Transient expression of visible marker genes in meristem cells of wheat embryos after ballistic micro-targeting |
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Authors: | Victor A Iglesias Andreas Gisel Roland Bilang Nathalie Leduc Ingo Potrykus Christof Sautter |
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Institution: | (1) Institute for Plant Sciences, Swiss Federal Institute of Technology, Universitätsstrasse 2, CH-8092 Zürich, Switzerland |
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Abstract: | Seven days after anthesis, the shoot apical meristem of immature embryos of wheat (Triticum aestivum L.) is not yet covered by the coleoptile or leaf primordia and provides an optimal target for ballistic micro-targeting. Gold particles 1.2 μm in diameter at a concentration of 5·105 particles per μl and propelled by 110-bar nitrogen penetrated up to four cell layers into embryo apical meristems but produced no deleterious effects on germination. The use of diaphragms with internal diameters of 100 or 200 μm restricted bombardment to meristem cells or also included surrounding tissues, respectively. The results of transient-expression experiments indicated successful delivery of foreign DNA into meristem cells. Cells of the central zone of the meristem or pro-meristem transiently expressed foreign genes driven by the Cauliflower mosaic virus (CaMV) 35S and rice actin1-D constitutive promoters. Partial plasmolysis before bombardment and slow recovery of normal turgor pressure increased transient-expression frequencies. Meristem cells transiently expressed foreign genes at frequencies 10-fold less than surrounding tissues under identical conditions. Transgenic sectors were observed in both coleoptiles and leaf primordia. |
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Keywords: | Direct gene transfer Microprojectile bombardment Micro-targeting Meristem Triticum |
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