Abstract
To improve the iron content of rice, we have transferred the entire coding sequence of the soybean ferritin gene into Oryza sativa (L. cv. Kita-ake) by Agrobacterium-mediated transformation. The rice seed-storage protein glutelin promoter, GluB-1, was used to drive expression of the soybean gene specifically in developing, self-pollinated seeds (T1 seeds) of transgenic plants, as confirmed by reverse transcription PCR analysis. Stable accumulation of the ferritin subunit in the rice seed was demonstrated by western blot analysis, and its specific accumulation in the endosperm by immunologic tissue printing. The iron content of T1 seeds was as much as threefold greater than that of their untransformed counterparts.
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References
World Health Organization (WHO). National Strategies for Overcoming Micronutrient Malnutrition. Document A45/3. (WHO, Geneva, Switzerland, 1992).
Craig, W.J. Iron status of vegetarians. Am. J. Clin. Nutr. 59, 1233S–1237S (1994).
Yip, R. Iron deficiency, contemporary scientific issues and international programmatic approaches. J. Nutr. 124, 1479S– 1490S (1994).
Anonymous. in Standard Tables of Food Composition (ed. Investigation Committee of Food Resources, Science and Technology Agency, Japan, 1992).
Gillooly, M. et al. The effect of organic acids, phytates and polyphenols on the absorption of iron from vegetables. Br. J. Nutr. 49, 331–342 (1983).
Inoue, K., Tashima, K., Sanada, K. & Yokota, H. Production of iron-enriched leaf vegetables using a soaking method. Jap. J. Soil Sci. Plant Nutr. 66, 527–534 ( 1995).
Theil, E.C. Ferritin: structure, gene regulation, and cellular function in animals, plants and microorganisms. Annu. Rev. Biochem. 56, 289–315 (1987).
Andrews, S. et al. Structure, function and evolution of ferritins. J. Inorg. Chem. 47, 161–174 ( 1992).
Ragland, M. et al. Evidence for a conservation of ferritin sequences among plants and animals and for a transit peptide in soybean. J. Biol. Chem. 265, 18339–18344 (1990).
Spence, M.J., Henzl, M.T. & Lammers, P.J. The structure of a Phaseolus vulgaris cDNA encoding the iron storage protein ferritin. Plant Mol Biol. 17, 499–504 (1991).
Lobréaux, S., Yewdall, S., Briat, J.F. & Harrison, P.M. Amino-acid sequence and predicted three-dimensional structure of pea seed (Pisum sativum ) ferritin. Biochem J. 288, 931– 939 (1992).
Lobréaux, S., Massenet, O. & Briat, J.F. Iron induces ferritin synthesis in maize plantlets. Plant Mol Biol. 19, 563– 575 (1992).
Theil, E.C. Regulation of ferritin and transferrin receptor mRNAs. J. Biol. Chem. 265, 4771–4774 ( 1990).
Van der Mark, F., Bienfait, F. & van den Ende, H. Variable amounts of translatable ferritin mRNA in bean leaves with various iron contents. Biochem. Biophys. Res. Commun. 115, 463–469 ( 1983).
Lescure, A.M. et al. Ferritin gene transcription is regulated by iron in soybean cell cultures. Proc. Natl. Acad. Sci. USA 88, 8222–8226 (1991).
Lobréaux, S. & Briat, J.F. Ferritin accumulation and degradation in different organs of pea (Pisum sativum) during development. Biochem. J. 274, 601–606 (1991).
Ragland, M. & Theil, E.C. Ferritin (mRNA. Protein) and iron concentration during soybean nodule development. Plant. Mol. Biol . 21, 555–560 ( 1993).
Theil, E.C., Burton, J.W. & Beard J.L. A sustainable solution for dietary iron deficiency through plant biotechnology and breeding to increase seed ferritin control. Eur. J. Clin. Nutr. 51, S28– S31 (1997).
Beard, J.L., Burton, J.W. & Theil E.C. Purified ferritin and soybean meal can be sources of iron for treating iron deficiency in rats. J. Nutr. 126, 154–160 (1996).
Takaiwa, F., Oono, K., Wing, D. & Kato, A. Sequences of three members and expression of a new major subfamily of glutelin gene from rice. Plant Mol. Biol. 17, 875– 885 (1991).
Proudhon, D., Briat, J.F. & Lescure, A.M. Iron induction of ferritin synthesis in soybean cell suspensions. >Plant Physiol. 90, 586– 590 (1989).
Kimata, Y. & Theil, E.C. Posttranscriptional regulation of ferritin during nodule development in soybean. Plant Physiol. 104, 263–270 (1994).
Van Wuytswinkel, O., Savino, G. & Briat, J.F. Purification and characterization of recombinant pea-seed ferritins expressed in Escherichia coli: influence of N-terminus deletions on protein solubility and core formation in vitro. Biochem. J. 305, 253–261 ( 1995).
