Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue

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

Research output: Contribution to journalArticlepeer-review

120 Citations (Scopus)

Abstract

A critical step in the development of Agrobacterium tumifaciens-mediated transformation is the establishment of optimal conditions for T-DNA delivery into tissue from which whole plants can be regenerated. The efficient transformation of inflorescence tissue from 'Baldus', a commercial wheat variety, using the Agrobacterium strain AGLI harbouring the binary vector pAL156 is reported here. The effects of various factors on delivery and the transient expression of the uidA gene were studied including the duration of preculture, vacuum infiltration, the effect of sonication treatments, and Agrobacterium cell density. Optimal T-DNA delivery (as measured by uidA activity) was obtained from inflorescence tissues precultured for 21d and sonicated. Increasing Agrobacterium cell density, the duration of inoculation/co-cultivation and vacuum pressure, up to a threshold, increased uidA expression. The investigation of factors that influence T-DNA delivery is an important first step in the utilization of Agrobacterium in the transformation of immature wheat inflorescence tissue.

Original languageEnglish
Pages (from-to)1135-1142
Number of pages8
JournalJournal of Experimental Botany
Volume52
Issue number358
DOIs
Publication statusPublished - May 2001

Keywords

  • Agrobacterium tumefaciens
  • wheat inflorescence
  • GUS
  • transformation
  • commercial variety
  • ORYZA-SATIVA-L
  • PLANT-REGENERATION
  • MICROPROJECTILE BOMBARDMENT
  • IMMATURE INFLORESCENCES
  • SOMATIC EMBRYOGENESIS
  • ARABIDOPSIS-THALIANA
  • HIGH-EFFICIENCY
  • GENE-TRANSFER
  • TRANSFORMATION
  • DNA

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