Updated on 2025/10/08

写真a

 
Tetsu Kinoshita
 
Organization
Graduate School of Nanobioscience Department of Life and Environmental System Science Professor
School of Science Department of Science
Title
Professor
Homepage
http://epigenome.jp/
External link

Degree

  • 博士（理学） ( 総合研究大学院大学 )

Research Interests

  • 胚乳DNAメチル化

  • DAN脱メチル化

  • Arabidopsis

  • DNAメチレーション

  • 野生イネ

  • インプリンティング

  • ゲノムインプリンティング

  • DNA脱メチル化

  • 生殖

  • バイオテクノロジー

  • 雑種

  • メチル化

  • 遺伝子

  • 遺伝学

  • 染色体

  • imprinting

  • ゲノム障壁

  • O. longistaminata

  • ゲノム

  • 生殖系列

  • タイリングアレイ

  • 突然変異

  • レーザーマイクロダイセクション

  • 組換え機構

  • 胚乳

  • エピジェネティクス

  • DNAメチル化

  • シロイヌナズナ

  • イネ

  • 生殖隔離

Research Areas

  • Life Science / Plant molecular biology and physiology

  • Environmental Science/Agriculture Science / Science in plant genetics and breeding

  • Life Science / Genetics

Research History

  • Yokohama City University   Professor

    2015

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  • Yokohama City University International College of Arts and Sciences, Life and Environmental Science Graduate School of Nanobioscience Department of Life and Environmental System Science   Professor

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Papers

  • Multilayered epigenetic control of persistent and stage-specific imprinted genes in rice endosperm. International journal

    Kaoru Tonosaki, Daichi Susaki, Hatsune Morinaka, Akemi Ono, Hiroki Nagata, Hiroyasu Furuumi, Ken-Ichi Nonomura, Yutaka Sato, Keiko Sugimoto, Luca Comai, Katsunori Hatakeyama, Taiji Kawakatsu, Tetsu Kinoshita

    Nature plants   10 ( 8 )   1231 - 1245   2024.8

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    Language：English   Publishing type：Research paper (scientific journal)  

    In angiosperms, epigenetic profiles for genomic imprinting are established before fertilization. However, the causal relationships between epigenetic modifications and imprinted expression are not fully understood. In this study, we classified 'persistent' and 'stage-specific' imprinted genes on the basis of time-course transcriptome analysis in rice (Oryza sativa) endosperm and compared them to epigenetic modifications at a single time point. While the levels of epigenetic modifications are relatively low in stage-specific imprinted genes, they are considerably higher in persistent imprinted genes. Overall trends revealed that the maternal alleles of maternally expressed imprinted genes are activated by DNA demethylation, while the maternal alleles of paternally expressed imprinted genes with gene body methylation (gbM) are silenced by DNA demethylation and H3K27me3 deposition, and these regions are associated with an enriched motif related to Tc/Mar-Stowaway. Our findings provide insight into the stability of genomic imprinting and the potential variations associated with endosperm development, different cell types and parental genotypes.

    DOI： 10.1038/s41477-024-01754-4

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  • Microtubules ensure transport of vegetative nuclei and sperm cells by fine-tuning their home positions

    Kazuki Motomura, Haruna Tsuchi, Marin Komojiri, Ayumi Matsumoto, Naoya Sugi, Daichi Susaki, Atsushi Takeda, Tetsu Kinoshita, Daisuke Maruyama

    2024.2

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  • Blue light irradiation induces pollen tube rupture in various flowering plants

    Naoya Sugi, Daichi Susaki, Yoko Mizuta, Tetsu Kinoshita, Daisuke Maruyama

    2023.12

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    Publisher：Cold Spring Harbor Laboratory  

    Abstract

    Pollen tubes exhibit one of the fastest apical growth rates among plant cells. Maintaining the proper balance between turgor pressure and cell wall synthesis at the pollen tube tip is crucial for this rapid growth, and any disruption can result in pollen tube rupture. In our study, we reveal that exposure to short-wavelength visible light, specifically blue light, induces pollen tube rupture. The frequency of pollen tube rupture increases in an intensity-dependent manner. Additionally, we observed Ca<sup>2+</sup>influx after blue light irradiation, accompanying with either pollen tube rupture or a temporary halt in elongation. These findings offer insights into the interplay between pollen tube integrity maintenance and Ca<sup>2+</sup>influx at the pollen tube tip, presenting a novel and efficient method to control pollen tube burst.

    Subject Areas

    (1) growth and development

    (11) new methodology

    DOI： 10.1101/2023.12.06.570347

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  • Polycomb repression of the asexual embryo. International journal

    Kaoru Tonosaki, Tetsu Kinoshita

    Nature plants   2023.10

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1038/s41477-023-01537-3

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  • Removal of the endoplasma membrane upon sperm cell activation after pollen tube discharge

    Naoya Sugi, Rie Izumi, Shun Tomomi, Daichi Susaki, Tetsu Kinoshita, Daisuke Maruyama

    Frontiers in Plant Science   14   2023.1

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    Publishing type：Research paper (scientific journal)   Publisher：Frontiers Media SA  

    In pollen and pollen tubes, immotile sperm cells are enclosed by an inner vegetative plasma membrane (IVPM), a single endomembrane originating from the vegetative-cell plasma membrane. It is widely believed that sperm cells must be removed from the IVPM prior to gamete associations and fusions; however, details of the timing and morphological changes upon IVPM dissociation remain elusive. Here, we report a rapid IVPM breakdown immediately before double fertilization in Arabidopsis thaliana. The IVPM was stably observed in coiling pollen tubes when pollen tube discharge was prevented using lorelei mutant ovules. In contrast, a semi-in vivo fertilization assay in wild-type ovules demonstrated fragmented IVPM around sperm nuclei 1 min after pollen tube discharge. These observations revealed the dynamic alteration of released sperm cells and provided new insights into double fertilization in flowering plants. With a summary of recent findings on IVPM lipid composition, we discussed the possible physiological signals controlling IVPM breakdown.

    DOI： 10.3389/fpls.2023.1116289

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  • Multi-layered epigenetic control of persistent and stage-specific imprinted genes in rice endosperm

    Kaoru Tonosaki, Akemi Ono, Hiroki Nagata, Hiroyasu Furuumi, Ken-ichi Nonomura, Yutaka Sato, Luca Comai, Katsunori Hatakeyama, Taiji Kawakatsu, Tetsu Kinoshita

    2023.1

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    Publisher：Research Square Platform LLC  

    Abstract

    In the offspring, imprinted genes show differential expression from the maternal and paternal alleles. Our time-course analysis in rice (Oryza sativa) revealed persistent and stage-specific classes of imprinted genes, including over a hundred novel genes, especially in the early stage of endosperm development. In the persistent maternally expressed genes (MEGs), CG and CHG sequences are heavily methylated, while these sites are less methylated in the stage-specific MEGs. Similarly, differences in H3K27me3 defined the two classes of paternally expressed genes (PEGs). The imprinting for MEGs involves activating silent genes. By contrast, many PEGs are active in various tissues and carry gene body DNA methylation (gbM); the maternally inherited alleles of these PEGs undergo silencing via gbM demethylation and deposition of H3K27me3. Stage-specific imprinting indicates that parental marks can be differentially interpreted during development. Its developmental variability contrasts the robustness of persistent imprinting, revealing the dynamic layering of parental effects and suggesting new mechanisms for evolution and manipulation of rice endosperm development.

    DOI： 10.21203/rs.3.rs-2403620/v1

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    Other Link： https://www.researchsquare.com/article/rs-2403620/v1.html

  • Elongation of Siliques Without Pollination 3 Regulates Nutrient Flow Necessary for Embryogenesis

    Hironori Takasaki, Miho Ikeda, Reika Hasegawa, Zhang Yilin, Shingo Sakamoto, Daisuke Maruyama, Nobutaka Mitsuda, Tetsu Kinoshita, Masaru Ohme-Takagi

    Plant and Cell Physiology   2022.10

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    Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

    Abstract

    Apomixis, defined as the transfer of maternal germplasm to offspring without fertilization, enables the fixation of F1-useful traits, providing advantages in crop breeding. However, most apomictic plants require pollination to produce endosperm. Endosperm is essential for embryogenesis and its development is suppressed until fertilization. We show that expression of a chimeric repressor of the Elongation of Siliques without Pollination 3 (ESP3) gene (Pro35S:ESP3-SRDX) induces ovule enlargement without fertilization in Arabidopsis thaliana. The ESP3 gene encodes a protein similar to the FWA homeodomain transcription factor containing a StAR-related lipid-transfer (START) domain. However, ESP3 lacks the homeobox-encoding region. Genes related to the cell cycle and sugar metabolism were upregulated in unfertilized Pro35S:ESP3-SRDX ovules similar as in fertilized seeds, while those related to autophagy were downregulated similar to fertilized seeds. Unfertilized Pro35S:ESP3-SRDX ovules partially nourished embryos when only the egg was fertilized, accumulating hexoses without central cell proliferation. ESP3 may regulate nutrient flow during seed development, and ESP3-SRDX could be a useful tool for complete apomixis that does not require pseudo-fertilization.

    DOI： 10.1093/pcp/pcac151

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  • F-actin regulates polarized secretion of pollen tube attractants in Arabidopsis synergid cell.

    Daichi Susaki, Rie Izumi, Takao Oi, Hidenori Takeuchi, Ji Min Shin, Naoya Sugi, Tetsu Kinoshita, Tetsuya Higashiyama, Tomokazu Kawashima, Daisuke Maruyama

    2022.6

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    Publisher：Cold Spring Harbor Laboratory  

    Pollen tube attraction is a key event of sexual reproduction in flowering plants. In the ovule, two synergid cells neighboring the egg cell control pollen tube arrival via the active secretion of attractant peptides such as AtLURE1 and XIUQIU from the filiform apparatus facing toward the micropyle. Distinctive cell polarity together with longitudinal F-actin and microtubules are hallmarks of the synergid cell in various species, though functions of these cellular structures are still unclear. In this study we used genetic and pharmacological approaches to elucidate the roles of cytoskeletal components in filiform apparatus formation and pollen tube guidance in Arabidopsis thaliana. Inhibition of microtubule formation reduced invaginations of the plasma membrane but did not abolish micropylar AtLURE1.2 accumulation. In contrast, the expression of a dominant-negative form of ACTIN8 induced disorganization of the filiform apparatus and loss of polar AtLURE1.2 distribution toward the filiform apparatus. Interestingly, after pollen tube reception, F-actin became unclear for a few hours in the persistent synergid cell, which may be involved in pausing and resuming pollen tube attraction during early polytubey block. Our data propose the central role of F-actin in the maintenance of cell polarity and function of male-female communication in the synergid cell.

