Plant Cell, Tissue and Organ Culture

, Volume 71, Issue 2, pp 147–155

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

Authors

  • Changhe Yu
    • Citrus Research and Education CenterUniversity of Florida
    • Department of HorticultureFujian Agricultural University
  • Shu Huang
    • Citrus Research and Education CenterUniversity of Florida
  • Chunxian Chen
    • Citrus Research and Education CenterUniversity of Florida
  • Zhanao Deng
    • Citrus Research and Education CenterUniversity of Florida
  • Paul Ling
    • Citrus Research and Education CenterUniversity of Florida
    • Citrus Research and Education CenterUniversity of Florida
Article

DOI: 10.1023/A:1019983107509

Cite this article as:
Yu, C., Huang, S., Chen, C. et al. Plant Cell, Tissue and Organ Culture (2002) 71: 147. doi:10.1023/A:1019983107509

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 transformationbacterium densitycitrusregenerationrooting in vitro

Copyright information

© Kluwer Academic Publishers 2002