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Molecular and physiological evaluation of transgenic tobacco plants expressing a maize phosphoenolpyruvate carboxylase gene under the control of the cauliflower mosaic virus 35S promoter

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Abstract

The expression of maize (Zea mays) phosphoenolpyruvate carboxylase (PEPC) gene constructs in transgenic tobacco plants (Nicotiana tabacum) was studied. Where transcription was under the control of a CaMV 35S promoter, maize PEPC transcripts of the correct size were detected. Western blot analysis indicated that the transgenic plants contained about twice as much PEPC as non-transformed plants. Furthermore, the enzymatic activity of PEPC in the leaves of these transgenic plants was up to twice as high as that in non-transformed plants. Two forms of PEPC with different kinetic properties were identified in leaf extracts of the transgenic plants: one form (the maize isoform) gave a high apparentK m value for phosphoenolpyruvate (PEP) and a high maximum activity, and the other (the tobacco isoform) exhibited a low apparentK m value for PEP and a low maximum activity. These biochemical differences resulted in several significant physiological changes in the transgenic plants: (1) the growth rate of the transgenic plants was lower than that of non-transgenic plants: (2) chlorophyll content per leaf area was relatively lower in the transgenic plants; and (3) the quantum yield of photosynthesis in the transgenic plants was not affected by changes in leaf temperature.

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Authors and Affiliations

  1. Department of Biological Science, Faculty of Science, Hokkaido University, 060, Sapporo, Japan

    Hiroyuki Kogami, Naoki Sentoku, Shigeyuki Tanifuji & Seiichi Toki

  2. Institute of Biological Science, University of Tsukuba, 305, Tsukuba, Japan

    Mariko Shono

  3. Hokkaido Research Center, Forestry and Forest Products Research Institute, 062, Sapporo, Japan

    Takayoshi Koike

  4. Department of Pure and Applied Science, College of Arts and Science, University of Tokyo, 153, Tokyo, Japan

    Syuuichi Yanagisawa

  5. Department of Agrobiology, Faculty of Agriculture, Kyoto University, 606-01, Kyoto, Japan

    Katsura Izui

  6. Institute of Molecular and Cellular Biosciences, University of Tokyo, 113, Tokyo, Japan

    Hirofumi Uchimiya

Authors
  1. Hiroyuki Kogami
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  2. Mariko Shono
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  3. Takayoshi Koike
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  4. Syuuichi Yanagisawa
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  5. Katsura Izui
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  7. Shigeyuki Tanifuji
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  8. Hirofumi Uchimiya
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  9. Seiichi Toki
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Kogami, H., Shono, M., Koike, T. et al. 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 Research 3, 287–296 (1994). https://doi.org/10.1007/BF01973588

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  • DOI: https://doi.org/10.1007/BF01973588

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