Establishment of cryopreservation method for plant and microorganism using two members culture
| Project/Area Number |
18K05697
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 39060:Conservation of biological resources-related
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| Research Institution | National Agriculture and Food Research Organization |
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Principal Investigator |
Tanaka Daisuke 国立研究開発法人農業・食品産業技術総合研究機構, 遺伝資源センター, 上級研究員 (60425593)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | 超低温保存 / ガラス化 / 二員ガラス化法 / 微生物 / 植物組織細胞 / 植物 / フタバネゼニゴケ / アーバスキュラー / 遺伝資源 |
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| Outline of Final Research Achievements |
Vitrification-based method was successfully established for storage at -196oC of Oomycetes. The optimal droplet vitrification method using Oomycetes for the storage at -196oC is as follows. Oomycete isolates were cultured with the rape seeds for 30 days at 25°C on solidified PDA medium. Rape seeds containing oomycete were placed in 15 ml petri dish and osmoprotected with LS (liquid medium supplemented with 1.0 M sucrose and 2.0 M glycerol) for 30 min. Then they were dehydrated PVS2 for 20 min at 25ºC. After dehydration placed in wells on an aluminum cryo-plate, and directly plunged into liquid nitrogen and maintained at -196 ºC. As results, three Oomycetes isolates were successfully cryopreserved with 100% viability. Vitrification-based protocol was cryopreservation method for reliable long-term preservation of Oomycetes resources.
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| Academic Significance and Societal Importance of the Research Achievements |
植物と共生する微生物には，驚くべき機能が期待される．微生物遺伝資源の多様性は，生態系と持続的農業環境の保全にとても重要である．宿主植物と二員でなければ生存が難しい微生物の長期保存は非常に困難である．本研究成果は，従来の保存法にとらわれずに植物や動物遺伝資源保存分野で用いられているガラス化保存技術を応用して，微生物の新規超低温保存技術を開発した．他の難保存性微生物の管理・系統保存技術の開発にも寄与するものと期待される．
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Report
(3 results)
Research Products
(1 results)
