卡那霉素在转基因芥菜中的应用-遗传.PDF

HEREDITAS (Beijing) 2008 4 , 30(4): 501―507 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2008.00501 赵爽, 雷建军, 陈国菊, 曹必好 , 510642 : 为了找出芥菜 (Brassica juncea Coss.) 遗传转化中最佳的卡那霉素(Kan)筛选浓度, 将芥菜的子叶接种于 含有不同浓度Kan 的分化培养基中, 当Kan 浓度达到 30 mg/L 时, 外植体的分化完全受到抑制。将芥菜种子播种 于含有不同浓度Kan 的培养基中, 当Kan 浓度达到200 mg/L 时, 长出的幼苗完全白化; 利用叶片涂抹方法, 将不 同浓度的Kan涂抹于田间生长的植株叶片上, 当Kan 浓度达到200 mg/L 时, 被处理的叶片完全变白。为了对转基 因芥菜后代中外源基因的分离情况进行遗传学分析, 分别用 200 mg/L 的Kan处理以npt- 基因为选择标记基因 的转基因芥菜的种子和转基因芥菜后代植株的叶片, 利用χ2 测验分析试验结果, 4 个含有单拷贝外源基因的转基 因株系后代, 对Kan 的抗感分离都符合3 ︰1 的分离规律; 而2 个含有双拷贝外源基因的转基因株系, 其中一个 对Kan 的抗感分离符合3 ︰1 而不符合15 ︰1, 另一个对Kan 的抗感分离既符合3 ︰1 也符合 15 ︰1, 双拷贝外 源基因在转基因芥菜中的整合方式有待进一步的研究。最后, 用PCR分析证实了该方法的准确性, 因此, 利用 Kan对转基因芥菜后代进行筛选是可行的。 : 卡那霉素; 芥菜; 转基因后代 Application of kanamycin in transgenic mustard(Brassica juncea Coss.) ZHAO Shuang, LEI Jian-Jun, CHEN Guo-Ju, CAO Bi-Hao Horticulture College of South China Agricultural University, Guangzhou 510642, China Abstract: In order to find the best screening kanamycin concentration in the genetic transformation of mustard (Brassica juncea Coss.), the seedling cotyledons of mustard were placed on bud-induced media supplemented with different kanamy- cin concentrations. The bud differentiation of explants was totally inhibited when the kanamycin concentration was greater than 30 mg/L. The seeds of mustard were placed on germination media supplemented with different concentrations of kanamycin. All the seedlings were white when kanamycin concentration was higher than 200 mg/L. The leaves of mustard in field were smeared with the solutions including di