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中国农学通报 ›› 2009, Vol. 25 ›› Issue (14): 22-26.

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

卡那霉素叶片涂抹法田间筛选转基因苜蓿的研究

燕丽萍,夏 阳,梁慧敏,王友平,,刘翠兰,王振猛,李 丽   

  • 收稿日期:2009-02-05 修回日期:2009-02-08 出版日期:2009-07-20 发布日期:2009-07-20

Study on the Field Screening for Transgenic Alfalfa by Smearing Kanamycin on the Leaves

Yan Liping, Xia Yang, Liang Huimin, WangYouping,, Liu Cuilan, Wang Zhenmeng, Li Li   

  • Received:2009-02-05 Revised:2009-02-08 Online:2009-07-20 Published:2009-07-20

摘要: 以田间生长的苜蓿为材料进行Kan涂抹叶片筛选转基因植株最佳浓度和适宜观察时间的研究,并对筛选的抗性植株进行了PCR检测验证。结果表明,Kan叶片涂抹田间筛选的最佳浓度为1000mg/L,观察时间以12d为宜;经PCR检测验证,Kan筛选抗性苗的阳性检测率达到93.8%。研究建立了一种在田间对转基因苜蓿进行大量筛选的简便方法,对于转基因苜蓿材料的选育具有重要利用价值。

关键词: 生长素, 生长素, 赤霉素, 油菜素内酯, 乙烯, 相互作用

Abstract: Kanamycin can be used to select the transformed plants containing the nptⅡgene. In order to assay the optimal screening concentration to nptⅡ gene, Kanamycin was smeared to the surface of the transformed alfalfa leaves. It showed that 1000mg/L Kanamycin with 12 days’ selecting was best. This method was compared with PCR as well, and the results suggested that the equivalent ratio was 93.8 % in the resistant plants. Accordingly, a simple method of filed screening gene transgenic alfalfa has been made. This result is very useful for the transgenic alfalfa to be screened and cultivated.