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GrainGenes Reference Report: JEB-52-865

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Reference	JEB-52-865
Title	Procedures allowing the transformation of a range of European elite wheat (Triticum aestivum L.) varieties via particle bombardment
Journal	Journal of Experimental Botany
Year	2001
Volume	52
Pages	865-874
Author	Rasco-Gaunt S Riley A Cannell M Barcelo P Lazzeri PA
Abstract	Summary: 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 T1 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
External Databases	Pubmed: 11413224
Keyword	[ Hide all but 1 of 48 ] ammonia ammonium assay bar bar gene bombardment co-transformation cotransformation culture decarboxylase escape fertile transgenic wheat frequencies gene gene activity gene sequence genetic transformation insertion line marker marker gene microprojectile bombardment modification particle bombardment pcr pcr analysis phosphinothricin progeny regeneration scutellar tissues segregation selectable marker selection sequence shoot stable transformation tissue tissue culture transformation transgene transgene integration transgene segregation transmission triticum triticum aestivum uida gene wheal transformation wheat varieties

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