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Genomic ecology of bradyrhizobia for denitrification systems to repress N_2O evolution

Research Project

Project/Area Number 14360037
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Plant nutrition/Soil science
Research InstitutionTohoku University

Principal Investigator

MINAMISAWA Kiwamu  Tohoku University, Graduate School of Life Sciences, Professor, 大学院・生命科学研究科, 教授 (70167667)

Co-Investigator(Kenkyū-buntansha) 鮫島 玲子  東北大学, 大学院・生命科学研究科, 日本学術振興会特別研究員
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥14,900,000 (Direct Cost: ¥14,900,000)
Fiscal Year 2004: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2003: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2002: ¥9,800,000 (Direct Cost: ¥9,800,000)
KeywordsBradyrhizobium japonicum / denitrofication / nitrous oxide(N_2O) / nosZ / symbiosis / soybean / 亜酸化窒素 / ^<15>N
Research Abstract

Denitrification is anaerobic respiration using nitrate (NO_3^-), nitrite (NO_2^-), nitric oxide (NO), and nitrous oxide (N_2O) as terminal electron acceptors. Nitrous oxide (N_2O) is a key atmospheric greenhouse gas that contributes to global climatic change through radiative warming and depletion of stratospheric ozone. Agricultural lands are one of major sources, and significantly contribute the net increase in atmospheric N_2O. We used N_2, which has a natural abundance of ^<15>N (0.366 atom%), as a carrier gas in a gas chromatograph with a thermal conductivity detector to detect ^<15>N-N_2 quantitatively. When Bradyrhizobium japonicum USDA110 was cultured anaerobically in medium containing ^<15>N-NO_3, ^<15>N-N_2 was detected stoichiometrically. Analysis of 63 bradyrhizobial isolates revealed that denitrification ability varied with field site. To examine whether nos gene cluster of Bradyrhizobium japonicum is responsible for N_2O reduction in symbiosis, we constructed a nosZ delet … More ion mutant of B.japonicum USDA110. The activity of N_2O reductase was determined by N_2O uptake and conversion of ^<15>N_2O into ^<15>N_2. Free-living cells of USDA110 showed N_2O reductase activity, while nosZ mutant did not. Complementation of cosmids containing nos gene cluster of B.japonicum USDA 110 to the nosZ mutant restored the N_2O rductase activity, indicating nosZ-dependent N_2O reductase activity and validity of the mutant. When the detached soybean nodules formed with wild-type USDA110 was fed with ^<15>N_2O, the nodules rapidly emitted ^<15>N_2 outside at a ratio of approximately 98.5% of ^<15>N-N_2O uptake. On the other hand, nosZ mutant-inoculated nodules did not mediate the conversion of ^<15>N_2O to ^<15>N_2 at all. Moreover, substantial decrease of N_2O concentration was detected in the rhizosphere of intact soybean plant inoculated with wild-type USDA110. These results demonstrate that the conversion of N_2O to N_2 depends exclusively to the function of nos gene cluster, and soybeans nodulated with B.japonicum carrying nos cluster are able to remove N_2O gas in rhizosphere environments Less

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (24 results)

All 2004 2003 2002 Other

All Journal Article (12 results) Publications (12 results)

  • [Journal Article] New method of denitrification analysis of Bradyrhizobium field isolates by gas chromatographic determination of ^<15>N-N_2.2004

    • Author(s)
      Sameshima-Saito, R., K.Chiba, K.Minamisawa
    • Journal Title

      Appl.Environ.Microbiol 70

      Pages: 2886-2891

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome.2004

    • Author(s)
      Uchiumi, T., T.Ohwada, et al.
    • Journal Title

      J.Bacteriol. 186

      Pages: 2439-2448

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] New method of denitrification analysis of Bradyrhizobium field isolates by gas chromatographic determination of ^<15>N-N_2.2004

    • Author(s)
      Sameshima-Saito, R., K.Chiba, K.Minamisawa
    • Journal Title

      Appl.Environ.Microbiol. 70

      Pages: 2886-2891

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome.2004

    • Author(s)
      Uchiumi, T., T.Ohwada, M.Itakura, H.Mitsui, N.Nukui, P.Dawadi, T.Kaneko, S.Tabata, T.Yokoyama, T.Tejima, K.Saeki, H.Omori, M.Hayashi, T.Maekawa, R.Sriprang, Y.Murooka, S.Tajima, K.Simomura, M.Nomura, A.Suzuki, Y.Shimoda, K.Sioya, M.Abe, d K.Minamisawa
    • Journal Title

      J.Bacteriol. 186

      Pages: 2439-2448

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] New method of denitrification analysis of Bradyrhizobium field isolates by gas chromatographic determination of ^<15>N-N_22004

    • Author(s)
      Sameshima-Saito, R., Chiba, K., Minamisawa, K.
    • Journal Title

      Appl.Environ.Microbiol. 70・5

      Pages: 2886-2891

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Expression islands clustered on symbiosis island of Mesorhizobium loti genome2004

