Culture method for synthesis and release of glucoraphanin and sulforaphane in hairy roots of broccoli
A kind of technology of sulforaphane and culture method, applied in the field of broccoli hairy root culture system
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Embodiment 1
[0030] Example 1 synthesis and release culture method of high glucoraphanin and glucoraphanin from hairy root of broccoli:
[0031] PCR was used to detect the gene of sterilized hairy root of broccoli. rol B and rol C , select exist. rol B and rol C Hairy root of the gene was used as raw material, which was inoculated into MS liquid culture medium with pH=6 and concentration of 0.1 mg / L according to the inoculation amount of 0.5 g / L, cultured at 25℃ and constant temperature of 110 r / min for 14 days, and then melatonin solution with concentration of 10 mmol / L was added according to the dosage of 20 μ mol / L., After 18 days of culture, it was found that the biomass of hairy roots in each bottle (300mL) was 4.5g dry weight, and the multiplication ratio was 50.7. The contents of glucoraphanin and glucoraphanin in hairy roots were 0.3 mg / g dry weight and 0.1mg / g dry weight, respectively, and the contents of glucoraphanin and glucoraphanin in culture medium were 61.5 mg / bottle and 7.3 ...
Embodiment 2
[0040] Example 2 synthesis and release of glucoraphanin and glucoraphanin from hairy root of broccoli culture method: PCR was performed on sterilized hairy root of broccoli to detect gene. rol B and rol C , select exist. rol B and rol C Hairy root of gene was used as raw material, which was inoculated into MS liquid culture medium containing methionine at pH=6 and concentration of 0.5 mg / L according to the inoculation amount of 0.5 g / L, and then cultured at 25℃ and constant temperature of 110 r / min for 16 days, and melatonin solution with concentration of 10mmol / L was added at the dosage of 60 μ mol / L., After 18 days of culture, it was found that the biomass of hairy roots in each bottle (300mL) was 6.2 g dry weight, the multiplication ratio was 70.4, the contents of glucoraphanin and glucoraphanin in hairy roots were 0.2 mg / g dry weight and 0.3 mg / g dry weight, and the contents of glucoraphanin and glucoraphanin in culture medium were 81.7 mg / bottle and 9.5 mg / bottle, respectiv...
Embodiment 3
[0043] Example 3 synthesis and release of glucoraphanin and glucoraphanin from hairy root of broccoli culture method: PCR was performed on sterilized hairy root of broccoli to detect gene. rol B and rol C , select exist. rol B and rol C Hairy root of the gene was used as raw material, which was inoculated into MS liquid culture medium with pH=6 and concentration of 0.3 mg / L according to the inoculation amount of 0.5 g / L, cultured at 25℃ and constant temperature of 110 r / min for 15 days, and then melatonin solution with concentration of 10mmol / L was added according to the dosage of 40 μmol / L, After 18 days of culture, it was found that the biomass of hairy roots in each bottle (300mL) was 3.4 g dry weight, and the multiplication ratio was 40.3. The contents of glucoraphanin and glucoraphanin in hairy roots were 0.2 mg / g dry weight and 0.2 mg / g dry weight, respectively. The contents of glucoraphanin and glucoraphanin in culture medium were 47.2 mg / bottle and 5.2 mg / bottle, respect...
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