Tissue culture breeding method for clonic fibraurea stem

Tissue culture breeding method for clonic fibraurea stem

  • CN 1,526,278 A
  • Filed: 09/23/2003
  • Published: 09/08/2004
  • Est. Priority Date: 09/23/2003
  • Status: Active Application
First Claim
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1. , herba fibraureae recisae clonal tissue culture propagation method is characterized in that:

  • 1) coppice spout explant collection prevents shattering of the sprout wound and the bud heart in the process of gathering the coppice spout explant, and to make the height of coppice spout explant be 15-40cm;

    2) sterilization of explant is adopted sterilization twice, at first, with washing powder explant is cleaned up, peel off 1-2 layer leaf sheath, with 75% alcohol disinfecting 15-20 second, 0.1% mercuric chloride sterilization 8-10 minute, rinsed with sterile water 1 time 5 minutes, second step;

    peel off unnecessary leaf sheath and be trimmed to the size that is suitable for cultivating, promptly coppice spout explant 3-4cm is long, terminal bud explant 5-6cm is long, again with 0.1% mercuric chloride sterilization 1-2 minute, rinsed with sterile water 3 times was respectively 5 minutes, 5 minutes, 10 minutes;

    3) prevent brown stain, adopt cysteine or ascorbic acid solution bubble to wash aforementioned coppice spout explant, or adopt cysteine or ascorbic acid mixed solution bubble to wash aforementioned coppice spout explant, can suppress the brown stain of bud explant effectively, the concentration of described cysteine and ascorbic acid is respectively 2mg/l and 10mg/l;

    4) the clump bud is induced and shoot proliferation, and the clump bud induces the culture medium prescription with shoot proliferation to be;

    KNO3, NH4NO3, CaCl2, MgSO4 and KH2PO4, and its proportioning is respectively;

    1.5MS, 0.5MS, 1.0MS, 1.0MS, 2.5MS;

    Clump bud induction duration 35~

    40 days, the cycle of successive transfer culture is 45~

    50 days;

    5) culture of rootage, height is fit to culture of rootage the most greater than the bud seedling of 2cm, root media is 1.0mg/L NAA+2.0mg/L IBA+0.33MS macroelement+20.g/L sucrose solution+1.0MS trace element+1.0MS vitamin and amino acid, the time of culture of rootage needs 50~

    60 days, after the bud seedling of not taking root removed outer leaf sheath, carry out the culture of rootage second time.

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