O'Kennedy, Maretha MBurger, JTBerger, DK2007-02-082007-06-072007-02-082007-06-072001-12O'Kennedy, MM, Burger, JT and Berger, DK. 2001. Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event. Plant Cell Reports, vol. 20(8), pp 721-7300721-7714http://hdl.handle.net/10204/1663http://hdl.handle.net/10204/1663Elite white maize lines W506 and M37W were transformed with a selectable marker gene (bar) and a reporter gene (uidA) or the polygalacturonase-inhibiting protein (pgip) gene after bombardment of cultured immature zygotic embryos using the particle inflow gun. Successful transformation with this device did not require a narrow range of parameters, since transformants were obtained from a wide range of treatments, namely pre-culture of the embryos for 4-6 days, bombardment at helium pressures of 700-900 kPa, selection-free culture for 2-4 days after bombardment and selection on medium containing bialaphos at 0.5-2 mg l(-1). However, bombardments with helium pressures below 700 kPa yielded no transformants. The culture of immature zygotic embryos of selected elite white maize lines on medium containing 2 mg l(-1) 2,4-dichlorophenoxyacetic acid and 20 MM L-proline proved to be most successful for the production of regenerable embryogenic calli and for the selection of putative transgenic calli on bialaphos-containing medium after transformation. Transgenic plants were obtained from four independent transformation events as confirmed by Southern blot analysis. Transmission of the bar and uidA genes to the T-4 progeny of one of these transformation events was demonstrated by Southern blot analysis and by transgene expression. In this event, the transgenes bar and uidA were inserted in tandem.139774 bytesapplication/pdfenCopyright: 2004 Springer-VerlagElite white maize transformationCerealsImmature embryosFertile transgenic plantsPlant regenerationGene expressionPlant sciencesArticleO'Kennedy, M. M., Burger, J., & Berger, D. (2001). Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event. http://hdl.handle.net/10204/1663O'Kennedy, Maretha M, JT Burger, and DK Berger "Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event." (2001) http://hdl.handle.net/10204/1663O'Kennedy MM, Burger J, Berger D. Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event. 2001; http://hdl.handle.net/10204/1663.TY - Article AU - O'Kennedy, Maretha M AU - Burger, JT AU - Berger, DK AB - Elite white maize lines W506 and M37W were transformed with a selectable marker gene (bar) and a reporter gene (uidA) or the polygalacturonase-inhibiting protein (pgip) gene after bombardment of cultured immature zygotic embryos using the particle inflow gun. Successful transformation with this device did not require a narrow range of parameters, since transformants were obtained from a wide range of treatments, namely pre-culture of the embryos for 4-6 days, bombardment at helium pressures of 700-900 kPa, selection-free culture for 2-4 days after bombardment and selection on medium containing bialaphos at 0.5-2 mg l(-1). However, bombardments with helium pressures below 700 kPa yielded no transformants. The culture of immature zygotic embryos of selected elite white maize lines on medium containing 2 mg l(-1) 2,4-dichlorophenoxyacetic acid and 20 MM L-proline proved to be most successful for the production of regenerable embryogenic calli and for the selection of putative transgenic calli on bialaphos-containing medium after transformation. Transgenic plants were obtained from four independent transformation events as confirmed by Southern blot analysis. Transmission of the bar and uidA genes to the T-4 progeny of one of these transformation events was demonstrated by Southern blot analysis and by transgene expression. In this event, the transgenes bar and uidA were inserted in tandem. DA - 2001-12 DB - ResearchSpace DP - CSIR KW - Elite white maize transformation KW - Cereals KW - Immature embryos KW - Fertile transgenic plants KW - Plant regeneration KW - Gene expression KW - Plant sciences LK - https://researchspace.csir.co.za PY - 2001 SM - 0721-7714 T1 - Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event TI - Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event UR - http://hdl.handle.net/10204/1663 ER -