    DOI： 10.1101/2022.06.14.496136

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  • Persistent directional growth capability in Arabidopsis thaliana pollen tubes after nuclear elimination from the apex International journal

    Kazuki Motomura, Hidenori Takeuchi, Michitaka Notaguchi, Haruna Tsuchi, Atsushi Takeda, Tetsu Kinoshita, Tetsuya Higashiyama, Daisuke Maruyama

    Nature Communications   12 ( 1 )   2331 - 2331   2021.12

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

    <title>Abstract</title>During the double fertilization process, pollen tubes deliver two sperm cells to an ovule containing the female gametes. In the pollen tube, the vegetative nucleus and sperm cells move together to the apical region where the vegetative nucleus is thought to play a crucial role in controlling the direction and growth of the pollen tube. Here, we report the generation of pollen tubes in <italic>Arabidopsis thaliana</italic> whose vegetative nucleus and sperm cells are isolated and sealed by callose plugs in the basal region due to apical transport defects induced by mutations in the WPP domain-interacting tail-anchored proteins (WITs) and sperm cell-specific expression of a dominant mutant of the CALLOSE SYNTHASE 3 protein. Through pollen-tube guidance assays, we show that the physiologically anuclear mutant pollen tubes maintain the ability to grow and enter ovules. Our findings provide insight into the sperm cell delivery mechanism and illustrate the independence of the tip-localized vegetative nucleus from directional growth control of the pollen tube.

    DOI： 10.1038/s41467-021-22661-8

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    Other Link： http://www.nature.com/articles/s41467-021-22661-8

  • Temporal Changes in Transcripts of MITE Transposable Elements during Rice Endosperm Development

    Hiroki Nagata, Akemi Ono, Kaoru Tonosaki, Taiji Kawakatsu, Kentaro Yano, Yuji Kishima, Tetsu Kinoshita

    2021.9

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    Publisher：Cold Spring Harbor Laboratory  

    The repression of transcription from transposable elements (TEs) by DNA methylation is necessary to maintain genome integrity and prevent harmful mutations. However, under certain circumstances, TEs are thought to escape from the host defense system and reactivate their transcription. In Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa), DNA demethylases target the sequences derived from TEs in the central cell, the progenitor cell for the endosperm in the female gametophyte. This genome-wide DNA demethylation is also observed in the endosperm after fertilization. In this study, we used a custom microarray to survey the transcripts generated from TEs during the rice endosperm development and at selected timepoints in the embryo as a control. The expression patterns of TE transcripts are dynamically up- and downregulated during endosperm development, especially for miniature inverted-repeat transposable elements (MITEs). Surprisingly, some TE transcripts were directionally controlled, while the other DNA transposons and retrotransposons were not. We also discovered the NF-Y binding motif, CCAAT, in the region near the 5′ terminal inverted repeat of Youren, one of the transcribed MITEs in the endosperm. Our results uncover dynamic changes in TE activity during endosperm development in rice.

    DOI： 10.1101/2021.09.27.461986

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  • Cubam receptor-mediated endocytosis in hindgut-derived pseudoplacenta of a viviparous teleost Xenotoca eiseni Reviewed International journal

    Atsuo Iida, Kaori Sano, Mayu Inokuchi, Jumpei Nomura, Takayuki Suzuki, Mao Kuriki, Maina Sogabe, Daichi Susaki, Kaoru Tonosaki, Tetsu Kinoshita, Eiichi Hondo

    Journal of Experimental Biology   224 ( 13 )   jeb242613   2021.6

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：The Company of Biologists  

    Nutrient transfer from mother to the embryo is essential for reproduction in viviparous animals. In the viviparous teleost Xenotoca eiseni belonging to the family Goodeidae, the intraovarian embryo intakes the maternal component secreted into the ovarian fluid via the trophotaenia. Our previous study reported that the epithelial layer cells of the trophotaenia incorporate a maternal protein via vesicle trafficking. However, the molecules responsible for the absorption were still elusive. Here, we focused on Cubam (Cubilin-Amnionless) as a receptor involved in the absorption, and cathepsin L as a functional protease in the vesicles. Our results indicated that the Cubam receptor is distributed in the apical surface of the trophotaenia epithelium and then is taken into the intracellular vesicles. The trophotaenia possesses acidic organelles in epithelial layer cells and cathepsin L-dependent proteolysis activity. This evidence does not conflict with our hypothesis that receptor-mediated endocytosis and proteolysis play roles in maternal macromolecule absorption via the trohotaenia in viviparous teleosts. Such nutrient absorption involving endocytosis is not a specific trait in viviparous fish. Similar processes have been reported in the larval stage of oviparous fish or the suckling stage of viviparous mammals. Our findings suggest that the viviparous teleost acquired trophotaenia-based viviparity from a modification of the intestinal absorption system common in vertebrates. This is a fundamental study to understand the strategic variation of the reproductive system in vertebrates.

    DOI： 10.1242/jeb.242613

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  • Epigenetics and plant reproduction: Multiple steps for responsibly handling succession

    Akemi Ono, Tetsu Kinoshita

    Current Opinion in Plant Biology   61   102032 - 102032   2021.6

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    Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

    DOI： 10.1016/j.pbi.2021.102032

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  • Mutation of the imprinted gene OsEMF2a induces autonomous endosperm development and delayed cellularization in rice

    Kaoru Tonosaki, Akemi Ono, Megumi Kunisada, Megumi Nishino, Hiroki Nagata, Shingo Sakamoto, Saku T Kijima, Hiroyasu Furuumi, Ken-Ichi Nonomura, Yutaka Sato, Masaru Ohme-Takagi, Masaki Endo, Luca Comai, Katsunori Hatakeyama, Taiji Kawakatsu, Tetsu Kinoshita

    The Plant Cell   2020.11

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    Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

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    In angiosperms, endosperm development comprises a series of developmental transitions controlled by genetic and epigenetic mechanisms that are initiated after double fertilization. Polycomb repressive complex 2 (PRC2) is a key component of these mechanisms that mediate histone H3 lysine 27 trimethylation (H3K27me3); the action of PRC2 is well described in Arabidopsis thaliana but remains uncertain in cereals. In this study, we demonstrate that mutation of the rice (Oryza sativa) gene EMBRYONIC FLOWER2a (OsEMF2a), encoding a zinc-finger containing component of PRC2, causes an autonomous endosperm phenotype involving proliferation of the central cell nuclei with separate cytoplasmic domains, even in the absence of fertilization. Detailed cytological and transcriptomic analyses revealed that the autonomous endosperm can produce storage compounds, starch granules, and protein bodies specific to the endosperm. These events have not been reported in Arabidopsis. After fertilization, we observed an abnormally delayed developmental transition in the endosperm. Transcriptome and H3K27me3 ChIP-seq analyses using endosperm from the emf2a mutant identified downstream targets of PRC2. These included &amp;gt;100 transcription factor genes such as type-I MADS-box genes, which are likely required for endosperm development. Our results demonstrate that OsEMF2a-containing PRC2 controls endosperm developmental programs before and after fertilization.

    DOI： 10.1093/plcell/koaa006

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    Other Link： http://academic.oup.com/plcell/advance-article-pdf/doi/10.1093/plcell/koaa006/36138656/koaa006.pdf

  • Sperm Entry into the Egg Cell Induces the Progression of Karyogamy in Rice Zygotes. Reviewed

    Yukinosuke Ohnishi, Iwao Kokubu, Tetsu Kinoshita, Takashi Okamoto

    Plant & cell physiology   60 ( 8 )   1656 - 1665   2019.8

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    Karyogamy is a prerequisite event for plant embryogenesis, in which dynamic changes in nuclear architecture and the establishment of appropriate gene expression patterns must occur. However, the precise role of the male and female gametes in the progression of karyogamy still remains elusive. Here, we show that the sperm cell possesses the unique property to drive steady and swift nuclear fusion. When we fertilized egg cells with sperm cells in vitro, the immediate fusion of the male and female nuclei in the zygote progressed. This rapid nuclear fusion did not occur when two egg cells were artificially fused. However, the nuclear fusion of two egg nuclei could be accelerated by additional sperm entry or the exogenous application of calcium, suggesting that possible increase of cytosolic Ca2+ level via sperm entry into the egg cell efficiently can facilitate karyogamy. In contrast to zygotes, the egg-egg fusion cells failed to proliferate beyond an early developmental stage. Our transcriptional analyses also revealed the rapid activation of zygotic genes in zygotes, whereas there was no expression in fused cells without the male contribution. Thus, the male sperm cell has the ability to cause immediate karyogamy and to establish appropriate gene expression patterns in the zygote.

    DOI： 10.1093/pcp/pcz077

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  • Salt Tolerance Improvement in Rice through Efficient SNP Marker-Assisted Selection Coupled with Speed-Breeding. Reviewed International journal

    Md Masud Rana, Takeshi Takamatsu, Marouane Baslam, Kentaro Kaneko, Kimiko Itoh, Naoki Harada, Toshie Sugiyama, Takayuki Ohnishi, Tetsu Kinoshita, Hiroki Takagi, Toshiaki Mitsui

    International journal of molecular sciences   20 ( 10 )   2019.5

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    Language：English   Publishing type：Research paper (scientific journal)  

    Salinity critically limits rice metabolism, growth, and productivity worldwide. Improvement of the salt resistance of locally grown high-yielding cultivars is a slow process. The objective of this study was to develop a new salt-tolerant rice germplasm using speed-breeding. Here, we precisely introgressed the hst1 gene, transferring salinity tolerance from "Kaijin" into high-yielding "Yukinko-mai" (WT) rice through single nucleotide polymorphism (SNP) marker-assisted selection. Using a biotron speed-breeding technique, we developed a BC3F3 population, named "YNU31-2-4", in six generations and 17 months. High-resolution genotyping by whole-genome sequencing revealed that the BC3F2 genome had 93.5% similarity to the WT and fixed only 2.7% of donor parent alleles. Functional annotation of BC3F2 variants along with field assessment data indicated that "YNU31-2-4" plants carrying the hst1 gene had similar agronomic traits to the WT under normal growth condition. "YNU31-2-4" seedlings subjected to salt stress (125 mM NaCl) had a significantly higher survival rate and increased shoot and root biomasses than the WT. At the tissue level, quantitative and electron probe microanalyzer studies indicated that "YNU31-2-4" seedlings avoided Na+ accumulation in shoots under salt stress. The "YNU31-2-4" plants showed an improved phenotype with significantly higher net CO2 assimilation and lower yield decline than WT under salt stress at the reproductive stage. "YNU31-2-4" is a potential candidate for a new rice cultivar that is highly tolerant to salt stress at the seedling and reproductive stages, and which might maintain yields under a changing global climate.

    DOI： 10.3390/ijms20102585

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  • DNA demethylation by ROS1a in rice vegetative cells promotes methylation in sperm. Reviewed International journal

    M Yvonne Kim, Akemi Ono, Stefan Scholten, Tetsu Kinoshita, Daniel Zilberman, Takashi Okamoto, Robert L Fischer

    Proceedings of the National Academy of Sciences of the United States of America   116 ( 19 )   9652 - 9657   2019.5

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    Epigenetic reprogramming is required for proper regulation of gene expression in eukaryotic organisms. In Arabidopsis, active DNA demethylation is crucial for seed viability, pollen function, and successful reproduction. The DEMETER (DME) DNA glycosylase initiates localized DNA demethylation in vegetative and central cells, so-called companion cells that are adjacent to sperm and egg gametes, respectively. In rice, the central cell genome displays local DNA hypomethylation, suggesting that active DNA demethylation also occurs in rice; however, the enzyme responsible for this process is unknown. One candidate is the rice REPRESSOR OF SILENCING1a (ROS1a) gene, which is related to DME and is essential for rice seed viability and pollen function. Here, we report genome-wide analyses of DNA methylation in wild-type and ros1a mutant sperm and vegetative cells. We find that the rice vegetative cell genome is locally hypomethylated compared with sperm by a process that requires ROS1a activity. We show that many ROS1a target sequences in the vegetative cell are hypomethylated in the rice central cell, suggesting that ROS1a also demethylates the central cell genome. Similar to Arabidopsis, we show that sperm non-CG methylation is indirectly promoted by DNA demethylation in the vegetative cell. These results reveal that DNA glycosylase-mediated DNA demethylation processes are conserved in Arabidopsis and rice, plant species that diverged 150 million years ago. Finally, although global non-CG methylation levels of sperm and egg differ, the maternal and paternal embryo genomes show similar non-CG methylation levels, suggesting that rice gamete genomes undergo dynamic DNA methylation reprogramming after cell fusion.

