| 研究課題/領域番号 |
23KF0288
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| 研究機関 | 沖縄科学技術大学院大学 |
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研究代表者 |
佐瀬 英俊 沖縄科学技術大学院大学, 植物エピジェネティクスユニット, 教授 (70510006)
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| 研究分担者 |
FURCI LEONARDO 沖縄科学技術大学院大学, 植物エピジェネティクスユニット, 外国人特別研究員
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| 研究期間 (年度) |
2023-11-15 – 2026-03-31
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| キーワード | Plant / Epigenome / rice / Pathogen response |
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| 研究実績の概要 |
We obtained transgenic rice plants for scalable DNA hypomethylation. We demonstrated estradiol-dependent activation of AtROS1 in rice using RT-PCR. Preliminary results using ELISA-based antibody quantification showed global DNA hypo-methylation in one rice sample treated with the highest concentration of estradiol. Analysis of DNA methylation level at previously identified regions though Chop-PCR confirmed DNA hypomethylation in estradiol-treated rice plants. These regions have previously been shown to dynamically change their DNA methylation status in response to abiotic stress. DNA hypomethylation was detected 2.5 weeks after initial estradiol treatment, indicating meta-stability of DNA hypomethylation during vegetative growth within the same generation. We completed an additional project on the role of RNA N6-adenine methylation in resistance against H. arabidopsidis infection in Arabidopsis.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
1: 当初の計画以上に進展している
理由
The project indicated three separated work packages (WP).
WP1 involved obtaining transgenic rice plants (T0) and validating AtROS1 accumulation and DNA de-methylation following estradiol treatment. Most tasks from WP1 have now been completed. AtROS1 activation followed estradiol treatment was confirmed through RT-PCR. DNA hypomethylation 2.5 weeks after estradiol treatment was assessed globally using ELISA-based colorimetric analysis, and at specific regions through Chop-PCR. More experiments will be carried on to further validate the activity of the vector..
The project is now moving forward into WP2, and WP3 in few months. We have obtained natural isolates of rice pathogens M. oryzae and X. oryzae, and successfully propagated them in preparation for WP2.
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| 今後の研究の推進方策 |
The project will now move into WP2, which will involve: a) isolating homozygous transgenic rice plants containing the XVE:ROS1 vector (T1), to be treated with various estradiol concentrations; b) producing transgenic plants displaying trans-generational meta-stable DNA hypomethylation. Both types of plant lines will be screened for resistance against pathogens M. oryzae and X. oryzae, as well as excess salt and drought stresses. Once resistant plant will be identified, the project will move to WP3: further characterization of causal epialleles.
If AtROS1-dependant DNA methylation displays trans-generational meta-stability, transgenic plants will be backcrossed to wild-type nipponbare to generate epigenetic recombinant inbred rice lines for further epialleles mapping.
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| 次年度使用額が生じた理由 |
The carry-over resources will be used to pay for Open Access publication fees for the upcoming paper currently under revision. Because of the revision, the paper is now due to come out in fiscal year 2024-2025.
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