摘要
嗜热链球菌γ-谷氨酰半胱氨酸谷胱甘肽合成酶Streptococcus thermophilusγ-glutamylcysteine synthetase-glutathione synthetase(StGCS-GS)是一类新型的非限速谷胱甘肽合成酶,其产物GSH在包括干旱在内的众多非生物逆境中发挥着重要作用。本研究利用Gateway系统构建了p GWB2重组植物表达载体。以农杆菌介导法对甜菜叶柄进行遗传操作,将编码该酶的StGCS-GS基因(Genbank accession no.GQ848551)转化到甜菜体内。研究结果表明,当农杆菌侵染浓度OD600值为0.6,侵染时间10 min,共培养4 d后,甜菜叶柄外植体经过农杆菌侵染,在筛选培养基上所获得的抗性芽比率相对最高。通过对潮霉素抗性植物基因组DNA的PCR检测,研究共获得7个转基因甜菜株系。
Streptococcus thermophilus γ-glutamylcysteine synthetase-glutathione synthetase （ StGCS-GS） is a novel enzyme without limit feedback inhibition involving in biosynthesis of glutathione （ GSH）, which plays an important role in abiotic stresses including drought. In this study, by using Gateway system, we introduced StGCS -GS gene （Genbank accession no. GQ848551） into plant expression vector --pGWB2. Petioles were transformed with aim gene via Agrobacterium-mediated transformation. The results showed that the highest rate of hygromycin resistant bud was acquired when Agrobacterium concentration of OD600 was 0.6, the infection time was 10 min, and co-culture of infection explants was 4 d. A total of 7 transgenic sugar beets were integrated with StGCS-GS successfully through the extraction of genomic DNA and PCR identification.
作者
刘大丽
马龙彪
LIU Da-li;MA Long-biao(Key Laboratory of Sugarbeet Genetics and Breeding,Colleges of Heilongjiang Province / Crop Academy o f Heilongjiang University,Harbin,150080,Heilongjiang;Key Laboratory of Resources and Application of Northern Sugar Crops/ Sugarbeet Research Institute,Chinese Academy o f Agricultural Sciences,Harbin,150080,Heilongjiang)
出处
《中国糖料》
2018年第5期18-21,共4页
Sugar Crops of China
基金
国家自然基金青年基金项目(31601229)
黑龙江大学学科青年学术骨干百人支持计划项目
哈尔滨市科技局创新人才项目(2013RFQYJ025)
黑龙江省高校基本科研业务费黑龙江大学专项资金项目(HDRCCX-201613)
农业部糖料现代产业技术体系建设项目(CARS-170204
CARS-170111)
关键词
甜菜
遗传转化
StGCS-GS
谷胱甘肽合成酶
干旱逆境
sugar beet （Beta vulgaris）
genetic transformation
StGCS-GS
glutathione synthetase
drought stress
