Updated on 2021/04/08

写真a

 
KURIHARA Daisuke
 
Organization
Institute of Transformative Bio-Molecules Designated lecturer
Title
Designated lecturer
External link

Degree 3

  1. 博士(工学) ( 2009.3   大阪大学 ) 

  2. 修士(工学) ( 2006.3   大阪大学 ) 

  3. 学士(工学) ( 2004.3   大阪大学 ) 

Research Interests 7

  1. 細胞運命決定

  2. 光顕微操作

  3. 植物雌性配偶体

  4. 植物胚発生

  5. 細胞分裂

  6. ライブイメージング

  7. 二光子顕微鏡

Research Areas 1

  1. Life Science / Morphology and anatomical structure

Current Research Project and SDGs 2

  1. 植物への長鎖DNA導入技術の開発

  2. 植物雌性配偶体発生・胚発生における細胞間コミュニケーションによる細胞運命制御機構の解明

Research History 7

  1. Nagoya University   Institute of Transformative Bio-Molecules   Designated lecturer

    2019.4

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  2. JST   さきがけ専任研究者

    2018.10

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    Country:Japan

  3. 名古屋大学トランスフォーマティブ生命分子研究所   招へい教員

    2018.10 - 2019.3

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    Country:Japan

  4. 名古屋大学大学院理学研究科   特任講師

    2017.4 - 2018.9

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    Country:Japan

  5. 名古屋大学大学院理学研究科   特任助教

    2011.1 - 2017.3

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    Country:Japan

  6. JST ERATO東山ライブホロニクスプロジェクト   光技術グループ   グループリーダー

    2011.1 - 2017.3

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    Country:Japan

  7. 名古屋大学大学院理学研究科   日本学術振興会特別研究員PD

    2009.4 - 2010.12

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    Country:Japan

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Education 3

  1. Osaka University   Graduate School, Division of Engineering

    - 2009.3

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    Country: Japan

  2. Osaka University   Graduate School, Division of Engineering

    - 2006.3

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    Country: Japan

  3. Osaka University   Faculty of Engineering

    - 2004.3

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    Country: Japan

Professional Memberships 3

  1. 日本植物形態学会

  2. 日本植物学会

  3. 日本植物生理学会

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Committee Memberships 1

  1. 日本植物形態学会   広報委員長  

    2020.1   

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    Committee type:学協会

Awards 6

  1. 2016年度JPR論文賞 Best Paper賞

    2016.9   The carboxyl-terminal tail of the stalk of Arabidopsis NACK1/HINKEL kinesin is required for its localization to the cell plate formation site

    Sasabe M, Ishibashi N, Haruta T, Minami A, Kurihara D, Higashiyama T, Nishihama R, Ito M, Machida Y

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    Award type:Honored in official journal of a scientific society, scientific journal 

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  2. 平瀬賞

    2016.9   ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging

    Kurihara D., Mizuta Y., Sato Y., Higashiyama T.

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    Award type:Award from Japanese society, conference, symposium, etc. 

  3. Honorable Mentions

    2015.12  

  4. 平瀬賞

    2015.9   Rapid Elimination of the Persistent Synergid through a Cell Fusion Mechanism

    Maruyama D., Volz R., Takeuchi H., Mori T., Igawa T., Kurihara D., Kawashima T., Ueda M., Ito M., Umeda M., Nishikawa S., Gross-Hardt R., Higashiyama T.

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    Award type:Award from Japanese society, conference, symposium, etc. 

  5. ITbM Research Award

    2013.10   Discovery of New Molecules that Control the Cell Cycle; Understanding the Mechanism of Animal and Plant

    Kurihara D., Kuwata K., Nambo M., Ohkawa T., Ueda M.

  6. 奨励賞

    2010.9  

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

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Papers 44

  1. Dynamics of the cell fate specifications during female gametophyte development in Arabidopsis Reviewed

    Daichi Susaki, Takamasa Suzuki, Daisuke Maruyama, Minako Ueda, Tetsuya Higashiyama, Daisuke Kurihara

    PLOS Biology   Vol. 19 ( 3 ) page: e3001123 - e3001123   2021.3

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Public Library of Science (PLoS)  

    The female gametophytes of angiosperms contain cells with distinct functions, such as those that enable reproduction via pollen tube attraction and fertilization. Although the female gametophyte undergoes unique developmental processes, such as several rounds of nuclear division without cell plate formation and final cellularization, it remains unknown when and how the cell fate is determined during development. Here, we visualized the living dynamics of female gametophyte development and performed transcriptome analysis of individual cell types to assess the cell fate specifications in <italic>Arabidopsis thaliana</italic>. We recorded time lapses of the nuclear dynamics and cell plate formation from the 1-nucleate stage to the 7-cell stage after cellularization using an in vitro ovule culture system. The movies showed that the nuclear division occurred along the micropylar–chalazal (distal–proximal) axis. During cellularization, the polar nuclei migrated while associating with the forming edge of the cell plate, and then, migrated toward each other to fuse linearly. We also tracked the gene expression dynamics and identified that the expression of <italic>MYB98pro</italic>::<italic>GFP–MYB98</italic>, a synergid-specific marker, was initiated just after cellularization in the synergid, egg, and central cells and was then restricted to the synergid cells. This indicated that cell fates are determined immediately after cellularization. Transcriptome analysis of the female gametophyte cells of the wild-type and <italic>myb98</italic> mutant revealed that the <italic>myb98</italic> synergid cells had egg cell–like gene expression profiles. Although in <italic>myb98</italic>, egg cell–specific gene expression was properly initiated in the egg cells only after cellularization, but subsequently expressed ectopically in one of the 2 synergid cells. These results, together with the various initiation timings of the egg cell–specific genes, suggest complex regulation of the individual gametophyte cells, such as cellularization-triggered fate initiation, MYB98-dependent fate maintenance, cell morphogenesis, and organelle positioning. Our system of live-cell imaging and cell type–specific gene expression analysis provides insights into the dynamics and mechanisms of cell fate specifications in the development of female gametophytes in plants.

    DOI: 10.1371/journal.pbio.3001123

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  2. ClearSeeAlpha: Advanced Optical Clearing for Whole-Plant Imaging Reviewed

    Daisuke Kurihara, Yoko Mizuta, Shiori Nagahara, Tetsuya Higashiyama

    Plant and Cell Physiology     2021

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Oxford University Press (OUP)  

    DOI: 10.1093/pcp/pcab033

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  3. ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging Reviewed

    Daisuke Kurihara, Yoko Mizuta, Yoshikatsu Sato, Tetsuya Higashiyama

    DEVELOPMENT   Vol. 142 ( 23 ) page: 4168 - 4179   2015.12

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:COMPANY OF BIOLOGISTS LTD  

    Imaging techniques for visualizing and analyzing precise morphology and gene expression patterns are essential for understanding biological processes during development in all organisms. With the aid of chemical screening, we developed a clearing method using chemical solutions, termed ClearSee, for deep imaging of morphology and gene expression in plant tissues. ClearSee rapidly diminishes chlorophyll autofluorescence while maintaining fluorescent protein stability. By adjusting the refractive index mismatch, whole-organ and whole-plant imaging can be performed by both confocal and two-photon excitation microscopy in ClearSee-treated samples. Moreover, ClearSee is applicable to multicolor imaging of fluorescent proteins to allow structural analysis of multiple gene expression. Given that ClearSee is compatible with staining by chemical dyes, the technique is useful for deep imaging in conjunction with genetic markers and for plant species not amenable to transgenic approaches. This method is useful for whole imaging for intact morphology and will help to accelerate the discovery of new phenomena in plant biological research.

    DOI: 10.1242/dev.127613

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  4. Live-Cell Imaging and Optical Manipulation of Arabidopsis Early Embryogenesis Reviewed

    Keita Gooh, Minako Ueda, Kana Aruga, Jongho Park, Hideyuki Arata, Tetsuya Higashiyama, Daisuke Kurihara

    DEVELOPMENTAL CELL   Vol. 34 ( 2 ) page: 242 - 251   2015.7

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:CELL PRESS  

    Intercellular communications are essential for cell proliferation and differentiation during plant embryogenesis. However, analysis of intercellular communications in living material in real time is difficult owing to the restricted accessibility of the embryo within the flower. We established a live-embryo imaging system to visualize cell division and cell fate specification in Arabidopsis thaliana from zygote division in real time. We generated a cell-division lineage tree for early embryogenesis in Arabidopsis. Lineage analysis showed that both the direction and time course of cell division between sister cells differed along the apical-basal or radial axes. Using the Arabidopsis kpl mutant, in which single-fertilization events are frequent, we showed that endosperm development is not required for pattern formation during early embryogenesis. Optical manipulation demonstrated that damage to the embryo initial cell induces cell fate conversion of the suspensor cell to compensate for the disrupted embryo initial cell even after cell fate is specified.

    DOI: 10.1016/j.devcel.2015.06.008

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  5. Mitochondrial dynamics and segregation during the asymmetric division of Arabidopsis zygotes International journal

    Yusuke Kimata, Takumi Higaki, Daisuke Kurihara, Naoe Ando, Hikari Matsumoto, Tetsuya Higashiyama, Minako Ueda

    Quantitative Plant Biology   Vol. 1   2020.11

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Cambridge University Press (CUP)  

    <title>Abstract</title>
    The zygote is the first cell of a multicellular organism. In most angiosperms, the zygote divides asymmetrically to produce an embryo-precursor apical cell and a supporting basal cell. Zygotic division should properly segregate symbiotic organelles, because they cannot be synthesized <italic>de novo</italic>. In this study, we revealed the real-time dynamics of the principle source of ATP biogenesis, mitochondria, in <italic>Arabidopsis thaliana</italic> zygotes using live-cell observations and image quantifications. In the zygote, the mitochondria formed the extended structure associated with the longitudinal array of actin filaments (F-actins) and were polarly distributed along the apical–basal axis. The mitochondria were then temporally fragmented during zygotic division, and the resulting apical cells inherited mitochondria at higher concentration compared to the basal cells. Further observation of postembryonic organs showed that these mitochondrial behaviours are characteristic of the zygote. Overall, our results showed that the zygote has spatiotemporal regulation that unequally distributes the mitochondria.

