摘要
以采用RNAi技术获得的HiⅡ、HiⅡA、HiⅡB和H99抗玉米矮花叶病转基因T1和T2代为材料,通过病毒接种、草丁膦涂抹、目的基因和标记基因PCR扩增,研究了利用RNAi原理构建的目的基因对玉米矮花叶病的抗性效果以及玉米转基因后代群体鉴定筛选技术。结果表明,90.00%的转基因株系抗病性超过非转基因株系,30.00%的转基因株系抗病株率超过了抗病对照黄早4,转基因株系的抗病性明显提高,说明利用RNAi技术选育抗矮花叶病转基因玉米株系是可行的。4种筛选鉴定方法比较结果表明,目的基因PCR和病毒接种鉴定方法最可靠,在转基因后代群体鉴定过程中,可在苗期用200mg/L的草丁膦筛选阳性植株基础上,再采用目的基因PCR鉴定,筛选出抗性转基因植株或株系,不仅可以减少鉴定的工作量,而且可以提高鉴定的准确性。
In the paper,four T1 or T2 generation materials of transgenic maize resistant to dwarf mosaic disease were obtained with RNAi technology,which were used to study the resistant effect of target gene constructed by RNAi principle to maize dwarf mosaic disease and the techniques to identify and screen transgenic progeny population through inoculating virus, daubing PPT and PCR amplification of the target gene and marker gene. The results showed that 90.00~ of transgenic plants had higher resistance than non-transgenic plants, and 30.00~ of transgenic plants were even better than CK（Huangzao 4）. The increase of disease resistance in transgenic lines indicated that it was feasible to breed the transgenic plants with resistance to dwarf mosaic disease by RNAi technique. The comparison among the four identifying and screening methods showed that PCR of target gene and virus inoculation were the most reliable. During the identifying and screening procedure of transgenic progenies, the positive plants should be first selected at seedling stage with 200 mg/L of PPT, from which the resistant transgenic plants or lines were confirmed using the target gene PCR identification in order to reduce the workload of identification and improve the accuracy of identification.
出处
《河南农业科学》
CSCD
北大核心
2014年第4期69-74,共6页
Journal of Henan Agricultural Sciences
基金
国家转基因重大专项(2008ZX08003-001)
河南省现代农业产业技术体系项目(S2010-02)
关键词
RNAI技术
玉米
矮花叶病
转基因
鉴定方法
RNAi technology
maize
dwarf mosaic disease
transgene
identification methods
