【作者】 郭晓琳； 孙东发； 谭振波；

【Author】 GUO Xiao-lin~1 SUN Dong-fa~(1*) TAN Zheng-bo~2 1.Huazhong Agricultural University,Hubei Wuhan 430070,China; 2.Dabeinong Academy of Agricultural Sicences,Beijing 100085,China

【机构】 华中农业大学植物科技学院； 大北农农业科技研究院；

【摘要】 以5个大麦品种的由成熟胚再生体系产生的胚性愈伤组织为受体材料,以构建好的 Bar 基因为选择基因的玉米淀粉分支酶基因表达载体为目的基因,用基因枪法对其进行了转化。在转化的大麦中,5个不同基因型品种的抗性愈伤获得率为10．32% ～17．13%,将抗性愈伤组织转移到分化培养基中进行分化,绿苗分化率为0%～14．29%,移栽到小花盆中的再生植株有28株。其中87-3175有11株,87-0053有9株,97-4010有3株,97-6004未分化出苗,208813-509有5株;移栽成活的87-3175有4株,87- 0053有5株,208813-509有3株。对10株再生植株进行了 PCR 检测,其中有7株扩增出0．5kb 的 Bar 基因特异条带,2株扩增出 2．4 kb 的 sbe2b 基因特异条带,3株扩增出2．5 kb 的 shel 基因特异条带。对 PCR 扩增的条带回收测序,测序结果与各自的基因序列相符合,说明外源基因已经整合到大麦基因组中。更多还原

【Abstract】 On the base of barley regeneration system,we transformed 5 barley cultivars with expression vectors carrying sbel,sbe2b gene and bar marker gene by particle bombardment.The results were as follow,the resistant calli rates of 5 barley cultivars were 10.32%- 17.13%,green plantlets rate were 0%-14.29%.Finally,28 regenerated independent transformants,including 11 independent trans- formanta of 87-3175,9 independent transformants of 87-0053,3 independent transformants of 97-4010 and 5 independent transformants of 208813-509,were transplanted to field.Only 4 independent transformants of 87-3175,5 independent transformants of 87-0053 and 3 in- dependent transformants of 208813-509 were transplanted successfully.PCR analysis of 10 regenerated plants showed that 7 plants pro- duced the same size of 0.5 kb specific DNA band as that produced from the vector p35 Scpbar by PCR,2 plants had the same size of 2.4 kb specific DNA band as that produced from vector prEBE2 by PCR,3 plants had the same size of 2.5 kb specific DNA bands as that from vector prEBE1 by PCR.The sequences of the PCR bands were identified with that of genes bar,sbe 2b and sbe1 respectively.It indicated that genes bar,sbe2b and sbe1 had been integrated into barley genome.更多还原