    DOI： 10.1073/pnas.1821435116

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  • A parental tug-of-war Reviewed

    Tetsu Kinoshita

    Nature Plants   4 ( 6 )   329 - 330   2018.6

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    Language：English   Publisher：Palgrave Macmillan Ltd.  

    DOI： 10.1038/s41477-018-0179-9

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  • Endosperm Development Reviewed

    Tonosaki K, Kinoshita T

    eLS   DOI: 10.1002/9780470015902.a00   2018.6

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  • FACT complex is required for DNA demethylation at heterochromatin during reproduction in Arabidopsis Reviewed

    Jennifer M. Frost, M. Yvonne Kim, Guen Tae Park, Ping-Hung Hsieh, Miyuki Nakamura, Samuel J. H. Lin, Hyunjin Yoo, Jaemyung Choi, Yoko Ikeda, Tetsu Kinoshita, Yeonhee Choi, Daniel Zilberman, Robert L. Fischer

    Proceedings of the National Academy of Sciences of the United States of America   115 ( 20 )   E4720 - E4729   2018.5

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：National Academy of Sciences  

    DOI： 10.1073/pnas.1713333115

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  • Optimized method of extracting rice chloroplast DNA for high-quality plastome resequencing and de Novo assembly Reviewed

    Takeshi Takamatsu, Marouane Baslam, Takuya Inomata, Kazusato Oikawa, Kimiko Itoh, Takayuki Ohnishi, Tetsu Kinoshita, Toshiaki Mitsui

    Frontiers in Plant Science   9   266   2018.2

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Frontiers Media S.A.  

    DOI： 10.3389/fpls.2018.00266

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  • Overcoming the species hybridization barrier by ploidy manipulation in the genus Oryza Reviewed

    Kaoru Tonosaki, Daisuke Sekine, Takayuki Ohnishi, Akemi Ono, Hiroyasu Furuumi, Nori Kurata, Tetsu Kinoshita

    Plant Journal   93 ( 3 )   534 - 544   2018.2

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Blackwell Publishing Ltd  

    DOI： 10.1111/tpj.13803

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  • Mutants in the Fe-S biogenesis component AtDRE2 develop twin embryos and are defective in DNA demethylation in the vegetative phase Invited Reviewed

    Diana Mihaela Buzas, Tetsu Kinoshita

    Molecular Life   doi: 10.26600/MolLife.1.2.4.20   2018

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  • A pharmacological study of Arabidopsis cell fusion between the persistent synergid and endosperm Reviewed

    Kazuki Motomura, Tomokazu Kawashima, Frédéric Berger, Tetsu Kinoshita, Tetsuya Higashiyama, Daisuke Maruyama

    Journal of Cell Science   131 ( 2 )   2018

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    Language：English   Publishing type：Research paper (scientific journal)   Publisher：Company of Biologists Ltd  

    DOI： 10.1242/jcs.204123

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  • Dormancy-specific imprinting underlies maternal inheritance of seed dormancy in Arabidopsis thaliana Reviewed

    Urszula Piskurewicz, Mayumi Iwasaki, Daichi Susaki, Christian Megies, Tetsu Kinoshita, Luis Lopez-Molina

    ELIFE   5   2016.12

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.7554/eLife.19573

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  • Fertilization-independent Cell-fusion between the Synergid and Central Cell in the Polycomb Mutant Reviewed

    Kazuki Motomura, Frederic Berger, Tomokazu Kawashima, Tetsu Kinoshita, Tetsuya Higashiyama, Daisuke Maruyama

    CELL STRUCTURE AND FUNCTION   41 ( 2 )   121 - 125   2016

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1247/csf.16010

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  • Possible roles for polycomb repressive complex 2 in cereal endosperm Reviewed

    Kaoru Tonosaki, Tetsu Kinoshita

    FRONTIERS IN PLANT SCIENCE   6   144   2015.3

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  • Rice SNF2 family helicase ENL1 is essential for syncytial endosperm development Reviewed

    Tomomi Hara, Hirokazu Katoh, Daisuke Ogawa, Yasuaki Kagaya, Yutaka Sato, Hidemi Kitano, Yasuo Nagato, Ryo Ishikawa, Akemi Ono, Tetsu Kinoshita, Shin Takeda, Tsukaho Hattori

    PLANT JOURNAL   81 ( 1 )   1 - 12   2015.1

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1111/tpj.12705

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  • Epigenetic Memory for Stress Response and Adaptation in Plants Reviewed

    Tetsu Kinoshita, Motoaki Seki

    PLANT AND CELL PHYSIOLOGY   55 ( 11 )   1859 - 1863   2014.11

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  • Epigenetic role for the conserved Fe-S cluster biogenesis protein AtDRE2 in Arabidopsis thaliana Reviewed

    Diana Mihaela Buzas, Miyuki Nakamura, Tetsu Kinoshita

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   111 ( 37 )   13565 - 13570   2014.9

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    Language：English   Publishing type：Research paper (scientific journal)  

    DOI： 10.1073/pnas.1404058111

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  • OsATG7 is required for autophagy-dependent lipid metabolism in rice postmeiotic anther development Reviewed

    Takamitsu Kurusu, Tomoko Koyano, Shigeru Hanamata, Takahiko Kubo, Yuhei Noguchi, Chikako Yagi, Noriko Nagata, Takashi Yamamoto, Takayuki Ohnishi, Yozo Okazaki, Nobutaka Kitahata, Daichi Ando, Masaya Ishikawa, Shinya Wada, Akio Miyao, Hirohiko Hirochika, Hiroaki Shimada, Amane Makino, Kazuki Saito, Hiroyuki Ishida, Tetsu Kinoshita, Nori Kurata, Kazuyuki Kuchitsu

    AUTOPHAGY   10 ( 5 )   878 - 888   2014.5

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    DOI： 10.4161/auto.28279

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  • Genomic Imprinting in Plants: What Makes the Functions of Paternal and Maternal Genes Different in Endosperm Formation? Reviewed

    Takayuki Ohnishi, Daisuke Sekine, Tetsu Kinoshita

    EPIGENETIC SHAPING OF SOCIOSEXUAL INTERACTIONS: FROM PLANTS TO HUMANS   86   1 - 25   2014

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    DOI： 10.1016/B978-0-12-800222-3.00001-2

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  • Dissection of two major components of the post-zygotic hybridization barrier in rice endosperm Reviewed

    Daisuke Sekine, Takayuki Ohnishi, Hiroyasu Furuumi, Akemi Ono, Toshihiro Yamada, Nori Kurata, Tetsu Kinoshita

    PLANT JOURNAL   76 ( 5 )   792 - 799   2013.12

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    DOI： 10.1111/tpj.12333

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  • The role of Arabidopsis thaliana NAR1, a cytosolic iron-sulfur cluster assembly component, in gametophytic gene expression and oxidative stress responses in vegetative tissue Reviewed

    Miyuki Nakamura, Diana Mihaela Buzas, Akira Kato, Masahiro Fujita, Nori Kurata, Tetsu Kinoshita

    NEW PHYTOLOGIST   199 ( 4 )   925 - 935   2013.9

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    DOI： 10.1111/nph.12350

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  • RNA-directed DNA methylation regulates parental genomic imprinting at several loci in Arabidopsis Reviewed

    Thiet Minh Vu, Miyuki Nakamura, Joseph P. Calarco, Daichi Susaki, Pei Qi Lim, Tetsu Kinoshita, Tetsuya Higashiyama, Robert A. Martienssen, Frédéric Berger

    Development (Cambridge)   140 ( 14 )   2953 - 2960   2013.7

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    DOI： 10.1242/dev.092981

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  • Epimutagenesis and its application for next generation breeding

    前川雅彦, 金澤章, 堤伸浩, 木下哲, 土生芳樹, 柴博史, 江面,浩

    Breeding Research   15 ( 2 )   42 - 50   2013.6

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    DOI： 10.1270/jsbbr.15.42

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    Other Link： https://jlc.jst.go.jp/DN/JALC/10021164544?from=CiNii

  • Opening the Door to Epigenetics in PCP Reviewed

    Tetsu Kinoshita, Steven E. Jacobsen

    PLANT AND CELL PHYSIOLOGY   53 ( 5 )   763 - 765   2012.5

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  • HMG Domain containing SSRP1 is required for DNA demethylation and genomic imprinting in arabidopsis Reviewed

    Yoko Ikeda, Yuki Kinoshita, Daichi Susaki, Yuriko Ikeda, Megumi Iwano, Seiji Takayama, Tetsuya Higashiyama, Tetsuji Kakutani, Tetsu Kinoshita

    Developmental Cell   21 ( 3 )   589 - 596   2011.9

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    DOI： 10.1016/j.devcel.2011.08.013

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  • The biotron breeding system: A rapid and reliable procedure for genetic studies and breeding in rice Reviewed

    Takayuki Ohnishi, Mihoko Yoshino, Hiromoto Yamakawa, Tetsu Kinoshita

    Plant and Cell Physiology   52 ( 7 )   1249 - 1257   2011.7

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    DOI： 10.1093/pcp/pcr066

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  • Rice interspecies hybrids show precocious or delayed developmental transitions in the endosperm without change to the rate of syncytial nuclear division Reviewed

    Ryo Ishikawa, Takayuki Ohnishi, Yuki Kinoshita, Mitsugu Eiguchi, Nori Kurata, Tetsu Kinoshita

    Plant Journal   65 ( 5 )   798 - 806   2011.3

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    DOI： 10.1111/j.1365-313X.2010.04466.x

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  • <I>ENDOSPERMLESS1</I> encodes an SNF2-like helicase essential for endosperm development in rice

    Katoh Hirokazu, Hara Tomomi, Sato Yutaka, Kitano Hidemi, Nagato Yasuo, Ishikawa Ryo, Kinoshita Tetsu, Takeda Shin, Hattori Tsukaho

    Plant and Cell Physiology Supplement   2011   0060 - 0060   2011

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    The rice <I>enl1</I> mutant bears seeds without endosperm. We mapped the <I>enl1</I> locus at a region close to the telomere of the Chromosome 4 long arm and identified a deletion of several nucleotides and substitutions in an exon of a predicted gene in this region. Transgenic introduction of a wild type genomic fragment containing this gene completely complemented the mutant phenotype, concluding this gene to be <I>ENL1</I>. <I>ENL1</I> encodes a protein belonging to the SNF2 class helicase family, which harbors an SNF2 domain and a HELICc domain. Orthologs of ENL1 are conserved among most of eukaryotes. The human ortholog, PICH, is reported to be required for chromosome condensation, chromosome arm architecture, and sister chromatid segregation. Laser confocal microscopic observation of PI-stained syncytial endosperms revealed the presence of giant endosperm nuclei with many nucleoli in the mutant. The abnormality of mutant nuclei appeared to derive from failure in chromosome separation. These results suggest that the errors in the chromosome cycle at the syncytial stage in the <I>enl1</I> mutant result in the inability of further endosperm development.

