发根农杆菌介导的大豆DAD基因沉默嵌合植株的获得

Agrobacterium-mediated GmDAD1 Gene Silence of Transgenic Hairy Roots in Chimeric Soybean

利用发根农杆菌介导的转化方法,获得了大豆DAD基因在根组织中沉默的嵌合植株。同时在pHells Gate12载体基础上插入一个GFP表达盒使得对转基因组织的筛选变得更加简便;通过对荧光组织进行q PCR检测发现转基因组织与对照相比,DAD基因转录水平降低45%~70%。以上结果表明大豆DAD基因在转基因根组织中已被成功沉默,并且利用改造后的p Hells Gate12：GFP载体通过荧光可以对转基因组织进行初步快速筛选。

Agrobacterium rhizogenes is able to transform plant genomes and induce the production of hairy roots. In this work,we described the use of A. rhizogenes in soybean to rapidly assess gene expression and function. We first cloned the soybean DAD gene into p Hellsgate12： GFP vector,which was reconstructed by inserted a 35S： GFP： nos expression cassette into Sac I digested p Hells Gate12,then transformed the DAD gene fragment into soybean by using this chimeric hairy root transgenic system. The transcription level of DAD in the fluorescence hairy roots were reduced to 45% ~ 70%. The results revealed that the DAD gene was silenced in transgenic hairy roots,and the reconstructed RNAi vector could simplify the progress of positive hairy roots screen.