    • Author(s)
      Uchiumi, T, Minamisawa K.(共著者24名)
    • Journal Title

      J.Bacteriol. 186・8

      Pages: 2439-2448

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Novel endophytic nitrogen-fixing clostridia from the grass Miscanthus sinensis as revealed by terminal restriction length polymorphism2004

    • Author(s)
      Miyamoto, T., Kawahara, M, Minamisawa, K.
    • Journal Title

      Appl.Environ.Microbiol. 70・11

      Pages: 6580-6586

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Evaluation of soil DNAs from arable lands in Japan with a modified direct extraction method2004

    • Author(s)
      Ikeda, S., Watanabe, K., Minamisawa, K., You, N.
    • Journal Title

      Microbes Environments. 19・4

      Pages: 301-309

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Phylogeny and distribution of extra-slow-growing Bradyrhizobium japonicum harboring high copy numbers of RSα, RSβ and IS16312003

    • Author(s)
      Sameshima, R., T.Isawa, M.J.Sadowsky, T.Hamada, H.Kosai, et al.
    • Journal Title

      FEMS Microbiol.Ecol. 44

      Pages: 191-202

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Phylogeny and distribution of extra-slow-growing Bradyrhizobium japonicum harboring high copy numbers of RSα, RSβ and IS1631.2003

    • Author(s)
      Sameshima, R., T.Isawa, M.J.Sadowsky, T.Hamada, H.Kosai, A.Shutsrirung, H.Mitsui, K.Minamisawa
    • Journal Title

      FEMS Microbiol.Ecol. 44

      Pages: 191-202

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110.2002

    • Author(s)
      Kaneko, T., Y.Nakamura, S.Sato, K.Minamisawa, et al.
    • Journal Title

      DNA Research. 9

      Pages: 189-197

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110.2002

    • Author(s)
      Kaneko, T., Y.Nakamura, S.Sato, K.Minamisawa, T.Uchiumi, S.Sasamoto, A.Watanabe, K.Idesawa, M.Iriguchi, K.Kawashima, M.Kohara, M.Matsumoto, S.Shimpo, H.Tsuruoka, T.Wada, M.Yamada, S.Tabata
    • Journal Title

      DNA Research. 9

      Pages: 189-197

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Sameshima, R., Isawa, T, Sadowsky, M.J., Hamada, T., Kasai, H., Shutsrirung, A, Mitsui, H, Minamisawa, K.: "Phylogeneny and distribution of extra-slow-growing Bradyrhizobium japonicum harboring high copy numbers of RSα, RSβ and IS1631"FEMS Microbiol.Ecol.. 44. 191-202 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Okazaki, S., Yuhashi, K., Minamisawa, K.: "Quantitative and time-course evaluation of nodulation competitiveness of rhizobitoxine-producing Bradyrhizobium elkanii"FMES Microbiol.Ecol.. 45(1). 155-160 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Okazaki, S., Sugawara, M., Minamisawa, K.: "Bradythizobium elkanii rtxC gene is required for expression of symbiotic phenotypes in the final step of rhizobitoxine biosynthesis"Appl.Environ.Microbiol.. 70(1). 535-541 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Uchiumi, T., Minamisawa, K.(共著者24名): "Expression islands clustered on symbiosis island of Mesorhizobium loti genome"J.Bacteriol.. (印刷中).

    • Related Report
      2003 Annual Research Report
  • [Publications] Sameshima-Saito, R., CHiba, K., Minamisawa, K.: "New Method of denitrification analysis of Bradyrhizobium field isolates by gaschromatographic determination of ^<15>N-N_2"Appl.Environ.Microbiol.. (印刷中).

    • Related Report
      2003 Annual Research Report
  • [Publications] Nukui, N., Ezura, H., Minamisawa, K.: "Transgenic Lotus japonicus with an ethylen receptor gene Cm-ERS1/H70A enhances formation of infection treads and nodule primordia"Plant Cell Physiol.. (印刷中).

    • Related Report
      2003 Annual Research Report
  • [Publications] Minamisawa K. et al.: "Horizontal transfer of nodulation genes in soils and microcosms from Bradyrhizobium japonicum to B. elkanii"Microbe. Environ.. 17. 82-90 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kaneko, T. et al.: "Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110"DNA Research. 9. 189-197 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sameshima, R. et al.: "Phylogeny and distribution of extra-slow-growing Bradyrhizobium japonicum harboring high copy numbers of RS-alpha, RS-beta, and IS 1631"FEMS Microbiol. Ecol.. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okazaki.S.et al.: "Quantitative and time-course evaluations on nodulation competitiveness of rhizobitoxine-producing Bradyrhizobium elknaii"FEMS Microbiol. Ecol.. (第1回審査後改訂稿作成中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kaneko, T. et al.: "Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110 (Supplement)"DNA Research. 9. 225-256 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kaneko, T., Saeki, T., Minamisawa, K.: "Genome analysis of Mesorhizobium loti : A symbiotic partner to Lotus japonicus. In Biotechnology in Agriculture and Forestry"Springer-Verlag, Heidelberg (in press). 203-216 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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