| Project/Area Number |
17K07661
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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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| Research Field |
Horticultural science
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| Research Institution | National Agriculture and Food Research Organization |
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Principal Investigator |
Nakajima Ikuko 国立研究開発法人農業・食品産業技術総合研究機構, 果樹茶業研究部門, 上級研究員 (80355362)
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| Project Period (FY) |
2017-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | ブドウ / ゲノム編集 / 巨峰 / 園芸学 |
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| Outline of Final Research Achievements |
Poor/less coloration in ‘Kyoho’ grape skin is a big problem, which can be frequently seen by global worming. ‘Kyoho’ has four MYB transcription factors (MYB-TFs), two of which are active types, but the other two are inactive ones, because a retrotransposon (Gret1) is inserted in the promotor of these inactive MYB-TFs. It has been known that the extent of grape skin coloration depends on the number of an active MYB-TF. Thus, we planned to delete Gret1 using genome editing, which cause reactivation of MYB-TFs, which would result in the stabilization of the skin color of ‘Kyoho’. Target sequence of guide RNA was selected in the LTR region. Then binary vectors harboring Cas9, and sgRNA were transformed to ‘Kyoho’ embryogenic calli (EC). Twelve transgenic plants obtained were analyzed by multiplex PCR to confirm Gret1 deletion. The results showed that no genome edited plants were unfortunately obtained.
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| Academic Significance and Societal Importance of the Research Achievements |
形質転換可能な二倍体品種「ネオ・マスカット」ではPDS遺伝子のゲノム編集に我々は成功している。しかしながら四倍体品種である「巨峰」について、これまでゲノム編集を行った例はない。「巨峰」の形質転換は「ネオ・マスカット」よりも難しく、また標的としたGret1が約8.8kbと長かった。そのため、Gret1の欠失を狙うためには「巨峰」の形質転換の効率化と、ゲノム編集の際のさらなる切断効率の改良の必要性が示唆された。
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