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The quantitative evaluation offixed nitrogen by the endophytic bacteria in sweet potato and sugarcane

Research Project

Project/Area Number 16380053
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Plant nutrition/Soil science
Research InstitutionUniversity of Miyazaki

Principal Investigator

AKAO Shoichiro  University of Miyazaki, Agriculture, Professor (40347050)

Co-Investigator(Kenkyū-buntansha) SAEKI Yuichi  University of Miyazaki, Agriculture, Associate professor (50295200)
Project Period (FY) 2004 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥10,960,000 (Direct Cost: ¥10,600,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2006: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2005: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2004: ¥4,600,000 (Direct Cost: ¥4,600,000)
KeywordsSugarcane / Endophyte / Nitrogen fixation / Non-leguminous plant / Sweet potato / Broccoli / 感染経路 / GFP標識 / 異宿主植物 / 接種 / 増殖 / 固定窒素寄与率 / ^<15>N同位体希釈法
Research Abstract

The nitrogen-fixing bacteria which inhabits the inside plant is being called nitrogen-fixation endophytic bacteria. The plant which fixes nitrogen is limited to the leguminous plants which have a symbiotic relationship with RhJzobia and about ten kinds of plants that have a symbiotic relationship with Frankia However, several twenty years ago ; the existence of the nitrogen-fixing endophitic bacteria was isolated from sugarcane in Brazil And also it was reported that the rate of fixed nitrogen to total nitrogen might reach 60-70% in the sugarcane that nitrogen- fixing endophytic bacteria inhabited. The rate of contribution of N2 fixation to total plant N is equal to the Rhizobium-legume symbiosis. As for the nitrogen fixing endophytic bacteria, they were isolated from wetland rice, wheat, maize, sorghum, and various forage grasses in addition to sugarcane. This showed the possibility that a nitrogen fixing function is given to the non-leguminous plants. But, there are some problems to … More solve to realize this. They were the selection of useful endophytic bacteria and a colonization to the inside plants. The infection and multiplication to the plant of the nitrogen fixing endophytic bacteria and the estimation method of the nitrogen fixed by endophytic diazotrophs was examined in this research.
A research period is four years from the 16th year of Heisei until 19. At the first half of research period, the inoculation examination by GFP-labeled Herbaspirillum sp. was conducted. And the isolation of the endophytic diazotrophs, are responsible for the observed N2 fixation were tried from sugarcane and sweet potato. Moreover the gene encoding green fluorescent protein (GFP) via conjugation and electotroporation At the latter half of research period, inoculation effectiveness and internal colonization ability as an introduced in the non host plants of endophytic diazotrophs isolated from sugarcane and sweet potato was examined. And also the method for the amount of nitrogen fixed a period of duration was examined.
As a result, we determined the relative abundance of N2-fixing enndophytic bacteria in the stem of sugarcane cvs. NiF8, Nil5, N7 and KTN94-88, and in the stem, tuber, leaf and root of sweet potato cvs. Koganesengan and Shiroyutaka and 39 isolates, 27 isolates from sugarcane and 12 isolates from sweet potato, were obtained from the above-mentioned materials. Herbaspirillum sp., Enterobacter sp. and Pantoea sp. used in this experiment could be established in plants other than their original host, suggesting a lack of host specificity, although the extent of colonization may vary depending on the combination of host and endophyte. And, higher bacterial numbers were detected in plant tissues inoculated with higher inoculum (10^8 cells/ml)concentration treatment.
We have used the root part of the micropropagated sugarcane plant to the target mainly as an inoculation part of the useful N_2-fixing endophytic bacteria in this study. However we got the conclusion that a suitable delivery method for introducing endophytic bacteria into the vegetative propagation plants such as sugarcane is the vacuum infiltration method to stem pieces. Less

Report

(5 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • 2005 Annual Research Report
  • 2004 Annual Research Report
  • Research Products

    (35 results)

All 2008 2007 2006 2005 2004

All Journal Article (24 results) (of which Peer Reviewed: 10 results) Presentation (11 results)

  • [Journal Article] Infection,multiplication and evaluation of the nitrogen-fixing ability of Herbaspirillum sp.B501gfp1 in sugarcane sterns inoculated by the vacuum infiltration method2008

