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Reinforcement of Silage-Making Lactobacilli and Their Utilization

Research Project

Project/Area Number 07556094
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 応用微生物学・応用生物化学
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

OGATA Seiya  Kyushu University, Faculty of Agriculture, Professor, 農学部, 教授 (20038277)

Co-Investigator(Kenkyū-buntansha) TANAKA Osamu  Ministry of Agriculture, Forestry and Fisheries, Researcher, 草地試験場, 研究員
OHMOMO Sadahiro  Ministry of Agriculture, Forestry and Fisheries, Researcher, 畜産試験場, 研究員
DOI Katsumi  Kyushu University, Faculty of Agriculture, Associate Professor, 農学部, 講師 (40253520)
MASUA Yasuhisa  Kyushu University, Faculty of Agriculture, Professor, 農学部, 教授 (60038263)
FUGIO Yusaku  Kyushu University, Faculty of Agricvltvre Professor, 農学部, 教授 (90038224)
田中 悟  農林水産省, 草地試験所, 研究員
五斗 一郎  九州大学, 農学部, 教授 (60038180)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1997: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1996: ¥4,500,000 (Direct Cost: ¥4,500,000)
KeywordsSilage / Silage inoculant / Lactic acid fermentation / Breeding / Phage resistant plasmid / Nitrate reductase / Recombinants / Utilization / クローニング / ファージ耐性
Research Abstract