    DOI： 10.14841/jspp.2011.0.0060.0

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  • Epigenetic programming: The challenge to species hybridization Reviewed

    Ryo Ishikawa, Tetsu Kinoshita

    Molecular Plant   2 ( 4 )   589 - 599   2009.7

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  • Abnormal development observed in the hybrid endosperm between cultivated and wild rice

    Ishikawa Ryo, Eiguchi Mitsugu, Shinzaki Yuki, Kurata Nori, Kinoshita Tetsu

    Plant and Cell Physiology Supplement   2009   0114 - 0114   2009

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    Abnormal endosperm development is observed in inter-specific or inter-ploidy crosses of many plant species. Several reports suggested that the endosperm failure is caused either by promotion or suppression of the endosperm development and this can be due to the functional difference between parental genomes. One of the possible molecular mechanisms underlying this phenomenon can be genomic imprinting, an epigenetic mechanism with parent-of-origin dependent mono-allelic gene expression. <br>We conducted inter-specific crosses between several cultivated and wild rice species and found abnormality in hybrid endosperm. In these crosses, the rate of nuclear division of the endosperm was not affected but the timing of cellularization was found to be delayed or advanced. We also carried out the microarray analysis using hybrid endosperm of <I>O. sativa</I> and <I>O. longistaminata</I>, and identified the candidate genes related to the endosperm failure. Possible involvement of these genes and their effects on the endosperm development will be discussed.

    DOI： 10.14841/jspp.2009.0.0114.0

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  • DNA demethylation: A lesson from the garden Reviewed

    Yoko Ikeda, Tetsu Kinoshita

    Chromosoma   118 ( 1 )   37 - 41   2009

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  • Genomic imprinting: A balance between antagonistic roles of parental chromosomes Reviewed

    Tetsu Kinoshita, Yoko Ikeda, Ryo Ishikawa

    Seminars in Cell and Developmental Biology   19 ( 6 )   574 - 579   2008.12

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  • MATERNALLY EXPRESSED PAB C-TERMINAL, a novel imprinted gene in Arabidopsis, encodes the conserved C-terminal domain of polyadenylate binding proteins Reviewed

    Sushma Tiwari, Reiner Schulz, Yoko Ikeda, Lindsay Dytham, Jaime Bravo, Lucille Mathers, Melissa Spielman, Plinio Guzmán, Rebecca J. Oakey, Tetsu Kinoshita, Rod J. Scotta

    Plant Cell   20 ( 9 )   2387 - 2398   2008.9

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    DOI： 10.1105/tpc.108.061929

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  • Evolution and control of imprinted FWA genes in the genus Arabidopsis Reviewed

    Ryo Fujimoto, Yuki Kinoshita, Akira Kawabe, Tetsu Kinoshita, Kazuya Takashima, Magnus Nordborg, Mikhail E. Nasrallah, Kentaro K. Shimizu, Hiroshi Kudoh, Tetsuji Kakutani

    PLoS Genetics   4 ( 4 )   2008.4

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    A central question in genomic imprinting is how a specific sequence is recognized as the target for epigenetic marking. In both mammals and plants, imprinted genes are often associated with tandem repeats and transposon-related sequences, but the role of these elements in epigenetic gene silencing remains elusive. FWA is an imprinted gene in Arabidopsis thaliana expressed specifically in the female gametophyte and endosperm. Tissue-specific and imprinted expression of FWA depends on DNA methylation in the FWA promoter, which is comprised of two direct repeats containing a sequence related to a SINE retroelement. Methylation of this element causes epigenetic silencing, but it is not known whether the methylation is targeted to the SINE-related sequence itself or the direct repeat structure is also necessary. Here we show that the repeat structure in the FWA promoter is highly diverse in species within the genus Arabidopsis. Four independent tandem repeat formation events were found in three closely related species. Another related species, A. halleri, did not have a tandem repeat in the FWA promoter. Unexpectedly, even in this species, FWA expression was imprinted and the FWA promoter was methylated. In addition, our expression analysis of FWA gene in vegetative tissues revealed high frequency of intra-specific variation in the expression level. In conclusion, we show that the tandem repeat structure is dispensable for the epigenetic silencing of the FWA gene. Rather, SINE-related sequence is sufficient for imprinting, vegetative silencing, and targeting of DNA methylation. Frequent independent tandem repeat formation events in the FWA promoter led us to propose that they may be a consequence, rather than cause, of the epigenetic control. The possible significance of epigenetic variation in reproductive strategies during evolution is also discussed. © 2008 Fujimoto et al.

    DOI： 10.1371/journal.pgen.1000048

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  • Evolution and control of imprinted FWA genes in the genus Arabidopsis Reviewed

    Ryo Fujimoto, Yuki Kinoshita, Akira Kawabe, Tetsu Kinoshita, Kazuya Takashima, Magnus Nordborg, Mikhail E. Nasrallah, Kentaro K. Shimizu, Hiroshi Kudoh, Tetsuji Kakutani

    PLOS GENETICS   4 ( 4 )   2008.4

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    A central question in genomic imprinting is how a specific sequence is recognized as the target for epigenetic marking. In both mammals and plants, imprinted genes are often associated with tandem repeats and transposon-related sequences, but the role of these elements in epigenetic gene silencing remains elusive. FWA is an imprinted gene in Arabidopsis thaliana expressed specifically in the female gametophyte and endosperm. Tissue-specific and imprinted expression of FWA depends on DNA methylation in the FWA promoter, which is comprised of two direct repeats containing a sequence related to a SINE retroelement. Methylation of this element causes epigenetic silencing, but it is not known whether the methylation is targeted to the SINE-related sequence itself or the direct repeat structure is also necessary. Here we show that the repeat structure in the FWA promoter is highly diverse in species within the genus Arabidopsis. Four independent tandem repeat formation events were found in three closely related species. Another related species, A. halleri, did not have a tandem repeat in the FWA promoter. Unexpectedly, even in this species, FWA expression was imprinted and the FWA promoter was methylated. In addition, our expression analysis of FWA gene in vegetative tissues revealed high frequency of intra-specific variation in the expression level. In conclusion, we show that the tandem repeat structure is dispensable for the epigenetic silencing of the FWA gene. Rather, SINE-related sequence is sufficient for imprinting, vegetative silencing, and targeting of DNA methylation. Frequent independent tandem repeat formation events in the FWA promoter led us to propose that they may be a consequence, rather than cause, of the epigenetic control. The possible significance of epigenetic variation in reproductive strategies during evolution is also discussed.

    DOI： 10.1371/journal.pgen.1000048

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  • Control of FWA gene silencing in Arabidopsis thaliana by SINE-related direct repeats Reviewed

    Yuki Kinoshita, Hidetoshi Saze, Tetsu Kinoshita, Asuka Miura, Wim J.J. Soppe, Maarten Koornneef, Tetsuji Kakutani

    Plant Journal   49 ( 1 )   38 - 45   2007.1

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    DOI： 10.1111/j.1365-313X.2006.02936.x

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  • Reproductive barrier and genomic imprinting in the endosperm of flowering plants Reviewed

    Tetsu Kinoshita

    Genes and Genetic Systems   82 ( 3 )   177 - 186   2007

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  • Maintenance of DNA methylation during the arabidopsis life cycle is essential for parental imprinting Reviewed

    Pauline E. Jullien, Tetsu Kinoshita, Nir Ohad, Frédéric Berger

    Plant Cell   18 ( 6 )   1360 - 1372   2006.6

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    DOI： 10.1105/tpc.106.041178

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  • One-Way Control of FWA Imprinting in Arabidopsis Endosperm by DNA Methylation Reviewed

    Tetsu Kinoshita, Asuka Miura, Yeonhee Choi, Yuki Kinoshita, Xiaofeng Cao, Steven E. Jacobsen, Robert L. Fischer, Tetsuji Kakutani

    Science   303 ( 5657 )   521 - 523   2004.1

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    DOI： 10.1126/science.1089835

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  • Control of development and transposon movement by DNA methylation in Arabidopsis thaliana Reviewed

    T. Kakutani, M. Kato, T. Kinoshita, A. Miura

    Cold Spring Harbor Symposia on Quantitative Biology   69   139 - 143   2004

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    DOI： 10.1101/sqb.2004.69.139

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  • Polycomb repression of flowering during early plant development Reviewed

    Tetsu Kinoshita, John J. Harada, Robert B. Goldberg, Robert L. Fischer

    Proceedings of the National Academy of Sciences of the United States of America   98 ( 24 )   14156 - 14161   2001.11

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    DOI： 10.1073/pnas.241507798

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  • Mutations in the FIE and MEA genes that encode interacting polycomb proteins cause parent-of-origin effects on seed development by distinct mechanisms Reviewed

    R. Yadegari, T. Kinoshita, O. Lotan, G. Cohen, A. Katz, Y. Choi, A. Katz, K. Nakashima, J. J. Harada, R. B. Goldberg, R. L. Fischer, N. Ohad

    Plant Cell   12 ( 12 )   2367 - 2381   2000

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    DOI： 10.1105/tpc.12.12.2367

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  • Imprinting of the MEDEA polycomb gene in the Arabidopsis endosperm Reviewed

    Tetsu Kinoshita, Ramin Yadegari, John J. Harada, Robert B. Goldberg, Robert L. Fischer

    Plant Cell   11 ( 10 )   1945 - 1952   1999.10

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    DOI： 10.1105/tpc.11.10.1945

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  • Vacuolar processing enzyme is up-regulated in the lytic vacuoles of vegetative tissues during senescence and under various stressed conditions.

    Tetsu Kinoshita, Tetsu Kinoshita, Kenji Yamada, Kenji Yamada, Nagako Hiraiwa, Maki Kondo, Mikio Nishimura, Mikio Nishimura, Ikuko Hara-Nishimura, Ikuko Hara-Nishimura

    Plant J.   19   43 - 53   1999.7

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  • The sequence and expression of the γ-VPE gene, one member of a family of three genes for vacuolar processing enzymes in arabidopsis thaliana

    Tetsu Kinoshita, Tetsu Kinoshita, Mikio Nishimura, Mikio Nishimura, Ikuko Hara-Nishimura, Ikuko Hara-Nishimura

    Plant and Cell Physiology   36 ( 8 )   1555 - 1562   1995.12

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  • Homologues of a vacuolar processing enzyme that are expressed in different organs in Arabidopsis thaliana

    Tetsu Kinoshita, Mikio Nishimura, Ikuko Hara-Nishimura

    Plant Molecular Biology   29 ( 1 )   81 - 89   1995.10

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    DOI： 10.1007/BF00019120

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  • Histological analysis of amyloid formation in EC1 peptides, sperm-cell-activating factors expressed in egg cell

    御調日向子, 永原史織, 後藤友美, 大井崇生, 武内秀憲, 武内秀憲, 豊岡公徳, 木下哲, 須崎大地, 丸山大輔

    日本植物生理学会年会(Web)   65th   2024

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  • How are patch-like extracellular structures between the egg cell and the central cell formed?

