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中国农学通报 ›› 2015, Vol. 31 ›› Issue (21): 130-135.doi: 10.11924/j.issn.1000-6850.casb15010233

所属专题: 玉米

• 生物技术科学 • 上一篇    下一篇

玉米茎尖再生体系优化与筛选物敏感性研究

曹士亮   

  1. 黑龙江省农业科学院玉米研究所
  • 收稿日期:2015-01-30 修回日期:2015-06-30 接受日期:2015-02-21 出版日期:2015-07-28 发布日期:2015-07-28
  • 通讯作者: 曹士亮
  • 基金资助:
    哈尔滨市青年科技创新人才项目 “农杆菌介导的 ODREB2B基因原位转化玉米萌动胚和茎尖的研究” (2013RFQYJ017)。

Research on the Optimization of Maize Stem Tip Regeneration System and Its Sensitivity to Selective Agent

  • Received:2015-01-30 Revised:2015-06-30 Accepted:2015-02-21 Online:2015-07-28 Published:2015-07-28

摘要: 玉米再生体系的优化及对筛选物敏感性的研究是玉米遗传转化的基础,为了明确玉米茎尖再生条件及其对筛选物的敏感性,以玉米骨干自交系郑58和K10为试材,对生长素和细胞分裂素的浓度进行了优化。在此基础上,研究了茎尖分生组织对4种筛选抗生素、一种抑菌抗生素和一种除草剂的敏感性,结果表明,玉米茎尖丛生芽诱导需要细胞分裂素的参与,诱导培养基为MS培养基+1.0mg/L2,4-D+2.0mg/L6-BA时具有相对较高的丛生芽诱导率,自交系K10的分化速度要快于郑58。茎尖分生组织对筛选物的敏感性顺序依次是遗传霉素G418>巴龙霉素Par>卡那霉素(Kan)>新霉素(Neo)。在4种筛选抗生素中,遗传霉素和巴龙霉素比较适合茎尖转化体系的筛选,遗传霉素的适宜浓度为15~30mg/L,巴龙霉素Par的浓度为30~50mg/L,抑菌抗生素头孢霉素(Cef)的适宜浓度为400~600mg/L,除草剂PPT筛选的适宜浓度为1~3mg/L。

关键词: 电极生物膜法, 电极生物膜法, 养殖废水, 氨氮, 反硝化, 水力停留时间

Abstract: For maize transformation, it is the basis to study the optimization of maize stem tip regeneration system and its sensitivity to selective agent. In this paper, two maize inbred lines K10 and Zheng 58 were used as materials to establish the maize stem tip regeneration system. Furthermore, maize stem tip of K10 was selected to research its sensitivity to four kinds of selective antibiotics, one antibacterial antibiotic and one herbicide. The results showed that cytokinin was necessary for multiple shoots induction of maize stem tip induction medium: MS+1.0 mg/L 2,4-D+ 2.0 mg/L 6-BA. The stem tip of K10 differentiated relatively faster than that of Zheng58. The order of maize stem tip to four kinds of selective antibiotics was geneticinG418 > paromomycin (Par) > kanamycin (Kan) > neomycin (Neo). geneticinG418 and paromomycin (Par) were more suitable to be used as selective agents among four kinds of antibiotics, and their suitable concentrations were 15-30 mg/L and 30-50 mg/L, respectively. The suitable concentration of antibacterial antibiotic Cef was 400- 600 mg/L. The suitable concentration of herbicide PPT was 1-3 mg/L.