棉花抗枯萎病相关GhB301基因对烟草的遗传转化及功能分析  

Genetic Transformation and Functional Analysis of Tobacco Transformed with Cotton Fusarium wilt Resistance Related GhB301 Gene

作　　者：何兰兰[1] 柴蒙亮[2] 韩泽刚 赵曾强 张薇[1] 

机构地区：[1]石河子大学农学院,新疆石河子832000 [2]石河子大学生命科学学院,新疆石河子832000

出　　处：《新疆农业科学》2015年第4期589-594,共6页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(31260358);农业部转基因生物新品种培育重大专项(2011ZX08005-005);石河子大学科学技术研究发展计划(gxjs2012-yz02)

摘　　要：【目的】ERF是乙烯信号传导途径中的重要转录因子,在植物与病原菌互作中发挥着重要的调控作用。棉花抗枯萎病相关Gh B301基因是从棉花中获得的ERF-B3亚组基因,探索该基因在烟草异位表达后的功能,为进一步研究其在棉花抗枯萎病中的功能奠定基础。【方法】以p PZP35S表达载体为基础,构建Gh B301的过表达载体;采用农杆菌介导法转化烟草并获得转基因植株。【结果】经卡那霉素筛选、PCR和RT-PCR检测,获得3株转基因阳性植株;q PCR分析显示,转基因阳性植株中部分病程相关蛋白基因的表达量较野生型明显升高。【结论】棉花抗枯萎病相关基因Gh B301在烟草中异位表达后可能会通过增强部分病程相关蛋白基因的表达来提高植株抗病性。[Objective] ERF are important transcription factors in the ethylene signal transduction pathway and play an important role in the regulation of plant pathogen interactions. Cotton Fttsarium wilt resistance related GhB301 belongs to ERF - B3 subgroup transcription factor gene. Through the study of GhB301 gene ectopic expression in tobacco, an foundation for the further function analysis of GhB301 in cotton Fusarium wilt resistance will be laid down. [ Method] An expression vector containing GhB301 was constructed on the basic expression vector pPZP35S. Using the method of Agrobacterium mediation, we conducted the genetic transformation of tobacco and transgenic plants were obtained. [ Result] Three transgenic plants were obtained after the screening of Kanamycin, PCR and RT - PCR. qPCR analysis indicated that partial pathogenesis - related proteins genes＇ expression in transgenic plants were obvious higher than those of the wild ones. [ Conclusion ] Dis- ease resistance of transgenie tobacco plants with cotton Fusarium wilt resistance related GhB301 gene could be improved through the enhancement of partial pathogenesis - related proteins genes＂ expression.

关 键 词：GhB301基因 遗传转化 病程相关蛋白基因 实时荧光定量PCR 

分 类 号：S561[农业科学—作物学]