Rapid Propagation Technique For Tetraploid Forsythia By Tissue Culture

In the present study, using the health and young shoots, leaves of tetraploid forsythiaas experimental material, established a tissue culture system of tetraploid forsythia. Thetest should include: disinfected explants, the primary culture, subculture, rooting andacclimatization, systematically studied the factors that affecting the tissue culture oftetraploid forsythia. The purpose of the test is conducive to industrialize breeding, andattempt to provide techniques and methods of biotechnology breeding tetraploid forsythia,further genetic manipulation studies and lay the foundation to expand its medicinalresources at the same time. The main contents and results are as follows:(1) explants disinfectionIn the study, using the shoot tips, stems and young leaves of tetraploid forsythia asexplants, to explore the effects of different microbicide. Selecting75%ethanol and2%sodium hypochlorite to disinfect. The results showed:2%sodium hypochlorite fordisinfection treatment duration tetraploid forsythia explant play a key role, needing strictlycontrol. The best approach for shoot tips and stems explants is75%alcohol treatment30s,2%sodium hypochlorite7min. The best way to deal with leaf explants tetraploid forsythiais75%ethanol treatment30s,2%sodium hypochlorite3min.(2) plant in vitro cultureIn the study, using shoot tips and stem segments with axillary buds of tetraploidforsythia as explants, to explore the effects of plantlets with different plant hormones6-BA,NAA concentrations of different ratios in vitro culture. The test results showed that:tetraploid forsythia stems should be chosen as explants in tissue culture. The suitablemedium for forsythia tetraploid axillary induced is MS+6-BA2.0mg/L+NAA0.2mg/L. The suitable medium formulation for stems, shoot tips proliferation of tetraploidforsythia is MS+6-BA9.0mg/L+NAA0.05mg/L.(3) callus cultureIn the study, using young leaves of tetraploid forsythia as explants, to explore theeffects of callus induction and proliferation with different plant hormones6-BA,2,4-D,and the ratio of different concentrations of6-BA and NAA. The test results showed that: The suitable medium for tetraploid forsythia leaves callus induction was MS+6-BA1.0mg/L+2,4-D1.0mg/L. According to the later observation showed that: leaf callus growslowly in the medium with different plant hormones and different the ratio of differentconcentrations, the color is yellow-green or dark green, and no adventitious buds producewere similar with small protrusions, after a period of time to continue, leaf callus werewither and die.(4) rooting cultureIn the study, using the single bud through induced proliferation of cultured as trainingmaterials to carry on rooting culture, to explore the effects of rooting culture with differentplant hormones IBA, IAA and the ratio of different concentrations of IBA and IAA. Thetest results showed that: in the test range, the suitable medium for rooting induction oftetraploidy forsythia is1/2MS+IBA0.4mg/L+IAA0.4mg/L, in this condition,rooting is good.(5) the acclimatization of plantletsIn the study, using the rooted and domesticated plantlets as transplanting materials, toexplore the effects of tetraploid forsythia survival rate with different time and differentmatrix. The test results showed that: the survival rate of transplanting plantlets existsignificant differences between hardening treatment and non-hardening treatment, and thesurvival rate of transplanting plantlets with hardening treatment was significantly higherthan that without hardening treatment. The suitable hardening treatment of tetraploidforsythia transplanting plantlets is opening mouth3d and open bottle of natural light4d.The suitable matrix of tetraploid forsythia transplanting plantlets is nutrition soil:vermiculite=3:1, at this time, the transplanting plantlets has dark green leaves, grow well.