Method for increasing in vitro induced mutation rate of stevia rebaudiana NaN3

Method for increasing in vitro induced mutation rate of stevia rebaudiana NaN3

  • CN 104,351,059 A
  • Filed: 12/05/2014
  • Published: 02/18/2015
  • Est. Priority Date: 12/05/2014
  • Status: Active Grant
First Claim
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1. one kind is improved stevia NaN 3the method of Vitro Mutation mutation rate, is characterized in that, it comprises following sport technique segment:

  • 1. culture medium prescription;

    add 30g/L sucrose, 2mg/L mutagen NaN in MS medium 3, 5 g/L agar and somatotropin 6-BA 1.0 mg/L, NAA 0.2 mg/L;

    2. medium packing;

    to be distributed into by the medium configured by above-mentioned formula in the tissue culture flasks of 250ml and to carry out autoclaving, often liter is packed as 24 bottles;

    3. inoculate;

    on superclean bench, stevia plantlet in vitro cut the stem section into ±

    0.5 cm length band 2 blade with scissors, with tweezers, stem section is inoculated in blake bottle, every bottle graft kind 5 stem sections;

    4. condition of culture;

    the tissue culture bottle inoculated is put into group training room and cultivate, cultivation temperature 25 ±

    1 DEG C, intensity of illumination ±

    1500 lx, light application time 12h/d;

    5. transit time;

    when two blades are withered in the stevia stem section top in medium and stem section, stem section moved into not add in the MS medium of any hormone and cultivate, condition of culture is the same;

    6. mutation rate detects;

    in stem section after axillalry bud restoration ecosystem two weeks, and get individual plant and carry out DNA extraction and SRAP Molecular Detection, mutation rate is up to 55-60%.

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