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1987 Fiscal Year Final Research Report Summary

Studies on haploid embryogenesis and its application in plant breeding

Research Project

Project/Area Number 61490006
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionInstitute of Biological Sciences, University of Tsukuba

Principal Investigator

HARADA Hiroshi  Institute of Biological Sciences, Univ. of Tsukuba, Professor, 生物化学系, 教授 (90015991)

Co-Investigator(Kenkyū-buntansha) KAMADA Hiroshi  Gene Experiment Center, Univ. of Tsukuba, Associate Professor, 遺伝子実験センター, 助教授 (00169608)
Project Period (FY) 1986 – 1987
KeywordsPollen Culture / Normal Development of Pollen / Starvation Treatment / Protein Phosphorylation / Tobacco / 半数性不定胚形成
Research Abstract

Binucleate pollen grains of tobacco were fractionated by percoll density gradient centrifugation and cultured in 0.4 M mannitol solution (starvation treatment).After therr days in culture the pollen grains were transferred to a modified MS medium containing mannitol, galactose, glutamine and ABA. About 80% of the pollen grains divided and about 40% of the dividing pollen grains developed into seedlings through haploid embryogenesis after transfer to an appropriate medium. On the other hand, when binucleate pollen grains obtained after Percoll density gradient centrifugation were cultured in basal medium with glutamine, most pollen grains underwent normal development of pollen.
By using these culture methods, we examined biochemical changes which occurred specifically during pollen dedifferentiation. Qualitative and quatitative changes in amino acids and nucleotides were observed during the dedifferentiation, but the changes were also observed during maturation of pollen grains. Changes in the rates of protein, RNA and DNA synthesis and in patterns of de nove protein synthesis were similar in both dedifferentiation and maturation pathways of pollen grains.
Patterns in protein phosphorylation were examined by using 32P inorganic phosphate. Four proteins (a-d) were phosphorylated specifically during pollen dedifferentiation. On the other hand, 5 other proteins (e-i) were phosphorylated spedifically during pollen maturation. We examined several factors affecting pollen dedifferentiation and protein phosphorylation such as glutamine, cytokinin, EDTA and developmental stage of pollen grains. Phosphorylation of proteins a - d was directly correlated with pollen dedifferentiation. Proteins a - d are localized in particular fractions such as mitochondria and chloroplasts. Now efforts are being made to elucidate role(s) of the proteins a - d in pollen dedifferentiation.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] M. Kyo and H. Harada: Planta. 168. 427-432 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H. Harada;M. Kyo and J. Imamura: Current Topics in Developmental Biology. 20. 397-408 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Z. M. Wei;M. Kyo and H. Harada: Theor, Appl, Genet.72. 252-255 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Z. M. Wei;M. Kyo and H. Harada: Proc. of Ciba Foundation Symposium No. 137 "Application of Plant Cell & Tissue Culture".

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M. Kyo and H. Harada: in preparation.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 鎌田博監修, 京正晴: "植物組織培養アトラス第1章第3節2"葯, 花粉培養"" R&Dプランニング社, 56-73 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M,Kyo and H,Harada: "Control of the developmetal pathway of tobacco pollen in vitro." Planta. 168. 427-432 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Harada, M.Kyo and J.Imamura: "Induction of embryogenesis and regulation of the developmental pathway in immature pollen of Nicotiana species" Current Topics in Developmental Biology. 20. 397-408 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Z.M.Wei, M.Kyo and H.Harada: "Callus formation and plant regeneration through direct culture of isolated pollen of Hordeum vulgare cv. Sabarlis." Theor. Appl. Genet.72. 252-255 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Harada, M.Kyo and J.Imamura: "The induction of embryogenesis in Nicotiana immature pollen in culture" Proc. of Ciba Foundation Symposium NO. 137. "Application of Plant Cell & Tissue Culture". in press. (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M,Kyo and H,Harada: "Changes in phosphory lated protein pattern of cultured pollen grains during early stage of pollen embryogenesis"

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M,Kyo: R & D Planning Inc.Plant Tissue Culture Atlas (ed. by H.Kamada) Chapter 1.3.2 "Anther and pollen culture", pp 56-73 (1987)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1989-03-30  

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