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Hormone-induced differentiation of BY-2 cultured tobacco cells ; development of a model system for differentiation of root-cap cells.

Research Project

Project/Area Number 12640634
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionNara Women's University

Principal Investigator

SAKAI Atsushi  Nara Women's University, Department of Biological Science, Associate Professor, 理学部, 助教授 (30235098)

Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 2002: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsAuxin / Cytokinin / Root cap / Tobacco / BY-2 / Secreted protein / Microarray / Peroxidase / 培養細胞
Research Abstract

In conventional medium containing auxin (2,4-D, 0.2mg1^<-1>), BY-2 cultured tobacco cells proliferate actively, forming simple cell clusters composed of 8 to 16 small cylindrical cells connected in tandem, and the plastids within the cells remain undifferentiated throughout the culture period. By contrast, in a modified medium that contain cytokinin (BA, 1mg1^<-1>) instead of auxin, BY-2 cells become large without active cell proliferation, and the plastids within the cells differentiate into amyloplasts. In addition to these characteristics (inactive cell proliferation, cell enlargement, amyloplast development), we found that the behavior of BY-2 cells in the auxin-depleted medium resembled to that of root-cap cells in that (i) they easily exfoliate with each other, (ii) exhibited short lifetime, and (iii) showed higher secretion activity. Four proteins specifically secreted by root-cap cell-like BY-2 cells were detected, and one of them (named SPCT44) was identified as secreted peroxidase by N-terminal peptide sequencing of the purified protein. For other three proteins, we could not determine N-terminal peptide sequence because N-terminal was blocked. Their identification by determining internal peptide sequence is now on progress. To analyze changes in the gene expression during differentiation of normal BY-2 cells to root-cap like ones, we performed ddRT-PCR and microarray analyses (collaboration with RIKEN), using mRNAs purified from BY-2 cells cultured 12 h in either conventional medium or modified medium, and identified several genes whose expression is activated according to differentiation of root-cap like cells. The results are opened in the BY-2 EST database by RIKEN (http://mrg.psc.riken.go.jp/strc/). I'd like to further analyze the similarity between root-cap cells and the root-cap cell-like BY-2 cells in the modified medium, by using molecular markers obtained through this research project.