    DOI: 10.1017/qpb.2020.4

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  6. A Peptide Pair Coordinates Regular Ovule Initiation Patterns with Seed Number and Fruit Size Reviewed

    Nozomi Kawamoto, Dunia Pino Del Carpio, Alexander Hofmann, Yoko Mizuta, Daisuke Kurihara, Tetsuya Higashiyama, Naoyuki Uchida, Keiko U. Torii, Lucia Colombo, Georg Groth, Rüdiger Simon

    Current Biology   Vol. 30 ( 22 ) page: 4352 - 4361.e4   2020.11

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

    DOI: 10.1016/j.cub.2020.08.050

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  7. Arabidopsis GEX1 Is a Nuclear Membrane Protein of Gametes Required for Nuclear Fusion During Reproduction Reviewed

    Shuh-ichi Nishikawa, Yuki Yamaguchi, Chiharu Suzuki, Ayaka Yabe, Yuzuru Sato, Daisuke Kurihara, Yoshikatsu Sato, Daichi Susaki, Tetsuya Higashiyama, Daisuke Maruyama

    Frontiers in Plant Science   Vol. 11   2020.10

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

    DOI: 10.3389/fpls.2020.548032

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  8. The formation of perinucleolar bodies is important for normal leaf development and requires the zinc-finger DNA-binding motif in Arabidopsis ASYMMETRIC LEAVES2. Reviewed International journal

    Lilan Luo, Sayuri Ando, Yuki Sakamoto, Takanori Suzuki, Hiro Takahashi, Nanako Ishibashi, Shoko Kojima, Daisuke Kurihara, Tetsuya Higashiyama, Kotaro T Yamamoto, Sachihiro Matsunaga, Chiyoko Machida, Michiko Sasabe, Yasunori Machida

    Plant Journal   Vol. 101 ( 5 ) page: 1118 - 1134   2020.3

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    In Arabidopsis, the ASYMMETRIC LEAVES2 (AS2) protein plays a key role in the formation of flat symmetric leaves via direct repression of the abaxial gene ETT/ARF3. AS2 encodes a plant-specific nuclear protein that contains the AS2/LOB domain, which includes a zinc-finger (ZF) motif that is conserved in the AS2/LOB family. We have shown that AS2 binds to the coding DNA of ETT/ARF3, which requires the ZF motif. AS2 is co-localized with AS1 in perinucleolar bodies (AS2 bodies). To identify the amino acid signals in AS2 required for formation of AS2 bodies and function(s) in leaf formation, we constructed recombinant DNAs that encoded mutant AS2 proteins fused to yellow fluorescent protein. We examined the subcellular localization of these proteins in cells of cotyledons and leaf primordia of transgenic plants and cultured cells. The amino acid signals essential for formation of AS2 bodies were located within and adjacent to the ZF motif. Mutant AS2 that failed to form AS2 bodies also failed to rescue the as2-1 mutation. Our results suggest the importance of the formation of AS2 bodies and the nature of interactions of AS2 with its target DNA and nucleolar factors including NUCLEOLIN1. The partial overlap of AS2 bodies with perinucleolar chromocenters with condensed ribosomal RNA genes implies a correlation between AS2 bodies and the chromatin state. Patterns of AS2 bodies in cells during interphase and mitosis in leaf primordia were distinct from those in cultured cells, suggesting that the formation and distribution of AS2 bodies are developmentally modulated in plants.

    DOI: 10.1111/tpj.14579

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  9. Live-Cell Imaging of Zygotic Intracellular Structures and Early Embryo Pattern Formation in Arabidopsis thaliana. Reviewed International journal

    Minako Ueda, Yusuke Kimata, Daisuke Kurihara

    Methods in Molecular Biology   Vol. 2122   page: 37 - 47   2020

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    Plant embryogenesis begins with fertilization and ends with the generation of the basic body plan of the future plant. Despite its importance, the dynamics of flowering plant ontogeny have long been a mystery, because the embryo develops deep in the maternal tissue. Recently, an embryonic live-cell imaging system was established in Arabidopsis thaliana by developing an in vitro ovule cultivation method and utilizing two-photon excitation microscopy (2PEM), which is suitable for deep imaging. This system enabled us to visualize intracellular dynamics during zygote polarization and monitor the cell division pattern during embryogenesis from the zygote until organ formation. In this chapter, we describe a method that allows for high-resolution imaging of cytoskeletal rearrangements in the zygote and long-term tracing of embryo patterning.

    DOI: 10.1007/978-1-0716-0342-0_4

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  10. Polar vacuolar distribution is essential for accurate asymmetric division of Arabidopsis zygotes. Reviewed International journal

    Kimata Y, Kato T, Higaki T, Kurihara D, Yamada T, Segami S, Morita MT, Maeshima M, Hasezawa S, Higashiyama T, Tasaka M, Ueda M

    Proc Natl Acad Sci U S A.   Vol. 116 ( 6 ) page: 2338 - 2343   2019.1

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  11. Spatiotemporal deep imaging of syncytium induced by the soybean cyst nematode Heterodera glycines Reviewed International journal

    Mina Ohtsu, Yoshikatsu Sato, Daisuke Kurihara, Takuya Suzaki, Masayoshi Kawaguchi, Daisuke Maruyama, Tetsuya Higashiyama

    PROTOPLASMA   Vol. 254 ( 6 ) page: 2107 - 2115   2017.11

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER WIEN  

    Parasite infections cause dramatic anatomical and ultrastructural changes in host plants. Cyst nematodes are parasites that invade host roots and induce a specific feeding structure called a syncytium. A syncytium is a large multinucleate cell formed by cell wall dissolution-mediated cell fusion. The soybean cyst nematode (SCN), Heterodera glycines, is a major soybean pathogen. To investigate SCN infection and the syncytium structure, we established an in planta deep imaging system using a clearing solution ClearSee and two-photon excitation microscopy (2PEM). Using this system, we found that several cells were incorporated into the syncytium; the nuclei increased in size and the cell wall openings began to be visible at 2 days after inoculation (DAI). Moreover, at 14 DAI, in the syncytium developed in the cortex, there were thickened concave cell wall pillars that resembled "Parthenon pillars." In contrast, there were many thick board-like cell walls and rarely Parthenon pillars in the syncytium developed in the stele. We revealed that the syncytia were classified into two types based on the pattern of the cell wall structures, which appeared to be determined by the position of the syncytium inside roots. Our results provide new insights into the developmental process of syncytium induced by cyst nematode and a better understanding of the three-dimensional structure of the syncytium in host roots.

    DOI: 10.1007/s00709-017-1105-0

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  12. In Vitro Ovule Cultivation for Live-cell Imaging of Zygote Polarization and Embryo Patterning in Arabidopsis thaliana Invited Reviewed International journal

    Daisuke Kurihara, Yusuke Kimata, Tetsuya Higashiyama, Minako Ueda

    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS   ( 127 ) page: .   2017.9

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    In most flowering plants, the zygote and embryo are hidden deep in the mother tissue, and thus it has long been a mystery of how they develop dynamically; for example, how the zygote polarizes to establish the body axis and how the embryo specifies various cell fates during organ formation. This manuscript describes an in vitro ovule culture method to perform live-cell imaging of developing zygotes and embryos of Arabidopsis thaliana. The optimized cultivation medium allows zygotes or early embryos to grow into fertile plants. By combining it with a poly(dimethylsiloxane) (PDMS) micropillar array device, the ovule is held in the liquid medium in the same position. This fixation is crucial to observe the same ovule under a microscope for several days from the zygotic division to the late embryo stage. The resulting live-cell imaging can be used to monitor the real-time dynamics of zygote polarization, such as nuclear migration and cytoskeleton rearrangement, and also the cell division timing and cell fate specification during embryo patterning. Furthermore, this ovule cultivation system can be combined with inhibitor treatments to analyze the effects of various factors on embryo development, and with optical manipulations such as laser disruption to examine the role of cell-cell communication.

    DOI: 10.3791/55975

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  13. Plant tissue clearing for fluorescence imaging Invited Reviewed International journal

    Plant Morphology   Vol. 29 ( 1 ) page: 81‐86(J‐STAGE) - 86   2017.7

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    Authorship:Lead author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)  

    DOI: 10.5685/plmorphol.29.81

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  14. Fluorescent Labeling of the Cyst Nematode Heterodera glycines in Deep-Tissue Live Imaging Reviewed International journal

    Mina Ohtsu, Daisuke Kurihara, Yoshikatsu Sato, Takuya Suzaki, Masayoshi Kawaguchi, Daisuke Maruyama, Tetsuya Higashiyama

    CYTOLOGIA   Vol. 82 ( 3 ) page: 251 - 259   2017.6

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:UNIV TOKYO CYTOLOGIA  

    Nematode infection of plant roots is a paradigm of host parasite interactions. Although nematodes can be labeled with fluorescent dyes, migration of the worms into the deep regions of host roots makes them difficult to track. Here we report the use of two fluorescent dyes, FM4-64 and SYBR green I, to intensely label the soybean cyst nematode (SCN) Heterodera glycines for one week in host plants. Continuous monitoring of the labeled SCN juveniles was achieved with two-photon microscopy. Additionally, we developed a transient transformation system consisting of the non-model leguminous plant (fabaceous) roots, Astragalus sinicus and Agrobacterium rhizogenes to observe the cellular structures of the plant during SCN infection. By the combined use of fluorescent dyes and two-photon microscopy, clear images of infecting SCNs were obtained even in deep regions of A. sinicus roots. The fluorescent labeling described herein can also be used in detailed monitoring of the infection processes of other non-model nematodes, as well as the associated morphological changes in the host plant roots.

    DOI: 10.1508/cytologia.82.251

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  15. Cytoskeleton dynamics control the first asymmetric cell division in Arabidopsis zygote Reviewed International journal

    Yusuke Kimata, Takumi Higaki, Tomokazu Kawashima, Daisuke Kurihara, Yoshikatsu Sato, Tomomi Yamada, Seiichiro Hasezawa, Frederic Berger, Tetsuya Higashiyama, Minako Ueda

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   Vol. 113 ( 49 ) page: 14157 - 14162   2016.12

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    The asymmetric cell division of the zygote is the initial and crucial developmental step in most multicellular organisms. In flowering plants, whether zygote polarity is inherited from the preexisting organization in the egg cell or reestablished after fertilization has remained elusive. How dynamically the intracellular organization is generated during zygote polarization is also unknown. Here, we used a live-cell imaging system with Arabidopsis zygotes to visualize the dynamics of the major elements of the cytoskeleton, microtubules (MTs), and actin filaments (F-actins), during the entire process of zygote polarization. By combining image analysis and pharmacological experiments using specific inhibitors of the cytoskeleton, we found features related to zygote polarization. The preexisting alignment of MTs and F-actin in the egg cell is lost on fertilization. Then, MTs organize into a transverse ring defining the zygote subapical region and driving cell outgrowth in the apical direction. F-actin forms an apical cap and longitudinal arrays and is required to position the nucleus to the apical region of the zygote, setting the plane of the first asymmetrical division. Our findings show that, in flowering plants, the preexisting cytoskeletal patterns in the egg cell are lost on fertilization and that the zygote reorients the cytoskeletons to perform directional cell elongation and polar nuclear migration.

    DOI: 10.1073/pnas.1613979113

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  16. Combination of Synthetic Chemistry and Live-Cell Imaging Identified a Rapid Cell Division Inhibitor in Tobacco and Arabidopsis thaliana. Reviewed

    Masakazu Nambo, Daisuke Kurihara, Tomomi Yamada, Taeko Nishiwaki-Ohkawa, Naoya Kadofusa, Yusuke Kimata, Keiko Kuwata, Masaaki Umeda, Minako Ueda

    Plant & cell physiology   Vol. 57 ( 11 ) page: 2255 - 2268   2016.11

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    Cell proliferation is crucial to the growth of multicellular organisms, and thus the proper control of cell division is important to prevent developmental arrest or overgrowth. Nevertheless, tools for controlling cell proliferation are still poor in plant. To develop novel tools, we focused on a specific compound family, triarylmethanes, whose members show various antiproliferative activities in animals. By combining organic chemistry to create novel and diverse compounds containing the triarylmethyl moiety and biological screens based on live-cell imaging of a fluorescently labeled tobacco Bright Yellow-2 (BY-2) culture cell line (Nicotiana tabacum), we isolated (3-furyl)diphenylmethane as a strong but partially reversible inhibitor of plant cell division. We also found that this agent had efficient antiproliferative activity in developing organs of Arabidopsis thaliana without causing secondary defects in cell morphology, and induced rapid cell division arrest independent of the cell cycle stage. Given that (3-furyl)diphenylmethane did not affect the growth of a human cell line (HeLa) and a budding yeast (Saccharomyces cerevisiae), it should act specifically on plants. Taking our results together, we propose that the combination of desired chemical synthesis and detailed biological analysis is an effective tool to create novel drugs, and that (3-furyl)diphenylmethane is a specific antiproliferative agent for plants.