    須崎大地, 大井崇生, 御調日向子, 武内秀憲, 武内秀憲, 永原史織, 杉直也, 木下哲, 丸山大輔

    日本植物生理学会年会(Web)   65th   2024

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  • 精細胞を覆う生体膜peri-germ cell membraneの選択的崩壊機構の解析

    杉直也, 須崎大地, 海老根一生, 海老根一生, 武内秀憲, 武内秀憲, 永原史織, 木下哲, WIDIEZ Thomas, 丸山大輔

    日本分子生物学会年会プログラム・要旨集(Web)   47th   2024

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  • 卵細胞外の斑点状構造はどのように形成されるか

    須崎大地, 大井崇生, 武内秀憲, 武内秀憲, 永原史織, 御調日向子, 杉直也, 木下哲, 丸山大輔

    日本植物学会大会研究発表記録(CD-ROM)   87th   2023

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  • Silencing mechanism by canonical and non-canonical RdDM pathway in egg cell

    八代惇, 鎌田千裕, 長谷川綾子, 金子澪, 須崎大地, 杉直也, 殿崎薫, 丸山大輔, 木下哲

    育種学研究   24   2022

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  • 精細胞輸送機構に欠損を示すシロイヌナズナ変異体の選抜

    池内麻妃, 杉直也, 土春菜, 須崎大地, 木下哲, 丸山大輔

    日本植物学会大会研究発表記録(CD-ROM)   86th   2022

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  • 卵細胞が制御する重複受精機構の解析

    須崎大地, 大井崇生, 武内秀憲, 武内秀憲, 永原史織, 杉直也, 木下哲, 丸山大輔

    日本植物学会大会研究発表記録(CD-ROM)   86th   2022

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  • Morphology, dynamics, and function of unique membrane surrounding sperm plasma membrane

    丸山大輔, 泉理恵, 武内秀憲, 東山哲也, 東山哲也, 木下哲, 元村一基

    日本植物生理学会年会(Web)   61st   2020

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  • シロイヌナズナ花粉管における生体膜の頑健性を修復系の解析

    西牧皐樹, 柳沢直樹, 時田(松浦)公美, 木下哲, 東山哲也, 東山哲也, 丸山大輔

    日本植物学会大会研究発表記録   83rd   2019

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  • 雌性配偶体による精細胞の放出方向制御の解析

    須崎 大地, 大井 崇生, 友実 駿, 榎本 早希子, 荒井 重勇, 木下 哲, 丸山 大輔

    日本植物学会第82回大会   2018.9

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  • 雌性配偶体の形態解析から精細胞の放出方向制御に迫る

    須崎 大地, 大井 崇生, 友実 駿, 榎本 早希子, 荒井 重勇, 木下 哲, 丸山 大輔

    日本植物形態学会第30回大会   2018.9

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  • How does female gametophyte regulate the destination of sperm cells released from pollen tube?

    Daichi Susaki, Takao Oi, Sakiko Enomoto, Shigeo Arai, Tetsu Kinoshita, Daisuke Maruyama

    25th ICSPR   2018.6

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  • Reception of pollen tube contents by the endosperm

    Daisuke Maruyama, Daichi Susaki, Tetsuya Higashiyama, Tetsu Kinoshita

    25th ICSPR   2018.6

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  • Synergid cell controls the destination of sperm cell discharge.

    須崎大地, 東山哲也, 木下哲, 丸山大輔

    Taiwan-Japan Plant Biology 2017   2017.11

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  • 助細胞は精細胞の放出先の位置を制御する微小流路として機能する

    須崎大地, 東山哲也, 東山哲也, 木下哲, 丸山大輔

    日本植物学会大会研究発表記録   81st   177   2017.9

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  • 伸長する花粉管における生体膜の頑健性の解析

    西牧皐樹, 柳沢直樹, 時田公美, 木下哲, 東山哲也, 丸山大輔

    日本植物学会大会研究発表記録   81st   225   2017.9

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  • シロイヌナズナ卵細胞被覆構造の解析

    丸山大輔, 大井崇生, 須崎大地, 東山哲也, 木下哲

    日本植物学会大会研究発表記録   81st   175   2017.9

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  • ヘテロ受精を利用した異種交配時の胚乳発達異常の回避

    前田愛, 木下哲, 丸山大輔

    日本植物学会大会研究発表記録   81st   246   2017.9

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  • Research on post-fertilization barriers in the context of plant breeding Invited

    19 ( 1 )   35 - 40   2017

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    Language：Japanese   Publishing type：Meeting report  

    DOI： 10.1270/jsbbr.19.35

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  • 次世代シーケンス解析に向けた高純度のイネ葉緑体DNA精製法

    高松壮, 大西孝幸, 猪俣拓也, 及川和聡, 木下哲, 三ツ井敏明

    日本農芸化学会大会講演要旨集(Web)   2016   2016

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  • 倍数性操作によるイネ胚乳の生殖的隔離の打破

    TONOSAKI KAORU, SEKINE DAISUKE, ONISHI TAKAYUKI, FURUUMI HIROYASU, KURATA NORI, KINOSHITA TETSU

    育種学研究   17   117   2015.3

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  • Mechanism of DNA demethylation in Arabidopsis thaliana

    Tetsu Kinoshita

    GENES & GENETIC SYSTEMS   89 ( 6 )   278 - 278   2014.12

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  • イネのバイオトロン栽培法の紹介

    ONISHI TAKAYUKI, YOSHINO MIHOKO, YAMAKAWA HIROMOTO, KINOSHITA TETSU

    応用糖質科学   4 ( 3 )   215 - 218   2014.8

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    Language：Japanese   Publisher：The Japanese Society of Applied Glycoscience  

    DOI： 10.5458/bag.4.3_215

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  • イネの質的形質遺伝子の単離と機能解明 第1章 おもな個別研究課題の成果 3 イネ種子の登熟や高温下での品質に係わる主要な質的形質遺伝子に関する研究(1)イネ胚乳の初期発生とゲノムインプリンティングの解析(IPG0017)

    KINOSHITA TETSU

    農林水産省農林水産技術会議事務局研究成果   ( 511 )   184 - 186   2014.3

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  • Induction of epigenetic modifications by RNA-directed DNA methylation (RdDM) and their inheritance to the next generation in plants(<Feature Articles>New gene modification techniques and useful plant production)

    Kinoshita Tetsu, Ono Akemi

    Regulation of Plant Growth & Development   48 ( 2 )   142 - 147   2013.12

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    Recent advances in the technologies for developing genetically modified crops have highlighted the importance of epigenetic controls in plants. Several studies have suggested that double-stranded RNA (dsRNA) can induce sequence-specific DNA methylation and silence expression of target genes. Moreover, DNA methylation-mediated gene silencing can be stably transmitted to the next generation without dsRNA trigger constructs. By exploring and elucidating the molecular mechanisms behind these phenomena, we have achieved a greater understanding of epigenetic molecular mechanisms and this knowledge has provided the foundation for new techniques, the so-called New Plant Breeding Techniques (NBT). Further advances in our understanding of the molecular mechanisms of RNA-directed DNA methylation (RdDM) will enable improvements to these new techniques.

    DOI： 10.18978/jscrp.48.2_142

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  • Rapid establishment of introgression lines using cytoplasmic male sterility and a restorer gene in Oryza sativa cv. Nipponbare

    Takayuki Ohnishi, Mihoko Yoshino, Kinya Toriyama, Tetsu Kinoshita

    MOLECULAR BREEDING   32 ( 4 )   831 - 839   2013.12

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  • FS-1 イネのバイオトロン栽培法の開発(第2回応用糖質フレッシュシンポジウム,日本応用糖質科学会平成25年度大会(第62回))

    大西 孝幸, 吉野 みほ子, 山川 博幹, 木下 哲

    応用糖質科学 : 日本応用糖質科学会誌   3 ( 3 )   B55   2013.8

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  • イネのバイオトロン栽培法の開発

    ONISHI TAKAYUKI, YOSHINO MIHOKO, YAMAKAWA HIROMOTO, KINOSHITA TETSU

    応用糖質科学   3 ( 3 )   55   2013.8

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  • バイオトロンブリーディング法:イネを1世代当たり2ケ月で育てる屋内栽培系

    ONISHI TAKAYUKI, YOSHINO MIHOKO, YAMAKAWA HIROMOTO, KINOSHITA TETSU

    日本植物生理学会年会要旨集   54th   111   2013.3

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  • Implications of parental genome imbalance for plant endosperm development

    Tetsu Kinoshita

    GENES & GENETIC SYSTEMS   87 ( 6 )   395 - 395   2012.12

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  • DNA de-methylation in the Arabidopsis thaliana female gametophyte: is DEMETER alone?

    Diana Mihaela Buzas, Yukiko Sugimoto, Yuki Kinoshita, Tetsu Kinoshita

    GENES & GENETIC SYSTEMS   87 ( 6 )   422 - 422   2012.12

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  • Control of genomic imprinting by a competent of the FACT histone chaperone

    Yoko Ikeda, Yuki Kinoshita, Daichi Susaki, Yuriko Ikeda, Megumi Iwano, SeijiTakayama, Tetsuya Higashiyama, Tetsuji Kakutani, Tetsu Kinoshita

    10th International Congress on Plant Molecular Biology   2012.10

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  • イネの胚乳発生・サイズを制御する父・母ゲノムの効果

    SEKINE DAISUKE, ONISHI TAKAYUKI, FURUUMI HIROYASU, KURATA NORI, KINOSHITA TETSU

    育種学研究   14   95   2012.3

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  • イネの倍数体間交雑における胚乳発生異常の解析

    SEKINE DAISUKE, ONISHI TAKAYUKI, FURUUMI HIROYASU, YOSHINO MIHOKO, KURATA NORI, KINOSHITA TETSU

    日本植物生理学会年会要旨集   53rd   182   2012.3

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  • 植物の生殖過程におけるリプログラミングとセクシャルコンフリクト

    池田陽子, 木下哲

    日本分子生物学会年会プログラム・要旨集(Web)   35th   1W1I-3 (WEB ONLY)   2012

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  • The biotron breeding system: a rapid and reliable procedure for genetic studies and breeding in rice

    Takayuki Ohnishi, Hiromoto Yamakawa, Tetsu Kinoshita

    GENES & GENETIC SYSTEMS   86 ( 6 )   399 - 399   2011.12

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  • HMGドメイン遺伝子SSRP1はシロイヌナズナにおけるDNA脱メチル化およびゲノムインプリンティングに必要である

    池田陽子, 木下哲

    細胞工学   30 ( 12 )   1296 - 1297   2011.11

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  • エピジェネティクス

    中村 みゆき, 木下 哲

    種生物学研究   34   109 - 122   2011.7

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  • ゲノムに刷り込まれた生殖隔離機構

    木下 哲

    種生物学研究   34   141 - 155   2011.7

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  • ヒストンシャペロン構成因子ALAC1のDNA脱メチル化及びゲノムインプリンティングへの関与

    池田陽子, 木下由紀, 池田有理子, 角谷徹仁, 木下哲

    日本植物生理学会年会要旨集   52nd   170   2011.3

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  • Interspecific hybridization barrier in the endosperm of flowering plants: what is the rule and underlying mechanism?

