摘要:
以油桃茎段为试验材料进行组织培养 ,诱导不定芽的增殖和进行愈伤组织诱导 ,获得茎段、叶片、愈伤组织等大量无菌材料 ,给原生质体制备及后续细胞融合工作提供了可重复性的试验材料 .初始培养的最适培养基为 :MS+1.5 mg/L BA+0.1 mg/L IBA ,诱导率达82.9% ;诱导不定芽增殖的最适培养基配方为MS+1.5 mg/L BA+0.2 mg/L IBA ;诱导愈伤组织的最佳培养基配方为MS+1.5 mg/L 2.4-D+0.5 mg/L NAA+0.3 mg/L BA .原生质体制备以嫩叶为分离原生质体的最佳材料 ,而最佳的分离酶组合为纤维素酶1% +果胶酶2% ,在此条件下 ,原生质体的产量和活性分别为9.2 × 107 个/g和80.4% .
Abstract:
Nectarine stems were used as explants for tissue culture ,and protoplasts from virus-free regenerated plantlets were obtained to be used in subsequent cell fusion .Stem explants were induced on MS medium added with different grow th regulators to produce adventitious buds and calli .Large numbers of stems ,leaves and calli were obtained and these could offer enough reproducible materials for protoplast preparation and cell fu-sion .The result indicated that the best medium for bud differentiation was MS +1.5 mg/L BA+0.1 mg/L IBA .A total of 30 buds were obtained on this medium ,the average induction rate being 82.9% ;the optimal medium for bud multiplication was MS+1.5 mg/L BA+0.2 mg/L IBA ;and the optimal medium for callus induction was MS medium supplemented with 1.5 mg/L 2 ,4-D+0.5 mg/L NAA+0.3 mg/L BA .Tender leaves were the most suitable materials for protoplast preparation ,and the best mixed enzyme was 1% cellu-lose+ 2% pectinase .Under this condition , 9.2 × 107 protoplasts per gram material were obtained , and 80.4% protoplasts were viable .
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