    DOI: 10.1093/pcp/pcw140

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  17. Plant Aurora kinases interact with and phosphorylate transcription factors Reviewed International journal

    Mai Takagi, Takuya Sakamoto, Ritsuko Suzuki, Keiichirou Nemoto, Takeshi Obayashi, Takeshi Hirakawa, Tomoko M. Matsunaga, Daisuke Kurihara, Yuko Nariai, Takeshi Urano, Tatsuya Sawasaki, Sachihiro Matsunaga

    JOURNAL OF PLANT RESEARCH   Vol. 129 ( 6 ) page: 1165 - 1178   2016.11

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER JAPAN KK  

    Aurora kinase (AUR) is a well-known mitotic serine/threonine kinase that regulates centromere formation, chromosome segregation, and cytokinesis in eukaryotes. In addition to regulating mitotic events, AUR has been shown to regulate protein dynamics during interphase in animal cells. In contrast, there has been no identification and characterization of substrates and/or interacting proteins during interphase in plants. The Arabidopsis thaliana genome encodes three AUR paralogues, AtAUR1, AtAUR2, and AtAUR3. Among them, AtAUR1 and AtAUR2 are considered to function redundantly. Here, we confirmed that both AtAUR1 and AtAUR3 are localized in the nucleus and cytoplasm during interphase, suggesting that they have functions during interphase. To identify novel interacting proteins, we used AlphaScreen to target 580 transcription factors (TFs) that are mainly functional during interphase, using recombinant A. thaliana TFs and AtAUR1 or AtAUR3. We found 133 and 32 TFs had high potential for interaction with AtAUR1 and AtAUR3, respectively. The highly AtAUR-interacting TFs were involved in various biological processes, suggesting the functions of the AtAURs during interphase. We found that AtAUR1 and AtAUR3 showed similar interaction affinity to almost all TFs. However, in some cases, the interaction affinity differed substantially between the two AtAUR homologues. These results suggest that AtAUR1 and AtAUR3 have both redundant and distinct functions through interactions with TFs. In addition, database analysis revealed that most of the highly AtAUR-interacting TFs contained a detectable phosphopeptide that was consistent with the consensus motifs for human AURs, suggesting that these TFs are substrates of the AtAURs. The AtAURs phosphorylated several highly interacting TFs in the AlphaScreen in vitro. Overall, in line with the regulation of TFs through interaction, our results indicate the possibility of phosphoregulation of several TFs by the AtAURs (280/300).

    DOI: 10.1007/s10265-016-0860-x

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  18. Haspin has Multiple Functions in the Plant Cell Division Regulatory Network Reviewed

    Elena Kozgunova, Takamasa Suzuki, Masaki Ito, Tetsuya Higashiyama, Daisuke Kurihara

    PLANT AND CELL PHYSIOLOGY   Vol. 57 ( 4 ) page: 848 - 861   2016.4

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    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS  

    Progression of cell division is controlled by various mitotic kinases. In animal cells, phosphorylation of histone H3 at Thr3 by the kinase Haspin (haploid germ cell-specific nuclear protein kinase) promotes centromeric Aurora B localization to regulate chromosome segregation. However, less is known about the function of Haspin in regulatory networks in plant cells. Here, we show that inhibition of Haspin with 5-iodotubercidin (5-ITu) in Bright Yellow-2 (BY-2) cells delayed chromosome alignment. Haspin inhibition also prevented the centromeric localization of Aurora3 kinase (AUR3) and disrupted its function. This suggested that Haspin plays a role in the specific positioning of AUR3 on chromosomes in plant cells, a function conserved in animals. The results also indicated that Haspin and AUR3 are involved in the same pathway, which regulates chromosome alignment during prometaphase/metaphase. Remarkably, Haspin inhibition by 5-ITu also led to a severe cytokinesis defect, resulting in binuclear cells with a partially formed cell plate. The 5-ITu treatment did not affect microtubules, AUR1/2 or the NACK-PQR pathway; however, it did alter the distribution of actin filaments on the cell plate. Together, these results suggested that Haspin has several functions in regulating cell division in plant cells: in the localization of AUR3 on centromeres and in regulating late cell plate expansion during cytokinesis.

    DOI: 10.1093/pcp/pcw030

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    Other Link: http://orcid.org/0000-0003-2703-0405

  19. Cytokinesis defect in BY-2 cells caused by ATP-competitive kinase inhibitors Invited Reviewed

    Elena Kozgunova, Tetsuya Higashiyama, Daisuke Kurihara

    PLANT SIGNALING & BEHAVIOR   Vol. 11 ( 10 ) page: e1238547   2016

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    Cytokinesis is last but not least in cell division as it completes the formation of the two cells. The main role in cell plate orientation and expansion have been assigned to microtubules and kinesin proteins. However, recently we reported severe cytokinesis defect in BY-2 cells not accompanied by changes in microtubules dynamics. Here we also confirmed that distribution of kinesin NACK1 is not the cause of cytokinesis defect. We further explored inhibition of the cell plate expansion by ATP-competitive inhibitors. Two different inhibitors, 5-Iodotubercidin and ML-7 resulted in a very similar phenotype, which indicates that they target same protein cascade. Interestingly, in our previous study we showed that 5-Iodotubercidin treatment affects concentration of actin filaments on the cell plate, while ML-7 is inhibitor of myosin light chain kinase. Although not directly, it indicates importance of actomyosin complex in plant cytokinesis.

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  20. Two-photon imaging with longer wavelength excitation in intact Arabidopsis tissues Reviewed

    Yoko Mizuta, Daisuke Kurihara, Tetsuya Higashiyama

    PROTOPLASMA   Vol. 252 ( 5 ) page: 1231 - 1240   2015.9

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    In vivo imaging of living organisms is an important tool to investigate biological phenomena. Two-photon excitation microscopy (2PEM) is a laser-scanning microscopy that provides noninvasive, deep imaging in living organisms based on the principle of multiphoton excitation. However, application of 2PEM to plant tissues has not been fully developed, as plant-specific autofluorescence, optically dense tissues, and multiple light-scattering structures diminish the clarity of imaging. In this study, the advantages of 2PEM were identified for deep imaging of living and intact Arabidopsis thaliana tissues. When compared to single-photon imaging, near-infrared 2PEM, especially at 1000 nm, reduced chloroplast autofluorescence; autofluorescence also decreased in leaves, roots, pistils, and pollen grains. For clear and deep imaging, longer excitation wavelengths using the orange fluorescent proteins (FPs) TagRFP and tdTomato gave better results than with other colors. 2PEM at 980 nm also provided multicolor imaging by simultaneous excitation, and the combination of suitable FPs and excitation wavelengths allowed deep imaging of intact cells in root tips and pistils. Our results demonstrated the importance of choosing both suitable FPs and excitation wavelengths for clear two-photon imaging. Further advances in in vivo analysis using 2PEM will facilitate more extensive studies in the plant biological sciences.

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  21. Live Imaging and Laser Disruption Reveal the Dynamics and Cell-Cell Communication During Torenia fournieri Female Gametophyte Development Reviewed

    Daichi Susaki, Hidenori Takeuchi, Hiroki Tsutsui, Daisuke Kurihara, Tetsuya Higashiyama

    PLANT AND CELL PHYSIOLOGY   Vol. 56 ( 5 ) page: 1031 - 1041   2015.5

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    The female gametophytes of many flowering plants contain one egg cell, one central cell, two synergid cells and three antipodal cells with respective morphological characteristics and functions. These cells are formed by cellularization of a multinuclear female gametophyte. However, the dynamics and mechanisms of female gametophyte development remain largely unknown due to the lack of a system to visualize directly and manipulate female gametophytes in living material. Here, we established an in vitro ovule culture system to examine female gametophyte development in Torenia fournieri, a unique plant species with a protruding female gametophyte. The four-nucleate female gametophyte became eight nucleate by the final (third) mitosis and successively cellularized and matured to attract a pollen tube. The duration of final mitosis was 28 +/- 6.5 min, and cellularization was completed in 54 +/- 20 min after the end of the third mitosis. Fusion of polar nuclei in the central cell occurred in 13.1 +/- 1.1 h, and onset of expression of LURE2, a pollen tube attractant gene, was visualized by a green fluorescent protein reporter 10.7 +/- 2.3 h after cellularization. Laser disruption analysis demonstrated that the egg and central cells were required for synergid cells to acquire the pollen tube attraction function. Moreover, aberrant nuclear positioning and down-regulation of LURE2 were observed in one of the two synergid cells after disrupting an immature egg cell, suggesting that cell specification was affected. Our system provides insights into the precise dynamics and mechanisms of female gametophyte development in T. fournieri.

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    Other Link: http://orcid.org/0000-0003-2703-0405

  22. Rapid Elimination of the Persistent Synergid through a Cell Fusion Mechanism Reviewed

    Daisuke Maruyama, Ronny Voelz, Hidenori Takeuchi, Toshiyuki Mori, Tomoko Igawa, Daisuke Kurihara, Tomokazu Kawashima, Minako Ueda, Masaki Ito, Masaaki Umeda, Shuh-ichi Nishikawa, Rita Gross-Hardt, Tetsuya Higashiyama

    CELL   Vol. 161 ( 4 ) page: 907 - 918   2015.5

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    In flowering plants, fertilization-dependent degeneration of the persistent synergid cell ensures one-on-one pairings of male and female gametes. Here, we report that the fusion of the persistent synergid cell and the endosperm selectively inactivates the persistent synergid cell in Arabidopsis thaliana. The synergid-endosperm fusion causes rapid dilution of pre-secreted pollen tube attractant in the persistent synergid cell and selective disorganization of the synergid nucleus during the endosperm proliferation, preventing attractions of excess number of pollen tubes (polytubey). The synergid-endosperm fusion is induced by fertilization of the central cell, while the egg cell fertilization predominantly activates ethylene signaling, an inducer of the synergid nuclear disorganization. Therefore, two female gametes (the egg and the central cell) control independent pathways yet coordinately accomplish the elimination of the persistent synergid cell by double fertilization.

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  23. The carboxyl-terminal tail of the stalk of Arabidopsis NACK1/HINKEL kinesin is required for its localization to the cell plate formation site Reviewed

    Michiko Sasabe, Nanako Ishibashi, Tsuyoshi Haruta, Aki Minami, Daisuke Kurihara, Tetsuya Higashiyama, Ryuichi Nishihama, Masaki Ito, Yasunori Machida

    JOURNAL OF PLANT RESEARCH   Vol. 128 ( 2 ) page: 327 - 336   2015.3

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    Plant cytokinesis is achieved by formation of cell plates in the phragmoplast, a plant-specific cytokinetic apparatus, which consists of microtubules (MTs) and microfilaments. During cytokinesis, the cell plate is expanded centrifugally outward from the inside of cells in a process that is supported by dynamic turnover of MTs. M-phase-specific kinesin NACK1, which comprises the motor domain at the amino-terminal half to move on MT bundles and the stalk region in the carboxyl-terminal half, is a key player in the process of MT turnover. That is, the specific region in the stalk binds the MAP kinase kinase kinase to activate the whole MAP kinase cascade, which stimulates depolymerization of MTs for the MT turnover. The stalk is also responsible for recruiting the activated kinase cascade to the mid-zone of the phragmoplast, which corresponds to the cell-plate formation site. It should be crucial to uncover roles of the NACK1 kinesin stalk as well as the motor domain in the formation of cell plates in order to understand the mechanisms of cell plate formation. Using dissected Arabidopsis NACK1 (AtNACK1/HINKEL) molecules and AtNACK1-fused GFP, we showed that the C-terminal tail of the stalk in addition to the motor domain is critical for its proper localization to the site of cell plate formation in the phragmoplast, probably by affecting its motility activity.

