摘要
实验构建了CaMV35S启动子控制下的大豆抗病相关基因SR1的正反义植物双元表达载体pBISR1(+)和pBISR1(-)。通过根癌农杆菌叶盘转化法,将正义和反义SR1 基因导入烟草Ha vana 425,经卡那霉素筛选,获得了抗性植株。经PCR和PCR-Southern印迹分析,证明抗性植株中整和了SR1基因,RT-PCR分析进一步表明正义和反义基因皆能转录为完整的mRNA,经疫霉根腐接种及抗病性鉴定表明,转反义基因株系和未转基因株系均轻微感病,而转正义基因株系始终没有出现感病症状。
Two plant expression vectors carrying sense or antisense soybean resistance-related gene SR1 under the regulation of cauliflower mosaic virus 35s promoter was constructed. Leaf segments of tobacco Havana 425 were infected by Agrobacterium tumefaciens LBA4404 with pBISR1(+) or pBISR1(-), from which kanamycin resistant plants were obtained. PCR and PCR-Southern analysis proved that the SR1 gene was integrated into the genomes of the tobacco plants, and RT-PCR analysis proved that sense or antisense gene was transcripted into a complete mRNA. The disease resistance assay showed that plants with antisense gene and control plants were slightly susceptible, and plants with sense gene were not susceptible.
出处
《大豆科学》
CAS
CSCD
北大核心
2005年第2期90-94,共5页
Soybean Science
基金
863计划(2003AA207060-4)资助项目
