利用卡那霉素筛选菊花转基因植株的方法研究  

Study on screening method of transgenic chrysanthemum using kanamycin

作　　者：刘鹏[1] 陈素梅[1] 房伟民[1] 蒋甲福[1] 管志勇[1] 陈发棣[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095

出　　处：《南京农业大学学报》2013年第5期45-50,共6页Journal of Nanjing Agricultural University

基　　金：江苏省科技支撑计划项目(BE2011325);国家农业科技成果转化项目(2010GB2360063);国家863计划项目(2011AA100208);教育部新世纪优秀人才支持计划项目(NCET-10-0492);江苏省高校科研成果产业化推进项目(JHB2011-8);国家自然科学基金项目(31272202)

摘　　要：将菊花品种‘神马’的叶盘和茎段分别接种含有0、5、7.5、10、15和20 mg·L-1卡那霉素(Kan)的再生培养基和生根培养基上,以建立简单有效的转基因菊花Kan抗性株系的筛选方法。结果表明:菊花叶盘不定芽分化筛选和生根筛选的Kan临界耐受质量浓度分别为10和7.5 mg·L-1。通过农杆菌介导法转化2 400个叶盘,经Kan筛选后得到358个抗性不定芽,经Kan生根筛选后得到65个生根株系。为了有效剔除假阳性植株,用500 mg·L-1的Kan溶液浸泡菊花转基因再生株系的叶片,通过性状观察筛选出6个Kan抗性株系,且6个抗性株系都通过PCR手段得到验证。表明成功建立了简单有效的筛选转基因菊花的方法。In order to establish simple and effective method of screening marker gene in chrysanthemum which was resistant to kanamycin（Kan）, the leaf-disks and shoots of a chrysanthemum cultivar＇ Jinba＇ were inoculated into regeneration medium and rooting medium with 0,5,7.5,10,15 and 20 mg·L^-1 of Kan respectively. The results showed that the critical concentration for shoot regeneration and rooting for chrysanthemum were 10 mg·L^-1and 7.5 mg·L^-1 Kan respectively. 358 Kan-resistant shoots were regenerated using the method of Agrobacterium-mediated transformation, and 65 rooted plants were obtained after rooting selection. In order to diminish escapes, the leaves of regenerated plants were immersed in the aqueous solution containing 500 mg·L^-1 Kan in petri dishes,and 6 Kan-resistant plants were selected by the treatment. The Kan-resistant plants were all confirmed by PCR analysis. Accordingly, a simple and effective method of screening transgenic chrysanthemum has been established.

关 键 词：转基因菊花 卡那霉素 抗性再生 筛选 

分 类 号：S682.1[农业科学—观赏园艺]