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 language | English |
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Pages (from-to) | 659-668 |
Number of pages | 10 |
Journal | Plant Cell Reports |
Volume | 21 |
Issue number | 7 |
Early online date | 16 Jan 2003 |
DOIs | |
Publication status | Published - 01 Mar 2003 |
Keywords
- Acetophenones
- DNA, Bacterial
- Glucuronidase
- Plants, Genetically Modified
- Regeneration
- Rhizobium
- Seeds
- Transformation, Genetic
- Triticum