Silage is a storable feed utilizing lactic fermentation. To develop a silage inoculant adaptable to the climatic conditions of south-west Japan such as Kyushu and Okinawa. we isolated many homofermentative lactobacilli strains with high lactic acid productivity under high temperature conditions. We then examined their properties and improved them to their greatest potential for use as silage inoculant.
(1) The strains isolated were classified into L.plantaraum, L.pentosus, L.casei and L.rhamnosus by the usual methods and sequencing of partial 16S rRNA genes and phosphofructokinase genes and phosphofrutokinase gene pfk. A phagetying method was also effective for their easy and quick classification. : (2) 25% of 77 silage samples tested contained phages, most of which were infectious to L.plantaraum followed by L.casei. The phage contamination of silage resulted in the deterioration of quality. : (3) A phage resistant plasmid pLKS (2.1 kb) isolated was a rheta-replicating type. The phage resistant activity of pLKS might be due to the similarity between the replication factors of pLKS and phages. : (4) Lactobacillus-E.coli shuttle vector pUCLSEm was constructed by ligating pLKS to pUC119. : (5) A strain No. 161 belonging to Bacillus licheniformis, which had a high activity of nitrate reductase, was isolated from a compost. This train was tried to use for the elimination of NO^-_ in the silage. Furthermore, nitrate reductase gene narH with significant homology to the E.coli narH was cloned. : (6) The recombinant strains of L.plantaraum and L.casei were constructed with alpha-amylase gene, pfk and narH using vector pUCLSEm which had a phage resistant activity.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] 緒方 靖哉 他: "サイレ-ジ乳酸菌の機能開発-ファージタイピングによる簡便な同定法の開発および高機能サイレ-ジ菌株作出-" 生物工学会誌. 75・5. 361-363 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 大桃 定洋 他: "サイレ-ジ発酵用乳酸菌に求められる能力とその探索" J.Jpn.Study G.LAB. 7・2. 72-78 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 大桃 定洋 他: "Effect of salt tolerant lactic acid bacteria and salt on the fermentation characteristics and aerobic stability of Silages" J.Appl.Microbiol.83. 307-313 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田中 治 他: "Lactic acid productivity of the selected strains of the Genus Lactobacillus in Laboratory-scale silages" Glassl.Sci. 43・3. 327-377 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 緒方 靖哉 他: "A conjugative linear plasmid in S.laurentii ATCC31255" Biosci.Biotech.Biochem.61・9. 1469-1473 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 緒方 靖哉 他: "サイレ-ジ乳酸菌の機能開発-付与有用形質の分離と応用-" 生物工学会誌. 76・11(印刷中). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 緒方 靖哉: "微生物とその利用" コロナ社, 194 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 大桃 定洋: "畜産総合辞典(p686〜694)" 朝倉書店, 9 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Doi, T.Eguchi, O.Tanaka, K.Mori, S.Ohmomo and S.Ogata: "Development of Silage-Making Lactobacilus sp.Identification for Lactic Acid Bacteria using Phage-Typing and of Improvement of Silage-Making Lactobacilli In Japanease." Seibutsu Kougaku. 75 (5). 361-363 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Ohmomo and O.Tanaka: "The Abilities Requested Lactic Acid Bacteria for Silage-Making and Its Screening In Japanease." J.Jpn. Study G.LAB.7 (2). 72-78 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Cai, S.Ohomomo, M.Ogawa and S.Kumai: "Effect of Salt-Tolerant Lactic Acid Bacteria and Salt on the Fermentation Characteristics and Aerobic Stability of Silages." J.Appl.Microbiol.83. 307-313 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] O.Tanaka and S.Ohmomo: "Lactic Acid Productivity of the Selected Strains of the Genus Lactobacillus in Laboratory-Scale Silages." Glassl.Sci.43・3. 327-377 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] C.Kinoshita-Iramina, M.Kitahara, K.Doi and S.Ogata: "A Conjugative Linear Plasmid in Streptomyces laurentii ATCC31255." Biosci. Biotech. Biochem.61 (9). 1469-1473 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Doi, T.Eguchi, N.Akiyoshi, F.Inagaki, O.Tanaka, Y.Fujio, S.Ohmomo and S.Ogata: "Development of Silage-Making Lactobacillus sp. Identification and Application of useful genes for Silage-Making Lactobacilli In Japanease." Seibutsu Kougaku. 76・11 (in printing.). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Ogata: Fundamental Approach for Microorganisum. Basic Microbiology and Its Applications. In Japanease.Koronasha, 1-194 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Ohmomo: Silage. In Encyclopedia of Animal Industry In Japanease.Asakura Shoten., 686-694 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 緒方靖哉 他: "サイレ-ジ乳酸菌の機能開発-ファージタイピングによる簡便な同定法の開発および高機能サイレ-ジ菌株作出-" 生物工学会誌. 75・5. 361-363 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 大桃定洋 他: "サイレ-ジ発酵用乳酸菌に求められる能力とその探索" J.Jpn.Study G.LAB. 7・2. 72-78 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 大桃定洋 他: "Effect of salt tolerant lactic acid bacteria and salt on the fermentation characteristics and aerobic stability of Silages" J.Appl.Microbiol.83. 307-313 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田中 治 他: "Lactic acid productivity of the selected strains of the Genus Lactobacillus in Laboratory-scale silages" Glassl.Sci. 43・3. 327-377 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 緒方靖哉 他: "A conjugative linear plasmid in S.laurentii ATCC 31255" Biosci.Biotech.Biochem.61・9. 1469-1473 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 緒方靖哉 他: "サイレ-ジ乳酸菌の機能開発-付与有用形質の分離と応用-" 生物工学会誌. 76・11(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 緒方靖哉: "微生物とその利用" コロナ社, 194 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 大桃定洋: "畜産総合事典" 朝倉書店, 8(686-694) (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 緒方 靖哉: "サイレ-ジ乳酸菌の機能開発" 日本生物工学会誌. 74・4. 294-297 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 緒方 靖哉: "生物生産における遺伝子ターゲッティング" 日本農芸化学会誌. 70・8. 883-884 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 大桃 定洋: "Acremonium cellulolyticus Y-94由来のセルラーゼの添加がアルファルファサイレ-ジの発酵品質に及ぼす影響" Grassland Science. 42・2. 159-162 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 大桃 定洋: "Acremonium cellulolyticus Y-94が生産するセルラーゼの添加がイネ科牧草サイレ-ジの発酵品質に及ぼす影響" Grassland Science. 42・2. 344-348 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 田中 治: "Lactic acid productivity of the Selected Strains of the Genus Lactobacillus in a Loboratory-Scale Silages" Grassland Science. 43・1(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 土居 克実: "サイレ-ジ乳酸菌の機能開発" 日本生物工学会乳酸菌工学研究部会講演会要旨集. 4 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 緒方 靖哉: "微生物とその利用" コロナ社, 206 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 大桃 定洋(共著): "乳酸菌の科学と技術" 学界出版センター, 4 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 緒方靖哉: "Relationship between fermentation quality of silage and presence of phage for silage-making Lactobacilli." 草地試験場研究報告. 51. 31-39 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 緒方靖哉: "サイレ-ジ乳酸菌のファージ耐性に関与するプラスミド" 日本農芸化学会誌(農林水産技術会議事務局). 69. 276- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 緒方靖哉: "高機能乳酸発酵菌の作出" 研究成果302. 34-50 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 土居克実: "サイレ-ジ乳酸菌の機能開発" 日本生物工学会乳酸菌工学研究部会講演会要旨集. 5-6 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 田中 治: "プラスチックフィルムを用いた小規模サイレ-ジ発酵試験法の開発" Grassland Science. 41. 55-59 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 大桃定洋: "Screening of lactic acid bacterria suitable for silage-making in tropical regions" JARQ. 29. 251-256 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 緒方靖哉(共編著): "微生物の生体防御「生物生産と生体防御」" コロナ社, 44(349) (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 緒方靖哉(共編著): "微生物の遺伝子ターゲッティング「遺伝子ターゲッティングの基礎と応用」" コロナ社, 38(234) (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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