl jasmonate (MeJA) solution to the medium, and set the final concentrations as 75 μmol / L MeJA (MJ-75), 150 μmol / L MeJA (MJ-150) and 300 μmol / L MeJA (MJ -300), each bottle of liquid medium was inoculated with 2g of Amaryllis lycoris tissue culture seedlings, each liquid medium of different treatment needs to inoculate 3 bottles, set 3 repetitions, at 27°C, 50μmol / m 2 / s light intensity for 12 hours, the oscillation frequency is 120r / min. After being cultured in liquid medium for 7d, 14d and 30d, they were taken out, and the contents of three alkaloids, galantamine, ricolamine and lycorine, were determined by a chromatograph connected in series with a secondary mass spectrometer.
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