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Planta

, Volume 219, Issue 1, pp 14–22 | Cite as

Formation of unidentified nitrogen in plants: an implication for a novel nitrogen metabolism

  • Hiromichi Morikawa
  • Misa Takahashi
  • Atsushi Sakamoto
  • Toshiyuki Matsubara
  • Gen-Ichiro Arimura
  • Yoshifumi Kawamura
  • Kazunari Fukunaga
  • Kounosuke Fujita
  • Naoki Sakurai
  • Toshifumi Hirata
  • Hiroshi Ide
  • Nobuaki Nonoyama
  • Hitomi Suzuki
Original Article

Abstract

Plants take up inorganic nitrogen and store it unchanged or convert it to organic forms. The nitrogen in such organic compounds is stoichiometrically recoverable by the Kjeldahl method. The sum of inorganic nitrogen and Kjeldahl nitrogen has long been known to equal the total nitrogen in plants. However, in our attempt to study the mechanism of nitrogen dioxide (NO2) metabolism, we unexpectedly discovered that about one-third of the total nitrogen derived from 15N-labeled NO2 taken up by Arabidopsis thaliana (L.) Heynh. plants was converted to neither inorganic nor Kjeldahl nitrogen, but instead to an as yet unknown nitrogen compound(s). We here refer to this nitrogen as unidentified nitrogen (UN). The generality of the formation of UN across species, nitrogen sources and cultivation environments for plants has been shown as follows. Firstly, all of the other 11 plant species studied were found to form the UN in response to fumigation with 15NO2. Secondly, tobacco (Nicotiana tabacum L.) plants fed with 15N-nitrate appeared to form the UN. And lastly, the leaves of naturally fed vegetables, grass and roadside trees were found to possess the UN. In addition, the UN appeared to comprise a substantial proportion of total nitrogen in these plant species. Collectively, all of our present findings imply that there is a novel nitrogen mechanism for the formation of UN in plants. Based on the analyses of the exhaust gas and residue fractions of the Kjeldahl digestion of a plant sample containing the UN, probable candidates for compounds that bear the UN were deduced to be those containing the heat-labile nitrogen–oxygen functions and those recalcitrant to Kjeldahl digestion, including organic nitro and nitroso compounds. We propose UN-bearing compounds may provide a chemical basis for the mechanism of the reactive nitrogen species (RNS), and thus that cross-talk may occur between UN and RNS metabolisms in plants. A mechanism for the formation of UN-bearing compounds, in which RNS are involved as intermediates, is proposed. The important broad impact of this novel nitrogen metabolism, not only on the general physiology of plants, but also on plant substances as human and animal food, and on plants as an integral part of the global environment, is discussed.

Keywords

Arabidopsis Kjeldahl nitrogen Nitrate Nitric oxide Nitrogen dioxide Reactive nitrogen species 

Abbreviations

NO

Nitric oxide

NO2

Nitrogen dioxide

RNS

Reactive nitrogen species

UN

Unidentified nitrogen

TNNAT, RNNAT, INNAT and UNNAT

Total, Kjeldahl, inorganic and unidentified nitrogen in naturally fed plants, respectively

TNNIT, RNNIT, INNIT and UNNIT

Total, Kjeldahl, inorganic and unidentified nitrogen derived from nitrate, respectively

TNNO2, RNNO2, INNO2 and UNNO2

Total, Kjeldahl, inorganic and unidentified nitrogen derived from NO2, respectively

Notes

Acknowledgements

We thank T. Sugiyama of Nagoya University and T. Yamaya of Tohoku University for discussions on nitrogen metabolism, and M. Kawahara of Hiroshima University for nitrogen analysis. This work was supported in part by the Research for the Future Program, Japanese Society for the Promotion of Science (JSPS-RFTF96L00604) and by a Grant-in-Aid for Creative Scientific Research (no.13GS0023) from the Japan Society for the Promotion of Science.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Hiromichi Morikawa
    • 1
    • 2
  • Misa Takahashi
    • 1
  • Atsushi Sakamoto
    • 1
  • Toshiyuki Matsubara
    • 2
  • Gen-Ichiro Arimura
    • 1
  • Yoshifumi Kawamura
    • 1
  • Kazunari Fukunaga
    • 1
  • Kounosuke Fujita
    • 3
  • Naoki Sakurai
    • 4
  • Toshifumi Hirata
    • 1
  • Hiroshi Ide
    • 1
  • Nobuaki Nonoyama
    • 5
  • Hitomi Suzuki
    • 5
  1. 1.Department of Mathematical and Life Sciences, Graduate School of ScienceHiroshima UniversityHigashi-HiroshimaJapan
  2. 2.Core Research for Evolutional Science and Technology (CREST)Japan Science and Technology Agency (JST)KawaguchiJapan
  3. 3.Graduate School of Biosphere Science, Faculty of Applied Biological ScienceHiroshima UniversityHigashi-HiroshimaJapan
  4. 4.Department of Environmental Studies, Faculty of Integrated Arts and SciencesHiroshima UniversityHigashi-HiroshimaJapan
  5. 5.Department of Chemistry, Faculty of ScienceKwansei Gakuin UniversityHyogoJapan

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