Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment.
Ten current European wheat varieties were transformed at efficiencies ranging from 1-17% (mean 4% across varieties) following modifications in particle bombardment and tissue culture procedures. All plants surviving phosphinothricin selection were screened for uidA and bar gene activity, and for the presence of marker gene sequences by PCR analysis. A minimum of 35% plant 'escape' frequency was achieved with selection on 4 mg l(-1) gluphosinate ammonium after shoot initiation. Mean co-transformation frequency with various genes-of-interest was 66%. The estimated number of insertions of the uidA gene in 25 lines were; 1-2 in 32%, 3-5 in 52%, and 6-8 in 16% of lines. In T(1) progenies, marker genes segregated in a Mendelian fashion in 50% of 39 lines analysed, as determined by transgene activity assays. Based on PCR analysis, it appeared that in some lines the occurrence of distorted segregation was due to poor transmission of the transgenes.
Rasco-Gaunt
S
S
Biochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, UK. Sonriza.Rasco-Gaunt@gbr.dupont.com
Riley
A
A
Cannell
M
M
Barcelo
P
P
Lazzeri
P A
PA
D001483:Base Sequence; D019470:Biolistics; D046508:Culture Techniques; D017931:DNA Primers; D017343:Genes, Plant; D030821:Plants, Genetically Modified / Q000235:genetics; D014170:Transformation, Genetic; D019076:Transgenes; D014908:Triticum / Q000235:genetics*
D017931:DNA Primers