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BASIC RESARCH FOR IMPROVEMENT OF COLD AND DESEASE RESISTANCE OF PERRENIAL WEED PLANTS BY GENETIC ENGENEERING

Research Project

Project/Area Number 12556044
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Zootechnical science/Grassland science
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

TAKEZAWA Daisuke  Inst., Inst. Low Temp. Sci., Hokkaido Univ., 低温科学研究所, 助手 (20281834)

Co-Investigator(Kenkyū-buntansha) ARAKAWA Keita  Inst., Inst. Low Temp. Sci., Hokkaido Univ., 低温科学研究所, 助手 (00241381)
OZAWA Shuji  Asso. Prof., Fac. Dairy Sci., Rakunou Gakuen Univ., 酪農学部, 助教授 (50204194)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥5,500,000 (Direct Cost: ¥5,500,000)
Keywordsalfalfa / disease resistance / freezing tolerance / genetic engineering / カルモジュリン / PRタンパク質 / 耐寒性 / 雪腐病 / トランスジェニック植物
Research Abstract

The goal of this research was to generate weeds that has increased tolerance to freezing and disease by genetic engineering. Experiments were focused on (1) isolation of genes that can confer plants with tolerance to freezing and disease, and (2) production of transgenic alfalfa plants by using the isolated genes.
As a candidate gene for production of transgenics, WAS-3 gene, which has similarity to antifreeze proteins and antifungal proteins was isolated from winter wheat that has high degree of winter-stress resistance. The WAS-3 protein produced in transgenic wheat cells efficiently blocked growth of hyphae of the snow mold, Microdochium nivale. In addition, transgenic plants expressing the WAS-3 protein exhibited a better resistance to Fusarium oxysporum than control non-transgenic plants. We also isolated genes encoding dehydrins and group III LEA from mulberry, which exhibits extremely high freezing tolerance in winter, and several ABA and cold-responsive genes from the moss Physcomitrella patens.
For production of transgenic alfalfa plants, we used Arabidopsis CBF3, a gene encoding a transcription factor that plays a critical role in the up-regulation of a number of cold-inducible genes. Agrobacterium tumefaciens carrying a construct for overexpression of the CBF3 gene was used for inoculation of leaf discs of alfalfa (Medicago sativa var. Regen-SY). Kanamycin-resistant calli were selected and shoots were regenerated by further cultivation. Nine transgenic plants were successfully regenerated. These transgenic plants showed normal growth indistinguishable from non-transgenics, suggesting they have less somatic mutations. These transgenic plants were being used for the freezing tests.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Takezawa, D.: "A rapid induction by elicitors of the mRNA encoding CCD-1, a 14 kDa Ca^<2+>-binding protein in wheat cultured cells"Plant Molecular Biology. 42. 807-817 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ukaji, H.: "Cold Acclimation-Induced WAP27 Localized in Endoplasmic Reticulum in Cortical Parenchyma Cells of Mulberry Tree Was Homologous to Group 3 Late Embryogenesis Abundant Proteins"Plant Physiology. 126. 1588-1592 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kuwabara, C.: "Abscisic acid-and cold-induced thaumatin-like protein in winter wheat has an antifungal activity against snow mould, Microdochium nivale"Physiologia Plantarum. 115. 101-110 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yamada, T.: "Roles of the plasma membrane and the cell wall in the responses of plant cells to freezing"Planta. 215. 770-778 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Minami, A.: "Abscisic acid-induced freezing tolerance in the moss Physcomitrella patens is accompanied by increased expression of stress-related genes"Journal of Plant Physiology. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Takezawa D.: "A rapid induction by elicitors of the mRNA encoding CCD-1, a 14 kDa Ca^<2+>-binding protein in wheat cultured cells"Plant Mol. Biol.. 42. 807-817 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ukaji H., Kuwabara, Takezawa D., Arakawa K., Fujikawa S.: "Cold acclimation-induced WAP27 localized in endoplasmic reticulum in cortical parenchyma cells of mulberry tree was homologous to group 3 late embryogenesis abundant protein"Planet Physiol. 126. 1588-1597 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kuwabara C., Takezawa D., Shimada T., Hamada T., Fujikawa S., Arakawa K.: "Abscisic acid-and cold-induced thaumatin-like protein in winter wheat has an antfungal activity against snow mould, Microdochium nivale"Physiol. Plant. 115. 101-110 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yamada T., Kuroda K., Jitsuyama Y., Takezawa D., Arakawa K., Fujikawa S.: "Roles of the plasma membrane and the cell wall in the responses of plant cells to freezing"Planta. 215. 770-778 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Minami A., Nagao M., Arakawa K., Fujikawa S., Takezawa D.: "Abscisic acid-induced freezing tolerance in the moss Physcomitrella patens is accompanied by increased expression of stress-related genes"J. Plant Physiol. in press (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yamada: "Roles of the plasma membrane and the cell wall in the responses of plant cells to freezing"Planta. 215. 770-778 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Minami: "Abscisic acid-induced freezing tolerance in the moss Physcomitrella patensis accompanied by increased expression of stress-related genes"Journal of Plant Physiology. (In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ukaji et al.: "Cold acclimation-induced WAP27 localized in endoplasmic reticulum in cortical parenchyma cells of mulberry tree was homologous to group 3 late-embryogenesis aboundant proteins"Plant Physiology. 126. 1588-1597 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takezawa, D.: "A rapid induction by elicitors of the mRNA encoding CCD-1, a 14 kDa Ca^<2+>-binding protein in wheat cultured cells."Plant Mol.Biol.. 42. 807-817 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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