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2004 Fiscal Year Final Research Report Summary

Development of Symbiotic Engineering for Phytoremediation

Research Project

Project/Area Number 15208032
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Boundary agriculture
Research InstitutionOsaka University

Principal Investigator

YAMASHITA Mitsuo  Osaka Univ., Eng., Assoc.Prof., 大学院・工学研究科, 助教授 (40220347)

Co-Investigator(Kenkyū-buntansha) ONO Hisao  Osaka Univ., Eng., Assis.Prof., 遺伝情報実験センター, 助手 (40224274)
HAYASHI Makoto  Osaka Univ., Eng., Assis.Prof., 大学院・工学研究科, 助手 (30291933)
Project Period (FY) 2003 – 2004
Keywordsphytoremediation / symbiotic engineering / Cd / metallothionein / Phytochelatin / heavy metal binding protein / ファイトレメディエーション
Research Abstract

In this research project, we have been developed a new bioremediation technology utilizing the symbiotic relationships of a leguminous plant and its rhizobia to clean up soils contaminated with heavy metals. In the rhizosphere, plant roots secrete various nutrients and attractants stimulating microbes to swarm plant roots. We believe that microbes have the ability to gather and carry metal ions to plant roots. The metal ions released by these microorganisms are taken in the plants. The phenomena gave me an idea for bioremediation using plants and their microbes.
Our major findings for this 2 year project included :
a)We constructed transgenic renge-soh (Astragalus sinicus) with tetrameric metallothionein (MTL4) gene for increased uptake of heavy metals by plants. To express a foreign gene in bacteroids in nodules, we chose the two promoters, PnifH and PnolB. The MTL4 gene expressed in nodules, and nodules accumulated 2 to 7 times more cadmium than nodules without MTL4.
b)To further improv … More e plant uptake of heavy metals, we introduced the phytochelatin synthetase(PCS) gene. Another naturally occurring metal-binding peptide, PCs are an attractive alternative to MTs because they offer the potential for enhanced affinity for, and selectivity of, heavy metals. We constructed vectors with Arabidopsis PCS gene under the control of nifH promoter. The gene expressed and catalyzed the synthesis of PCn, with values of n from 2 to 7.
c)We found expression of the AtPCS gene in M.huakuii subsp.rengei B3 along with the increased ability of bacterial cells to bind Cd 10to20 fold. The B3 strain carrying the AtPCS gene was used to infect A.sinicus roots, and nodules formed. The AtPCS gene expressed in nodules, and we found 1.5 times the increased accumulation of Cd in these engineered nodules.
d)Although recombinant free-living bacteria accumulated 20 times more Cd than wild type bacteria, bacteroids in plant root nodules increased Cd accumulation less than 2 times. We expected that there might be a limitation of Cd transportation into nodules. Thus, we isolated the AsIRT1 gene fro Arabidopsis and transferred it together with AtPCS to the B3 strain. The AsIRY1 and AtPCS genes expressed in the bacteroid in plant nodules. Our results show bacterial cells having the IRT1 gene stimulated the incorporation of Cd. However, no increased accumulation of Cd in nodules was observed. Interestingly, we found that Cd accumulated more in roots infected with B3 cells carrying the AtPCS and AtIRT1 genes.
e)We cultivated A.sinicus carrying the MT and AtPCS genes in rice field soil containing cadmium. Cadmium ions accumulated significantly in roots and nodules. Our results showed about 9 % Cd accumulation for this engineered plant. Less

  • Research Products

    (19 results)

All 2005 2004 2003

All Journal Article (18 results) Book (1 results)

  • [Journal Article] Root hair abundant genes LjRH101 and LjRH102 encode peroxidase and xyloglucan endotransglycosylase in Lotus japonicus2005

    • Author(s)
      T.Maekawa
    • Journal Title

      Journal of Bioscience and Bioengineering 99(1)

      Pages: 84-86

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots.2005

    • Author(s)
      H.Imaizumi-Anraku
    • Journal Title

      Nature 433

      Pages: 527-531

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] EspB from enterohaemorrhagic Escherichia coli is a natively partially folded protein.2005

    • Author(s)
      D.Hamada
    • Journal Title

      FEBS Journal 272

      Pages: 756-768

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Expression of a Major House Dust Mite Allergen Gene from Dermatophagoides farinae in Lotus japonicus accession Miyakojima MG-202005

    • Author(s)
      T.Kato
    • Journal Title

      Journal of Bioscience and Bioengineering 99(2)

      Pages: 165-168

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Lotus burttii takes a position of the third corner in the Lotus molecular genetics triangle2005

    • Author(s)
      Kawaguchi, M
    • Journal Title

      DNA Reseach 99(2)(in press)

