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Please use this identifier to cite or link to this item: http://hdl.handle.net/10204/1663

Title: Transformation of elite white maize using the particle inflow gun and detailed analysis of a low-copy integration event
Authors: O'Kennedy, MM
Burger, JT
Berger, DK
Keywords: Elite white maize transformation
Immature embryos
Fertile transgenic plants
Plant regeneration
Gene expression
Plant sciences
Issue Date: Dec-2001
Publisher: Springer-Verlag
Citation: O'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 721-730
Abstract: 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.
URI: http://hdl.handle.net/10204/1663
ISSN: 0721-7714
Appears in Collections:General science, engineering & technology
Agroprocessing and chemical technology

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