Factors influencing successful Agrobacterium-mediated genetic transformation of wheat

H. Wu, C. Sparks, B. Amoah, H. D. Jones

Research output: Contribution to journalArticlepeer-review

195 Citations (SciVal)

Abstract

The development of a robust Agrobacterium-mediated transformation protocol for a recalcitrant species like bread wheat requires the identification and optimisation of the factors affecting T-DNA delivery and plant regeneration. We have used immature embryos from range of wheat varieties and the Agrobacterium strain AGL1 harbouring the pGreen-based plasmid pAL156, which contains a T-DNA incorporating the bar gene and a modified uidA (beta-glucuronidase) gene, to investigate and optimise major T-DNA delivery and tissue culture variables. Factors that produced significant differences in T-DNA delivery and regeneration included embryo size, duration of pre-culture, inoculation and co-cultivation, and the presence of acetosyringone and Silwet-L77 in the media. We fully describe a protocol that allowed efficient T-DNA delivery and gave rise to 44 morphologically normal, and fully fertile, stable transgenic plants in two wheat varieties. The transformation frequency ranged from 0.3% to 3.3%. Marker-gene expression and molecular analysis demonstrated that transgenes were integrated into the wheat genome and subsequently transmitted into progeny at Mendelian ratios.

Original languageEnglish
Pages (from-to)659-668
Number of pages10
JournalPlant Cell Reports
Volume21
Issue number7
Early online date16 Jan 2003
DOIs
Publication statusPublished - 01 Mar 2003

Keywords

  • Acetophenones
  • DNA, Bacterial
  • Glucuronidase
  • Plants, Genetically Modified
  • Regeneration
  • Rhizobium
  • Seeds
  • Transformation, Genetic
  • Triticum

Fingerprint

Dive into the research topics of 'Factors influencing successful Agrobacterium-mediated genetic transformation of wheat'. Together they form a unique fingerprint.

Cite this