| Project/Area Number |
16H06182
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| Research Category |
Grant-in-Aid for Young Scientists (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Science in genetics and breeding
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| Research Institution | Tohoku University |
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Principal Investigator |
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| Research Collaborator |
TORIYAMA kinya
ARIMURA shin-ich
Nakamura takahiro
ITO masako
OGATA ikuya
KINOSHITA nozomi
SASAKI miho
OOMUKAI shiho
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| Project Period (FY) |
2016-04-01 – 2019-03-31
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| Project Status |
Completed (Fiscal Year 2018)
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| Budget Amount *help |
¥24,180,000 (Direct Cost: ¥18,600,000、Indirect Cost: ¥5,580,000)
Fiscal Year 2017: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2016: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
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| Keywords | 植物ミトコンドリア / 細胞質雄性不稔性 / 次世代育種技術 / オルガネラ / ミトコンドリア / ゲノム編集 / 育種学 |
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| Outline of Final Research Achievements |
In this study, we attempted to develop artificial fertility restorer genes using the next plant breeding techniques (NBT), TALEN (transcription activator-like effecter nuclease) and PPR (pentatricopeptide repeat protein). First, we introduced mitochondria-targeted TALENs, which are bound to a CMS-associated gene of BT-type CMS rice, orf79. In this case, we obtained orf79-depleted plants. Those plants showed fertility restoration. Interestingly, the knockout of orf79 caused unexpected changes in the mitochondrial genome by homologous recombination via short sequence, possibly as a result of some unusual feature of plant mitochondrial genome. Next, we constructed artificial PPR protein-coding genes and introduced them into BT-type CMS. Those proteins were confirmed to bind to orf79 RNA in vitro, however, we could not obtained fertility-restored transgenic plants. This indicates that not only binding activity but other activities are required to create functional artificial PPR proteins.
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| Academic Significance and Societal Importance of the Research Achievements |
植物ミトコンドリアゲノムには，呼吸に関与するタンパク質をコードする遺伝子だけでなく，細胞質雄性不稔性の原因遺伝子もコードしている．しかし，植物ミトコンドリアは形質転換ができないため，遺伝子の機能解析は限定的にしか行われてきていない．そこで，人工制限酵素TALENと配列特異的RNA結合タンパク質PPRを用いて，ミトコンドリア遺伝子の発現制御を行うことを目的とした．本研究でミトコンドリア遺伝子の発現制御が可能になれば，基礎科学としてのインパクトも高い．さらに，農業上重要な形質である細胞質雄性不稔性の稔性回復遺伝子を人為的に合成することが可能となり，多種育種の基盤に大きく貢献できると考えられる．
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