Abstract
Using an Agrobacterium -mediated transformation system, we have introduced the intact gene of maize phospho enol pyruvate carboxylase (PEPC), which catalyzes the initial fixation of atmospheric CO 2 in C 4 plants into the C 3 crop rice. Most transgenic rice plants showed high-level expression of the maize gene; the activities of PEPC in leaves of some transgenic plants were two- to threefold higher than those in maize, and the enzyme accounted for up to 12% of the total leaf soluble protein. RNA gel blot and Southern blot analyses showed that the level of expression of the maize PEPC in transgenic rice plants correlated with the amount of transcript and the copy number of the inserted maize gene. Physiologically, the transgenic plants exhibited reduced O 2 inhibition of photosynthesis and photosynthetic rates comparable to those of untransformed plants. The results demonstrate a successful strategy for installing the key biochemical component of the C 4 pathway of photosynthesis in C 3 plants.
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References
Bowes, G. 1996. Photosynthetic responses to changing atmospheric carbon dioxide concentration, pp. 387–407, in Advances in photosynthesis, photosynthesis and the environment, vol 5. Baker, N.R. (ed.). Kluwer Academic Publisher, Dordrecht, The Netherlands.
Ogren, W.L. 1984. Photorespiration: pathways, regulation, and modification. Annu. Rev. Plant Physiol. 35: 415– 442.
Somerville, C.R. 1990. The biochemical basis for plant improvement, pp. 490–501, in Plant physiology and plant molecular biology. Dennis, D.T., Turpin, D.H. (eds.). Longman Group, Essex, UK.
Edwards, G.E. and Walker, D.A. 1983. C 3, C4: mechanisms, and cellular and environmental regulation of photosynthesis. Blackwell Science Publishers, London, UK.
Hatch, M.D. 1987. C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochim. Biophys. Acta. 895: 81–106.
Furbank, R.T. and Taylor, W.C. 1995. Regulation of photosynthesis in C3 and C4 plants: a molecular approach. Plant Cell 7: 797–807.
Ku, M.S.B., Kano-Murakami, Y., and Matsuoka, M. 1996. Evolution and expression of C4 photosynthesis genes. Plant Physiol. 111: 949–957.
Dai, Z., Ku, M.S.B., and Edwards, G.E. 1993. C4 photosynthesis: the CO2 concentration mechanism and photorespiration. Plant Physiol. 103: 83–90.
Brown R.H. and Bouton, J.H. 1993. Physiology and genetics of interspecific hybrids between photosynthetic types. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44: 435– 456.
Fushimi, T., Umeda, M., Shimazaki, T., Kato, A., Toriyama, K., and Uchimiya, H. 1994 . Nucleotide sequence of a rice cDNA similar to a maize NADP-dependent malic enzyme. Plant Mol. Biol. 24: 965– 967.
Walter, M.H., Grima-Pettenati, J., and Feuillet, C. 1994. Characterization of a bean (Phaseolus vulgaris L.) malic enzyme. Eur. J. Biochem. 224: 999–1009.
Schaaf, J., Walter, M.H., and Hess, D. 1995. Primary metabolism in plant defense. Plant Physiol. 108: 949– 960.
Drincovich, M.F., Casati, P., Andreo, C.S., Donahue, R., and Edwards, G.E. 1988. UV-B induction of NADP-malic enzyme in etiolated and green maize seedlings. Plant Cell Environ. 21: 63–70.
Hudspeth, R.L., Grula, J.W., Dai, Z., Edwards, G.E., and Ku, M.S.B. 1991. Expression of maize phosphoenolpyruvate carboxylase in transgenic tobacco. Effects on biochemistry and physiology. Plant Physiol. 98: 458– 464.
Kogami, H., Shono, M., Koike, T., Yanagiswa, S., Izui, K., Sentoku, N. et al. 1994. Molecular and physiological evaluation of transgenic tobacco plants expressing a maize phosphoenolpyruvate carboxylase gene under the control of the cauliflower mosaic virus 35S promoter. Transgenic Res. 3: 287–296.
Gehlen, J., Panstruga, R., Smets, H., Merkelbach, S., Kleines, M., Porsch, P. et al. 1996. Effects of altered phosphoenolpyruvate carboxylase activities on transgenic C3 plant Solanum tuberosum . Plant Mol. Biol. 32: 831– 848.
Gallardo, F., Miginiac-Maslow, M., Sangwan, R.S., Decottignies, P., Keryer, E., Dubois, F. et al. 1995. Monocotyledonous C4 NADP-malate dehydrogenase is efficiently synthesized, targeted to chloroplasts and processed to an active form in transgenic plants of the C3 dicotyledonous tobacco. Planta 197: 324– 332.
Toki, S. 1997. Rapid and efficient Agrobateriun-mediated transformation in rice. Plant Mol. Biol. Reporter 15: 16 –21.
Chan, M.T., Chang, H.H., Ho, S.L., Tong, W.F., and Yu, S.M. 1993. Agrobacterium-mediated production of transgenic rice plants expressing a chimeric α-amylase promoter/β-glucuronidase gene. Plant Mol. Biol. 22: 491– 506.
