Potato stem fragment tissue culture medium and culture method thereof
The technology of a tissue culture medium and a culture method is applied in the field of one-step seedling culture of potato test-tube seedlings and stems, and the field of tissue culture medium of potato stems. The effect of short cycle, reduced economic cost and simple training procedure
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Embodiment 2
[0061] Utilize described potato one-step seedling-forming medium, make potato stem section tissue culture one-step seedling, comprise the following steps: (1), add 1mg 6-benzylaminopurine, 0.01mg naphthaleneacetic acid, 2.5 to every liter of MS medium mg gibberellin, 50 mg inositol, 25 g edible sugar, and 3 g agar; the pH value was adjusted to 5.8; after aliquoting, it was sterilized by high-pressure steam. Put the sterilized culture medium into the inoculation chamber and let it cool down to solidify.
[0062] (2) Select aseptic test-tube seedlings of the Atlantic potato variety with a seedling age of 20-30 days, cut 0.5-1 cm stem segments without axillary buds under sterile conditions, and inoculate them on the above-mentioned medium. Place the inoculated culture bottle on a lighting culture rack for cultivation, the temperature is 21±2°C, the light intensity is 3500LX, and the light time is 16h / d.
[0063] (3) Cultivate for 28 days to obtain regenerated potato plants.
Embodiment 3
[0065] Add 3mg 6-benzylaminopurine, 0.1mg naphthaleneacetic acid, 7.5mg gibberellin, 100mg inositol, 30g edible sugar and 4.5g agar to each liter of MS medium; After the agar is completely dissolved, adjust the pH value to 6.0; after dispensing, high-pressure steam sterilization, steam temperature 120 ° C, pressure 1.5Mpa conditions for 15 to 20 minutes, the sterilized medium is left to cool down and solidify; under sterile conditions , cut 0.5cm ~ 1cm stem section without axillary buds, and inoculate it in the medium of step B; place the medium inoculated with the stem section in step C under the conditions of a temperature of 21±2°C, a light intensity of 4000LX, and a light time of 16h / day Under cultivation; the potato variety LK99 with a seedling age of 20 to 30 days was selected, and the one-step seedling growth medium was prepared and cultivated in the same way as above, and cultivated for 40 days to obtain potato regenerated plants.
Embodiment 4
[0067] Add 3mg 6-benzylaminopurine, 0.01mg naphthaleneacetic acid, 5mg gibberellin, 100mg inositol, 30g edible sugar and 4.5g agar to each liter of MS medium; After complete dissolution, adjust the pH value to 6.0; after subpackaging, high-pressure steam sterilization, steam temperature 120 ° C, pressure 1.5Mpa conditions for 15 to 20 minutes, the sterilized culture medium is cooled and solidified; under sterile conditions, cut out 0.5cm ~ 1cm stem section without axillary buds, inoculated in the medium of step B; the medium inoculated with the stem section of step C was cultured under the conditions of temperature 21±2°C, light intensity 4000LX, and light time 16h / day ;
[0068] The potato variety Longshu No. 7 with a seedling age of 20-30 days was selected, and the one-step seedling growth medium was prepared and cultured in the same way as above, and cultured for 32 days to obtain potato regenerated plants.
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