    Tetsu Kinoshita, Takayuki Ohnishi, Ryo Ishikawa, Yuki Kinoshita, Mitsugu Eiguchi, Nori Kurata

    GENES & GENETIC SYSTEMS   85 ( 6 )   449 - 449   2010.12

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  • マイクロアレイを用いた新規インプリント遺伝子の探索

    池田陽子, 木下由紀, 池田有理子, 木下哲

    日本植物学会大会研究発表記録   73rd   115   2009.9

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  • ALARM CLOCK1はDNA脱メチル化を介してゲノムインプリンティングの確立に関与する

    池田陽子, 木下由紀, 角谷徹仁, 木下哲

    日本植物生理学会年会要旨集   50th   217   2009.3

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  • Control of genomic imprinting by ALARM CLOCK1 gene in Arabidopsis

    Yoko Ikeda, Yuki Kinoshita, Tetsuji Kakutani, Tetsu Kinoshita

    GENES & GENETIC SYSTEMS   83 ( 6 )   534 - 534   2008.12

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  • ゲノムインプリンティングの制御に関わるALARM CLOCK FOR FWA IMPRINTING 1の解析

    池田陽子, 木下由紀, 角谷徹仁, 木下哲

    日本植物学会大会研究発表記録   72nd   244   2008.9

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  • シロイヌナズナのゲノムインプリンティング制御に関与する変異体の解析

    池田陽子, 木下由紀, 角谷徹仁, 木下哲

    日本遺伝学会大会プログラム・予稿集   80th   63   2008.8

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  • Control of plant life cycle by polycomb complex in Arabidopsis(<Feature Articles>Regulation of plant development and differentiation by epigenetics)

    Kinoshita Tetsu, Ikeda Yoko, Ishikawa Ryo

    Regulation of Plant Growth & Development   43 ( 1 )   29 - 34   2008.5

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    DOI： 10.18978/jscrp.43.1_29

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  • イネの胚乳における生殖隔離機構とゲノムインプリンティング

    石川亮, 永口貢, 池田陽子, 倉田のり, 木下哲

    日本植物生理学会年会要旨集   49th   225 - 0488   2008.3

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    Defect of the endosperm development is observed in inter-specific crosses of many plant species. Abnormal endosperm development causes seed abortion, therefore, this phenomenon is known to be a reproductive barrier after fertilization. Several reports suggested that defect of endosperm development is caused by the functional difference between parental genome. One of the possible molecular mechanisms underlying this phenomenon can be genomic imprinting, an epigenetic mechanism resulting in mono-allelic gene expression by parent-of-origin dependent manner. Recent advances of the study of genomic imprinting in <I>Arabidopsis</I> are applied to understand the reproductive barrier of hybrid endosperm in rice.<br>To begin with, we analyzed how imprinted genes are involved in the development of hybrid endosperm. We conducted inter-specific cross between cultivated and wild rice and observed developmental defect in the hybrid endosperm. We also investigated the expression of imprinted gene. Possible involvement of genomic imprinting in the hybrid endosperm will be presented.

    DOI： 10.14841/jspp.2008.0.0488.0

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  • ゲノムインプリンティング制御因子の単離と機能解析

    池田陽子, 木下由紀, 角谷徹仁, 木下哲

    日本植物生理学会年会要旨集   49th   256   2008.3

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  • Evolution of imprinted FWA gene in Arabidopsis

    Fujimoto Ryo, Kawabe Akira, Kinoshita Yuki, Takashima Kazuya, Kinoshita Tetsu, Miura Asuka, Kakutani Tetsuji

    GENES & GENETIC SYSTEMS   82 ( 6 )   549 - 549   2007.12

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  • Young Investigator Award - Study on genomic imprinting in flowering plant

    Tetsu Kinoshita

    PLANT AND CELL PHYSIOLOGY   48   S16 - S16   2007

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  • A change in the CACTA transposon mobility in Arabidopsis thaliana.

    Miyuki Nakamura, Asuka Miura, Yuki Kinoshita, Tetsu Kinoshita, Masaomi Kato, Tetsuji Kakutani

    PLANT AND CELL PHYSIOLOGY   48   S90 - S90   2007

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  • Mechanism of FWA imprinting in Arabidopsis endosperm

    T Kinoshita, A Miura, Y Kinoshita, T Kakutani

    PLANT AND CELL PHYSIOLOGY   45   S72 - S72   2004

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  • Parent-of-origin-dependent FWA gene expression in the endosperm of Arabidopsis

    T Kinoshita, A Miura, Y Kinoshita, T Kakutani

    PLANT AND CELL PHYSIOLOGY   44   S28 - S28   2003

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  • Epigenetic regulation of a homeodomain gene FWA

    T Kinoshita, A Miura, T Kakutani

    PLANT AND CELL PHYSIOLOGY   43   S132 - S132   2002

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  • Isolation and Characterization of Mouse VPE, a Novel Lysosomal Cysteine Protease.

    SHIRAHAMA Kanae, KINOSHITA Tetsu, HARA-NISHIMURA Ikuko, NISHIMURA Mikio

    21   577 - 577   1998.12

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  • シロイヌナズナにおける液胞プロセシング酵素発現細胞の微細構造

    山本 陽子, 森田 淳子, 木下 哲, 西村 いくこ, 西村 幹夫, 野口 哲子

    日本植物学会大会研究発表記録 = Proceedings of the annual meeting of the Botanical Society of Japan   62   179 - 179   1998.9

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  • γVPE mRNA Level Is Increased by Plant Hormones, Wounding and Senescence

    YAMADA Kenji, KINOSHITA Tetsu, HARA-NISHIMURA Ikuko, NISHIMURA Mikio

    39   S138 - S138   1998.5

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  • Physiological Functions of Vacuoles in Hydathodal Cells of Arabidopsis thaliana

    KINOSHITA Tetsu, NOGUCHI Tetsuko, HARA-NISHIMURA Ikuko, NISHIMURA Mikio

    39   S54 - S54   1998.5

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  • 液胞プロセシング酵素の発現からみたシロイヌナズナ排水組織の構造と機能

    木下 哲, 西村 いくこ, 西村 幹夫

    日本植物学会大会研究発表記録 = Proceedings of the annual meeting of the Botanical Society of Japan   61   151 - 151   1997.9

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  • 花粉成熟過程における液胞プロセシング酵素の発現と液胞の機能

    木下 哲, 西村 いくこ, 西村 幹夫

    日本植物学会大会研究発表記録 = Proceedings of the annual meeting of the Botanical Society of Japan   60   198 - 198   1996.10

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  • TEMPORAL AND SPATIAL EXPRESSION OF THREE GENES ENCODING VACUOLAR PROCESSING ENZYME IN Arabidopsis thaliana

    KINOSHITA Tetsu, HARA-NISHIMURA Ikuko, NISHIMURA Mikio

    37   99 - 99   1996.3

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Research Projects

  • Genomic dynamics underlying the plastic hermaphroditism in plants: the basis of exploratory reproductive adaptations

    Grant number：22H05172  2022.6 - 2027.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Transformative Research Areas (A)  Grant-in-Aid for Transformative Research Areas (A)

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    Grant amount：\317590000 （ Direct Cost: \244300000 、 Indirect Cost：\73290000 ）

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  • 自殖と他殖を規定する胚乳のエピゲノム制御

    Grant number：22H05175  2022.6 - 2027.3

    日本学術振興会  科学研究費助成事業 学術変革領域研究(A)  学術変革領域研究(A)

    木下 哲, 遠藤 真咲

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    Grant amount：\103350000 （ Direct Cost: \79500000 、 Indirect Cost：\23850000 ）

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  • サカナ（胎生発生真骨魚類）にゲノムインプリンティングは存在するか？

    Grant number：21K19237  2021.7 - 2023.3

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)  挑戦的研究(萌芽)

    木下 哲, 飯田 敦夫

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    Grant amount：\6500000 （ Direct Cost: \5000000 、 Indirect Cost：\1500000 ）

    胎生発生する生物を対象とした興味深い学説として、『コンフリクト仮説』がある。母体内での胎仔の成長に関して、オス親の利益が自分の子供を大きくすることであるのに対し、メス親の利益は、栄養資源を節約し自分の子供を小さくすることであると説明される。本仮説では、この利害の対立から、オス親とメス親から受け継いだ対立遺伝子が、その子供で異なる発現を示すことが想定されている。これらの遺伝子は『インプリント遺伝子』と定義され、オス・メスの利害のせめぎ合いは『コンフリクト仮説』として説明される。この仮説は、胎盤を持つ哺乳動物と胚乳を持つ被子植物で検証されているが、それ以外の胎生生物での検証は進んでいない。令和３年度は、胎生魚類にインプリント遺伝子が存在することで仮説の検証に取り組むため、ハイランドカープからRNA-seq解析を進めること、SNPsが存在するであろう２系統の交配準備を進めた。予備的には、胎仔への栄養供給を担っていると考えられる栄養リボンで発現する遺伝子セットを同定している。この中にインプリント遺伝子が複数含まれると予想される。

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  • イネ胚乳の生殖的隔離における「綱引き」モデルの分子実態解明

    Grant number：23K21188  2021.4 - 2025.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    木下 哲

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    Grant amount：\17290000 （ Direct Cost: \13300000 、 Indirect Cost：\3990000 ）

    胚乳における生殖的隔離や最終的な胚乳の大きさは、エピジェネティックな制御によりその初期発生期に決定されることが理解されつつある。穀類胚乳の生殖的隔離では、オス・メスゲノムの「綱引き」とも表現される仕組みが古くから想定されているが、その分子実態は未だに明らかではない。イネを用いた解析により、「綱引き」の候補因子として、ポリコーム複合体構成因子のOsEMF2aと、その直接の標的でありH3K27me3のヒストン修飾を受けるOsMADS77を含む幾つかのType-I MADS-box遺伝子群が浮上している。両遺伝子ともにそれぞれメス由来ゲノム、オス由来ゲノム特異的に発現するインプリント遺伝子であり、まさに「綱引き」を担いうる遺伝子である。本研究計画では、ゲノム編集による機能解析ならびにエピゲノム解析により、「綱引き」の分子カスケードの全容を明らかにし、古くから提唱されている胚乳における生殖的隔離の分子実態を解明する。

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  • イネ胚乳の生殖的隔離における「綱引き」モデルの分子実態解明

    Grant number：21H02170  2021.4 - 2025.3

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    木下 哲

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    Grant amount：\17290000 （ Direct Cost: \13300000 、 Indirect Cost：\3990000 ）

    胚乳における生殖的隔離や最終的な胚乳の大きさは、エピジェネティックな制御によりその初期発生期に決定されることが理解されつつある。穀類胚乳の生殖的隔離では、オス・メスゲノムの「綱引き」とも表現される仕組みが古くから想定されているが、その分子実態は未だに明らかではない。イネを用いた申請者らの解析により、「綱引き」の候補因子として、ポリコーム複合体構成因子のOsEMF2aと、その直接の標的でありH3K27me3のヒストン修飾を受けるOsMADS77を含む幾つかのType-I MADS-box遺伝子群が浮上している。興味深いことに、両遺伝子ともにそれぞれメス由来ゲノム、オス由来ゲノム特異的に発現するインプリント遺伝子であり、まさに「綱引き」を担いうる遺伝子である。令和3年度は、既に作成していたOsEMF2a変異体を用いた詳細な表現型解析、ならびにその標的と考えられるMADS-box変異体の変異体胚乳における詳細な表現型解析を行った。MADSの変異体に関しては、現在のところ単独変異や二重変異体においては明確な表現型を得られておらず、さらなる多重変異体の作出と表現型解析を行っている。OsEMF2a変異体を用いて、ChIP-seqの解析を行い、直接の標的遺伝子群に関してその候補を明らかにしているが、さらにオスメスの対立遺伝子を区別するため、日本晴とキタアケを交配しChIP-seqを行う準備を進めた。

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  • Determining principles in the birth of new plant species: elucidation of lock-and-key molecular systems in sexual reproduction

    Grant number：21H00458  2021.4 - 2022.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

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    Grant amount：\3900000 （ Direct Cost: \3000000 、 Indirect Cost：\900000 ）

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  • Elucidation of the molecular mechanisms of apomixis in plants

    Grant number：17H06256  2017.6 - 2020.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Challenging Research (Pioneering)  Challenging Research (Pioneering)

    TAKAGI MASARU

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    Grant amount：\26000000 （ Direct Cost: \20000000 、 Indirect Cost：\6000000 ）

    Using the newly developed gene silencing technology (CRES-T), we isolated chimeric repressors that frequently induces adventitious embryos for the purpose of artificially inducing the apomixis phenomenon. As a result of expression in the embryonic endothelium, which is a maternal 2n tissue, we succeeded in isolating a transformant having two torpedo-type embryos, which are considered to be a fertilized embryo and an apomixis embryo, in one ovule at the same time. We also found a chimeric repressor for five transcription factors named ESP, which enlarges the endosperm without fertilization. It was revealed that rice plants expressing a chimeric repressor for these rice orthologs could be produced endosperm formation without fertilization.