    • Author(s)
      Zakria, M., Udonishi K, Yamamoto A, Saeki Y, Akao S
    • Journal Title

      Microbes and Environments 23

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Influence of inoculation technique on the endophytic colonization of rice by Pantoea sp.isolated from sweet potato and Enterobacter sp.from sugarcane2008

    • Author(s)
      M. Zakria, K. Udonishi, T. Ogawa, A. Yamamoto, Y. Saeki, S. Akao
    • Journal Title

      Soil Science and Plant Nutrition 54

      Pages: 224-236

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Colonization and Nitrogen-Fixing Ability of Herbaspirillum sp.Strain B501gfp1 and Assessment of Its Growth-Promoting Ability in Cultivated Rice2007

    • Author(s)
      Muhammad Zakria, Joyce Njoloma, Yuichi Saeki, Shoichiro Akao
    • Journal Title

      Microbes and Environments 22

      Pages: 197-206

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Colonization and Nitrogen-Fixing Ability of Herbaspirillum sp. Strain B501gfpl and Assessment of Its Growth-Promoting Ability in Cultivated Rice2007

    • Author(s)
      Zakria, M., J., Njoloma, Y., Saeki, S., Akao
    • Journal Title

      Microbes and Environments 22(3)

      Pages: 197-206

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium,Herbaspirillum sp.B501gfp12006

    • Author(s)
      J. Njoloma, K. Tanaka, T. Shimizu, T. Nishiguchi, Z. Muhammad, R. Akashi, M. Oota, S. Akao.
    • Journal Title

      Biology and Fertility of Soils 43

      Pages: 137-143

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Incorporation of DNA sequence encording GFP into the isolates from sugercane and sweetpotato and colonization abihty and GFP-tagged bacteria2006

    • Author(s)
      K. Tanaka, T. Shimizu, M. Zakria, J. Njoloma, Y. Saeki, M. Sakai, T. Yamakawa, K. Minamisawa, S. Akao
    • Journal Title

      Microbes and Environment 21

      Pages: 122-128

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Colonization ability of Herbaspirillum sp.B501gfp1 in sugarcane,a non-host plant in the presence of indigenous diazotrophic endophytes2006

    • Author(s)
      Njoloma, J. P., M. Oota, K. Taroura, Y. Saeki, S. Akao
    • Journal Title

      African Journal of Biotechnology 5

      Pages: 836-841

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] サトウキビに接種した窒素固定エンドファイトHerbaspirillumの動態追跡のための緑色蛍光タンパク質の利用2006

    • Author(s)
      清水 友・豊里 学・久綱 泰代・佐伯 雄一・南澤 究・中西 康博・Joyce Nioloma・明石 良・赤尾 勝一郎
    • Journal Title

      土と微生物 60

      Pages: 3-9

    • NAID

      110009468401

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfpl2006

    • Author(s)
      Njoloma, J., K., Tanaka, T., Shimizu, T., Nishiguchi, Z., Muhammad, R., Akashi
    • Journal Title

      Biology and Fertility of Soils 43

      Pages: 137-143

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Incorporation of DNA sequence encording GFP into the isolates from sugercane and sweetpotato and colonization ability and GFP-tagged bacteria2006

    • Author(s)
      Tanaka, K., T., Shimizu, M., Zakria, J., Njoloma, Y., Saeki, M., Sakai, T., Yamakawa, K., Minamisawa, S., Akao
    • Journal Title

      Microbes and Environment 21(2)

      Pages: 122-128

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Colonization ability of Herbaspirillum sp. B501gfpl in sugarcane, a non-host plant in the presence of indigenous diazotronhic endoohvtes2006

    • Author(s)
      Njoloma, J.P., M., Oota, K., Taroura, Y., Saeki, S., Akao
    • Journal Title

      African Journal of Biotechnology 5(10)

      Pages: 836-841

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfp1.2006

    • Author(s)
      Joyce NJOLOMA, Kazunori TANAKA, Tasuku SHIMIZU, Tomohiro NISHIGUCHI, Zakria MUHAMMAD, Ryo AKASHI, Moriya OOTA, Shoichiro AKAO.
    • Journal Title

      Biology and Fertility of Soil 43

      Pages: 137-143

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Incorporation of DNA sequence encording GFP into the isolates from sugercane and sweetpotato and colonization ability and GFP-tagged bacteria2006

