Study On Tissue Culture Propagation Of Pulsatill Dahurica(Fisch.) Spreng

The aseptic seedling was obtained through tissue culture of seed explants inpulsatilla dahurica first. Then the induction of callus, the induction and proliferationof adventitious buds, the induction of adventitious buds and roots as well as thetransplantation of hardening-seedling were conducted on the leaves, petioles and budsof aseptic seedling as explants. In this way, the rapid propagation system of pulsatilladahurica in vitro was established. To expand the utilization of pulsatilla dahuricaresources, this dissertation built the optimal propagation system to provide theoreticalbasis for its industrialized production through studying the tissue culture of pulsatilladahurica.1.Sterilization of explantsThe experiment results showed that the germination rate of pulsatilla dahuricaseed sterilization at4min reached the highest of48.67%, and it was found that themost suitable culture medium for pulsatilla dahurica seeds was MS+0.2mg/L IAA.2. Induction of callusThe most suitable culture media for pulsatilla dahurica leave callus inductionwere MS+0.15mg/L2and4-D+1.0mg/L6-BA, whose induction rate reached96.7%.As for the callus induction of pulsatilla dahurica petioles, the optimal culture mediafor pulsatilla dahurica petiole callus induction were MS+0.2mg/L2and4-D+0.5mg/L6-BA, whose induction rate was86.7%. Besides, the optimal culture media forpulsatilla dahurica buds callus induction were MS+0.3mg/L2and4-D+0.1mg/L6-BA,whose induction rate reached80.0%.3.Subculture and propagation of pulsatill dahurica（Fisch．）SprengThe most appropriate culture medium for the callus induction of pulsatilladahurica adventitious buds was MS+6-BA1.5mg/L+0.2mg/L NAA, whose inductionrate was62.47%. When sucrose was30g, the adventitious buds grew best and robustly.The culture medium of MS+1.5mg/LIAA+0.5mg/L ZT was optimal for thepropagation of pulsatilla dahurica adventitious buds with the multiplicationcoefficient of3.08. The optimal bud quantity for adventitious buds was three or four,and the optimal planting quantity were three or four. On the other hand, the optimalsubculture cycle for the propagation of adventitious buds was20d and the bestsubculture times were5generations. 4. Rooting and strengtheningThe optimal culture medium for pulsatilla dahurica tube seedling wasMS+2.0mg/LNAA, whose rooting rate was80.0%. The most appropriate sucroseconcentration for pulsatilla dahurica tube seedling was25mg/L.5.Transplantation of hardening-seedlingThe optimal transplantation time was3d. When turf: garden soil=1:1, thepulsatilla dahurica grew best with green leaves and robust roots.