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林业科学 ›› 2008, Vol. 44 ›› Issue (4): 54-59.doi: 10.11707/j.1001-7488.20080411

• 论文 • 上一篇    下一篇

香樟胚性愈伤组织遗传转化体系建立*

杜丽1 庞振凌1 周索1 曾晓慧1 包满珠2   

  1. (1. 南阳师范学院生命科学与技术学院 南阳473000; 2. 华中农业大学园艺林学学院 教育部园艺植物生物学重点实验室 武汉430070)
  • 收稿日期:2007-09-19 修回日期:1900-01-01 出版日期:2008-04-25 发布日期:2008-04-25
  • 通讯作者: 包满珠

Establishment of Agrobacterium Mediated Transformation System of Embryogenic Calli of Cinnamomum camphora

Du Li1,Pang Zhenling1,Zhou Suo1,Zeng Xiaohui1,Bao Manzhu2   

  1. (1. Shool of Life Science and Technology, Nanyang Normal University Nanyang 473000; 2. Key Laboratory of Horticultural Plant Biology of Ministry of Education College of Horticulture and Forestry Sciences, Huazhong Agricultural University Wuhan 430070)
  • Received:2007-09-19 Revised:1900-01-01 Online:2008-04-25 Published:2008-04-25

摘要:

通过根癌农杆菌介导的遗传转化方法,建立香樟胚性愈伤组织遗传转化体系。结果表明:以胚性愈伤组织为受体,能取得比较理想的转化效果;50 mg·L-1的潮霉素对胚性愈伤组织有很好的筛选效果; 根癌农杆菌0D600为0.6时,能获得较高的转化率; 侵染时间提高为40 min时,对提高转化效率有显著的效果; 共培养3 d能够得到较好的转化效率。对转基因香樟愈伤组织进行PCR检测,结果表明GUS基因已整合到香樟胚性愈伤组织的基因组中。

关键词: 香樟, 根癌农杆菌, 遗传转化, PCR

Abstract:

A genetic transformation system of embryogenic calli (Cinnamomum camphora) mediated by Agrobacterium tumefaciens was established. Experimental result indicated that efficient transformation was obtained with embryogenic calli as acceptor. Hygromycin of 50 mg\5L-1 was efficient in screening embryogenic calli. When the OD600 value of Agrobacterium was 0.6, a high transformation rate was gained. The infection time of 40 min, combined with co culture time of 3 d, remarkably enhanced transformation rate. Results of PCR proved that the GUS gene had integrated into embryogenic calli of C. camphora genome.

Key words: Cinnamomum camphora, Agrobacterium tumefaciens, genetic transformation, PCR