金钗石斛类原球茎诱导及增殖的正交试验

The orthogonal test of induction and proliferation of Dendrobium nobile protocrom-like bodies (PLBs)

通过正交试验,筛选了金钗石斛Dendrobiumnobile类原球茎(protocrom likebodies,PLBs)诱导及增殖适宜的培养基.以1/2MS附加8g·L-1琼脂为基本培养基,诱导以NAA、TDZ(噻二唑苯基脲,thidiazuron)、CPPU[N(2氯4吡啶基)N1苯脲]做三因子四水平试验,增殖则以NAA、BA、蔗糖、pH值做四因子四水平试验,根据L16(45)正交表设计,分别以PLBs诱导率和增质量率为指标,筛选了适宜于PLBs诱导和增殖的培养基.结果显示:1/2MS+30g·L-1蔗糖+8g·L-1琼脂+0.01mg·L-1TDZ+0.005mg·L-1CPPU,pH5.6的培养基适宜于PLBs诱导,1/2MS+20g·L-1蔗糖+8g·L-1琼脂+20g·L-1香蕉汁+1.0mg·L-1NAA+0.2mg·L-1BA,pH5.6的培养基增殖效果较好;诱导率和增质量率分别为46.00%和756.74%.在各因子中,NAA对PLBs诱导率的影响最大,并对PLBs的诱导起抑制作用;蔗糖质量浓度对PLBs增质量率影响最大,且较低的质量浓度更为适宜;筛选出的PLBs诱导和增殖培养基经重复试验,结果相对稳定.

Medium of induction and multplication of Dendrobium nobile protocrom-like bodies (PLBs) was screened out by orthogonal test. Added 8 g·L^(-1) agar in the basic medium of 1/2 MS, NAA, TDZ and CPPU were selected as factors in PLBs induction, NAA, 6-BA, sucrose and pH value were selected as factors in proliferation. Experiment was designed by the orthogonal test table of L_(16)(4~5). The results showed that the optimal medium for induction was 1/2 MS+30 g·L^(-1) sucrose+8 g·L^(-1) agar + 0.01 mg·L^(-1) TDZ+0.005 (mg·L^(-1)) CPPU and 1/2 MS+1.0 mg·L^(-1) NAA+0.2 mg·L^(-1) BA+20 g·L^(-1) sucrose+8 g·L^(-1) agar+20 (g·L^(-1)) banana juice for proliferation. The rate of induction was 46.00 % and muitiplication was 756.74 %. NAA had inhibtory effect on PLBs induction and sucrose concentration was the most important factor in PLBs proliferation. The effect of selected medium was confirmed by the repeated experments.