马倩倩;宋跃;孙毅;张绍铃;崔贵梅;吴俊;

• 1:南京农业大学梨工程技术研究中心
• 2:山西省农业科学院生物技术研究中心

摘要(Abstract)：

【目的】建立超声波处理‘黄花’梨花粉介导插入外源基因的技术体系。【方法】以‘黄花’梨花粉为试材,检测不同蔗糖溶液中梨花粉破损率;通过中心组合设计方法,对超声波处理功率、处理时间、间隙时间、处理次数4个因子进行优化;利用含荧光蛋白基因的表达载体pCAMBIA 1304检测转化效率。【结果】梨花粉的适宜等渗溶液为15g.L-1蔗糖溶液。超声波处理的梨花粉受体最佳参数为:超声波处理功率160¹70 W,处理时间6170 W,处理时间6⁷ s,间隙时间7 s,处理次数7次。采用加入含GFP(green fluorescent protein)基因的pCAMBIA 1304表达载体质粒检测优化的超声波处理组合,梨花粉外源基因的插入效率为3%。【结论】研究确定了超声波处理‘黄花’梨花粉遗传转化体系。

关键词(KeyWords)： ‘黄花’梨;花粉介导;超声波处理;转化体系;GFP基因

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(31171928);; 江苏省科技支撑(BE2011320)

作者(Authors): 马倩倩;宋跃;孙毅;张绍铃;崔贵梅;吴俊;

DOI: 10.13925/j.cnki.gsxb.2013.04.012

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