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Establishment of experimental platform for physiological and pathological studies based on insect midgut stem cell culture

Research Project

Project/Area Number 17H03944
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Insect science
Research InstitutionYamaguchi University

Principal Investigator

Kobayashi Jun  山口大学, 大学院創成科学研究科, 教授 (70242930)

Co-Investigator(Kenkyū-buntansha) 東 政明  鳥取大学, 農学部, 教授 (20175871)
伊藤 雅信  京都工芸繊維大学, 応用生物学系, 教授 (60221082)
森 肇  京都工芸繊維大学, 応用生物学系, 教授 (80201812)
Project Period (FY) 2017-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2020: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2019: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2018: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Keywords中腸幹細胞 / カイコ / Bacillus thuringiensis / Cry毒素 / トランスクリプトーム / Bacillus thringiensis / 昆虫病原ウイルス
Outline of Final Research Achievements

In order to establish a stable in vitro differentiation system of the insect midgut stem cell and to verify its physiological and pathological functions, we have developed a simple protocol for stem cell preparation from larvae and pupae of the silkworm, Bombyx mori, and have investigated effectiveness of supplementation to the primary stem cell culture for survival rate improvement, together with the development of a short-term intact stem cell preserving method at 4℃. We have also revealed the possibility that factors other than hormones are predominantly involved in changes in gene expression of the stem cell during the growth of B. mori. Furthermore, we have succeeded to make protein crystals containing Bacillus thuringiensis Cry toxins showing specific insecticidal effects on B. mori and Aedes albopictus, respectively, which are useful not only for elucidation of detailed mechanisms for pathogenic changes in midgut cells, but also for pest management.

Academic Significance and Societal Importance of the Research Achievements

本研究課題では,数多くの農業害虫を含むチョウ目幼虫の中腸細胞の分化に伴う機能変化や病原微生物の侵入過程を分子レベルで明らかにするための世界初となる中腸幹細胞の培養分化系の構築を目指し,その基盤となる幹細胞の分離調製法が確立され,発育ステージやホルモンが幹細胞の遺伝子発現に及ぼす影響が明らかにされ,また,害虫防除への応用が期待されるBacillus thuringiensisの数種Cry毒素内包多角体が構築され,それらの中腸細胞に対する種特異的な殺虫効果が明らかにされるなど,基礎ならびに応用昆虫学上意義ある成果が得られ,今後,農業や関連産業に役立つ技術開発につながる可能性が期待される。

Report

(5 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (22 results)

All 2021 2020 2019 2018 2017 Other

All Int'l Joint Research (2 results) Journal Article (9 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 6 results,  Open Access: 4 results,  Acknowledgement Compliant: 1 results) Presentation (10 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results) Book (1 results)

  • [Int'l Joint Research] Monash University(オーストラリア)

    • Related Report
      2018 Annual Research Report
  • [Int'l Joint Research] Cell Guidance Systems Ltd(英国)

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Isolation and primary culture of stem cells from the midgut epithelium of the silkworm <i>Bombyx mori</i>2020

    • Author(s)
      N. Fujiwara, M. Azuma, M. Itoh, J. Kobayashi
    • Journal Title

      Journal of Insect Biotechnology and Sericology

      Volume: 89 Issue: 3 Pages: 3_055-3_062

    • DOI

      10.11416/jibs.89.3_055

    • NAID

      130007987477

    • ISSN
      1346-8073, 1884-7978
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 特集「昆虫工場の現場から」にあたって ~その光と影,そして新たな展開に向けて~2020

    • Author(s)
      小林淳
    • Journal Title

      Sanshi-Konchu Biotec

      Volume: 89 Issue: 2 Pages: 2_049-2_050

    • DOI

      10.11416/konchubiotec.89.2_049

    • NAID

      130008091004

    • ISSN
      1881-0551, 1884-7943
    • Related Report
      2020 Annual Research Report
  • [Journal Article] 起業15年を迎えるバキュロテクノロジーズ2020

    • Author(s)
      馬縞景,小林淳
    • Journal Title

      Sanshi-Konchu Biotec

      Volume: 89 Issue: 2 Pages: 2_085-2_089

    • DOI

      10.11416/konchubiotec.89.2_085

    • NAID

      130008090990

    • ISSN
      1881-0551, 1884-7943
    • Related Report
      2020 Annual Research Report
  • [Journal Article] Structure of in cell protein crystals containing organometallic complexes2018

    • Author(s)
      Abe Satoshi、Atsumi Kohei、Yamashita Keitaro、Hirata Kunio、Mori Hajime、Ueno Takafumi
    • Journal Title

      Physical Chemistry Chemical Physics

      Volume: 20 Issue: 5 Pages: 2986-2989

    • DOI

      10.1039/c7cp06651a

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Supramolecular protein cages constructed from a crystalline protein matrix2018