Report

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

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Sakai, et al.: "Organelle-nuclei in higher plants : Their structure, composition, function and evolution"International Review of Cytology. (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai, et al.: "Studies on dynamic changes of organelles using tobacco BY-2 as the model plant cell line"Biotechnology in Agriculture and Forestry. 53. 192-216 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa, et al.: "Activation of cell proliferation by brassinolide application in tobacco BY-2 cells : effects of brassinolide on cell multiplication, cell-cycle-related gene expression, and organellar DNA contents."Journal of Experimental Botany. 54. 2669-2678 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamura, et al.: "Activation of organelle DNA synthesis during the initial phase of proliferation of BY-2 cultured tobacco cells after medium renewal."Plant Morphology. 14. 16-28 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa, et al.: "Isolation and expression of a novel starch-storing cell-specific gene containing the KH RNA binding domain from tobacco-cultured cells BY-2."Journal of Experimental Botany. 53. 2451-2452 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa, et al.: "Differential regulation of starch synthesis gene expression during amyloplast development in cultured tobacco BY-2 cells."Journal of Plant Physiology. 158. 1077-1084 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai: "In vitro transcription/DNA synthesis using isolated organelle-nuclei : application to the analysis of the mechanisms that regulate organelle genome function."Journal of Plant Research. 114. 199-211 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai, et al.: "Isolation of chloroplasts and chloroplast-nuclei(nucleoids) from Chlamydomonas reinhardtii."Plant Morphology. 12. 2-9 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa, et al.: "Organellar protein synthesis controls amyloplast formation independent of starch synthesis gene expression."Cytologia. 65. 435-442 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 酒井 敦: "朝倉植物生理学講座(1)植物細胞(5.1プラスチドの部分pp.111-124を執筆担当)(西村幹夫 編集)"朝倉書店. 179 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 酒井 敦: "光合成事典(一部を執筆担当)(日本光合成研究会 編)"学会出版センター. 430 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai et al.: "Organelle-nuclei in higher plants : Their structure, composition, function and evolution."International Review of Cytology. (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai et al.: "Studies on dynamic changes of organelles using tobacco BY-2 as the model plant cell line."Biotechnology in Agriculture and Forestry. 53. 192-216 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa et al.: "Activation of cell proliferation by brassinolide application in tobacco BY-2 cells : effects of brassinolide on cell multiplication, cell-cycle-related gene expression, and organellar DNA contents."Journal of Experimental Botany. 54. 2669-2678 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Okamura et al.: "Activation of organelle DNA synthesis during the initial phase of proliferation of BY-2 cultured tobacco cells after medium renewal."Plant Morphology. 14. 16-28 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa et al.: "Isolation and expression of a novel starch-storing cell-specific gene containing the KH RNA binding domain from tobacco-cultured cells BY-2."Journal of Experimental Botany. 53. 2451-2452 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa et al.: "Differential regulation of starch synthesis gene expression during amyloplast development in cultured tobacco BY-2 cells."Journal of Plant Physiology. 158. 1077-1084 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai: "In vitro transcription/DNA synthesis using isolated organelle-nuclei : application to the analysis of the mechanisms that regulate organelle genome function."Journal of Plant Research. 114. 199-211 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sakai et al.: "Isolation of chloroplasts and chloroplast-nuclei (nucleoids) from Chlamydomonas reinhardtii."Plant Morphology. 12. 2-9 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa et al.: "Organellar protein synthesis controls amyloplast formation independent of starch synthesis gene expression."Cytologia. 65. 435-442 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Miyazawa et al.: "Isolation and expression of a novel starch-storing cell specific gene containing the KH RNA binding domain from tobacco-cultured cells BY-2"J.Exp.Bot.. 53. 2451-2452 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Okamura et al.: "Activation of organelle DNA synthesis during the initial phase of proliferation of BY-2 cultured tobacco cells after medium renewal"Plant Morph.. 14. 16-28 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Inada et al.: "Three-dimensional progression of programmed death in the rice coleoptile"Int.Rev.Cytol. 218. 221-258 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakai et al.: "Studies on dynamic change of organelles using BY-2 as the model plant cell line"Biotech Agricul.Forest.. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] Miyazawa Y et al.: "Differential regulation of starch synthesis gene expression during amyloplast development in cultured to bacco BY-2 cells"J. Plant Physiol. 158. 1077-1084 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Sakai A: "In vitro transcription/DNA synthesis system using isolated organelle-nuclei : analysis on the mechanisms that regulate the organelle genome function"J. Plant Res.. 114. 199-211 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takahara M et al.: "A putative mitochondrial ftsZ gene is present in the unicellular primitive red alga Cyanidioschyzon merolae."Mol.Gen.Genet.. 264. 452-460 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sakai A et al.: "Isolation of chloroplasts and chloroplast-nuclei (nucleoids) from Chlamydomonas reinhardtii."Plant Morph.. (in press).

    • Related Report
      2000 Annual Research Report
  • [Publications] Saito C et al.: "Behavior of plastid nucleoids during male gametogenesis in Plumbago auriculata."Protoplasma. (in press).

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyazawa Y et al.: "Organellar protein synthesis controls amyloplast formation independent of starch synhesis gene expression."Cytologia. 65. 435-442 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Takahara M et al.: "Isolation, characterization, and chromosomal mapping of an ftsZ gene from the unicellular primitive red alga Cyanidium caldarium RK-1."Curr.Genet.. 37. 143-151 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Saito C et al.: "Unequal distribution of DNA-containing organelles in generative and sperm cells of Erythrina crista-galli (Fabaceae)."Sex.Plant Reprod.. 12. 296-301 (2000)

    • Related Report
      2000 Annual Research Report

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

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