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  24. Increase in Invaginated Vacuolar Membrane Structure Caused by Plant Cell Expansion by Genotoxic Stress Induced by DNA Double-Strand Breaks Reviewed

    Junko Hasegawa, Takumi Higaki, Yuki Hamamura, Daisuke Kurihara, Natsumaro Kutsuna, Tetsuya Higashiyama, Seiichiro Hasezawa, Sachihiro Matsunaga

    CYTOLOGIA   Vol. 79 ( 4 ) page: 467 - 474   2014.12

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    Vacuoles occupy 80-90% of a mature plant cell and mainly contribute to all types of cell expansion. Zeocin, an inducer of DNA double-strand breaks, causes cell expansion with endoreduplication. The vacuolar structure after zeocin treatment was examined in tobacco (Nicotiana tabacum) cultured cells expressing GFP fused to a vacuole membrane protein. We found that the genotoxic stress induced the cell expansion with subdivision of the vacuolar lumen by cytoplasmic strands. When a femtosecond laser was used to cut off the cytoplasmic strand, mitochondrial transport along the strand stopped. This suggested that in the elongated cells under the genotoxic stress, the transport of subcellular materials was activated for DNA repair within the damaged cell nucleus by the construction of a network of cytoplasmic strands in the vacuolar lumen.

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  25. Live imaging of calcium spikes during double fertilization in Arabidopsis Reviewed

    Yuki Hamamura, Moe Nishimaki, Hidenori Takeuchi, Anja Geitmann, Daisuke Kurihara, Tetsuya Higashiyama

    NATURE COMMUNICATIONS   Vol. 5   2014.8

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    Ca2+ waves and oscillation are key signalling elements during the fertilization process of animals, and are involved, for example, in egg activation. In the unique double fertilization process in flowering plants, both the egg cell and the neighbouring central cell fuse with a sperm cell each. Here we succeeded in imaging cytosolic Ca2+ in these two cells, and in the two synergid cells that accompany the gametes during semi-in vivo double fertilization. Following pollen tube discharge and plasmogamy, the egg and central cells displayed transient Ca2+ spikes, but not oscillations. Only the events in the egg cell correlated with the plasmogamy. In contrast, the synergid cells displayed Ca2+ oscillations on pollen tube arrival. The two synergid cells showed distinct Ca2+ dynamics depending on their respective roles in tube reception. These Ca2+ dynamics in the female gametophyte seem to represent highly specific signatures that coordinate successful double fertilization in the flowering plants.

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  26. Fabrication of microcage arrays to fix plant ovules for long-term live imaging and observation Reviewed

    Jongho Park, Daisuke Kurihara, Tetsuya Higashiyama, Hideyuki Arata

    SENSORS AND ACTUATORS B-CHEMICAL   Vol. 191   page: 178 - 185   2014.2

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    The long-term observation of plant ovules is important to investigate embryogenesis and the development occurring inside. However, it has been difficult to observe the ovule consistently in a specific position because ovules are randomly dispersed on a substrate with conventional methods. In this work, we fabricated PDMS microcage arrays to fix plant ovules from Arabidopsis thaliana and observed them over a long period. Microcages were designed based on experimental data of ovule growth so that ovules could be positioned inside the cage and observed in real time. First, we performed the sieving test to investigate the relationship between ovule size and different microcage widths. Next, we performed ovule culture for a week using microcage arrays developed based on the results of sieving tests. We successfully positioned and fixed ovules inside the microcages and confirmed that the ovules did not move from their original positions even though the culture dish was frequently moved for observation and incubation. Moreover, we observed that the ovules grew inside the cages without being affected due to the flexibility of PDMS. Thus, we confirmed the validity of microcage arrays as the useful tool to fix and culture plant ovules for long-term observation. We expect that microcage arrays could be widely used as the essential device for pursuing scientific findings in further researches of plant embryogenesis and reproduction. (C) 2013 Elsevier B.V. All rights reserved.

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  27. Independent Control by Each Female Gamete Prevents the Attraction of Multiple Pollen Tubes Reviewed

    Daisuke Maruyama, Yuki Hamamura, Hidenori Takeuchi, Daichi Susaki, Moe Nishimaki, Daisuke Kurihara, Ryushiro D. Kasahara, Tetsuya Higashiyama

    DEVELOPMENTAL CELL   Vol. 25 ( 3 ) page: 317 - 323   2013.5

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    In flowering plants, double fertilization is normally accomplished by the first pollen tube, with the fertilized ovule subsequently inhibiting the attraction of a second pollen tube. However, the mechanism of second-pollen-tube avoidance remains unknown. We discovered that failure to fertilize either the egg cell or the central cell compromised second-pollen-tube avoidance in Arabidopsis thaliana. A similar disturbance was caused by disrupting the fertilization-independent seed (FIS) class polycomb-repressive complex 2 (FIS-PRC2), a central cell- and endosperm-specific chromatin-modifying complex for gene silencing. Therefore, the two female gametes have evolved their own signaling pathways. Intriguingly, second-pollen-tube attraction induced by half-successful fertilization allowed the ovules to complete double fertilization, producing a genetically distinct embryo and endosperm. We thus propose that each female gamete independently determines second-pollen-tube avoidance to maximize reproductive fitness in flowering plants.

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  28. Live-cell analysis of plant reproduction: Live-cell imaging, optical manipulation, and advanced microscopy technologies Invited Reviewed

    Daisuke Kurihara, Yuki Hamamura, Tetsuya Higashiyama

    DEVELOPMENT GROWTH & DIFFERENTIATION   Vol. 55 ( 4 ) page: 462 - 473   2013.5

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    Sexual reproduction ensures propagation of species and enhances genetic diversity within populations. In flowering plants, sexual reproduction requires complicated and multi-step cell-to-cell communications among male and female cells. However, the confined nature of plant reproduction processes, which occur in the female reproductive organs and several cell layers of the pistil, limits our ability to observe these events in vivo. In this review, we discuss recent live-cell imaging in in vitro systems and the optical manipulation techniques that are used to capture the dynamic mechanisms representing molecular and cellular communications in sexual plant reproduction.

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  29. Arabidopsis ASYMMETRIC LEAVES2 protein required for leaf morphogenesis consistently forms speckles during mitosis of tobacco BY-2 cells via signals in its specific sequence Reviewed

    Lilan Luo, Sayuri Ando, Michiko Sasabe, Chiyoko Machida, Daisuke Kurihara, Tetsuya Higashiyama, Yasunori Machida

    JOURNAL OF PLANT RESEARCH   Vol. 125 ( 5 ) page: 661 - 668   2012.9

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    Leaf primordia with high division and developmental competencies are generated around the periphery of stem cells at the shoot apex. Arabidopsis ASYMMETRIC-LEAVES2 (AS2) protein plays a key role in the regulation of many genes responsible for flat symmetric leaf formation. The AS2 gene, expressed in leaf primordia, encodes a plant-specific nuclear protein containing an AS2/LOB domain with cysteine repeats (C-motif). AS2 proteins are present in speckles in and around the nucleoli, and in the nucleoplasm of some leaf epidermal cells. We used the tobacco cultured cell line BY-2 expressing the AS2-fused yellow fluorescent protein to examine subnuclear localization of AS2 in dividing cells. AS2 mainly localized to speckles (designated AS2 bodies) in cells undergoing mitosis and distributed in a pairwise manner during the separation of sets of daughter chromosomes. Few interphase cells contained AS2 bodies. Deletion analyses showed that a short stretch of the AS2 amino-terminal sequence and the C-motif play negative and positive roles, respectively, in localizing AS2 to the bodies. These results suggest that AS2 bodies function to properly distribute AS2 to daughter cells during cell division in leaf primordia; and this process is controlled at least partially by signals encoded by the AS2 sequence itself.

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  30. Programmed induction of endoreduplication by DNA double-strand breaks in Arabidopsis Reviewed

    Sumiko Adachi, Kazunori Minamisawa, Yoko Okushima, Soichi Inagaki, Kaoru Yoshiyama, Youichi Kondou, Eli Kaminuma, Mika Kawashima, Tetsuro Toyoda, Minami Matsui, Daisuke Kurihara, Sachihiro Matsunaga, Masaaki Umeda

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   Vol. 108 ( 24 ) page: 10004 - 10009   2011.6

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    Genome integrity is continuously threatened by external stresses and endogenous hazards such as DNA replication errors and reactive oxygen species. The DNA damage checkpoint in metazoans ensures genome integrity by delaying cell-cycle progression to repair damaged DNA or by inducing apoptosis. ATM and ATR (ataxia-telangiectasia-mutated and -Rad3-related) are sensor kinases that relay the damage signal to transducer kinases Chk1 and Chk2 and to downstream cell-cycle regulators. Plants also possess ATM and ATR orthologs but lack obvious counterparts of downstream regulators. Instead, the plant-specific transcription factor SOG1 (suppressor of gamma response 1) plays a central role in the transmission of signals from both ATM and ATR kinases. Here we show that in Arabidopsis, endoreduplication is induced by DNA double-strand breaks (DSBs), but not directly by DNA replication stress. When root or sepal cells, or undifferentiated suspension cells, were treated with DSB inducers, they displayed increased cell size and DNA ploidy. We found that the ATM-SOG1 and ATR-SOG1 pathways both transmit DSB-derived signals and that either one suffices for endocycle induction. These signaling pathways govern the expression of distinct sets of cell-cycle regulators, such as cyclin-dependent kinases and their suppressors. Our results demonstrate that Arabidopsis undergoes a programmed endoreduplicative response to DSBs, suggesting that plants have evolved a distinct strategy to sustain growth under genotoxic stress.

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  31. Identification and characterization of plant Haspin kinase as a histone H3 threonine kinase Reviewed

    Daisuke Kurihara, Sachihiro Matsunaga, Tomohiro Omura, Tetsuya Higashiyama, Kiichi Fukui

    BMC PLANT BIOLOGY   Vol. 11   2011.4

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    Background: Haspin kinases are mitotic kinases that are well-conserved from yeast to human. Human Haspin is a histone H3 Thr3 kinase that has important roles in chromosome cohesion during mitosis. Moreover, phosphorylation of histone H3 at Thr3 by Haspin in fission yeast, Xenopus, and human is required for accumulation of Aurora B on the centromere, and the subsequent activation of Aurora B kinase activity for accurate chromosome alignment and segregation. Although extensive analyses of Haspin have been carried out in yeast and animals, the function of Haspin in organogenesis remains unclear.
    Results: Here, we identified a Haspin kinase, designated AtHaspin, in Arabidopsis thaliana. The purified AtHaspin phosphorylated histone H3 at both Thr3 and Thr11 in vitro. Live imaging of AtHaspin-tdTomato and GFP alpha tubulin in BY-2 cells showed that AtHaspin-tdTomato localized on chromosomes during prometaphase and metaphase, and around the cell plate during cytokinesis. This localization of AtHaspin overlapped with that of phosphorylated Thr3 and Thr11 of histone H3 in BY-2 cells. AtHaspin-GFP driven by the native promoter was expressed in root meristems, shoot meristems, floral meristems, and throughout the whole embryo at stages of high cell division. Overexpression of a kinase domain mutant of AtHaspin decreased the size of the root meristem, which delayed root growth.
    Conclusions: Our results indicated that the Haspin kinase is a histone H3 threonine kinase in A. thaliana. AtHaspin phosphorylated histone H3 at both Thr3 and Thr11 in vitro. The expression and dominant-negative analysis showed that AtHaspin may have a role in mitotic cell division during plant growth. Further analysis of coordinated mechanisms involving Haspin and Aurora kinases will shed new light on the regulation of chromosome segregation in cell division during plant growth and development.

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  32. Live-Cell Imaging Reveals the Dynamics of Two Sperm Cells during Double Fertilization in Arabidopsis thaliana Reviewed

    Yuki Hamamura, Chieko Saito, Chie Awai, Daisuke Kurihara, Atsushi Miyawaki, Tsuyoshi Nakagawa, Masahiro M. Kanaoka, Narie Sasaki, Akihiko Nakano, Frederic Berger, Tetsuya Higashiyama

    CURRENT BIOLOGY   Vol. 21 ( 6 ) page: 497 - 502   2011.3

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    Flowering plants have evolved a unique reproductive process called double fertilization, whereby two dimorphic female gametes are fertilized by two immotile sperm cells conveyed by the pollen tube [1, 2]. The two sperm cells are arranged in tandem with a leading pollen tube nucleus to form the male germ unit and are placed under the same genetic controls [3]. Genes controlling double fertilization have been identified [4-6], but whether each sperm cell is able to fertilize either female gamete is still unclear [7-9]. The dynamics of individual sperm cells after their release in the female tissue remain largely unknown. In this study, we photolabeled individual isomorphic sperm cells before their release and analyzed their fate during double fertilization in Arabidopsis thaliana. We found that sperm delivery was composed of three steps. Sperm cells were projected together to the boundary between the two female gametes. After a long period of immobility, each sperm cell fused with either female gamete in no particular order, and no preference was observed for either female gamete. Our results suggest that the two sperm cells at the front and back of the male germ unit are functionally equivalent and suggest unexpected cell-cell communications required for sperm cells to coordinate double fertilization of the two female gametes.

