
%0 Journal Article
%A Weir, Brian 
%A Gu, Xu 
%A Wang, Mingbo 
%A Upadhyaya, Narayana 
%A Elliott, Adrian R. 
%A Brettell, Richard I. S. 
%T <emph type="2">Agrobacterium tumefaciens</emph>-mediated transformation of wheat using suspension cells as a model system and green fluorescent protein as a visual marker
%J Functional Plant Biology
%D 2001/08/06
%V 28
%N 8
%P 807-818
%K Agrobacterium tumefaciens, green fluorescent protein, wheat transformation.
%R https://doi.org/10.1071/PP99211
%U https://www.publish.csiro.au/paper/PP99211 
%X Conditions for Agrobacterium-mediated transformation of wheat (Triticum aestivum L.) were defined using wheat suspension cells as a model system and green fluorescent protein (GFP) as a visual marker. Different strains of Agrobacterium tumefaciens were compared using established wheat cell suspension cultures, where the frequency of cell clusters showing transient activity of GFP ranged from 2 to 52%. High levels of transient GFP activity and stable transformed callus lines were obtained with plasmid pTO134 containing a gfp gene with an enhanced CaMV 35S promoter and a bar gene with a 35S promoter in combination with Agrobacterium strain AGL0. These results suggest that the important variables in Agrobacterium-mediated transformation of wheat cells include media composition, Agrobacterium strain, plasmid vector and the addition of virulence-inducing agents such as acetosyringone. The conditions deemed optimal for transformation of wheat suspension cell lines were applied to scutella isolated from immature embryos and scutella-derived calli. Transient GFP expression in these tissues ranged from 10 to 75% and, while quite variable among and within cultivars, stably transformed scutellum-derived callus was obtained. Further studies with scutellum-derived calli suggested that variables such as duration of pre-inoculation culture and co-cultivation, as well as co-cultivation temperature, were also important. Optimisation of these variables resulted in the recovery of transformed wheat plants at a transformation frequency of 1.8%, which is comparable with other reports. 
