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1993 Fiscal Year Final Research Report Summary

Fusion of Protoplasts from Wild Type of Bangia atropurpurea and Green Type of Porphyra tenera

Research Project

Project/Area Number 04660203
Research Category

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

Allocation TypeSingle-year Grants
Research Field General fisheries
Research InstitutionMie University

Principal Investigator

ARAKI Toshiyoshi  Mie University. Faculty of Bioresources. Associate Professor, 生物資源学部, 助教授 (40091368)

Project Period (FY) 1992 – 1993
KeywordsBangia atropurpurea / Protoplast / beta-1, 4-Mannanase / beta-1, 3-Xylanase / Porphyranase / ポルフィラナーゼ
Research Abstract

Three kinds of enzymes, beta-1, 4-mannanase, beta-1, 3-xylanase, and agarase, required for isolation of protoplats from a red alga, Bangia atropurpurea, were prepared from bacterial culture fluids of Vibrio sp. MA-138, Alcaligenes sp. XY-234, and Vibrio sp. P0-303, isolated from sea environment. The optimal pHs of all the crude enzymes were around 7.5. The suitable condition for protoplast isolation was examined. The treatment of the fronds with papain solution (20 mM MES buffer, pH 7.5, containing 2 % papain and 0.5 M mannitol) have contributed to isolatating protoplasts. When razorcut fragments of the fronds (about 200 mg in weight) dipped in the cell wall-digestive enzyme solution (1 unit each of 1, 4-beta-mannanase, 1, 3-beta-xylanase, and agarase, and 0.5 M mannitol in 20 mM MES buffer, pH 7.5) were incubated at 17 ゚C for 90 min with gentle agitation, 7.1 x 10^6 of protoplasts were released from them.
For carrying out cell fusion polyethylen glycol (PEG) was added into the suspension of protoplasts from the wild type of B.atropurpurea and the green varient of Porphyra tenera. Homo- and hetero-plasmic adhesion of protoplasts were observed. The PEG-treated protoplasts were cultured in artificial light of about 48 muEm^<-2>s^<-1> on a 9 h light/15 h dark cycle at 17 ゚C.They grew into calli and plantrets of brown or green color after 6-week culture. A pair of protoplast which parts of their menbrans adhesed also developed into plantlets.

  • Research Products

    (2 results)

All Other

All Publications (2 results)

  • [Publications] Toshiyoshi Araki: "β-1,4-Mannanases from Marine Bacteria,Vibrio SPP.MA-129 and MA-138" J.Gen.Appl.Microbiol. 38. 343-351 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshiyoshi Araki: "beta-1, 4-Mannanases from marine bacteria, Vibrio spp. MA-129 and MA-138" J.Gen. Appl. Microbiol.38. 343-351 (1992)

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

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Published: 1995-03-27  

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