Plant Cell, Tissue and Organ Culture

, Volume 71, Issue 2, pp 147–155 | Cite as

Factors affecting Agrobacterium-mediated transformation and regeneration of sweet orange and citrange

  • Changhe Yu
  • Shu Huang
  • Chunxian Chen
  • Zhanao Deng
  • Paul Ling
  • Fred G. GmitterJr.
Article

Abstract

Epicotyl explants of sweet orange and citrange were infected with Agrobacterium strain EHA101 harboring binary vector pGA482GG, and factors affecting the plant regeneration and transformation efficiency were evaluated. Increasing the wounded area of explants by cutting longitudinally into two halves, and optimization of inoculation density, dramatically enhanced both regeneration and transformation frequency. Inclusion of 2,4-dichlorophenoxyacetic acid (2,4-D) in the explant pretreatment medium and the co-culture medium improved the transformation efficiency by decreasing the escape frequency. More than 90% rooting frequency of transformed citrange shoots was achieved by two-step culture: first on media supplemented with auxins, and then on media without hormones. Inclusion of 20 mg l−1 kanamycin in rooting medium efficiently discriminated transformed shoots from non-transgenic escaped shoots. Shoot grafting in vitro was used to regenerate transformed plants, due to the slow growth of most sweet orange shoots.

Agrobacterium-mediated transformation bacterium density citrus regeneration rooting in vitro 

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Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Changhe Yu
    • 1
    • 2
  • Shu Huang
    • 1
  • Chunxian Chen
    • 1
  • Zhanao Deng
    • 1
  • Paul Ling
    • 1
  • Fred G. GmitterJr.
    • 1
  1. 1.Citrus Research and Education CenterUniversity of FloridaLake AlfredUSA
  2. 2.Department of HorticultureFujian Agricultural UniversityFuzhouPR China

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