摘要
该实验通过RT-PCR获得了番茄八氢番茄红素脱氢酶(Phytoene desaturase,PDS)基因的部分序列,双酶切PDS片段和烟草脆裂病毒载体(pTV00),构建重组载体pTV00-PDS,经农杆菌GV3101介导侵染番茄叶片并观察植株表型变化。结果显示,被侵染的番茄表现出明显的光漂白现象。半定量RT-PCR检测表明,PDS的mRNA被显著降解。该沉默体系的建立为下一步大规模验证番茄基因功能奠定了基础。
The tomato phytoene desaturase （PDS） gene fragment was obtained by using RT-PCR from tomato. Tobacco rattle virus vector （pTV00） and PDS fragment were digested by double restriction enzymes and recombinant pTVOO-PDS vector was constructed. The constructed vector was transformed into Agrobacterium GV3101 to further inoculate tomato plants. The infected plants showed obvious photobleaching symptom,while semi-quantitative RT-PCR result demonstrated that the decline of PDS mRNA redundancy during infection. The established VIGS system will benefit the high-throughout function analysis of tomato genes in the following experiments.
出处
《西北植物学报》
CAS
CSCD
北大核心
2009年第8期1531-1537,共7页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金项目(30600413)
教育部科学技术研究重点项目(207064)
留学回国人员科研启动基金(教外司留[2007]1108号)
河南高校杰出人才科研创新工程项目(2007KYCX017)
关键词
番茄
八氢番茄红素脱氢酶
病毒诱导基因沉默
烟草脆裂病毒
omato（Solanum esculentum）
phytoene desaturase
virus induced gene silencing
tobacco rattle virus
