Plasmolysis of precultured immature embryos improves Agrobacterium mediated gene transfer to rice (Oryza sativa L.) |
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| Institution: | 1. Department of Biomedical Engineering, Case Western Reserve University School of Medicine, 10900 Euclid Ave., Cleveland, OH 44106, USA;2. Department of Radiology, Case Western Reserve University School of Medicine, 10900 Euclid Ave., Cleveland, OH 44106, USA;3. Department of Materials Science and Engineering, Case Western Reserve University School of Engineering, 10900 Euclid Ave., Cleveland, OH 44106, USA;4. Department of Macromolecular Science and Engineering, Case Western Reserve University School of Engineering, 10900 Euclid Ave., Cleveland, OH 44106, USA;5. Division of General Medical Sciences-Oncology, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH 44106, USA;6. Department of Molecular Biotechnology, RWTH-Aachen University, Aachen 52064, Germany;1. Biological and Agricultural Engineering Department, Texas A&M University, College Station, TX, USA;2. Texas A&M AgriLife Research & Extension Center, Texas A&M University System, 2415 E. Highway 83, Weslaco TX 78596, USA;3. Department of Plant Pathology, University of California, Davis, Davis, CA, USA;4. Department of Agriculture, Agribusiness, and Environmental Sciences, Texas A&M University-Kingsville, Citrus Center, Weslaco, TX, USA;5. Department of Biochemistry and Biophysics, Texas A&M University, 2128 TAMU, College Station, TX, USA;6. Department of Plant Pathology and Microbiology, Texas A&M University, 2132 TAMU, College Station, TX, USA;7. Institute for Advancing Health Through Agriculture, Texas A&M AgriLife, College Station, TX, USA;1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada-ku, Kobe 657-8501, Japan;2. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan;3. RIKEN Center for Sustainable Resource Science, 1-7-22 Suehirocho, Turumi-ku, Yokohama, Kanagawa 230-0045, Japan;4. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan |
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| Abstract: | Efficiency of Agrobacterium mediated gene transfer was improved in rice by microprojectile pretreatment. Fertile transgenic plants were easily recovered using this protocol. Since an osmotic treatment is part of the bombardment protocol, we studied the effect of plasmolysis alone (i.e. without any microprojectiles) on transient expression of the gus gene transferred by Agrobacterium to precultured rice embryos. We report here for the first time that plasmolysis alone as a single pretreatment, yielded an even higher number of cells expressing the marker gene than the combination of microprojectile bombardment and osmotic treatment. This indicates that the effect of the osmoticum on gene transfer is independent from bombardment. |
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