Title
Elimination of a selectable marker gene in barley: via segregation of uncoupled TDNAs in populations of doubled haploids,Used
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Barley represents one of the economically most important and widely distributed crops worldwide, and genetic engineering is expected to play a crucial role in its further improvement. However, once transgenic plants are obtained, the selectable marker gene is not necessary anymore or even unwanted, and thus viable means to circumvent the use of selectable markers or to remove them from transgenic plants are required. In the present study, a novel strategy has been pursued to generate selectable markerfree transgenic lines instantly homozygous for the geneofinterest. Primary cotransgenic plants (T0) containing both the selectable marker gene (hygromycin phosphotransferase) and a model geneofinterest (glucuronidase) were produced via Agrobacteriummediated gene transfer using two independent TDNAs. Both removal of the selectable marker and establishment of instant homozygosity of the geneofinterest was then achieved at a time through segregation of uncoupled TDNAs in populations of doubled haploid (DH) lines produced from the T0 plants via embryogenic pollen culture.
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