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Protoplast and pollen culture and their cell fusion of self-incompatible cruciferous vegetables

Research Project

Project/Area Number 02660032
Research Category

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

Allocation TypeSingle-year Grants
Research Field 園芸・造園学
Research InstitutionOkayama University, Department of Agriculture

Principal Investigator

MATSUBARA Sachiko  岡山大学, 農学部, 教授 (90026378)

Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1991: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1990: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsCruciferous vegetables / Protoplast Culture / Electric cell fusion / カルス培養 / 細胞融合 / アブラナ科野菜
Research Abstract

Protoplast culture and electric cell fusion of cruciferous vegetables were studied. Protoplast culture : The most effective conditions for protoplast isolation were to shake sliced 5day-old cotyledons at 27゚C for 2 h in the enzyme solution containing 2% Cellulase Y-C)0.2% Pectolyase Y-23, CPW salts and 0.3 M mannitol. Isolated protoplasts were cultured on the initial plating medium, Pelettier's B medium which supplemented 1 mg/l NAA and BA, 0.25 mg/l 2, 4-D, 0.5 M mannitol and 20g/l glucose. Then C medium supplemented with 0.2 mg/l NAA, 1 mg/ BA, 0.1 mg/ 2, 4-D, 0.02 mg/l GA_3 and 2 mg/l zeatin was added every 7-10 days. Colonies were formed after 4 weeks, and transferred to E medium. Colonies developed to calli after 1 month, and they were transferred to E medium supplemented 1 mg/l BA. Regenerated shoots were excited and transferred to MS medium, and most of shoots rooted. Shoot regeneration have not achieved in all radish cultivars except 'Moriguchi' and in turnip cultivar 'Shogoin Ookabu'. Other Brassica species showed shoot formation. Root formation occurred in all cultivars.
Electrofusion : The best condition for alignment of 2 protoplasts was AC electric field from 10V for 19sec to 25V for 5sec in radish, 10V for 43sec to 25V for 15sec in cabbage and 10V for 75sec to 25V for 15sec in kairan.
The condition for membrane breakdown was to apply 2 pulses with 500V DC electric field. In these treatments, the medium was supplied with 1mM Ca.
On the other hand, pollen culture was not formed colony, since protoplasts from pollen grains showed only 2 or 3 cell divisions.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] 松原 幸子,千田 泰義,村上 賢治: "アブラナ科植物の胚軸からのカルス形成及び植物体再分化" 園芸学会雑誌別冊. 60. 212-213 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hegazi,H.Hegazi Sachiko Matsubara: "Callus formation and plant regeneration from protoplast derived from cotyledons and hypocotyls of radish (Raphanus sativus L.) and other cruciferous plants." J.Japan.Soc.Hort.Sci.61. (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Matsubara, S., H. Senda and K. Murakami: "Plantlet regeneration from hypocotyl callus in cruciferous plants" Jour. Japan. Soc. Hort. Sci.60 (Suppl. 1). 212-213 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Hegazi, H. Hegazi and Sachiko Matsubara: "Callus formation and plant regeneration from protoplast derived from cotyledons and hypocotyls of radish (Raphanus sativus L.) and other cruciferous plants." J. Japan. Soc. Hort. Sci.61. 1992

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 松原 幸子,千田 泰義,村上 賢治: "アブラナ科植物の胚軸からのカルス形成及び植物体再分化" 園芸学会雑誌別冊. 60. 212-213 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Hegazi,H.Hegazi;Sachiko Matsubara: "Callus formation and plant regeneration from protoplast derived from cotyledons and hypocotyls of radish(Raphanus Sativus L.)and other cruciferous plants." J.Japan.Soc.Hort.Sci. 61. (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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