棉花学报2007，Vol.19Issue(2)：134-138,5.
含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法
Three Methods for Rapid Selection of Transgenic Cotton Plants in Laboratory with Kanamycin as Indirect Marker
摘要
Abstract
Three methods were developed to rapidly identify Bt-transgenic cotton plants with kanamycin-resistant gene as indirect selective marker. The first method was to culture uncoated seeds on the kanamycin containing medium (kan-medium), the difference between transgenic and non-transgenic plants was revealed after seeds germinated on the medium. According to the color of the cotyledons, kan-resistant plants were discovered at optimal concentration of 0.75 g·L-1. The second method was to paint kanamycin solution (kan-solution) on the cotyledons directly with optimal concentration at 4.0 g·L-1, by which after 4-7 days, plants with cotyledon normal green were considered as kan-resistant plants. The third one was that a pore was dug on cotyledons and dropped kan-solution in . The best concentration of the kan-solution was 2.0 g·L-1. All the plants could be rapidly screened out in 4-7 days after kanamycin treatment by the three methods. The efficacy of above methods for transgenic plant indirect identification were tested using PCR analysis with Bt-gene specific primers, giving a examination ratio of more than 90%. Moreover, all the seeds decorticated could be more easily identified with kanamycin treatments.关键词
棉花/转基因植株/卡那霉素/鉴定Key words
cotton/kanamycin/transgenic plants/rapid detection in the laboratory分类
农业科技引用本文复制引用
王彦霞,吴立柱,王省芬,马峙英..含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法[J].棉花学报,2007,19(2):134-138,5.基金项目
河北省自然科学基金重点项目(C2005000209) （C2005000209）
