正交设计在欧洲菘蓝组织培养中的应用

Application of orthogonal design in tissue culture of Isatis tinctoria

以欧洲菘蓝下胚轴和子叶为外植体,应用正交设计方法,对愈伤组织的诱导和分化培养基进行了优化筛选.分析结果表明,6-BA和2,4-D的交互作用对下胚轴愈伤组织的诱导有显著影响,结合直观分析确定下胚轴的愈伤组织最佳诱导培养基为MS+6-BA 0.5 mg.L-1(后同)+2,4-D 0.5;6-BA和NAA的交互作用对子叶愈伤组织诱导有极显著影响,结合直观分析确定子叶最佳诱导培养基为MS+6-BA 0.5+NAA 0.2.在不同培养基对欧洲菘蓝愈伤组织分化能力的影响试验中,各种因子在所试浓度范围内的作用均不显著,有待进一步试验.根据直观分析我们认为MS+6-BA 2+NAA 1诱导的愈伤组织更有利于分化.不定芽在转移到MS+NAA 0.5一个月后80%以上都能长出很好的根系.生根后的组培苗经过3天炼苗后移栽至花盆后80%以上可以存活.

Orthogonal design was used to study callus induction and and differentiation from cotyledons and hypocotyls of I. tinctoria . Our results indicated that the interaction between 6 - BA and 2,4 - D had significant effect on the callus induction from hypocotyls and the best medium for callus induction from hypocotyls was MS ＋ 6 - BA 0.5 mg·L^-1 （ the same as follows） ＋ 2, 4 - D 0.5. Interaction between 6 - BA and NAA had highly significant effect on callus induction from cotyledons and the best medium for callus induction from cotyledons was MS ＋ 6 - BA 0.5 ＋ NAA 0.2. The hormone concentrations tested had no significant effect on the regeneration ability of the calli and further experiment needs to be carried out. Based on direct analysis we suggested that calli induced on MS ＋ 6 - BA2 ＋ NAA1 were easier to be regenerated. The shoots regenerated were induced to form roots on MS ＋ NAA0.5 at a frequency higher than 80 %.