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含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法(英文)

标题: 含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法(英文)
英文标题: Three Methods for Rapid Selection of Transgenic Cotton Plants in Laboratory with Kanamycin as Indirect Marker
作者: 王彦霞,吴立柱,王省芬,马峙英
英文作者: WANG Yan-xia,WU Li-zhu,WANG Xing-fen,MA Zhi-ying*(Agricultural University of Hebei,Key Laboratory of Crop Germplasm Resources of Hebei,Baoding 071001,China)
出版时间: 2007-03-15
机构: 河北农业大学棉花研究所河北省作物种质资源重点实验室,河北农业大学棉花研究所河北省作物种质资源重点实验室,河北农业大学棉花研究所河北省作物种质资源重点实验室,河北农业大学棉花研究所河北省作物种质资源重点实验室 保定071001,保定071001,保定071001,保定071001
关键词: 棉花,转基因植株,卡那霉素,鉴定
英文关键词: cotton,kanamycin,transgenic plants,rapid detection in the laboratory
刊名: 棉花学报
英文刊名: 棉花学报
ISSN: 1002-7807
期号: 02
国内刊号: 41-1163/S
基金: 河北省自然科学基金重点项目(C2005000209)
主题: 含NPTⅡ标记基因的转基因抗虫棉室内快速鉴定方法(英文)
页码: 56-60
分类号: S562;
出版单位: 棉花学报
数据来源: CNKI
是否SCI: false
是否EI: false
是否核心: true
摘要: 通过试验研究,提出了3种室内快速筛选和鉴定含NPTⅡ标记基因的转基因抗虫棉的方法。一是待检测棉子去种皮,在含卡那霉素培养基培养,根据幼苗子叶颜色和棉苗状态来辨别是否是转基因材料,卡那霉素最适浓度为0.75 g.L-1;二是去皮种子培养在无卡那霉素的萌苗培养基上,在棉苗子叶上涂抹卡那霉素溶液进行鉴定,最适浓度为4.0 g.L-1;三是在待测棉苗子叶上打孔、滴加一定浓度的卡那霉素,根据打孔并滴加卡那霉素部位的叶片颜色变化,鉴别并统计转基因植株,其最适浓度为2.0 g.L-1。3种方法均可在卡那霉素处理后4-7d内快速鉴定出转基因植株。同时,对转基因阳性植株进行PCR鉴定,结果表明3种卡那霉素方法鉴定的准确率均在90%以上。
英文摘要: 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.