    • Author(s)
      K.Tanaka, T.Shimizu, M.Zakria, J.Njoloma, Y.Saeki, M.Sakai, T.Yamakawa, K.Minamisawa, S.Akao
    • Journal Title

      Microbes and Environment 21(2)

      Pages: 122-128

    • Related Report
      2006 Annual Research Report
  • [Journal Article] サトウキビに接種した窒素固定エンドファイトHerbaspirillumの動態追跡のための緑色蛍光タンパク質の利用2006

    • Author(s)
      清水友, 豊里学, 久綱泰代, 佐伯雄一, 南澤究, 中西康博, Joyce Njoloma, 明石良, 赤尾勝一郎
    • Journal Title

      土と微生物 60(1)

      Pages: 3-9

    • NAID

      110009468401

    • Related Report
      2006 Annual Research Report
  • [Journal Article] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfp1.2006

    • Author(s)
      Joyce NJOLOMA, Kazunori TANAKA, Tasuku SHIMIZU, Tomohiro NISHIGUCHI, Zakria' MUHAMMAD, Ryo AKASHI, Moriya OOTA, Shoichiro AKAO.
    • Journal Title

      Biology and Fertility of Soils (In press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] N2 fixation in sugarcane and population of N2-fixing endophytes in stem apoplast solution2005

    • Author(s)
      Asis Jr. Constancio A., katsuki Adachi, Shoichiro Akao
    • Journal Title

      Philippine Journal of Crop Science 29

      Pages: 45-56

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] 15N同位体希釈法によるサトウキビの固定窒素量の推定2005

    • Author(s)
      西口 友広・清水 友・Joyce Njoloma・大田 守也・佐伯 雄一・赤尾 勝一郎
    • Journal Title

      宮崎大学農学部研究報告 51

      Pages: 53-62

    • NAID

      110004634132

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues2005

    • Author(s)
      J. Njoloma, M. Oota, Y. Saeki, S. Akao
    • Journal Title

      African Journal of Biotechnology Vol.4

      Pages: 1372-1377

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] N_2fixation in sugarcane and population of N_2-fixing endophytes in stem apoplast solution2005

    • Author(s)
      Constancio, A., Katsuki, Adachi, Shoichiro, Akao
    • Journal Title

      Philippine Journal of Crop Science 29(2)

      Pages: 45-56

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] The Estimation of the amount of nitrogen fixation in the sugarcane by ^<15>N dilution technique.2005

    • Author(s)
      Nisiguchi, T., Shimizu, T., Njoloma, J., Oota, M., Saeki, Y., Akao, S.
    • Journal Title

      Bull. Faculty Agric. Univ. Miyazaki 51

      Pages: 53-62

    • NAID

      110004634132

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues2005

    • Author(s)
      Njoloma, J., M., Oota, Y., Saeki, S., Akao
    • Journal Title

      African Journal of Biotechnology 4(12)

      Pages: 1372-1377

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Detection of gfp expression from gfp-labelled bacteria spot inoculated onto sugarcane tissues2005

    • Author(s)
      Joyce Prisca, NJOLOMA, Moriya OOTA, Yuichi SAEKI, Shoichiro AKAO.
    • Journal Title

      African Journal of Biotechnology. Vol.4・12

      Pages: 1372-1377

    • Related Report
      2005 Annual Research Report
  • [Journal Article] N2 fixation in sugarcane and population of N2-fixing entophytes in stem apoplast solution2005

    • Author(s)
      Asis Jr., Constancio A., Katsuki Adachi, Shoichiro Akao
    • Journal Title

      Philippine Journal of Crop Science 29(2)

      Pages: 45-56

    • Related Report
      2005 Annual Research Report
  • [Journal Article] ^<15>N同位体希釈法によるサトウキビの固定窒素量の推定2005

    • Author(s)
      西口, 清水, ジョイス, 大田, 佐伯, 赤尾
    • Journal Title

      宮崎大学農学部研究報告 51(1・2)

      Pages: 53-62

    • NAID

      110004634132

    • Related Report
      2004 Annual Research Report
  • [Presentation] Colonization and growth promotion characteristics of Herbaspirillum sp.B501 and Enterobacter sp.for Brasica oleracea2007

