| 摘要: |
| 为了优化84K杨的再生和遗传转化体系，以84K杨树叶片为外植体，采用正交试验研究了不同培养基和激素配比对芽和芽苗生根的影响，并从预培养时间、浸染菌液浓度、浸染时间、共培养时间及添加AS（乙酰丁香酮）5个方面，以卡那霉素抗性苗频率为衡量指标，研究了各因素对84K杨遗传转化率的影响。结果表明，84K杨叶片最适的芽诱导培养基及激素配比为MS＋6-BA 0.5 mg/L＋NAA 0.05m/L，芽苗最适诱导生根培养基及激素配比为1/2MS＋NAA 0.05 mg/L＋IBA 0.05 mg/L；优化筛选的最适转化体系为：农杆菌活化菌液用MS液体培养基稀释10倍，浸染5 min，共培养4 d。采用上述优化的再生和遗传转化体系，84K杨每片叶平均生芽数可达20个，芽苗生根率可达99％，生根数为6条，叶盘转化率可达35％。 |
| 关键词: 84 K杨 再生系统 遗传转化 转化率 |
| DOI： |
| 分类号: |
| 基金项目:科技部国家转基因植物研究与产业化专项（JY03-B-26-02） |
|
| Prioritization of the system of regeneration and genetic transformation of Populus 84 K |
|
|
| Abstract: |
| To prioritize the system of regeneration and genetic transformation of Populus 84 K,influences of different medium and proportion of hormone on bud and root of leaf-explant were studied by orthogonal designing.The impact of five aspects(pre-culture time,the concentration of agro-infection,infect time,co-culture time and addition of AS) on genetic transformation frequency of Populus 84 K was studied,with the rate of kanamycin resistance seedling as the measure index.The results demonstrated that the optimal differentiation medium and proportion of hormone for leaf-explant was MS +BA 0.5 mg/L +NAA 0.05 mg/L,and the optimal rooting medium and proportion of hormone for shoot was 1/2 MS +NAA 0.05 mg/L +IBA 0.05 mg/L.The optimal prioritization optimal system was as follows:to use MS dilute activated agro-infection for 10 times,to infect 5 minutes and to co-culture for 4 days.By adopting above prioritization system of regeneration and genetic transformation,every leaf of populus 84 K could be induced 20 buds on average,rooting rate achieved 99%,rooting number was 6,and transforming rate of leaf disc was 35%. |
| Key words: Populus 84 K regeneration system, genetic transformation transforming rate |
