摘要
通过对转基因大白菜进行TuMV-C4病毒摩擦接种鉴定、ELSIA检测、提取转基因大白菜DNA进行PCR检测,并通过PCR-Southern杂交、RT-PCR检测转基因大白菜,构建了针对无选择标记转基因大白菜的系统检测方法。试验结果表明:转基因大白菜通过摩擦接种TuMV-C4病毒抗病性鉴定,188株表现为抗性,初步筛选率为19.46%;进行ELISA检测抗病植株中TuMV含量,有63株明显低于非转化植株,筛选率达到26.25%;对抗TuMV植株直接进行目的基因PCR检测,有25株呈阳性,筛选率达到13.30%。经PCR-Southern杂交检测和RT-PCR检测,表明转基因大白菜中不仅整合了Nib基因,而且获得了表达。这种逐级递进的检测系统可以高效地获得无选择标记的转基因植株。
A systematic detection method for nonselective marker transgenic Chinese cabbage was constructed by the identification of TuMV-C4 virus friction inoculation of transgenic Chinese cabbage,ELSIA detection,extraction and PCR detection of the transgenic Chinese cabbage DNA,PCR-Southern hybridization and RT-PCR detection of the transgenic Chinese cabbage.The results showed that 188 strains of transgenic Chinese cabbage were resistant to TuMV-C4 virus inoculated by friction,and the preliminary screening rate was 19.46%.Secondly,the content of TuMV in resistant plants was detected by ELISA,and 63 of them were significantly lower than those of nontransformed plants.The screening rate was 26.25%,and then the target gene PCR was detected directly against TuMV plants,25 of them were positive and the screening rate was 13.30%.Finally,PCR-Southern hybridization and RT-PCR analysis were carried out,which proved that Nib gene was not only integrated but also expressed in transgenic Chinese cabbage.This stepbystep detection system can efficiently obtain nonselective marker transgenic plants.
作者
佟容
于占东
吴春燕
乔宏宇
乔建磊
王彧彧
张新影
TONG Rong;YU Zhandong;WU Chunyan;QIAO Hongyu;QIAO Jianlei;WANG Yuyu;ZHANG Xinying(College of Horticulture,Jilin Agricultural University,Changchun 130118,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2020年第4期364-369,共6页
Journal of Jilin Agricultural University
基金
吉林省教育厅“十三五”科技项目(JJKH20170314KJ)。
