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High-level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants

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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Figure 1: Schematic representation of the intact maize pepc gene and the selective antibiotic resistance gene (hygromycin phosphotransferase, HPT II) used for rice transformation.
Figure 2: The activities of PEPC in primary transgenic rice plants.
Figure 3: (A) Composition of leaf soluble protein extracted from maize (50 μg), Kitaake, and primary (T1) transgenic rice plants derived from Kitaake (35 μg).
Figure 4: (A) Composition of leaf soluble protein, (B) RNA gel blot, and (C) Southern blot analyses of six independent transgenic rice plants (lane 1–6) with maize (lane M) and untransformed rice (lane R).
Figure 5: (A) Oxygen inhibition of photosynthesis for Kitaake (), Nipponbare (), and transgenic rice plants as a function of PEPC activity.

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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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Correspondence to Maurice S.B. Ku.

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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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