Genetic engineering of nitrite reductase gene improves uptake and assimilation of nitrogen dioxide by Rhaphiolepis umbellata (Thunb.) Makino

  • Shigeto Jun
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
  • Yoshihara Sanae
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
  • Adam Suaad E. H.
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University
  • Sueyoshi Kuni
    Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University
  • Sakamoto Atsushi
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
  • Morikawa Hiromichi
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
  • Takahashi Misa
    Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)

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Abstract

Hypocotyl sections of Rhaphiolepis umbellata (Thunb.) Makino were transformed by Agrobacterium tumefaciens bearing a binary vector pIG121-AtNiR, which contains cDNA of the nitrite reductase gene from Arabidopsis thaliana (Atni) under the control of a modified cauliflower mosaic virus 35S promoter and chimeric hygromycin phosphotransferase gene (hph). A 4% of the hypocotyl explants transfected with Agrobacterium formed hygromycin resistant adventitious shoots, and most of them rooted upon root induction treatment. The presence and expression of the introduced transgene in putative transgenic plants (12 months after transfection) were respectively confirmed by polymerase chain reaction (PCR) analysis using primers specific to Atni and by western blot analysis using anti NiR antibody. A total of 37 transgenic plant lines were obtained. Plants 33 months after the transfection were fumigated with 200±50 ppb 15NO2 under the natural light for one week in the fumigation chamber in a confined glasshouse. The amount of total nitrogen derived from NO2 (reflecting uptake of NO2) and that of Kjeldahl nitrogen derived from NO2 (reflecting assimilation of NO2) were determined using a mass spectrometer. One of the 9 transgenic plants tested was 1.6–2.0 times higher both in the uptake and assimilation of NO2 than non-transformed wild-type ones.

Journal

  • Plant Biotechnology

    Plant Biotechnology 23 (1), 111-116, 2006

    Japanese Society for Plant Biotechnology

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