A method for obtaining stevia callus
A callus and callus induction technology, applied in the field of stevia callus acquisition, can solve the problems of no report, loss of division ability, etc., and achieve the unique technology, short establishment period, shortened selection and cultivation time. Effect
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Embodiment 1
[0097] This example is used to illustrate the method for obtaining the Stevia rebaudiana callus of the present invention.
[0098] (1) Acquisition of epidermal fragments (containing stomatal guard cells) of Stevia rebaudiana
[0099] Stevia epidermal fragments (containing stomatal guard cells) were obtained as follows:
[0100] Stevia rebaudiana seeds are cultured and germinated in a cultivation medium to obtain stevia rebaudiana sterile seedlings. Select robust stevia aseptic seedling plants with vigorous growth, thick stems and thick leaves, large and flat leaves, and bright green leaves, and transfer them to a new cultivation medium for the subculture of the first subculture cycle (28 days) , then transfer to a new cultivation medium to continue the subculture of the second subculture cycle (28 days), then transfer to a new cultivation medium to continue the third subculture cycle (28 days) ) subculture, select the stevia tissue culture sterile plant that adopts in the pr...
Embodiment 2
[0127] This example is used to illustrate the method for obtaining the Stevia rebaudiana callus of the present invention.
[0128] Except the following operating parameters, all the other are the same as Example 1.
[0129] (1) Acquisition of epidermal fragments (containing stomatal guard cells) of Stevia rebaudiana
[0130] The skimmer operates for 1 second in "low speed" mode and for 30 seconds in "high speed" mode.
[0131] (2) Induction culture
[0132] The induction medium is: based on the B5 basic medium, adding sucrose to a final weight percentage of 3%, adding 6-BA to a final concentration of 0.5 mg / L, adding NAA to a final concentration of 0.5 mg / L, Glutamine was added to a final concentration of 100 mg / L.
[0133] The division rate of stomatal guard cells can reach 3.0%; the number of cells contained in the obtained small cell mass is 10; the rate of planting plate is 0.15%.
[0134] (3) Suspension culture
[0135] After the induced culture product (containing s...
Embodiment 3
[0142] This example is used to illustrate the method for obtaining the Stevia rebaudiana callus of the present invention.
[0143] Except the following operating parameters, all the other are the same as Example 1.
[0144] (1) Acquisition of epidermal fragments (containing stomatal guard cells) of Stevia rebaudiana
[0145] The skimmer operates for 1 second in "low speed" mode and for 30 seconds in "high speed" mode.
[0146] (2) Induction culture
[0147] The induction medium is: based on the B5 basic medium, adding sucrose to a final weight percentage of 3%, adding 6-BA to a final concentration of 5.0 mg / L, adding NAA to a final concentration of 0.05 mg / L, Add glutamine to a final concentration of 200 mg / L.
[0148] The first addition of the liquid medium: the volume ratio of the liquid medium to the original medium is 0.5:10;
[0149] Add liquid medium for the second time: the volume ratio of liquid medium and original medium is 1:10.
[0150] The division rate of sto...
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