A protocol for consistent, large-scale production of fertile transgenic rice plants |
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| Authors: | L Chen S Zhang R N Beachy C M Fauquet |
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| Institution: | (1) International Laboratory for Tropical Agricultural Biotechnology (ILTAB/ORSTOM-TSRI), Division of Plant Biology-BCC 206, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA e-mail: iltab@scripps.edu, US |
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| Abstract: | A protocol for consistent production of fertile transgenic rice plants was established utilizing microparticle bombardment
of embryogenic tissues (Oryza sativa L. japonica cv. Taipei 309). This system has been employed to produce several thousand independently transformed plant lines
carrying the hygromycin phosphotransferase (hph) gene and various genes of interest. The most efficient target tissue was highly embryogenic callus or suspension cell aggregates,
when they were given an osmotic pre- and post-transformation treatment of 0.6 m carbohydrate. By optimizing the age of the tissue at the time of gene transfer and applying an improved selection procedure,
transgenic plants were recovered in 8 weeks from the time of gene transfer, at an average of 22.3±9.7 per 100 calli and 22.4±8.0
plant lines per dish of suspension cell aggregates. This system has facilitated a number of studies using rice as a model
for genetic transformation and will enable the large-scale production of transgenic rice plants for genomic studies.
Received: 12 March 1998 / Revision received: 5 May 1998 / Accepted: 15 May 1998 |
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| Keywords: | Rice Microparticle bombardment Genetic transformation Embryogenic tissues |
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