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Expression of Maize Gene Encoding C4-Pyruvate Orthophosphate Dikinase (PPDK) and C4-Phosphoenolpyruvate Carboxylase (PEPC) in Transgenic Arabidopsis

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Abstract

To improve the efficiency of CO2 fixation in C3 photosynthesis, C4-cycle genes were overexpressed in Arabidopsis plants. In this study, a full-length cDNA for pyruvate orthophosphate dikinase (PPDK) with 971 amino acids was isolated from Zea mays using LA-PCR. The PPDK gene (GenBank accession number GU363532) and the phosphoenolpyruvate carboxylase (PEPC) gene (GenBank accession number: FJ415327) derived from a previous study were introduced into wild-type Arabidopsis both individually and in combination. The vector pCAMBIA3301 (p3301) was introduced into Arabidopsis as a control using an Agrobacterium-mediated system. The integration, transcription, and translation of the PPDK and the PEPC genes were confirmed by Southern blot, quantitative real-time PCR, and Western blot analysis. The average enzyme activity of C4-PPDK and C4-PEPC peaked at 4.63- and 5.81-fold compared to that of controls, respectively. The insertion of PPDK was found to increase the activity of PEPCase and vice versa. The photosynthetic rates of C4-PPDK, C4-PEPC, and C4-PKC(PPDK+PEPC) transgenic lines increased to 123.3, 136.4, and 146.6 % compared to that of controls, respectively.

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Acknowledgments

We would like to thank Dr. Zhenhua Wang for donating the inbred maize lines and Drs. Diane Hu and Huiwei Wang for the critical reading of this manuscript. This work was supported by the Natural Science Fund of China (30971785), the Genetically Modified Organism Breeding Major Projects of China (2011ZX08002-003), and the Henan Province Outstanding Person Plan Program (104100510021).

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Authors and Affiliations

  1. Henan Agricultural University, Zhengzhou, 450002, Henan, China

    Yu-Min Wang & Xi-He Du

  2. Wheat Research Center, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, Henan, China

    Wei-Gang Xu, Lin Hu, Lei Zhang & Yan Li

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  1. Yu-Min Wang
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  2. Wei-Gang Xu
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  4. Lei Zhang
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Correspondence to Wei-Gang Xu.

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Wang, YM., Xu, WG., Hu, L. et al. Expression of Maize Gene Encoding C4-Pyruvate Orthophosphate Dikinase (PPDK) and C4-Phosphoenolpyruvate Carboxylase (PEPC) in Transgenic Arabidopsis . Plant Mol Biol Rep 30, 1367–1374 (2012). https://doi.org/10.1007/s11105-012-0451-5

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