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GrainGenes Reference Report: PRT-19-1069

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Reference	PRT-19-1069
Title	Green fluorescent protein as a visual marker for wheat transformation
Journal	Plant Cell Reports
Year	2000
Volume	19
Pages	1069-1075
Author	Jordan MC
Abstract	Summary: Wheat (Triticum aestivum L.) transformation via particle bombardment is now established in many laboratories, but transformation efficiencies are still largely low and the highest efficiencies can only be obtained with certain genotypes. For rapid optimization and improvement of wheat transformation protocols, a non-destructive marker which permits early detection of transformed cells is needed. We have assessed the ability of a modified version of the Aequorea victoria green fluorescent protein (GFP) to act as a marker for detecting transformed cells and tissues of wheat. Multicellular clusters emitting green fluorescence were observed 14 days after particle bombardment with a sGFPS65T gene construct, and gfp-expressing shoots (often with expressing roots) could be observed as early as 21 days after bombardment. These shoots can be removed from the callus and grown further until they are ready to transfer to soil. Transgenic wheat plants could be selected on the basis of gfp expression alone although the inclusion of antibiotic resistance as a selectable marker could improve the efficiency. Using sgfpS65T as a marker gene in an experiment comparing bombardment parameters allowed the rapid identification of variables that could be targeted for optimization
Keyword	[ Hide all but 1 of 31 ] bombardment callus cell detection fertile transgenic wheat fluorescence gene genotype gfp green fluorescent protein improvement marker microprojectile bombardment optimization parameters particle bombardment protocol resistance root selectable marker shoot soil tissue transformation transgenic transgenic wheat triticum aestivum victoria vital marker wheat plants wheat transformation

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