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  33. Chemical Visualization of an Attractant Peptide, LURE Reviewed

    Hiroaki Goto, Satohiro Okuda, Akane Mizukami, Hitoshi Mori, Narie Sasaki, Daisuke Kurihara, Tetsuya Higashiyama

    PLANT AND CELL PHYSIOLOGY   Vol. 52 ( 1 ) page: 49 - 58   2011.1

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    The pollen tube attractant peptide LUREs of Torenia fournieri are diffusible peptides that attract pollen tubes in vitro. Here, we report a method enabling the direct visualization of a LURE peptide without inhibiting its attraction activity by conjugating it with the Alexa Fluor 488 fluorescent dye. After purifying and refolding the recombinant LURE2 with a polyhistidine tag, its amino groups were targeted for conjugation with the Alexa Fluor dye. Labeling of LURE2 was confirmed by its fluorescence and mass spectrometry. In our in vitro assay using gelatin beads, Alexa Fluor 488-labeled LURE2 appeared to have the same activity as unlabeled LURE2. Using the labeled LURE2, the relationship between the spatiotemporal change of distribution and activity of LURE2 was examined. LURE2 attracted pollen tubes when embedded in gelatin beads, but hardly at all when in agarose beads. Direct visualization suggested that the significant difference between these conditions was the retention of LURE2 in the gelatin bead, which might delay diffusion of LURE2 from the bead. Direct visualization of LURE peptide may open the way to studying the spatiotemporal dynamics of LURE in pollen tube attraction.

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  34. Live cell imaging reveals plant Aurora kinase has dual roles during mitosis Reviewed

    Daisuke Kurihara, Sachihiro Matsunaga, Susumu Uchiyama, Kiichi Fukui

    PLANT AND CELL PHYSIOLOGY   Vol. 49 ( 8 ) page: 1256 - 1261   2008.8

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    The proper segregation of chromosomes during mitosis is required for accurate distribution of genetic information by two daughter cells. Here, we used live cell imaging of microtubules and kinetochores after treatment with an Aurora kinase inhibitor, hesperadin, in tobacco BY-2 cells to analyze the function of plant Aurora kinase during mitosis. Hesperadin treatment induced the delay of CenH3 alignment on the spindle equator. Furthermore, two types of dynamics of lagging CenH3s were observed during chromosome segregation. The findings indicate that the plant Aurora kinase has dual roles; correction of aberrant kinetochoremicrotubule attachment and dissociation of cohesin during chromosome alignment and segregation.

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  35. Visualization of mitotic HeLa cells by advanced polarized light microscopy Reviewed

    Akihiro Morimoto, Sachihiro Matsunaga, Daisuke Kurihara, Kiichi Fukui

    MICRON   Vol. 39 ( 5 ) page: 635 - 638   2008.7

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    The conventional polarized light imaging system can observe sub-microscopic molecular order non-destructively, quantitatively and without labeling or staining. Recently, a more sophisticated version, Abrio (TM) imaging system, than the conventional polarized light imaging system, was developed. This advanced polarized light imaging system has simplified the process of birefringent imaging, higher sensitivity and accuracy irrespective of sample orientation. By performing time-lapse observations using the advanced polarized light microscopy, we visualized mitotic spindles, especially kinetochore microtubules, of HeLa cells. Moreover, we successfully visualized the detailed structure of several filament bundles, which possibly are components of the contractile ring. Here, we report the potential of advanced polarized light imaging systems for imaging mitotic HeLa cells and the new insights obtained during this microscopic Study. (c) 2007 Elsevier Ltd. All rights reserved.

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  36. The Arabidopsis SDG4 contributes to the regulation of pollen tube growth by methylation of histone H3 lysines 4 and 36 in mature pollen Reviewed

    Joyce A. Cartagena, Sachihiro Matsunaga, Motoaki Seki, Daisuke Kurihara, Masami Yokoyama, Kazuo Shinozaki, Satoru Fujimoto, Yoshitaka Azumi, Susumu Uchiyama, Kiich Fukui

    DEVELOPMENTAL BIOLOGY   Vol. 315 ( 2 ) page: 355 - 368   2008.3

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    Plant SET domain proteins are known to be involved in the epigenetic control of gene expression during plant development. Here, we report that the Arabidopsis SET domain protein, SDG4, contributes to the epigenetic regulation of pollen tube growth, thus affecting fertilization. Using an SDG4-GFP fusion construct, the chromosomal localization of SDG4 was established in tobacco BY-2 cells. In Arabidopsis, sdg4 knockout showed reproductive defects. Tissue-specific expression analyses indicated that SDG4 is the major ASH1-related gene expressed in the pollen. Immunological analyses demonstrated that SDG4 was involved in the methylation of historic H3 in the inflorescence and pollen grains. The significant reduction in the amount of methylated historic H3 K4 and K36 in sdg4 pollen vegetative nuclei resulted in suppression of pollen tube growth. Our results indicate that SDG4 is capable of modulating the expression of genes that function in the growth of pollen tube by methylation of specific lysine residues of the histone H3 in the vegetative nuclei. (C) 2007 Elsevier Inc. All rights reserved.

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  37. Selective labeling of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein Reviewed International journal

    Wataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Daisuke Kurihara, Shin-Ichi Arimura, Nobuhiro Tsutsumi, Kiichi Fukui, Kazuyoshi Itoh

    Progress in Biomedical Optics and Imaging - Proceedings of SPIE   Vol. 6860   2008

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    We present space-selective labeling of organelles by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. Two-photon excitation of photoconvertible fluorescent-protein, Kaede, enables space-selective labeling of organelles. We alter the fluorescence of target mitochondria in a tobacco BY-2 cell from green to red by focusing femtosecond laser pulses with a wavelength of 750 nm.

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  38. Selective labeling of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein Reviewed International journal

    Wataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Daisuke Kurihar, Shin-ichi Arimura, Nobuhiro Tsutsumi, Kiichi Fukui, Kazuyoshi Itoh

    MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES VIII   Vol. 6860   2008

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    We present space-selective labeling of organelles by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. Two-photon excitation of photoconvertible fluorescent-protein, Kaede, enables space-selective labeling of organelles. We alter the fluorescence of target mitochondria in a tobacco BY-2 cell from green to red by focusing femtosecond laser pulses with a wavelength of 750 nm.

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    Other Link: http://orcid.org/0000-0002-9537-1626

  39. PHB2 protects sister-chromatid cohesion in mitosis Reviewed

    Hideaki Takata, Sachihiro Matsunaga, Akihiro Morimoto, Nan Ma, Daisuke Kurihara, Rika Ono-Maniwa, Masatoshi Nakagawa, Takachika Azuma, Susumu Uchiyama, Kiichi Fukui

    CURRENT BIOLOGY   Vol. 17 ( 15 ) page: 1356 - 1361   2007.8

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    Cohesion between sister chromatids is essential for proper chromosome segregation in mitosis. In vertebrate mitotic cells, most cohesin is removed from the chromosome arms [1-4], but centromeric cohesin is protected by shugoshin until the onset of anaphase [5]. However, the mechanism of this protection of centromeric cohesion is not well understood. Here, we demonstrate that prohibitin 2 (PHB2) is involved in the regulation of sister-chromatid cohesion during mitosis in HeLa cells. PHB2 is an evolutionarily conserved protein in eukaryotes and has multiple functions, such as transcriptional regulation and cell viability and development [6-8]. However, its functions in mitosis have not yet been determined. We show that depletion of PHB2 by RNA interference (RNAi) causes premature sister-chromatid separation and defects in chromosome congression accompanied by mitotic arrest by spindle-checkpoint activation. In the absence of PHB2, cohesin is dissociated from centromeres during early mitosis, although the centromeric localization of shugoshin is preserved. Thus, our findings suggest that, in addition to the shugoshin, PHB2 is also required to protect the centromeric cohesion from phosphorylation by PIk1 during early mitosis and that its function is essential for proper mitotic progression.

    DOI: 10.1016/j.cub.2007.07.009

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  40. Single-organelle tracking by two-photon conversion Reviewed

    Wataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Daisuke Kurihara, Kiichi Fukui, Shin-ichi Arimura, Nobuhiro Tsutsumi, Keisuke Isobe, Kazuyoshi Itoh

    OPTICS EXPRESS   Vol. 15 ( 5 ) page: 2490 - 2498   2007.3

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    Spatial and temporal information about intracellular objects and their dynamics within a living cell are essential for dynamic analysis of such objects in cell biology. A specific intracellular object can be discriminated by photoactivatable fluorescent proteins that exhibit pronounced light-induced spectral changes. Here, we report on selective labeling and tracking of a single organelle by using two-photon conversion of a photoconvertible fluorescent protein with near-infrared femtosecond laser pulses. We performed selective labeling of a single mitochondrion in a living tobacco BY-2 cell using two-photon photoconversion of Kaede. Using this technique, we demonstrated that, in plants, the directed movement of individual mitochondria along the cytoskeletons was mediated by actin filaments, whereas microtubules were not required for the movement of mitochondria. This single-organelle labeling technique enabled us to track the dynamics of a single organelle, revealing the mechanisms involved in organelle dynamics. The technique has potential application in direct tracking of selective cellular and intracellular structures. (c) 2007 Optical Society of America.

    DOI: 10.1364/OE.15.002490

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  41. Characterization of a splicing variant of plant Aurora kinase Reviewed

    Daisuke Kurihara, Akira Kawabe, Sachihiro Matsunaga, Katsuyuki Nakagawa, Satoru Fujimoto, Susumu Uchiyama, Kiichi Fukui

    PLANT AND CELL PHYSIOLOGY   Vol. 48 ( 2 ) page: 369 - 374   2007.2

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS  

    Aurora kinases play a key role in chromosome segregation and cytokinesis. In plants, three Aurora kinases (AtAUR1-AtAUR3) have been identified in Arabidopsis thaliana. Here, we report an AtAUR2 splicing variant (AtAUR2S), which lacks the fourth exon encoding a part of the kinase domain of AtAUR2. AtAUR2S was shown to have lost its kinase activity to phosphorylate histone H3 at Ser10; however, it maintained its ability to bind to histone H3. The localization pattern of AtAUR2S was the same as that of AtAUR2. The findings suggest that AtAUR2S affects cell division by competing with AtAUR2.

