Obtained transgenic wheat expressing pac1 mediated by Agrobacterium is resistant against Barley yellow dwarf virus-GPV  

作　　者：YAN Fei ZHENG Yinying ZHANG Wenwei XIAO Hong LI Shifang CHENG Zhuomin 

机构地区：[1]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Chinese Academy of Agricultural Sciences, Beijing 100094, China

出　　处：《Chinese Science Bulletin》2006年第19期2362-2368,共7页

摘　　要：In fission yeast (Schizosaccharomyces pombe), pac1 gene was cloned with 99.3% nucleo- tide sequence similarity with published pac1 in GenBank. In pET-5α expression system, the expres- sion product of cloned pac1 in E. coli showed activity to degrade the double-strand RNA. Harboring the binary vector pBI121, which contains pac1 gene, Agrobacterium tumefaciens strain LBA4404 was used to transform the wheat immature embryos pre- cultured 7―10 d. After preregeneration, regeneration and selection culture stage, totally 41 G418 resistant plants were obtained, in which 25 lines were proved to integrate with transgene and express transgene normally by PCR, Dot blot, RT-PCR and ELISA de- tection. Antivirus test carried out on 25 positive lines with high dose of Barley yellow dwarf virus-GPV re- vealed that 12 lines had resistance to BVDV-GPV in low level, another 12 lines had resistance to BVDV- GPV in middle level, and 1 line showed resistance to BVDV-GPV in high level. However, both low and middle level of resistance plants showed no symp- toms when infected by viruses at low dose, which suggested the dose-dependent effect of the resis- tance mediated by pac1 to BYDV-GPV.In fission yeast （Schizosaccharomyces pombe）, pacl gene was cloned with 99.3% nucleotide sequence similarity with published pacl in GenBank. In pET-5α expression system, the expression product of cloned pacl in E. coil showed activity to degrade the double-strand RNA. Harboring the binary vector pBI121, which contains pacl gene, Agrobacterium tumefaciens strain LBA4404 was used to transform the wheat immature embryos precultured 7-10 d. After preregeneration, regeneration and selection culture stage, totally 41 G418 resistant plants were obtained, in which 25 lines were proved to integrate with transgene and express transgene normally by PCR, Dot blot, RT-PCR and ELISA detection. Antivirus test carried out on 25 positive lines with high dose of Barley yellow dwarf virus-GPV revealed that 12 lines had resistance to BVDV-GPV in low level, another 12 lines had resistance to BVDV- GPV in middle level, and 1 line showed resistance to BVDV-GPV in high level. However, both low and middle level of resistance plants showed no symptoms when infected by viruses at low dose, which suggested the dose-dependent effect of the resistance mediated by pacl to BYDV-GPV.

关 键 词：转基因小麦 pac1 土壤杆菌 抗病毒素 大麦黄矮病 

分 类 号：S435.12[农业科学—农业昆虫与害虫防治]