早开堇菜组织培养及植株再生体系的建立

Tissue culture in vitro and plant regeneration of Violaprionantha

以野生早开堇菜为材料,研究了不同外植体类型(子叶节、叶片、叶柄)和不同激素配比对其不定芽诱导、愈伤组织诱导和分化的影响,成功建立了早开堇菜组织培养植株再生体系。结果表明,1)诱导子叶节和叶柄不定芽诱导的最适培养基为MS+2.0mg/L 6-BA+0.1mg/L NAA,叶片不适合作为直接诱导不定芽的外植体。2)3种外植体均能诱导愈伤组织,子叶节愈伤组织诱导的时间最短,其次为叶柄,叶片最晚。子叶节和叶柄愈伤组织诱导的最适培养基为MS+1.0mg/L 6-BA+0.5mg/L 2,4-D,诱导率分别为98.3%和96.7%;叶片愈伤组织诱导的最适培养基为MS+1.0mg/L 6-BA+0.5mg/L 2,4-D+0.05mg/L NAA,诱导率可达88.3%。3)最适愈伤组织分化培养基为:MS+1.5 mg/L 6-BA+0.1 mg/L NAA,分化率为100%。4)不定芽增殖的最佳培养基为:MS+1.5mg/L 6-BA+0.075mg/L NAA,增殖率为4.68。5)再生苗移植到添加1.0mg/L 6-BA的1/2 MS培养基上,生根率92.3%。6)组培苗移栽到珍珠岩∶河沙∶腐殖质(1∶2∶2)的混合基质时,100%成活,且植株长势好。

The effects of various hormone ratios on adventitious buds,callus inductions and differentiation were investigated using the cotyledon node,leaf blade and petiole of Viola prionantha as explants.Finally,a plant regeneration system via tissue culture was established.The results showed that the optimal medium for adven-titious bud induction from the cotyledon node and petiole was MS+2.0 mg/L 6-BA+0.1 mg/L NAA,but the leaf blade was not suitable as an explant.Three kinds of explants could induce callus formation.The optimal medium for callus induction from the cotyledon node and petiole was MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D with the highest callus induction rate being 98.3% and 96.7%,respectively.The optimal medium for callus induction from the leaf blade was MS+1.0 mg/L 6-BA+0.5 mg/L 2,4-D+0.05 mg/L NAA with the highest callus induction rate being 88.3%.The best medium for callus differentiation was MS+1.5 mg/L 6-BA+0.1 mg/L NAA with the highest callus differentiation rate being 100%.The optimal medium for the proliferation of adventitious buds was MS+1.5 mg/L 6-BA+0.075 mg/L NAA,the highest proliferation times being 4.68. The rooting rate of regenerated plants was up to 92.3% on half strength MS medium supplemented with 1.0 mg/L 6-BA.Rooted plantlets were grown on mixture with perlite:sand:humus (1:2:2);survival rate was up to 100% with good growth.