    DOI: 10.1093/pcp/pcl064

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  42. Tracking a single organelle with two-photon protein conversion Reviewed International journal

    Wataru Watanabe, Tomoko Shimada, Sachihiro Matsunaga, Daisuke Kurihara, Shin-Ichi Arimura, Nobuhiro Tsutsumi, Kiichi Fukui, Kazuyoshi Itoh

    Optics and Photonics News   Vol. 18 ( 12 ) page: 20   2007

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    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:Optical Society of American (OSA)  

    DOI: 10.1364/OPN.18.12.000020

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    Other Link: http://orcid.org/0000-0002-9537-1626

  43. Aurora kinase is required for chromosome segregation in tobacco BY-2 cells Reviewed

    Daisuke Kurihara, Sachihiro Matsunaga, Akira Kawabe, Satoru Fujimoto, Masanori Noda, Susumu Uchiyama, Kiichi Fukui

    PLANT JOURNAL   Vol. 48 ( 4 ) page: 572 - 580   2006.11

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL PUBLISHING  

    Post-translational modifications of core histone tails play crucial roles in chromatin structure and function. Although phosphorylation of Ser10 and Ser28 (H3S10ph and H3S28ph) of histone H3 is ubiquitous among eukaryotes, the phosphorylation mechanism during the cell cycle remains unclear. In the present study, H3S10ph and H3S28ph in tobacco BY-2 cells were observed in the pericentromeric regions during mitosis. Moreover, the Aurora kinase inhibitor Hesperadin inhibited the kinase activity of Arabidopsis thaliana Aurora kinase 3 (AtAUR3) in phosphorylating both Ser10 and Ser28 of histone H3 in vitro. Consistently, Hesperadin inhibited both H3S10ph and H3S28ph during mitosis in BY-2 cells. These results indicate that plant Aurora kinases phosphorylate not only Ser10, but also Ser28 of histone H3 in vivo. Hesperadin treatment increased the ratio of metaphase cells, while the ratio of anaphase/telophase cells decreased, although the mitotic index was not affected in Hesperadin-treated cells. These results suggest that Hesperadin induces delayed transition from metaphase to anaphase, and early exit from mitosis after chromosome segregation. In addition, micronuclei were observed frequently and lagging chromosomes, caused by the delay and failure of sister chromatid separation, were observed at anaphase and telophase in Hesperadin-treated BY-2 cells. The data obtained here suggest that plant Aurora kinases and H3S10ph/H3S28ph may have a role in chromosome segregation and metaphase/anaphase transition.

    DOI: 10.1111/j.1365-313X.2006.02893.x

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  44. Characterization of plant Aurora kinases during mitosis Reviewed

    A Kawabe, S Matsunaga, K Nakagawa, D Kurihara, A Yoneda, S Hasezawa, S Uchiyama, K Fukui

    PLANT MOLECULAR BIOLOGY   Vol. 58 ( 1 ) page: 1 - 13   2005.5

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    The Aurora kinase family is a well-characterized serine/threonine protein kinase family that regulates different processes of mitotic events. Although functions of animal and yeast Aurora kinases have been analyzed, plant aurora kinases were not identified and characterized. We identified three Aurora kinase orthologs in Arabidopsis thaliana and designated these as AtAUR1, AtAUR2, and AtAUR3. These AtAURs could phosphorylate serine 10 in histone H3, in vitro. Dynamic analyses of GFP-fused AtAUR proteins revealed that AtAUR1 and AtAUR2 localized at the nuclear membrane in interphase and located in mitotic spindles during cell division. AtAUR1 also localized in the cell plates. AtAUR3 showed dot-like distribution on condensed chromosomes at prophase and then localized at the metaphase plate. At late anaphase, AtAUR3 is evenly localized on chromosomes. The localization of AtAUR3 during mitosis is very similar to that of phosphorylated histone H3. Interestingly, an overexpression of AtAUR3 induces disassembly of spindle microtubules and alteration of orientation of cell division. Our results indicate that plant Aurora kinases have different characters from that of Aurora kinases of other eukaryotes.

    DOI: 10.1007/s11103-005-3454-x

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Books 21

  1. 透明化技術を用いた植物組織蛍光観察のすすめ

    栗原大輔, 水多陽子( Role: Joint author)

    Plant Morphology  2017.7 

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    Language:Japanese Book type:Scholarly book

  2. イメージング技術を駆使して植物寄生性線虫の感染を捉える

    大津美奈, 栗原大輔, 東山哲也( Role: Joint author)

    BSJ-Review  2017.7 

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  3. 植物を丸ごと透明化し,中まで蛍光観察する新技術を開発 細胞レベルでの個体全体の観察を目指して

    栗原大輔( Role: Sole author)

    化学と生物  2016.10 

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    Language:Japanese Book type:General book, introductory book for general audience

  4. 植物組織透明化試薬ClearSeeの開発

    栗原大輔( Role: Sole author)

    和光純薬時報  2016.10 

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    Language:Japanese Book type:Scholarly book

  5. 植物研究における2光子励起顕微鏡の活用

    栗原大輔( Role: Sole author)

    BSJ-Review  2016.5 

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    Language:Japanese Book type:Scholarly book

  6. 植物の透明化試薬「クリアシー」で線虫の侵入状況など観察可能に

    栗原大輔( Role: Sole author)

    ニューカントリー 5月号  2016.4 

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    Language:Japanese Book type:General book, introductory book for general audience

  7. 透明にすることによって見えてくるもの

    栗原大輔( Role: Sole author)

    現代化学  2016.2 

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    Language:Japanese Book type:General book, introductory book for general audience

  8. Visualization of Plant Sexual Reproduction in the Whole-mount Pistil by ClearSee

    ( Role: Joint author)

    2016.1 

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    Language:English Book type:Scholarly book

  9. 植物の細胞板形成を支えるM期キネシンNACK1とMAPKカスケード

    笹部美知子, 桧垣匠, 栗原大輔, 栗原大輔, 東山哲也, 東山哲也, 東山哲也, 馳澤盛一郎, 町田泰則( Role: Joint author)

    バイオイメージング  2015.9 

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    Language:Japanese Book type:General book, introductory book for general audience

  10. 2光子励起顕微鏡を用いた生体深部イメージングと光顕微操作

    水多陽子, 栗原大輔( Role: Joint author)

    植物の生長調節  2014.12 

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  11. Kinetochore and microtubule dynamics during cell division of tobacco BY-2 cells by live-cell imaging

    ( Role: Joint author)

    2014.9 

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  12. 細胞分裂期において分裂期キナーゼが制御する染色体動態

    栗原大輔( Role: Sole author)

    Plant Morphology  2011.5 

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  13. 植物染色体の最前線 染色体動態―染色体の動き,動物と植物の共通点と違い

    栗原大輔, 松永幸大( Role: Joint author)

    生物の科学 遺伝  2009.5 

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    Language:Japanese Book type:General book, introductory book for general audience

  14. 染色体動態-染色体の動き、動物と植物の共通点と違い

    栗原大輔, 松永幸大( Role: Joint author)

    生物の科学遺伝  2009 

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  15. Plant Aurora kinase has dual roles on chromosome alignment and segregation during mitosis

    ( Role: Joint author)

    2008 

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  16. ASURAは体細胞分裂で姉妹染色分体の接着を保護する

    松永幸大, 高田英昭, 森本晃弘, MA Nan, 栗原大輔, 光島正浩, 内山進, 福井希一( Role: Joint author)

    生化学  2007 

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  17. 可視化技術による植物Auroraキナーゼの機能解析

    栗原大輔, 内山進, 松永幸大, 福井希一( Role: Joint author)

    生化学  2007 

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  18. 植物のヒストン翻訳後修飾研究の幕開け 分化発生,形態形成,ストレス応答を制御する“第2のコード”の意味とは?

    松永幸大, 栗原大輔, 中園幹生( Role: Joint author)

    化学と生物  2006.12 

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    Language:Japanese Book type:General book, introductory book for general audience

  19. Chromosome dynamics in Arabidopsis

    ( Role: Joint author)

    2006 

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    Language:English Book type:Scholarly book

  20. 植物におけるヒストン翻訳後修飾研究の幕開け

    松永幸大, 栗原大輔, 中園幹生( Role: Joint author)

    化学と生物  2006 

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    Language:Japanese Book type:General book, introductory book for general audience

  21. Dynamics of the plant Aurora kinase AtAUR3 during mitosis and spindle abnormality induced by AtAUR3 overexpression

    ( Role: Joint author)

    2005 

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MISC 1

  1. Dynamics of the cell fate specifications during female gametophyte development in Arabidopsis International journal

    Daichi Susaki, Takamasa Suzuki, Daisuke Maruyama, Minako Ueda, Tetsuya Higashiyama, Daisuke Kurihara

    bioRxiv     2020.4

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    Authorship:Corresponding author   Language:English   Publisher:Cold Spring Harbor Laboratory  

    <title>ABSTRACT</title>The female gametophytes of angiosperms contain cells with distinct functions, such as those that enable reproduction via pollen tube attraction and fertilization. Although the female gametophyte undergoes unique developmental processes, such as several rounds of nuclear division without cell plate formation, and the final cellularization, it remains unknown when and how the cell fate is determined during their development. Here, we visualized the living dynamics of female gametophyte development and performed transcriptome analysis of its individual cell types, to assess the cell fate specifications in <italic>Arabidopsis thaliana</italic>. We recorded time lapses of the nuclear dynamics and cell plate formation from the one-nucleate stage to the seven-cell stage after cellularization, using the <italic>in vitro</italic> ovule culture system. The movies showed that the nuclear division occurred along the micropylar–chalazal axis. During cellularization, the polar nuclei migrated while associating with forming edge of the cell plate. Then, each polar nucleus migrated to fuse linearly towards each other. We also tracked the gene expression dynamics and identified that the expression of the <italic>MYB98pro∷GFP</italic>, a synergid-specific marker, was initiated before cellularization, and then restricted to the synergid cells after cellularization. This indicated that cell fates are determined immediately after cellularization. Transcriptome analysis of the female gametophyte cells of the wild type and <italic>myb98</italic> mutant, revealed that the <italic>myb98</italic> synergid cells had the egg cell-like gene expression profile. Although in the <italic>myb98</italic>, the egg cell-specific gene expressions were properly initiated only in the egg cells after cellularization, but subsequently expressed ectopically in one of the two synergid cells. These results, together with the various initiation timings of the egg cell-specific genes suggest the complex regulation of the individual gametophyte cells, such as cellularization-triggered fate initiation, MYB98-dependent fate maintenance, cell morphogenesis, and organelle positioning. Our system of live-cell imaging and cell-type-specific gene expression analysis provides insights into the dynamics and mechanisms of cell fate specifications in the development of female gametophytes in plants.

    DOI: 10.1101/2020.04.07.023028

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Presentations 147

  1. トマト果実におけるGCaMP6用いたカルシウムイメージング

    堀千秋, 栗原大輔, 大村道明, 西山学, 金山喜則, 加藤一幾

    園芸学会令和3年度春季大会  2021.3.27 

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    Event date: 2021.3.27 - 2021.3.28

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  2. シロイヌナズナ核融合欠損株で観察される受精後の胚発生異常の解析

    西川周一, 高木祐理, 佐藤譲, 栗原大輔, 佐藤良勝, 東山哲也, 丸山大輔

    第62回日本植物生理学会年会  2021.3.14 

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    Event date: 2021.3.14 - 2021.3.16

    Language:English   Presentation type:Oral presentation (general)  

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  3. 植物初期発生過程における細胞運命制御機構の解明 Invited

    栗原大輔

    『配偶子インテグリティの構築』『全能性プログラム』合同公開シンポジウム  2020.12.21 

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    Event date: 2020.12.21 - 2020.12.23

    Language:English   Presentation type:Oral presentation (general)  

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  4. シロイヌナズナ雌性配偶体細胞における運命決定のダイナミクス Invited

    栗原大輔, 須崎大地, 海老根一生, 鈴木孝征, 丸山大輔, 植田美那子, 東山哲也

    日本植物学会第84回大会  2020.9.19 

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    Event date: 2020.9.19 - 2020.9.21

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  5. 動物培養細胞を用いた動植物の受精因子群の接着性の検証

    中島耕大, Clari Valansi, 栗原大輔, 佐々木成江, Benjamin Podbilewicz, 東山哲也

    日本植物学会第84回大会  2020.9.19 

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    Event date: 2020.9.19 - 2020.9.21

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  6. シロイヌナズナ雌性配偶体細胞のRNA-seq解析

