| Project/Area Number |
20K15553
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 40010:Forest science-related
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| Research Institution | The University of Tokyo |
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Principal Investigator |
LI CHAOFENG 東京大学, 大学院農学生命科学研究科(農学部), 特任研究員 (30866687)
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| Project Period (FY) |
2020-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Keywords | MiRNA sequencing / Degradome sequencing / Ectomycorrhiza / Populus tomentosa / RT-qPCR / Phenylpropanoid / Cenococcum geophilum / miRNA / 菌根形成 / 分子メカニズム / 遺伝子 / マイクロRNA |
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| Outline of Research at the Start |
外生菌根菌と宿主樹木との菌根形成の分子メカニズムを明らかにするため、近年植物とECM菌の相互作用に重要な役割を果たすことが示唆されたmiRNAの菌根形成における生体内での機能を解明する。ポプラのmiRNA配列をデータベースから取得し、候補のmiRNAおよびターゲットとなる遺伝子を探索する。形質転換、CRISPR/Cas9ゲノム編集を用い、候補のmiRNAを過剰発現およびノックダウンしたポプラ変異体を作成する。これらポプラ変異体へのECM菌の接種、遺伝子発現解析、Degradation-Seq、タンパク質相互作用の解析等を駆使し、各miRNAが菌根形成にどのような役割を果たすかを明らかにする。
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| Outline of Final Research Achievements |
We used the miRNA sequencing to identify 51 differentially expressed miRNAs and corresponding 2043 targets in the roots of Populus tomentosa colonized by Cenococcum geophilum. Then we predicted the target sequences of 51 miRNAs with psRNATarget. The prediction of miRNAs-seq and psRNATarget were further confirmed by degradome sequencing and RT-qPCR analysis. GO and KEGG enrichment analysis indicated that the target genes of some differentially expressed miRNAs regulate the ECM symbiosis through the pathways of phenylpropanoid biosynthesis, isoquinoline alkalold biosynthesis, and apoplast, etc. This work indicates that Populus miRNAs and their target genes are an important part of the regulatory network leading to ECM symbiosis development.
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| Academic Significance and Societal Importance of the Research Achievements |
The research demonstrates that Populus miRNAs and their target sequences are essential to ectomycorrhizal symbiosis, which plays key roles in forest ecosystems and carbon cycle. The results may supply some new insights about how to manage and protect forests.
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