    • Author(s)
      Zakria M., 大迫 明日香・佐伯 雄一・山本 昭洋・赤尾 勝一郎
    • Organizer
      日本土壌肥料学会九州支部会
    • Place of Presentation
      九州大学
    • Year and Date
      2007-04-25
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Colonization and growth promotion characteristics of Herbaspirillum sp. B501 and Enterobacter sp. for Brasica oleracea2007

    • Author(s)
      Zakria, M., A., Ohsako, Y., Saeki, A., Yamamoto, S., Akao
    • Organizer
      Japanese Society of Soil and Plant
    • Place of Presentation
      University nof Kyusyu
    • Year and Date
      2007-04-25
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Inoculation of Herbaspirillum spp.B501gfp1 on sugarcane under unsterile condition2006

    • Author(s)
      Njoloma J., M. Zakria, Y. Saeki, S. Akao
    • Organizer
      日本土壌肥料学会九州支部会
    • Place of Presentation
      宮崎市JAアズムホール
    • Year and Date
      2006-05-12
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Inoculation of Herbaspirillum spp. B501gfpl on sugarcane under unsterile condition2006

    • Author(s)
      Njoloma, J., M., Zakria, Y., Saeki, S., Akao
    • Organizer
      Japanese Society of Soil and Plant Nutrition
    • Place of Presentation
      JAAmuzu hallMiyazaki
    • Year and Date
      2006-05-11
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixillg endophytic bacterium,Herbaspirillum sp.B501 GFP12005

    • Author(s)
      Njoloma, J., K. Tanaka, T. Shimizu, T. Nishiguchi, M. Zakria, Y. Saeki, M. Oota, K. Minamizawa, S. AKAO
    • Organizer
      日本土壌肥料学会九州支部学会
    • Place of Presentation
      佐賀大学
    • Year and Date
      2005-05-12
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 窒素固定エンドファイトへのGFP(緑色蛍光タンパク質)遺伝子の導入方法の検討2005

    • Author(s)
      田中 和憲・清水 友・佐伯 雄一・山川 武夫^1・南沢 究^2・赤尾 勝一郎
    • Organizer
      日本土壌肥料学会九州支部会
    • Place of Presentation
      佐賀大学
    • Year and Date
      2005-05-12
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Studies on endophytic colonization,diazotrophic and growth promoting abilities of Herbaspirillum sp.B501gfp1,Pantoea sp.And Klebsiella sp.In rice cultivar(Nipponbare)2005

    • Author(s)
      Zakria M. J. Njoloma, T. Ogawa, S. Akao
    • Organizer
      日本土壌肥料学会九州支部会
    • Place of Presentation
      佐賀大学
    • Year and Date
      2005-05-12
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] The infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501 GFP12005

    • Author(s)
      Njoloma, J., K., Tanaka, T., Shimizu, T., Nishiguchi, M., Zakria, Y., Saeki, M., Oota, K., Minamizawa, S., AKAO
    • Organizer
      Japanese Society of Soil and Plant Nutrition
    • Place of Presentation
      University of Saga
    • Year and Date
      2005-05-12
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Studies on endophytic colonization, diazotrophic and growth promoting abilities of Herbaspirillum sp. B5Olgfpl, Pantoea sp. And Klebsiella sp. In rice cultivar(Nipponbare)2005

    • Author(s)
      Zakria, M.J., Njoloma, T., Ogawa, S., Akao
    • Organizer
      Japanese Society of Soil and Plant Nutrition
    • Place of Presentation
      University of Saga
    • Year and Date
      2005-05-12
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] サツマイモ及びサトウキビに内生する窒素固定エンドファイト細菌の分離と同定2004

    • Author(s)
      清水 友・佐伯 雄一・Joyce Njoloma・Constancio A. Asis・安達 克樹・中西 康博・赤尾 勝一郎
    • Organizer
      植物微生物研究会
    • Place of Presentation
      広島大学
    • Year and Date
      2004-06-05
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] 15N同位体希釈法によるサトウキビの窒素固定量の推定2004

    • Author(s)
      西口 友広・清水 友・Joyce Nloloma・大田 守也・佐伯 雄一・赤尾 勝一郎
    • Organizer
      植物微生物研究会
    • Place of Presentation
      広島大学
    • Year and Date
      2004-06-05
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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