Method for preserving hemerocallis fulva tissue culture seedlings at low temperature

Method for preserving hemerocallis fulva tissue culture seedlings at low temperature

  • CN 105,766,651 A
  • Filed: 04/14/2016
  • Published: 07/20/2016
  • Est. Priority Date: 04/14/2016
  • Status: Active Application
First Claim
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1. the method for a Hemerocallis fulva L. tissue cultured seedling cryopreservation, it is characterised in that follow these steps to intoOK:

  • (1) by the bennet of Hemerocallis fulva L. 0.5-0.8g/L liquid detergent aqueous cleaning surface,Again by tap water flow wash 30min, the most aseptically with denseDegree is the ethanol surface sterilization 30s of 75%, then goes out in 0.1% mercuric chlorideBacterium 10-15min, then uses aseptic water washing 4-5 time.(2) the Hemerocallis fulva L. bennet after described step (1) processes is cut into cutting of 1cmThe outer implant of Duan Zuowei, is inoculated on calli induction media and carries out induction trainingSupport 30d, obtain callus.(3) callus in described step (2) is inoculated in division culture medium enterprisingThe induction of row adventitious bud.(4) by the Multiple Buds that induces in described step (3) at described differentiation cultureOn base, subculture rises in value twice, it is thus achieved that Multiple Buds.(5) by the Multiple Buds with wound healing strong for growing way in described step (4),It is positioned in cold closet and carries out low temperature and see that light preserves, after 3 months, move on toIn culturing room, under the conditions of 25 DEG C, carry out slow Seedling successive transfer culture 25-30 days.(6) tissue cultured seedling of the robust growth of subculture in described step (5) is being taken rootCultivate 20 days in culture medium.

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