摘要
油菜耐热基因(BnTR1)克隆自油菜,编码一种E3连接酶.为验证它的抗逆功能,本研究用携带BnTR1和新霉素磷酸转移酶(neomycin phosphotransferase,nptⅡ)基因的植物表达载体pCAMBIA2301-TR1的根癌农杆菌EHA105,对多年生黑麦草丛生芽进行遗传转化.经共培养和巴龙霉素筛选培养,获得了175株抗性植株.通过PCR和RT-PCR检测,获得62株转基因植株.结果表明,外源BnTR1基因已整合到转基因植株基因组中并在转录水平上表达.初步抗旱检测表明,BnTR1基因能明显提高转基因植株的抗逆能力.
The rape heat-resistant gene （BnTR1） encodes a E3 Ligase. To detect its function and breed perennial ryegrass of stress tolerance, the clump shoots from perennial ryegrass were transformed via Agrobactrium tumefaciens EHA105 carrying expression vector pCAMBIA2301-TR1 containing BnTR1 and nptII genes. After co-cultured and selected with paromomycin, 175 plants showed resistance. Among them, 62 transgenic plants were obtained by PCR and RT-PCR. The results presented that the BnTR1 was integrated in perennial ryegrass genome and expressed at the transcriptional level. And drought tolerance test implied that BnTR1 gene could obviously elevate the stress tolerance ability of transgenic plants.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
2013年第4期881-886,共6页
Journal of Sichuan University(Natural Science Edition)
基金
国家自然科学基金(31171586)
国家863计划项目(2008AA10Z409
2009AA10Z108)
关键词
多年生黑麦草
油菜耐热蛋白基因
遗传转化
丛生芽
根癌农杆菌
perennial ryegrass
rape heat-resistant protein gene
genetic transformation
shoot clumps
Agrobacterium tumefaciens
