大麦转玉米淀粉分支酶基因的初步研究  

Obtainment of transgenic barley with amylopeetase genes(sbe1 and sbe2b) from maize

作　　者：郭晓琳[1] 孙东发[1] 谭振波 

机构地区：[1]华中农业大学植物科技学院,湖北武汉430070 [2]大北农农业科技研究院,北京100085

出　　处：《西南农业学报》2005年第5期638-642,共5页Southwest China Journal of Agricultural Sciences

基　　金："863"项目(2002AA241201)资助

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sbe1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.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 transformants of 87-3175,9 independent transformants of 87-0053,3 independent transformants of 97-4010 and 5 independent transforrnants of 208813-509, were transplanted to field. Only 4 independent transfomaants of 87-3175,5 independent transformants of 87-0053 and 3 independent transformants of 208813-509 were transplanted successfully. PCR analysis of 10 regenerated plants showed that 7 plants produced 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, she 2b and sbel respectively. It indicated that genes bar, sbe2b and sbel had been integrated into barley genome.

关 键 词：大麦 淀粉分支酶 sbel sbe2b 基因枪 遗传转化 

分 类 号：S512.3[农业科学—作物学]