Research On Optimization Of The Tissue Culture Of Lilium Brownii Var. Viridulum

The experiment were designed by regression optimum design to optimize lilium brownii var.viridulum tissue culture system,and to analyze the effect of each factor on lilium brownii var.viridulum tissue culture,then find the best condition of culture.Explants of experiment were terminal bud, young leaf,stem segment and squama,and the inducement of bud and bulblet, subculture,the inducement of root and prouducing bulb and transplant of tissue culture seedling had done.The results were as follows:1.Initial inducing cultureThe effect of 6-BA and NAA on squama,squama of bulblet,stem segment and leaves were researched by saturation D-optimum design,and the datas were analyzed and multinomial regression equations were established by SAS 8.2,and single effect and interaction effect of 6-BA and NAA had been done,then used Qbasic to optimize,then the best culture were find.The best culture of inducing bulblet of squama is MS+6-BA1.5 mg/L+NAA0.5 mg/L,and the best culture of inducing the fasciculate-buds of squama of bulblet is MS+6-BA2.0 mg/L+NAA0.25 mg/L,and the best culture of inducing the fasciculate-buds of stem segment is MS+6-BA2.40 mg/L+NAA0.25 mg/L,and the best culture of inducing fasciculate-buds of leaves is MS+6-BA2.50 mg/L+NAA0.25 mg/L.By inducing terminal buds,MS and the different concentration 6-BA and NAA as culture,the better culture were found,being MS+6-BA2.0 mg/L+NAA0.2 mg/L.2.The subculture multiplication of budThe experiment found that the culture including 6-BA was the best for subculture multiplication of bud,MS as the basic culture medium,by adding 6-BA,KT and both 6-BA and NAA to the culture.The regression equation were established by Qbasic,then found the best concentration of 6-BA was 1.0 mg/L,and the multiplication coefficient 5.02/30 d at this concentration of 6-BA.3.The culture of inducing root and prouducing bulb1/2MS as the basic culture,the experiment found that,the concentration of sucrose was positively orrelative to producing bulb,and was negatively correlative to root,but the high concentration of sucrose decreased the percent of survival by addition different concentrations of sucrose to culture.The experiment proved that the number of bulb,when the concentration of sucrose is 6%.The experiment,the culture 1/2MS with 6%sucrose were from MS of different concentration KH₂PO₄,found that when the concentration of KH₂PO₄ is 5.(?)g/L,KH₂PO₄ improve the inducing root and prouducing bulb, and decreased the time of producing the bulb,the diameter of bulb is more.The experiment,the culture is mend 1/2MS as the basic culture,added 6%sucrose,was designed by 311-A optimum mixing design of the concentration of NAA,IBA and IAA.The experiment results were used to establish the equation by SAS 8.2,and did the analysis of single effect and interaction effect by Excel,then optimized by Qbasic sofeware.The best culture of inducing root and prouducing bulb is 1/2MS+NAA0.33 mg/L+IBA0.27 mg/L+IAA0.30 mg/L+6%sucrose.