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots.2005

    • Author(s)
      H.Imaizumi-Anraku^*, N.Takeda^*, M.Charpentier, J.Perry, H.Miwa, Y.Umehara, H.Kouchi, Y Murakami, L.Mulder, K.Vickers, J.Pike, A.Downie, T.Wang, S.Sato, E.Asamizu, S.Tabata, M.Yoshikawa, Y.Murooka, G.-J.Wu, M.Kawaguchi, S.Kawasaki, M.Parniske, M.Hayashi (^*These authors contributed equally to this work)
    • Journal Title

      Nature 433

      Pages: 527-531

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Root hair abundant genes LjRH101 and LjRH102 encode peroxidase and xyloglucan endglycosylase in Lotus japonicus.2005

    • Author(s)
      Maekawa T, Hayashi M, Murooka Y.
    • Journal Title

      J.Biosci.Bioeng. 99(1)

      Pages: 84-86

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] EspB from enterohaemorrhagic Escherichia coli is a natively partially folded protein.2005

    • Author(s)
      D.Hamada, T.Kato, T.Ikegami, K.N.Suzuki, M.Hayashi, Y.Murooka, T.Honda, I.Yanagihara.
    • Journal Title

      FEBS Journal 272

      Pages: 756-768

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Lotus burttii takes a position of the third corner in the Lotus molecular genetics triangle.2005

    • Author(s)
      Kawaguchi, M.^*, Pedrosa-Harand, A.^*, Yano, K.^*, Hayashi, M., Murooka, Y, Saito, K., Namai, K., Nishida, H., Shibata, D., Sato, S., Tabata, S., Sandal, N, Stougaard, J., Bachmair, A., Grant, WF.(^*These authors contributed equally to this work)
    • Journal Title

      DNA Res. (in press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Expression of a Major House Dust Mite Allergen Gene from Dermatophagoides farinae in Lotus japonicus accession Miyakojima MG-20.2005

    • Author(s)
      T.Kato, Y.Goto, M.Hayashi, K.Ono, Y.Murooka
    • Journal Title

      J.Biosci.Bioeng. (in press)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Pollen development and tube growth are affected in the symbiotic mutant of Lotus japonicus, crinkle.2004

    • Author(s)
      M.Tansengco
    • Journal Title

      Plant and Cell Physiology 45(5)

      Pages: 511-520

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Molecular analysis of promoter elements from Propionibacterium freudenreichii.2004

    • Author(s)
      Y-Z.Piao, N.Kawaraichi, R.Asegawa, P.Kiatpapan, H.Ono, M.Yamashita, Y.Murooka.
    • Journal Title

      J.Biosci.Bioeng. 97(5)

      Pages: 310-316

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Production of vitamin B_<12> in genetically engineered Propionibacterium freudenreichii.2004

    • Author(s)
      Y.-Z.Piao, M.Yamashita, N.Kawaraichi, R.Asegawa, H.Ono, Y.Murooka.
    • Journal Title

      J.Biosci.Bioeng. 98(3)

      Pages: 167-173

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Effect of expression of hemA and hemB genes on production of porphyrin in Propionibacterium freudenreichii.2004

    • Author(s)
      Y-Z.Piao, P.Kiatpapan, M.Yamashita, Y.Murooka.
    • Journal Title

      Appl.Enviroment.Microbiol. 70(12)

      Pages: 7561-7566

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

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

      J.Bacteriol. 186(8)

      Pages: 2439-2448

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Pollen development and tube growth are affected in the symbiotic mutant of Lotus japonicus, crinkle.2004

    • Author(s)
      M.Tansengco, H.Imaizumi-Anraku, M.Yoshikawa, S.Takagi, M.Kawaguchi, M.Hayashi, Y.Murooka.
    • Journal Title

      Plant Cell Physiol. 45(5)

      Pages: 511-520

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Enhanced accumulation of Cd^<2+> by a Mesorhizobium sp.transformed with a gene from Arabidopsis thaliana coding for phytochelatin synthase.2003

    • Author(s)
      R.Sriprang, M.Hayashi, H.Ono, M.Takagi, K.Hirata, Y.Murooka.
    • Journal Title

      Appl.Environ.Microbiol. 69(3)

      Pages: 1791-1796

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] crinkle, a novel symbiotic mutant that affects the infection thread growth and alters the root hair, trichome, and seed development in Lotus japonicus.2003

    • Author(s)
      M.Tansengco, M.Hayashi, M.Kawaguchi, H.Imaizumi, Y.Murooka.
    • Journal Title

      Plant Physiol. 131

      Pages: 1054-1063

    • Description
      「研究成果報告書概要(欧文)」より
  • [Book] Biotechnology in Agriculture and Forestry, Vol.52 (eds. Nagata/Tabata)2003

    • Author(s)
      K.Harada (共著)
    • Total Pages
      167-182
    • Publisher
      Springer-Verlag, Berlin, Heidelberg
    • Description
      「研究成果報告書概要(和文)」より

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Published: 2006-07-11  

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