热带作物学报2013，Vol.34Issue(7)：1282-1287,6.DOI:10.3969/j.issn.1000-2561.2013.07.016
nptⅡ::mgfp5融合基因的构建及其在柑桔遗传转化中的应用
Construction and Application of a Bifunctional Gene nptⅡ::mgfp5 for Citrus Genetic Transformation
摘要
Abstract
Early,no damage,easy operation detection methods for positive shoots are very useful and helpful to citrus genetic transformation and regeneration.In this paper,we designed and constructed a bifunctional fusion gene,npt Ⅱ::mgfp5,which could be used for Agrobacterium-mediated transformation,with citrus epicotyl segments transformation system as a model.The npt Ⅱ::mgfp5 fusion gene,consisted of 1 578 bp,encoding 525 amino acids,was cloned in the vector p1301NG as a selectable marker and reporter gene.Epicotyl segments from Jingcheng orange (citrus sinensis) were co-cultivated with A.tumefaciens EHA105 carrying the binary vector p1301NG and transferred to shoot-induction and selective medium.High-level GFP expression was detected in regeneration shoots on selection medium for 9 days,and 10～15 days was the peak period of fluorescent buds formation.Green fluorescent shoots were selected,plants were regenerated,and confirmed by GUS assay and PCR analysis.These results showed that npt Ⅱ::mgfp5 could be used to improve the survival rate of regenerated plantlets,and increase citrus transformation efficiency.关键词
nptⅡ::mgfp5融合基因/柑橘遗传转化/GFP荧光/转化效率Key words
npt Ⅱ::mgfp5 fusion gene/ Citrus transformation/ GFP fluorescent/ Transformation efficiency分类
农业科技引用本文复制引用
刘小丰,彭爱红,许兰珍,邹修平,朱世平,赵晓春,陈善春..nptⅡ::mgfp5融合基因的构建及其在柑桔遗传转化中的应用[J].热带作物学报,2013,34(7):1282-1287,6.基金项目
重庆市自然科学基金项目(No.CSTC,2010BB1153) （No.CSTC,2010BB1153）
现代农业技术体系资助项目(No.CARS-27). （No.CARS-27）