    須崎大地, 栗原大輔, 鈴木孝征, 丸山大輔, 植田美那子, 東山哲也

    日本植物学会第84回大会  2020.9.21 

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    Event date: 2020.9.19 - 2020.9.21

    Language:English   Presentation type:Poster presentation  

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  7. 受精卵の内部動態から迫る植物の体軸形成機構 Invited

    植田 美那子, 木全 祐資, 松本 光梨, 檜垣 匠, 栗原 大輔, 東山 哲也

    日本植物学会第84回大会  2020.9.19 

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    Event date: 2020.9.19 - 2020.9.21

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  8. 葉形成に関与するAS2タンパク質の動態変化と機能の関係

    笹部美知子, 雪森桃花, 吉田みのり, 三石萌, 小島晶子, 栗原大輔, 東山哲也, 町田千代子, 町田泰則

    日本植物学会第84回大会  2020.9.20 

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    Event date: 2020.9.19 - 2020.9.21

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  9. 花粉不発芽を引き起こすDPL遺伝子の機能解析

    水多陽子, 栗原大輔

    日本植物学会第84回大会  2020.9.19 

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    Event date: 2020.9.19 - 2020.9.21

    Language:English   Presentation type:Oral presentation (general)  

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  10. GCaMP6を用いたカルシウムイメージングのトマトへの応用

    堀千秋, 伊藤輝, 栗原大輔, 石田宏幸, 大村道明, 金山喜則, 加藤一幾

    園芸学会令和2年度春季大会  2020.3.21 

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    Event date: 2020.3.21 - 2020.3.22

    Language:English   Presentation type:Oral presentation (general)  

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  11. Live-cell analysis of female gametophyte development in Arabidopsis International conference

    Daisuke Kurihara, Daichi Susaki, Tetsuya Higashiyama

    Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture  2019.3.18 

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    Event date: 2019.3.18

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  12. シロイヌナズナにおける初期胚のライブイメージング

    植田美那子, 木全祐資, 田中小百合, 加藤壮英, 桧垣匠, 栗原大輔, 山田朋美, 安藤奈央惠, 森田(寺尾)美代, 瀬上紹嗣, 前島正義, 馳澤盛一郎, 桑田啓子, 佐藤綾人, 鈴木孝征, 東山哲也, 田坂昌生

    第60回日本植物生理学会年会  2019.3.14 

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    Country:Japan  

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  13. 花粉への一過的遺伝子導入系による花粉管及び精細胞の動態解析

    永原史織, 栗原大輔, 東山哲也, 水多陽子

    第60回日本植物生理学会年会  2019.3.14 

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    Event date: 2019.3.14

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    Country:Japan  

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  14. 花粉管をベクターとした生殖細胞の遺伝子改変と植物生殖機構の解明

    水多陽子, 永原史織, 栗原大輔, 東山哲也

    第60回日本植物生理学会年会  2019.3.13 

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    Event date: 2019.3.13

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Country:Japan  

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  15. 光ピンセットを用いた細胞操作による受精因子解析の試み

    中島耕大, 永原史織, 佐々木妙子, Clari Valansi, 栗原大輔, 佐藤良勝, 佐々木成江, Benjamin Podbilewicz, 東山哲也

    日本植物学会第82回大会  2018.9.15 

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    Event date: 2018.9.15

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  16. シロイヌナズナ胚のパターン形成におけるHD-ZIP IV転写因子群の機能の解析

    田中小百合, 栗原大輔, 柳沢直樹, 東山哲也, 植田美那子

    日本植物学会第82回大会  2018.9.15 

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    Event date: 2018.9.15

    Language:English   Presentation type:Oral presentation (general)  

    Country:Japan  

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  17. 植物透明化試薬ClearSeeの改良

    栗原大輔, 水多陽子, 永原史織, 東山哲也

    日本植物学会第82回大会  2018.9.15 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  18. さらなる植物透明化へ向けて

    栗原大輔, 水多陽子, 永原史織, 東山哲也

    日本植物形態学会第30回大会  2018.9.13 

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    Event date: 2018.9.13

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  19. 光細胞操作技術を用いて,受精因子の機能を探る

    中島耕大, 永原史織, 佐々木妙子, Clari Valansi, 栗原大輔, 佐藤良勝, 佐々木成江, Benjamin Podbilewicz, 東山哲也

    日本植物形態学会第30回大会  2018.9.13 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  20. Live-cell imaging of the axis formation in Arabidopsis zygote Invited International conference

    Minako Ueda, Yusuke Kimata, Takehide Kato, Takumi Higaki, Daisuke Kurihara, Tomomi Yamada, Shoji Segami, Miyo Terao Morita, Masayoshi Maeshima, Seiichiro Hasezawa, Tetsuya Higashiyama, Masao Tasaka, Naoe Ando

    the 25th International Congress on Sexual Plant Reproduction  2018.6.14 

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    Event date: 2018.6.14

    Language:English   Presentation type:Oral presentation (invited, special)  

    Country:Japan  

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  21. Imaging Analysis of mRNA decay mutants at early plant development International conference

    Kazuki Motomura, Daisuke Maruyama, Daisuke Kurihara, Naoyoshi Kumakura, Yuichiro Watanabe, Tetsuya Higashiyama

    the 25th International Congress on Sexual Plant Reproduction  2018.6.13 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  22. Calcium dynamics of the pollen tube in ovular guidance International conference

    Kumi Matsuura-Tokita, Yoko Mizuta, Daisuke Kurihara, Tetsuya Higashiyama

    the 25th International Congress on Sexual Plant Reproduction  2018.6.13 

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    Event date: 2018.6.13

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  23. Intracellular dynamics controlling Arabidopsis zygote polarization International conference

    Yusuke Kimata, Takehide Kato, Takumi Higaki, Daisuke Kurihara, Tomomi Yamada, Shoji Segami, Miyo Terao Morita, Masayoshi Maeshima, Seiichiro Hasezawa, Tetsuya Higashiyama, Masao Tasaka, Minako Ueda

    the 25th International Congress on Sexual Plant Reproduction  2018.6.12 

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    Event date: 2018.6.12

    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  24. Live-cell analysis of female gametophyte development in Arabidopsis International conference

    Daisuke Kurihara, Daichi Susaki, Tetsuya Higashiyama

    the 25th International Congress on Sexual Plant Reproduction  2018.6.12 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  25. Two-photon imaging reveals spatio-temporal regulation on the one-to-one pollen tube guidance International conference

    Yoko Mizuta, Daisuke Kurihara, Shiori Nagahara, Tetsuya Higashiyama

    the 25th International Congress on Sexual Plant Reproduction  2018.6.12 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  26. Synthetic biological approach to understand plant gametic interactions by micromanipulation techniques International conference

    Kohdai Nakajima, Taeko Sasaki, Shiori Nagahara, Clari Valansi, Daisuke Kurihara, Yoshikatsu Sato, Narie Sasaki, Benjamin Podbilewicz, Tetsuya Higashiyama

    the 25th International Congress on Sexual Plant Reproduction  2018.6.12 

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    Language:English   Presentation type:Poster presentation  

    Country:Japan  

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  27. 植物組織における透明化イメージング Invited International conference

    栗原大輔

    レーザ顕微鏡研究会講演会抄録集  2018.1.19 

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    Event date: 2018.1.19

    Language:Japanese   Presentation type:Poster presentation  

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  28. 生殖段階における個々のRNA分解経路のイメージング解析 International conference

    元村一基, 丸山大輔, 栗原大輔, 熊倉直祐, 渡邊雄一郎, 東山哲也, 東山哲也

    日本植物生理学会年会(Web)  2018 

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    Language:Japanese   Presentation type:Poster presentation  

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  29. 雌性配偶体形成過程における細胞運命制御機構の解析 International conference

    栗原大輔, 須崎大地, 東山哲也, 東山哲也

    日本植物学会大会研究発表記録  2017.9.9 

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    Event date: 2017.9.9

    Language:Japanese   Presentation type:Poster presentation  

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  30. シロイヌナズナ花粉管誘引のライブイメージング International conference

    時田公美, 水多陽子, 水多陽子, 栗原大輔, 東山哲也, 東山哲也

    日本植物学会大会研究発表記録  2017.9.1 

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  31. 時空間的制御に着目したRNA分解経路の比較解析 International conference

    元村一基, 丸山大輔, 栗原大輔, 熊倉直祐, 渡邊雄一郎, 東山哲也

    日本植物学会大会研究発表記録  2017.9.1 

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  32. 受精卵の不等分裂を制御する細胞骨格ダイナミクス International conference

    木全祐資, 檜垣匠, 河島友和, 河島友和, 栗原大輔, 栗原大輔, 佐藤良勝, 山田朋美, 山田朋美, 馳澤盛一郎, BERGER Frederic, 東山哲也, 東山哲也, 東山哲也, 植田美那子, 植田美那子

    日本植物学会大会研究発表記録  2017.9.1 

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  33. 受精による細胞極性の破壊と再構成~ライブイメージングで迫る受精前後の細胞内変化~ International conference

    植田美那子, 木全祐資, 加藤壮英, 檜垣匠, 山田朋美, 栗原大輔, 森田(寺尾)美代, 馳澤盛一郎, 東山哲也, 田坂昌生

    日本植物学会大会研究発表記録  2017.9.1 

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    Language:Japanese   Presentation type:Poster presentation  

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  34. Live imaging and optical manipulation of plant reproduction at a single cell level Invited

    Daisuke Kurihara

    Live imaging and optical manipulation of plant reproduction at a single cell level  2017.3.18 

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    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  35. ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging International conference

    Daisuke Kurihara, Yoko Mizuta, Yoshikatsu Sato, Tetsuya Higashiyama

    The 1st ABiS Symposium - Towards the Future of Advanced Bioimaging for Life Sciences -  2017.2.19 

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    Language:Japanese   Presentation type:Poster presentation  

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  36. パーティクルボンバードメント法による植物生殖細胞へのCRISPR/Cas9の導入 International conference

    永原史織, 栗原大輔, 東山哲也, 水多陽子

    日本生化学会大会(Web)  2017 

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  37. 植物のからだを覗いてみよう Invited International conference

    栗原大輔

    名古屋大学出前授業in豊橋  2016.12.4 

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  38. 今、顕微鏡技術で観えるもの Invited International conference

    栗原大輔

    第12回関西創農薬研究会  2016.11.11 

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    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  39. 植物深部に侵入した微生物を光学顕微鏡で観察する Invited

    栗原大輔, 大津美奈, 水多陽子, 東山哲也

    日本植物学会第80回大会  2016.9.17 

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  40. ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging Invited

    2016.9.15 

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    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  41. ClearSeeで観る植物生殖のメカニズム

    水多陽子, 栗原大輔, 東山哲也

    日本植物学会大会研究発表記録  2016.9.1 

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  42. 二光子励起イメージング技術を駆使したシストセンチュウが誘導する合胞体形成過程の観察

    大津美奈, 栗原大輔, 佐藤良勝, 丸山大輔, 東山哲也

    日本植物学会大会研究発表記録  2016.9.1 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  43. 植物受精卵のライブイメージング:細胞極性とは何か?