    • Author(s)
      Negishi Hashiru、Abe Satoshi、Yamashita Keitaro、Hirata Kunio、Niwase Kento、Boudes Marion、Coulibaly Fasseli、Mori Hajime、Ueno Takafumi
    • Journal Title

      Chemical Communications

      Volume: 54 Issue: 16 Pages: 1988-1991

    • DOI

      10.1039/c7cc08689j

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Raman spectroscopy insight into Norovirus encapsulation in Bombyx mori cypovirus cubic microcrystals2018

    • Author(s)
      Mori Hajime、Oda Naoki、Abe Satoshi、Ueno Takafumi、Zhu Wenliang、Pernstich Chris、Pezzotti Giuseppe
    • Journal Title

      Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy

      Volume: 203 Pages: 19-30

    • DOI

      10.1016/j.saa.2018.05.066

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Water influx via aquaporin directly determines necrotic cell death induced by the Bacillus thuringiensis Cry toxin.2017

    • Author(s)
      Endo H, Azuma M, Adegawa S, Kikuta S, Sato R
    • Journal Title

      FEBS Letters

      Volume: 591 Issue: 1 Pages: 56-64

    • DOI

      10.1002/1873-3468.12506

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Comparative Analyses of Deletion Mutations and the Pathological Effects of Nucleopolyhedroviruses Isolated from Saturniid Wild Silkworms2017

    • Author(s)
      Kuni Sasaki, Zenta Kajiura, Jun Kobayashi
    • Journal Title

      Journal of Insect Biotechnology and Sericology

      Volume: 86 Issue: 3 Pages: 3_067-3_075

    • DOI

      10.11416/jibs.86.3_067

    • NAID

      130006223745

    • ISSN
      1346-8073, 1884-7978
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] バキュロウイルスを利用したバイオテクノロジー2017

    • Author(s)
      小林淳
    • Journal Title

      アグリバイオ

      Volume: 1 Pages: 1456-1461

    • Related Report
      2017 Annual Research Report
  • [Presentation] カイコ幼虫消化管幹細胞の初代培養とトランスクリプトーム解析2021

    • Author(s)
      吉永侑生, 東政明, 小林淳
    • Organizer
      第65回日本応用動物昆虫会
    • Related Report
      2020 Annual Research Report
  • [Presentation] カイコ(Bombyx mori)消化管幹細胞トランスクリプトームの幼虫脱皮に伴う変化2020

    • Author(s)
      吉永侑生, 東政明, 小林淳
    • Organizer
      日本分子生物学会第43回年会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 昆虫培養細胞の樹立と利用2019

    • Author(s)
      小林 淳
    • Organizer
      第27回病害動物の生理分子生物談話会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] チョウ目昆虫の中腸幹細胞の分離・培養とトランスクリプトーム解析2019

    • Author(s)
      吉永侑生, 小林淳, 東政明
    • Organizer
      西日本応用動物昆虫研究会・中国地方昆虫学会 令和元年度合同例会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Molecular characterization, tissue distribution and transmission of nucleopolyhedrovirus in oak tasar silkworm, Antheraea proylei2019

    • Author(s)
      Kangayam M. Ponnuvel, Sinam Subharani Devi, Rakesh K. Mishra, Olle Terenius, Jun Kobayashi
    • Organizer
      The 25th International Congress on Sericulture and Silk Industry
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research
  • [Presentation] チョウ目昆虫中腸幹細胞の分離・培養とトランスククリプトーム解析2019

    • Author(s)
      吉永侑生, 東政明, 小林淳
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] エリサンとシンジュサンの雑種系統作出と遺伝生理学的解析2018

    • Author(s)
      岩本帆乃か,吉永侑生,小林淳
    • Organizer
      第41回日本分子生物学会年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ゲノム配列に基づくアジア産ヤママユガ亜科核多角体病ウイルスのゲノム分類2018

    • Author(s)
      佐々木邦,梶浦善太,K. M. Ponnuvel,小林淳
    • Organizer
      第24回日本野蚕学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 東~南アジアに分布するヤママユガ科野蚕核多角体病ウイルスのゲノム配列と構造に関する進化的考察2017

    • Author(s)
      佐々木邦,梶浦善太,小林淳
    • Organizer
      第32回中国四国ウイルス研究会
    • Related Report
      2017 Annual Research Report
  • [Presentation] 東~南アジアに分布するヤママユガ科野蚕核多角体病ウイルスの比較ゲノム解析2017

    • Author(s)
      佐々木邦,梶浦善太,小林淳
    • Organizer
      2017年度生命科学系学会合同年次大会(ConBio2017)
    • Related Report
      2017 Annual Research Report
  • [Book] カイコの科学2020

    • Author(s)
      小林淳,東政明,伊藤雅信ほか,日本蚕糸学会編
    • Total Pages
      210
    • Publisher
      朝倉書店
    • ISBN
      9784254420432
    • Related Report
      2020 Annual Research Report

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Published: 2017-04-28   Modified: 2022-01-27  

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