Isolation of transgenic progeny of maize by embryo rescue under selective conditions |
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Authors: | Kris Weymann Katharina Urban Daniel M Ellis Roseann Novitzky Erik Dunder Susan Jayne Gary Pace |
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Institution: | (1) CIBA-GEIGY Agricultural Biotechnology, P. O. Box 12257, 27709 Research Triangle Park, North Carolina |
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Abstract: | Summary Fertile transgenic maize plants (T0) and progeny (T1) were obtained using microprojectile bombardment and callus selection on hygromycin B. To quickly identify progeny expressing
the transgene, embryos from T3 generation kernels were excised 20 days after pollination and exposed to different concentrations
of hygromycin B. Surviving and non-surviving embryos were assayed for the presence of the hygromycin phosphotransferase (aphIV) gene using polymerase chain reaction. Embryos that germinated and survived on 25, 50, or 100 mg/liter hygromycin possessed
theaphIV gene. Embryos that did not germinate lacked the gene. Progeny surviving selection were transferred to the greenhouse and
tested for expression of the gene using a leaf disc assay. The results demonstrated that the gene construct was expressed
in both embryo and leaf tissue and that selection during germination successfully eliminated progeny lacking the gene of interest.
This method is also useful for rapid-cycling of maize generations. |
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Keywords: | embryo rescue selection hygromycin transgenic maize |
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