Van Wuytswinkel, O. & Briat, J.F. Conformational changes and in vitro core-formation modifications induced by site-directed mutagenesis of the specific N-terminus of pea seed ferritin. Biochem. J. 305, 959–965 ( 1995).
Sturm, A., Voelker, T.A., Herman, E.M. & Chrispeels, M.J. Correct glycosylation, golgi-processing, and targeting to protein bodies of the vacuolar protein phytohemaggletinin in transgenic tobacco. Planta. 175, 170–183 ( 1988).
Ohtani, T., Wallace, J.C., Thompson, G.A., Galili, G. & Larkins, B.A. Normal and lysine containing zeins are unstable in transgenic tobaccoseeds. Plant Mol. Biol. 16, 117–128 (1990).
Takaiwa, F. et al. High level accumulation of soybean glycinin in vacuole-derived protein bodies in the endosperm tissue of transgenic tobacco seed. Plant Sci. 111, 39–49 ( 1995).
Takaiwa, F. et al. Characterization of common cis-regulatory elements responsible for the endosperm-specific expression of members of the rice glutelin multigene family. Plant Mol. Biol. 30, 1207– 1221 (1996).
Wu, C.-Y., Suzuki, A., Washida, H. & Takaiwa, F. The GCN4 motif in a rice glutelin gene is essential for endosperm-specific expression and is activated by Opaque-2 in transgenic rice plants. Plant J. 14, 673–683 (1998).
Utsumi, S. et al. Synthesis, processing and accumulation of modified glycinins of soybean in the seeds, leaves and stems of transgenic tobacco. Plant Sci. 92, 191–202 (1993).
Shirsat, A.H., Wilford, N. & Croy, R.R.D. 1989.Gene copy number and levels of expression in transgenic plants of a seed specific gene. Plant Sci. 61, 75–80.
Goto, F., Yoshihara, T. & Saiki, H. Iron accumulation in tobacco plants expressing soyabean ferritin gene. Transgenic Res. 7, 173– 180 (1998).
Delhaize, E. A metal-accumulator mutant of Arabidopsis thaliana. Plant Physiol. 111, 849–855 ( 1996).
Becker, D., Kemper, E., Schell, J. & Masterson, R. New plant binary vectors with selectable markers located proximal to the left T-DNA border. Plant Mol. Biol. 20, 1195– 1197 (1992).
Chu, C., Wang, C. & Sun, C. Establishment of an efficient medium for anther culture of rice through comparative experiment on the nitrogen sources. Scientia Sinica 18: 659-668 (1975).
Murashige, T. & Skoog, F. A revised medium for rapid growth and bio assays with tobacco tissue culture. Physiol. Plantarum. 15, 473–497 ( 1962).
Shure, M., Wessler, S. & Fedoroff, N. Molecular identification and isolation of Waxy locus in maize. Cell 35, 225– 233 (1983).
Enger-Blum, G., Meier, M., Frank, J. & Müller, G.A. Reduction of background problems in non-radioactive northern and Southern blot analyses enables higher sensitivity than 32P-based hybridizations. Anal. Biochem. 210, 235–244 (1993).
Bradfold, M.M. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 ( 1976).
Sambrook, J., Fritsch, E.F. & Maniatis, T. in Molecular Cloning 2nd ed.(eds. Ford, N. & Nolan, C.) 18.67–75. (Cold Spring Harbor Laboratory Press, New York, 1989).
Kamachi, K., Yamaya, T., Hayakawa, T., Mae, T. & Ojima, K. Vascular bundle–specific localization of cytosolic glutamine synthetase in rice leaves. Plant Physiol. 99, 1481– 1486 (1992).
Anonymous. in Standard Methods for the Examination of Water and Wastewater. 18th edn (eds Greenberg, A.E., Clesceri, L.S., & Eaton, A.D.) 3-66–3–68 (American Public Health Association, Washington, DC, 1992).
Acknowledgements
We wish to express our gratitude to J. Guerin, CSIRO (Commonwealth Scientific and Industrial Research Organization) for critically reading this paper and D. Becker (Max-Planck Institute) for providing the binary vector pGPTV-bar. We also thank Ms. Kusaka, Ms. Inoue, Ms. Miyake, and Ms. Tsunokawa (CRIEPI) for their technical assistance. This research was supported, in part, by research grants from the Enhancement Center-of-Excellence, the Special Coordination Funds for promoting the Science and Technology and from Bio-oriented Technology Research Advancement Institute (PRO-BRAIN) to F.T.
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Goto, F., Yoshihara, T., Shigemoto, N. et al. Iron fortification of rice seed by the soybean ferritin gene. Nat Biotechnol 17, 282–286 (1999). https://doi.org/10.1038/7029
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DOI: https://doi.org/10.1038/7029
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