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  • ミトコンドリアに由来するエピジェネティック経路の解明と育種利用

    Grant number：17H03749  2017.4 - 2021.3

    日本学術振興会  科学研究費助成事業 基盤研究(B)  基盤研究(B)

    木下 哲

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    Grant amount：\18200000 （ Direct Cost: \14000000 、 Indirect Cost：\4200000 ）

    育種においては、母本に用いる細胞質の由来に応じて、開花時期や耐病性などの様々な育種形質に影響が現れることが知られている。この現象には、核とオルガネラゲノムの相互作用、エピジェネティックなコミュニケーションが想定されているが、その実体は殆ど明らかではない。我々は、DNA脱メチル化制御に関与する細胞質の鉄・硫黄クラスター生合成経路をすでに明らかにしている。興味深いことに、この経路はミトコンドリア内膜のトランスポーターの機能に依存している。本研究計画では、イネをモデルにミトコンドリアに由来するエピジェネティック経路の詳細と育種形質への影響を明らかにし、将来の育種基盤技術創出に資することを目的としている。

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  • 植物新種誕生の原理 -国際的研究中心形成に向けた国際活動支援センター-

    Grant number：16K21727  2016.6 - 2021.3

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    東山 哲也, 木下 哲, 瀬々 潤, 渡辺 正夫, 辻 寛之

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    Grant amount：\58370000 （ Direct Cost: \44900000 、 Indirect Cost：\13470000 ）

    国際活動支援班では、国内外の人的交流による国際共同研究、ブレインストーミング、ミーティングを実施することを通して、「植物新種誕生原理」のために国外研究者が結集・交流できる国際的研究拠点を我が国に構築することを目的とする。特に領域代表が強力にリーダーシップをとり、本領域が推進する3つのブレークスルーテクノロジー、すなわちライブセルイメージング、構造生物学、生命科学と有機合成化学の融合における優位性を通じて生殖研究を国際的にリードするための活動を実施した。本年度は欧州・北米海外拠点への若手の派遣に加え、特に、領域代表の東山がプレジデントを務め、計画研究の木下が大会長を務める植物生殖科学の最大の国際学会を運営した。国際学会の前後には名古屋大と横浜市大の2拠点でそれぞれ生殖科学とエピジェネティクスにフォーカスしサテライトワークショップを開催した。世界的にも顕著な業績を上げた有力研究者を我が国に多数招へいして本領域の研究者と交流し、国際共同研究をより活性化するための体制を強固にすると同時に、本領域の研究成果を積極的に発信することで国際的なプレゼンスを向上させた。
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    国際植物生殖会議：領域代表の東山が学会長を、また計画班の木下が大会長を務めるThe 25th International Congress on Sexual Plant Reproductionにて、本領域が共催した。参加者は目標の300人を大きく超え335名で、そのうち海外からの参加者は236名（約30か国）と参加者全体の70%を越えた。本領域が、東山が招待講演を行ったセッションを共催セッションとするなど、本領域を大きくアピールできた。

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  • 植物新種誕生の原理 -生殖過程の鍵と鍵穴の分子実態解明を通じて-

    Grant number：16H06464  2016.6 - 2021.3

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    東山 哲也, 瀬々 潤, 辻 寛之, 木下 哲, 高山 誠司, 渡辺 正夫

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    Grant amount：\305240000 （ Direct Cost: \234800000 、 Indirect Cost：\70440000 ）

    総括班を、研究代表者、研究分担者、連携研究者の14 名によって構成した。また、異分野融合研究支援センターが中心となって、領域内の異分野融合研究を推進した。本センターは、ブレークスルーテクノロジーをはじめ特定の研究者のみ実施可能な解析手法を領域内で簡単に共有する枠組みをつくり、共同研究を強力に推進することを目的とした。
    ・領域運営：領域代表・東山を中心に円滑な領域運営を行い、公募研究を加えた領域全体が一体となって研究を進めた。中間評価のあった本年度は、後半の更なる発展につながるよう、計画の遂行と、想定を超えた展開を目指した。これまでに構築してきた領域全体の研究方針、共同研究の支援の枠組み、具体的な共同研究体制を活用して領域の研究を推進した。このために、事務局長・辻の運営によって領域会議（班会議）、総括班会議、若手の会を開催した。若手の会については、際立った盛り上がりを見せる学生たちの主体性が発揮できるよう心掛けた。
    ・共同研究：計画研究班が進めてきた研究を基盤に、公募研究も交えて、領域内での様々な異分野融合共同研究を推進した。領域会議、若手の会、各拠点が中心となって行ってきた異分野融合研究ミーティングなどを通して構築したネットワークを通じて、共同研究の推進を促した。さらに、リサーチコーディネーターが積極的に関わることで、一層の共同研究をはかった。
    ・情報発信・国民への還元：領域ウェブサイトを活用し、論文発表などの研究成果を速やかに発信した。計画通りに本年度日本で開催する国際植物生殖会議2018を通して、世界に向けて本領域およびその研究を発信した。また、計画研究班員および公募研究班員が中心となり、アウトリーチ活動(小中高での出前講義など)を積極的に展開し、植物科学の研究成果を国民へ還元した。アウトリーチ活動についても実施内容・評価等をホームページで公開した。

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  • 胚乳における種の障壁：エピゲノム制御の鍵分子機構

    Grant number：16H06471  2016.6 - 2021.3

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    木下 哲, 赤木 剛士

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    Grant amount：\115960000 （ Direct Cost: \89200000 、 Indirect Cost：\26760000 ）

    植物の生殖過程では、「他の植物種と交雑することなく自らのゲノムの維持するシステム」が生殖障壁として機能している。このシステムは、受精に至る多段階に配置される「鍵と鍵穴」の分子認証の総体として捉えることができる。興味深いことに、受精前の分子認証を突破した場合でも、受精後の胚乳において、極めて鋭敏な異種ゲノムを感知する分子認証システムが配置されている。ここでは、両親ゲノムの塩基配列レベルの違いのみならず、可塑性の高いエピジェネティックな制御機構が重要な位置をしめていることを、我々は科研費を中心とした研究費支援により一連の分子機構を明らかにしてきた。本研究計画では、植物新種誕生原理の解明のため、（１）受精後の胚乳の生殖障壁をエピジェネティックに誘導するDNA脱メチル化の役割と（２）受精後の胚乳が自らを崩壊させることにより異種ゲノムを排除する機構を明らかにすることを目的とする。

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  • Search for paternally expressed imprinted genes regulating endosperm development

    Grant number：16K14903  2016.4 - 2018.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research  Grant-in-Aid for Challenging Exploratory Research

    TAKAYAMA Seiji, WADA Yuko, TSUCHIMATSU Takashi, KINOSHITA Tetsu, ITO Toshiro

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    Grant amount：\3770000 （ Direct Cost: \2900000 、 Indirect Cost：\870000 ）

    This project aims to identify paternally expressed imprinted genes that regulate endosperm development. We searched for these genes through the following strategies.
    1. Search based on the conflict between maternal and paternal genomes: When Arabidopsis thaliana Col-0 strain was crossed with other natural A. thaliana strains, the obtained F1 seeds exhibited high phenotypic variation in size. A genome-wide association study of seed size identified some associated loci, in which some paternally expressed genes were located.
    2. Search based on the gene expression dependent on DNA methylation: The expression of target genes is suggested to be dependent on DNA methylation. The comparative pollen transcriptome analysis of wild type and a DNA methylation mutant identified several candidates genes. Furthermore, the analysis of F1 hybrids between wild type and epiRIL liens identified some loci that affect seed size dependent on DNA methylation.

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  • Development studies on high temperature and high CO2 tolerant rice during seed ripening stage

    Grant number：15H02486  2015.4 - 2019.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)  Grant-in-Aid for Scientific Research (A)

    MITSUI Toshiaki

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    Grant amount：\41600000 （ Direct Cost: \32000000 、 Indirect Cost：\9600000 ）

    In this study, we performed proteome and starch glycome analyses of high temperature ripened brown rice, and concluded that the chalking of grain was caused by the imbalance between starch synthesis and degradation. In addition, when the effects of strong light ＆ high CO2 concentration and high temperature＆ high CO2 concentration on appearance quality of rice grains were examined, the sensitivity was shown to be high in the early stage of the ripening period from flowering. However, it was found that high CO2 promotes the effects of high temperature stress, although remarkable chalkiness of brown rice does not occur only under high CO2 conditions. Mn-type superoxide dismutase (MSD1) was identified as a key enzyme involved in high temperature tolerance of rice, and it was demonstrated that high temperature ripening tolerance is improved by the constitutively high expression of MSD1 gene, while the suppression of MSD1 gene markedly enhanced the high temperature susceptibility.

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  • Study on Fe-S cluster assembly genes in Arabidopsis thaliana

    Grant number：26291063  2014.4 - 2017.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    Kinoshita Tetsu

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    Authorship：Principal investigator  Grant type：Competitive

    Grant amount：\14560000 （ Direct Cost: \11200000 、 Indirect Cost：\3360000 ）

    DNA demethylation plays important roles for many aspects of plant development and gene expression control related to environmental adaptation. In this study, we identified the cytosolic iron-sulfur cluster assembly (CIA) genes that are required for activation and DNA demethylation of the maternally expressed imprinted genes in the central cell of the femalegametophyte in A. thaliana. Our detailed analysis using one of the mutation in the CIA components revealed that this pathway also controls genes for plant immunity and stress tolerance in vegetative tissues.

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  • Mechanisms of hybridization barriers in rice endosperm

    Grant number：23113003  2011.4 - 2016.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

    Kinoshita tetsu

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    Authorship：Principal investigator  Grant type：Competitive

    Grant amount：\138450000 （ Direct Cost: \106500000 、 Indirect Cost：\31950000 ）

    In plants, the endosperm is a site for hybridization barrier, where maternally- and paternally derived genomes have distinct roles for their development.
    Classical studies predicted that the maternal genome represses endosperm growth and the paternal genome promote it. In this study, we conducted detailed analyses of the hybrid endosperms in interspecific or interploidy cross of rice or combination of those (Plant J. 2011, 2013, and in preparation). These analysis revealed that developmental transition under the control of Polycomb Repressive Complex 2 (PRC2) is a key determinant for this phenomena. We also identified several components for DNA demethylation that is required for activation of imprinted genes in Arabidopsis thaliana (Dev Cell 2011, New Phytologist 2013, PNAS 2014).

