针对目前众多的棉花转基因方法存在转化过程复杂、成本高、效率较低等问题,实验对转基因方法 -喷花法进行优化,明确了在棉花育种科研中应用条件,达到快速、高效转化外源基因的目的。实验选用了5个陆地棉品种(系)和2个海岛棉品种为转化受体材料,转化了2个外源基因-抗虫(Bt+CpTI)和增强纤维品质基因(NPTIIGhCAD6),并对转化时等渗液各成分的比例、转化的时间段、温度及湿度条件等因素对转化率的影响进行研究。结果表明,转化效率最高时等渗液各成分的含量:陆地棉为1/2MS+10%蔗糖+0.5g/L MES+0.01mg/L 6-BA+200μl/L表面活性剂,pH=6菌株浓度OD600=0.5;海岛棉为1/2MS+15%蔗糖+0.5g/L MES+0.01mg/L 6-BA+200μl/L表面活性剂,pH=6菌株浓度OD600=0.5;转化效率最高时转化的时间段为11∶00~13∶30,相应的温度为22~29℃、湿度为70%~85%;转化后的T0代受体材料,首先在田间进行抗生素检测,然后在获得的阳性植株中,转抗虫基因材料用金标BT-Cry I Ab/Ac试纸条定性检测,转纤维品质基因材料做PCR电泳检测,经检测最...
【英文摘要】
Concerning lots of problems about the complex process,high cost and low efficiency in numerous cotton gene transformation methods at present,this experiment was to optimize the application condition of an injection method,and the aims were to clear about the application conditions in the cotton breeding research and achieve rapid and effective transformation methods of exogenous genes. Five upland cotton varieties( or lines) and two sea island cotton varieties were used as transformation materials in this e...
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