植物学报 ›› 2009, Vol. 44 ›› Issue (06): 728-734.DOI: 10.3969/j.issn.1674-3466.2009.06.011

• 技术方法 • 上一篇    下一篇

胡杨转基因体系的建立

范源伟1, 刘挨枝2, 王华芳 1   

  1. 1 北京林业大学生物科学与技术学院, 林木育种国家工程实验室, 林木、花卉遗传育种教育部重点实验室, 国家林业局树木花卉育种生物工程重点开放性实验室, 北京 100083; 2 内蒙古阿拉善盟农牧业局, 巴彦浩特 750306
  • 收稿日期:2009-03-06 修回日期:2009-09-03 出版日期:2009-11-01 发布日期:2009-11-01
  • 通讯作者: 王华芳

Transformation of Populus euphratica

Yuanwei Fan1, Aizhi Liu2, Huafang Wang1   

  1. 1National Engineering Laboratory for Tree Breeding, Ministry of Education, College of Biological Sciences and Biotechnology, Beijing Forestry University; 2Agriculture and Animal Husbandry Bureau of Alashan League of Inner Mongolia, Bayanhot
  • Received:2009-03-06 Revised:2009-09-03 Online:2009-11-01 Published:2009-11-01
  • Contact: Huafang Wang

摘要: 胡杨(Populus euphratica)是唯一能在沙漠里生长的高大乔木树种, 建立其转基因体系可为胡杨抗逆分子机制与应用技术研究提供基本方法。通过研究农杆菌介导的胡杨转GUS基因的技术体系得出以下结论: (1) 胡杨再生植株体系, 叶片在附加1.0 mmol.L-1 6-BA和5.0 mmol.L-1 NAA的1/2MS培养基上不定芽诱导率较高; (2) 转基因体系, 胡杨叶片在含100 mmol.L-1 乙酰丁香酮的OD600值为0.4-0.6的根癌农杆菌菌液中浸染15分钟, 共培养2天, GUS基因转化效率较高; (3) 转基因植株抗生素筛选, 转GUS基因胡杨叶片用300 mg.L-1头孢霉素抑制农杆菌生长, 在含9 mg.L-1 G418的培养基上诱导不定芽以获得转基因的抗性植株。

Abstract: The uniquearbor tree species Populus euphratica can grow well in arid desert. We established an Agrobacterium based transformation system for this species using a GUS gene as the reporter. Shoots of P. euphratica were regenerated from leaves placed on 1/2 MS supplemented with 1.0 mmol.L-1 6-BA and 5.0 mmol.L-1 NAA, leaves were infected for 15 min in a solution of A. tumefaciens with 0.4-0.6 OD600 and 100 mmol.L-1 AS, and then co-cultured for 2 days. A. tumefaciens was inhibited with 300 mg.L-1 Cef, and transgenic tissues and shoots were selected with 9 mg.L-1 G418.