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  • Correlative gene system: Establishing next-generation genetics

    Grant number：23113001  2011.4 - 2016.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

    TAKAYAMA Seiji, KINOSHITA Tetsu, SUZUKI Go, TAKAHASHI Aya, MATSUOKA Makoto, KITANO Jun, MATSUDA Youichi, TERAUCHI Ryouhei, WATANABE Masao, OKADA Norihiro, KAKUTANI Tetsuji, TORIYAMA Kinya, SEKI Motoaki

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    Grant amount：\93600000 （ Direct Cost: \72000000 、 Indirect Cost：\21600000 ）

    Genomes and genes are a blue print of many organisms. However, the structural organization and variation of those are often complex in natural population, and therefore, it is difficult to uncover its principle and common mechanisms by using model organisms, although many of those simple genome sequences are identified recently. In this research group, a Grant-in-Aid for Scientific Research on Innovative Areas, “Correlative Gene System: Establishing Next-Generation Genetics”, have focused on the conflict of genes and genomes within natural population, sexual pair and interactions of organisms such as parasite and host. To facilitate collaboration and discussion about principle mechanism underling the genetic conflict, we held annual meeting and international symposium. We also released our achievements to public through our homepage and outreach activities.

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  • QTL analysis of hybridization barrier in rice endosperm

    Grant number：23380005  2011.4 - 2014.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    TETSU Kinoshita

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    Authorship：Principal investigator  Grant type：Competitive

    Grant amount：\16380000 （ Direct Cost: \12600000 、 Indirect Cost：\3780000 ）

    We identified QTL region located on the long arm of chr.1. The locus causes hybridization barrier of rice endosperm in the cross of female O. sativa cv. Nipponbare and male O. longistaminata. In addition, we investigated and compared detailed endosperm development in interploidy and interspecific crosses. As a result, both cases show very similar endosperm phenotypes and patterns of developmental transitions, but in the nuclear division rate of endosperm.

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  • Reverse genetics and recombination

    Grant number：22370001  2010 - 2012

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    IIDA Shigeru, KINOSHITA Tetsu, ISHIZAKI Kimitsune, KOBAYASHI Hirokazu, NOGUCHI Hiroshi, WATANABE Kenji

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    Grant amount：\18850000 （ Direct Cost: \14500000 、 Indirect Cost：\4350000 ）

    To facilitate reverse genetic approaches, which include homologous recombination-mediated gene targeting using positive-negative selection and gene tagging employing endogenous nDart1-related DNA transposons, we were attempting to elucidate recombination mechanisms and their regulation mechanisms associated with gene targeting and tagging. We also characterized the functions of the genes identified by gene targeting and tagging.

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  • シロイヌナズナのインプリント遺伝子活性化におけるクロマチン機能とDNA脱メチル化

    Grant number：09F09311  2009 - 2011

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    木下 哲, DIANA Buzas

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    Grant amount：\2100000 （ Direct Cost: \2100000 ）

    植物のインプリント遺伝子がどのようにDNA脱メチル化を受け、どのように片一方の対立遺伝子のみが活性化されるかは多くの部分が不明である。この過程では、DNA脱メチル化を受ける領域の認識・決定、クロマチンリモデリング、ヒストン修飾、塩基除去修復系によるDNA脱メチル化、転写のステップ等が少なくとも考えられる。しかしながら、これらがどのような順番で制御されるかに関してさえ未だ不明である。本研究では、シロイヌナズナを用いて、DNAメチル化を伴って遺伝子発現不活化状態にあるFWAインプリント遺伝子をモデルにこれらの問題に切り込むことを目的としている。FWAはDNAの脱メチル化を受ける時期と細胞、シスエレメント等、多くが明らかになっている数少ないモデル遺伝子である。これまでに、インプリント遺伝子FWAの遺伝子発現を指標にして、ゲノムインプリンティングの制御に関わる変異体を複数単離している。外国人特別研究員のDiana Buzasさんの解析から、alac4変異体は受精前の中央細胞においてFWA-GFPリポーターを活性化できないとともに、他のインプリント遺伝子も活性化できないため、インプリンティングの確立に深く関わる因子と考えられる。また、原因遺伝子は、人から酵母まで保存されているが、その機能が未解明な低分子タンパク質をコードする遺伝子が原因であることを明らかにしている。さらにALAC4遺伝子が中央細胞や胚乳を取り囲む細胞層で強く発現することなどを明らかにしつつある。

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  • Analysis of DNA methylation in the reproductive cells using a tiling array

    Grant number：19380004  2007 - 2009

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

    NAKAZONO Mikio, TSUTSUMI Nobuhiro, KINOSHITA Tetsu, ISHIMARU Tsutomu

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    Grant amount：\20410000 （ Direct Cost: \15700000 、 Indirect Cost：\4710000 ）

    In this research, we tried to investigate DNA methylation pattern in laser microdissection (LMD)-isolated embryo and endosperm by a tiling array. We examined optimal conditions for bisulfite sequencing using total DNA extracted from some tissues in rice. We succeeded in the bisulfite sequencing when we used enough amounts of DNA, but failed in the sequencing using small amounts of DNA extracted from the LMD isolated tissues. As a result, we did not succeed in solution of this problem, and thus were not able to carry out the tiling array. We are continuing to examine the optimal conditions for the bisulfite sequencing using small amounts of DNA.

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  • Genomic barriers during endosperm development

    Grant number：18075010  2006 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas  Grant-in-Aid for Scientific Research on Priority Areas

    KINOSHITA Tetsu

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    Authorship：Principal investigator  Grant type：Competitive

    Grant amount：\78800000 （ Direct Cost: \78800000 ）

    In plants, interspecific cross often display hybrid incompatibility in the endosperm. We speculated that epigenetic mechanisms such as genomic imprinting contribute to this phenomena (GGS 2007). In this project, we identified the fact that SSRP1, know as a component of FACT histone chaperon component, is required for DNA demethylation and activation of imprinted genes (Dev. Cell 2011). We also investigated endosperm development in the hybrid endosperm in rice, and identified that developmental alteration is the cause of hybrid incompatibility (Plant J. 2011).

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  • Gene expression Profiling in plant reproductive process

    Grant number：18075012  2006 - 2010

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas  Grant-in-Aid for Scientific Research on Priority Areas

    KURATA Nori, WATANABE Masao, TSUTSUMI Nobuhiro, ITO Yukihiro, TORIYAMA Kinya, MATSUOKA Makoto, HATTORI Tsukaho, KINOSHITA Tetsu

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    Grant amount：\98300000 （ Direct Cost: \98300000 ）

    We revealed the gene expression profiles throughout whole reproductive stages/tissues of rice, and found many genes which were expressed in the tissues as a developmental stage-dependent manner. The genes which showed tissue-/stage-specific expression were supposed to have specialized function in plant reproduction steps, such as development of gametophytes (pollen grains and egg cells), pollination, fertilization, and early developmental stages of embryos. We also established methods of the genome-wide gene expression analysis for small amount of tissues. Based on the established methods, the gene expression profiles in developing microspores and egg cells were revealed.

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  • Epigenetic controls of plant development and genome structure

    Grant number：14GS0321  2002 - 2006

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Creative Scientific Research  Grant-in-Aid for Creative Scientific Research

    KAKUTANI Tetsuji, KINOSHITA Tetsu, SHIBAHARA Keiichi, ARAKI Takashi

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    Grant amount：\337350000 （ Direct Cost: \278130000 、 Indirect Cost：\59220000 ）

    FWA has been identified as the gene responsible for a late-flowering epigenetic trait (Mol Cell 6, 791-). In this project, we examined expression of the FWA gene during normal development. The FWA gene was expressed specifically in endosperm in an imprinted manner. The tissue-specific and imprinted FWA expression depends on DNA methyltransferase MET1 and DNA demethylase DEMETER (Science 303, 521-). It was also shown that FT is the inducer of flowering (Science 309, 1052-), and the FWA affects flowering by inhibiting the FT function at the protein level (Plant Cell Physiol 48, 205-).
    We have previously shown that Arabidopsis CACTA transposons are silent in wild type, but they are mobilized in mutant of a chromatin remodeling gene DDM1 (decrease in DNA methylation) (Nature 411, 212-). In this project, we showed that the CACTA transposons transposed in the double mutants of CG methylase MET1 and non-CG methylase CMT3. The results suggest that DNA methylation is necessary for immobilization of this class of transposons (Curr Biol 13, 421-).
    Transcription of CACTA was de-repressed by mutations in the MET1 or a chromatin assembly factor FAS (Curr Biol 13, 421-, Genes to Cells 18, 153-). Heterochromatin in CACTA locus or other peri-centromeric sequences was disrupted by the ddml mutation, and this effect was heritable even in the wild type background (EMBO J 21, 6549-). Using this system, we also showed that each of CACTA transposition was not targeted to heterochromatin (Genetics 168, 961-). It is interesting how the transposons accumulate to form the heterochromatin.

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  • 植物の新規インプリンティング遺伝子の網羅的検索

    Grant number：14654168  2002 - 2003

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    木下 哲

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    Authorship：Principal investigator  Grant type：Competitive

    Grant amount：\3500000 （ Direct Cost: \3500000 ）

    ゲノムインプリンティングは胎生発生をする動植物に共通に見られ、マウスではすでに50以上の遺伝子が明らかになっている。被子植物では胚乳においてゲノムインプリンティングが観察される。胚乳では、母由来のゲノムは胚乳発生に抑制的に働き、父由来のゲノムは促進的に働くことがわかっている。その原因となるインプリント遺伝子の解析は胚乳発生を理解する上で必要不可欠である。本研究ではシロイヌナズナのFWA遺伝子が二つ目インプリント遺伝子であることを明らかにした。FWA遺伝子の発現を詳細に検討し、受精前の中央細胞と受精後の胚乳において発現していることを明らかにした。また雄性配偶体と雌性配偶体由来のどちらのアレルが発現しているかを検討した結果より、FWA遺伝子は雌性配偶体由来アレルのみ発現し、維持型DNAメチル化酵素遺伝子とDNAグリコシラーゼ遺伝子の機能によって発現調節されることが明らかになった(Science 2004 303:521-523)。
    また、シロイヌナズナを用いて新規インプリント遺伝子の変異体の選抜をおこなった。変異体の選抜方法には、インプリント遺伝子はDNAのメチル化により制御されていることを利用した。維持型DNAメチル化酵素遺伝子の変異体を用い、新たに低メチル化等のエピジェネティックな変異を誘導した。その個体の中から、インプリント遺伝子MEDEAを抑圧し、その抑圧の表現形にparent-of-originがある遺伝子座を選抜し候補変異体を得た。マッピングにより、候補遺伝子は2番染色体に座乗する結果を得た。

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  • 液胞プロセシング酵素の組織特異的発現からみた液胞機能のダイナミズム

    Grant number：96J01663  1998

    日本学術振興会  科学研究費助成事業 特別研究員奨励費  特別研究員奨励費

    木下 哲

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    Grant amount：\1200000 （ Direct Cost: \1200000 ）

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