Hiei, Y., Ohata, S., Komari, T., and Komashiro, T. 1994 . Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6: 271–282.
Rashid, H., Yokoi, S., Toriyama, K., and Hinata, K. 1996. Transgenic plant production mediated by Agrobacterium in Indica rice. Plant Cell Report 15: 727– 730.
Ku, M.S.B. and Edwards, G.E. 1977. Oxygen inhibition of photosynthesis. I. Temperature dependence and relation to CO 2/O2 solubility ratio. Plant Physiol. 59: 986–990.
Dai, Z., Ku, M.S.B., and Edwards, G.E. 1995. C4 photosynthesis. The effects of leaf development on the CO2-concentrating mechanism and photorespiration in maize. Plant Physiol. 107: 815–825.
Matsuoka, M., Kyozuka, J., Shimamoto, K., and Kano-Murakami, Y. 1994. The promoters of two carboxylases in a C 4 plant (maize) direct cell-specific, light-regulated expression in a C3 plant (rice). Plant J. 6: 311–319.
Matsuoka, M., Tada, Y., Fujimura, T., and Kano-Murakami, Y. 1993. Tissue-specific light-regulated expression directed by the promoter of a C4 gene, maize pyruvate, orthophosphate dikinase, in a C3 plant, rice. Proc. Natl. Acad. Sci. USA 90: 9586–9590.
Eckes, P., Schmitt, P., Daub, W., and Wengenmayer, F. 1989. Overexpression of alfalfa glutamine synthetase in transgenic tobacco plants. Mol. Gen. Genet. 217: 263 –268.
Birch, R.G. 1997. Plant transformation: problems and strategies for practical application. Ann. Rev. Plant Physiol. Plant Mol. Biol. 48: 297–326.
Simpson, G.G., and Filipowicz, W. 1996 . Splicing of precursors to messenger RNA in higher plants: mechanism, regulation and sub-nuclear organization of the spliceosomal machinery. Plant Mol. Biol. 32: 1–41.
Koziel, M.G., Carozzi, N.B., and Desai, N. 1996. Optimizing expression of transgenes with an emphasis on post-transcriptional events. Plant Mol. Biol. 32: 393–405.
Cogoni, C., and Macino, G. 1997. Conservation of transgene-induced post-transcriptional gene silencing in plants and fungi. Trends in Plant Sci. 2: 438– 443.
Matzke, M., and Matzke, A.M. 1995. How and why do plants inactivate homologous (trans)genes? Plant Physiol . 107: 679–685.
Callis, J., Fromm, M., and Walbot, V. 1987. Introns increase gene expression in cultured maize cells. Genes Dev.. 1: 1183 –1200.
Keith, B. and Chua, N-H. 1986. Monocot and dicot pre-mRNAs are processed with different efficiencies in transgenic tobacco. EMBO J. 5: 2419– 2425.
Goodall, G.J. and Filipowicz, W. 1991. Different effects of intron nucleotide composition and secondary structure on pre-mRNA splicing in monocot and dicot plants. EMBO J. 10: 2635–2644.
Harris, G.C., Cheesbrough, J.K., and Walker, D.A. 1983. Effects of mannose on photosynthetic gas exchange in spinach leaf discs. Plant Physiol. 71: 108–111.
Harley, P.C., and Sharkey, T.D. 1991. An improved model of C3 photosynthesis at high CO2: reversed O2 sensitivity explained by lack of glycerate reentry into the chloroplast. Photosynth. Res. 27: 169– 178.
Matsuoka, M. and Minami, E. 1989. Complete structure of the gene for phosphoenolpyruvate carboxylase from maize. Eur. J. Biochem. 181: 593– 598.
Kanai, R. and Edwards, G.E. 1973. Separation of mesophyll protoplasts and bundle heath cells from maize leaves for photosynthetic studies. Plant Physiol. 51: 1133– 1137.
Dai, Z., Ku, M.S.B., Zhang, D., and Edwards, G.E. 1994 . Effects of growth regulators on the induction of Crassulacean acid metabolism in the facultative halophyte Mesembryanthemum crystallinum L. Planta 192: 287– 294.
Murray, M.G. and Tompson W.F. 1980. Rapid isolation of high molecular weight plant DNA. Nucl. Acids Res. 8: 4321–4325.
McGookin, R. 1984. RNA extraction by the guanidine thiocyanate procedure, pp. 113–116, in Walker, J.M. (ed.). Methods in molecular biology, Vol. 2, Humana Press, Totawa, NJ.
Acknowledgements
This work was supported in part by a PROBRAIN grant to M.M. and M.M. from the Bio-Oriented Technology Research Advancement Institution (BRAIN) of Japan.
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Ku, M., Agarie, S., Nomura, M. et al. High-level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants. Nat Biotechnol 17, 76–80 (1999). https://doi.org/10.1038/5256
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DOI: https://doi.org/10.1038/5256
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