花粉管通道法遗传转化核桃的研究  

Studies on Gene Transformation via Pollen Tube Pathway in Walnut

作　　者：师校欣[1] 杜国强[1] 王晓蔓[1] 裴东[2] 

机构地区：[1]河北农业大学园艺学院,河北保定071001 [2]中国林业科学研究院林业研究所,北京100091

出　　处：《园艺学报》2012年第7期1243-1252,共10页Acta Horticulturae Sinica

基　　金：国家‘863’计划项目(2007AA10Z182);河北省自然科学基金项目(C2009000635;C2012204091)

摘　　要：为了提高核桃花粉管通道法转化成功率,以‘清香’核桃(Juglans regia L.)为试材,研究了外源DNA导入受体时间、DNA浓度和导入方法对导入效率的影响。结果表明:外源DNA滴加处理的适宜时间为授粉后14～20h;外源 DNA 注射处理的适宜时间为授粉后24h。综合考虑外源DNA浓度对坐果率和GUS检测结果的影响,认为350～500μg .mL-1DNA适宜转化。采用切割柱头滴加、柱头直接滴加和子房注射等3种导入外源DNA的方法均获得了转化成功的果实,其中以切割柱头滴加法获得的转化果实率最高(36.4%),其次是柱头直接滴加法(20.7%),子房注射法因易造成机械损伤,坐果率最低(10.9%)。In order to enhance the efficiency of gene transformation via the pollen tube pathway, the timing of DNA inoculation, the concentration of DNA, and the different methods of DNA inoculation were studied in ＇Qingxiang＇ walnut （Juglans regia L.） . The results showed that a period of 14 to 20 hours after pollination was the optimum time for the treatment of dripping exogenous DNA on the pistils. And a period of 24 hours was suitable for the treatment of injection. A DNA concentration of 350 - 500 μg .mL^-1 was optimal for gene transformation according to the fruit set rates and GUS analysis. The fruits carrying the exogenous gene were obtained by using three methods of dripping DNA with or without the pistil cut and of the ovule injection, while the dripping DNA without the pistil cut treatment had the highest transformation rate （36.4%）, followed by the dripping DNA with the pistil cut treatment （ 20.7% ）. The ovule injection treatment had the lowest fruit set rate （10.9%） due to mechanical damage during injection.

关 键 词：核桃 外源DNA 花粉管通道 柱头滴加 转化效率 

分 类 号：S664.1[农业科学—果树学]