    木全祐資, 桧垣匠, 河島友和, 栗原大輔, 栗原大輔, 佐藤良勝, 山田朋美, 山田朋美, 馳澤盛一郎, BERGER Frederic, 東山哲也, 東山哲也, 東山哲也, 植田美那子, 植田美那子

    日本植物学会大会研究発表記録  2016.9.1 

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    Language:Japanese   Presentation type:Oral presentation (invited, special)  

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  44. 顕微鏡を基盤とした初期発生におけるmRNA分解機構の比較解析

    元村一基, 丸山大輔, 栗原大輔, 栗原大輔, 熊倉直祐, 渡邊雄一郎, 東山哲也, 東山哲也, 東山哲也

    日本植物学会大会研究発表記録  2016.9.1 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  45. Cell fate specification in Arabidopsis female gametophyte development International conference

    the 24th International Congress on Sexual Plant Reproduction  2016.3.18 

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  46. Live cell analysis and deep imaging for plant development Invited International conference

    2016.3.8 

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  47. 二光子顕微鏡を用いたシストセンチュウが誘導する合胞体形成時の細胞壁再構成過程の観察

    大津美奈, 寿崎拓哉, 佐藤良勝, 栗原大輔, 川口正代司, 丸山大輔, 東山哲也

    日本植物病理学会大会プログラム・講演要旨予稿集  2016.3.3 

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  48. 顕微鏡技術で切り拓く植物発生研究 Invited

    栗原 大輔

    第41回植物バイテクシンポジウム  2016.1.22 

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    Language:English   Presentation type:Oral presentation (invited, special)  

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  49. シロイヌナズナ花粉管の膜輸送のライブイメージング解析

    時田公美, 水多陽子, 栗原大輔, 栗原大輔, 東山哲也, 東山哲也, 東山哲也

    日本細胞生物学会大会(Web)  2016 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  50. 深部観察でWhole plant imagingを目指す Invited

    栗原大輔, 栗原大輔, 水多陽子, 水多陽子, 佐藤良勝, 東山哲也, 東山哲也, 東山哲也

    日本植物学会大会研究発表記録  2015.9.6 

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  51. M期特異的キネシンNACK1のC末端領域は細胞板形成部位への局在に必要である

    笹部美知子, 桧垣巧, 西田結花, 石橋奈々子, 栗原大輔, 東山哲也, 西浜竜一, 伊藤正樹, 馳澤盛一郎, 町田泰則

    日本植物学会大会研究発表記録  2015.9.1 

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  52. 一本の花粉管が一つの胚珠にたどりつくには?~深部イメージングで探る花粉管ガイダンスの謎~

    水多陽子, 栗原大輔, 東山哲也

    日本植物学会大会研究発表記録  2015.9.1 

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  53. シロイヌナズナ受精卵のイメージングとケミカルスクリーニング:細胞極性と不等分裂を制御する仕組みとは?

    植田美那子, 木全祐資, 栗原大輔, 山田朋美, 南保正和, 大川(西脇)妙子, 桑田啓子, 梅田正明, 梅田正明, 東山哲也

    日本植物学会大会研究発表記録  2015.9.1 

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  54. Live-cell imaging and optical manipulation of Arabidopsis early embryogenesis Invited International conference

    Daisuke Kurihara

    International ERATO Higashiyama Live-Holonics Symposium 2015  2015.8.27 

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    Event date: 2015.8.27

    Language:English   Presentation type:Oral presentation (invited, special)  

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  55. シロイヌナズナ雌性配偶体発生における細胞個性獲得変異体の解析

    栗原大輔, 東山哲也

    Plant Morphology  2015.4 

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  56. 生体深部イメージングによる花粉管ガイダンスの「形」と「通り道」の解析

    水多陽子, 栗原大輔, 東山哲也

    Plant Morphology  2015.4 

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  57. アポミクシス性特異的遺伝子の機能解析―組換えシロイヌナズナでのASG‐1遺伝子の時空的な発現

    CHEN L, 西村佳子, 吉田薫, 鉄村琢哉, 杉田亘, 栗原大輔, 東山哲也

    育種学研究  2015.3.21 

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  58. 受精卵の極性化動態~植物の体軸形成のしくみ~

    木全祐資, 栗原大輔, 東山哲也, 植田美那子

    日本生化学会大会(Web)  2015 

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  59. アポミクシス特異的遺伝子の機能解析―ASG‐1遺伝子組換えシロイヌナズナでの多胚嚢形成と多胚形成現象の解析―

    CHEN L, 西村佳子, 吉田薫, 鉄村琢哉, 杉田亘, 栗原大輔, 東山哲也

    育種学研究  2014.9.26 

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  60. アポミクシス特異的遺伝子の機能解析―イネ科植物へのASG‐1遺伝子導入

    西村佳子, 吉田薫, 杉田亘, 栗原大輔, 東山哲也, CHEN L

    育種学研究  2014.9.26 

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  61. シロイヌナズナ初期胚における細胞運命決定機構の解析

    栗原大輔, 牛王啓太, 有賀花奈, 植田美那子, 東山哲也

    日本植物学会第78回大会  2014.9.12 

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  62. シロイヌナズナ雌性配偶体発生における細胞個性獲得変異体の解析

    栗原 大輔, 東山 哲也

    日本植物形態学会第26回総会・大会  2014.9.11 

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  63. Live-cell analysis of Arabidopsis early embryogenesis Invited International conference

    Daisuke Kurihara

    International ERATO Higashiyama Live-Holonics Symposium 2014  2014.9.9 

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    Language:English   Presentation type:Oral presentation (invited, special)  

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  64. 2光子顕微鏡を用いた花粉管ガイダンスのin vivoライブイメージング

    水多陽子, 栗原大輔, 東山哲也

    日本植物学会大会研究発表記録  2014.9.1 

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  65. シロイヌナズナ初期胚における細胞運命決定機構の解析

    栗原大輔, 牛王啓太, 有賀花奈, 植田美那子, 東山哲也

    日本植物学会大会研究発表記録  2014.9.1 

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  66. Live-cell analysis to reveal the cell fate determination during early embryogenesis in Arabidopsis thaliana International conference

    Daisuke Kurihara, Keita Gooh, Minako Ueda, Tetsuya Higashiyama

    23rd International Congress on Sexual Plant Reproduction  2014.7.14 

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  67. The functional analysis of apomixis specific genes: Plant regeneration and its gene expression of HSP::ASG-1::GFP transgenic Arabidopsis

    西村佳子, 吉田薫, 鉄村琢哉, 杉田亘, 栗原大輔, 東山哲也, CHEN Lanzhuang

    南九州大学研究報告 自然科学編  2014.4.1 

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  68. In vivo live-cell imaging in plant tissues by two-photon excitation microscopy

    水多陽子, 栗原大輔, 東山哲也

    Plant Morphol  2014.4 

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  69. シロイヌナズナ胚発生過程のライブイメージング―マイクロデバイスを用いたアプローチ

    栗原大輔, 牛王啓太, 朴鍾淏, 新田英之, 東山哲也

    Plant Morphol  2014.4 

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  70. 細胞周期を制御するオーロラキナーゼのイメージング解析

    北原英里奈, 坂本卓也, 伊藤正樹, 松永朋子, 栗原大輔, 松永幸大

    Plant Morphol  2014.4 

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  71. 花粉管のin vivoイメージングで観えてきた植物生殖の実態―2光子顕微鏡を用いたアプローチ―

    水多陽子, 栗原大輔, 東山哲也

    Plant Morphol  2014.4 

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  72. アポミクシス性特異的遺伝子の機能解析―ギニアグラスの完熟種子由来カルスを用いたASG‐1組換え体作出の諸条件検討―

    西村佳子, 鉄村琢哉, 吉田薫, 栗原大輔, 東山哲也, 杉田亘, CHEN L

    育種学研究  2014.3.21 

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  73. 多胚嚢胚珠および多胚形成現象がASG‐1遺伝子組換えシロイヌナズナで現れる

    CHEN L, 西村佳子, 鉄村琢哉, 吉田薫, 栗原大輔, 東山哲也, 杉田亘

    育種学研究  2014.3.21 

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  74. シロイヌナズナの葉の向背軸分化に関わるAS2タンパク質の分裂期における核小体周縁部局在の解析

    町田千代子, LILAN Luo, 石橋奈々子, 栗原大輔, 氣多澄江, 町田泰則

    日本分子生物学会年会プログラム・要旨集(Web)  2014 

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  75. 光顕微操作で植物胚発生に迫る Invited

    栗原 大輔

    シンポジウム「細胞を創る操る」  2013.11.28 

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  76. 長期間観察を目的にする胚珠固定用のマイクロケージアレイの作製

    朴鍾淏, 栗原大輔, 東山哲也, 新田英之

    日本機械学会マイクロ・ナノ工学シンポジウム講演論文集  2013.11.4 

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  77. アポミクシス性特異的遺伝子の機能解析―ASG‐1遺伝子組換えシロイヌナズスでの乾燥耐性評価―

    西村佳子, 鉄村琢哉, 吉田薫, 杉田亘, 栗原大輔, 東山哲也, LI L, CHEN L

    育種学研究  2013.10.12 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  78. マイクロデバイスを用いたシロイヌナズナ胚発生過程のライブイメージング

    栗原大輔, 牛王啓太, 朴鍾淏, 新田英之, 東山哲也

    日本植物学会大会研究発表記録  2013.9.13 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  79. シロイヌナズナ胚発生過程のライブイメージング―マイクロデバイスを用いたアプローチ

    栗原大輔, 牛王啓太, 朴鍾淏, 新田英之, 東山哲也

    日本植物形態学会第25回総会・大会  2013.9.12 

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    Language:English   Presentation type:Poster presentation  

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  80. 2光子顕微鏡による深部イメージングで観えてきた植物生殖の実態

    水多陽子, 栗原大輔, 東山哲也

    日本植物学会大会研究発表記録  2013.8.20 

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    Event date: 2013.8.20

    Language:Japanese   Presentation type:Oral presentation (general)  

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  81. オーロラキナーゼの細胞周期のイメージング解析

    北原英里奈, 坂本卓也, 伊藤正樹, 松永朋子, 小牧伸一郎, 栗原大輔, 杉本慶子, 松永幸大

    日本植物学会大会研究発表記録  2013.8.20 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  82. Live-cell analysis of embryogenesis in Arabidopsis thaliana International conference

    Daisuke Kurihara, Keita Gooh, Tetsuya Higashiyama

    the 24th International Conference on Arabidopsis Research  2013.6.25 

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    Event date: 2013.6.25

    Language:English   Presentation type:Poster presentation  

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  83. 光顕微操作とライブイメージングで捉える植物胚発生

    栗原大輔

    2013年度細胞周期合同セミナー  2013.6.7 

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    Language:English   Presentation type:Oral presentation (general)  

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  84. 長時間ライブイメージングを実現する胚珠配列用マイクロデバイス

    朴鍾淏, 栗原大輔, 東山哲也, 新田英之

    化学とマイクロ・ナノシステム学会研究会講演要旨集  2013.5.23 

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    Event date: 2013.5.23

    Language:Japanese   Presentation type:Oral presentation (general)  

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  85. シロイヌナズナin vitro胚発生系の開発を基盤とした光顕微操作による胚発生過程の解析

    牛王啓太, 栗原大輔, 栗原大輔, 浜村有希, 東山哲也, 東山哲也

    Plant Morphol  2013.4 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  86. ホロニックコミュニケーションを担うシグナリング分子の可視化に向けて

    栗原大輔, 東山哲也

    Plant Morphol  2013.4 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  87. シロイヌナズナにおけるオーロラキナーゼのイメージング解析

    北原英里奈, 松永朋子, 伊藤正樹, 坂本卓也, 小牧伸一郎, 石田喬志, 栗原大輔, 杉本慶子, 松永幸大

    Plant Morphol  2013.4 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  88. アポミクシス性特異的遺伝子の機能解析―ASG1::GFP遺伝子組換えシロイヌナズナでの蛍光発現―

    西村佳子, 鉄村琢哉, 杉田亘, 長田龍太郎, 吉田薫, 栗原大輔, 東山哲也, CHEN L

    育種学研究  2013.3.27 

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    Event date: 2013.3.27

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  89. アポミクシス性特異的遺伝子の機能解析―ASG1::GFP遺伝子組換えシロイヌナズナを用いたPCR解析と変異体検出―

    内藤琢仁, 西村佳子, 鉄村琢哉, 杉田亘, 長田龍太郎, 吉田薫, 栗原大輔, 東山哲也, CHEN L

    育種学研究  2013.3.27 

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  90. シロイヌナズナsemi‐in vivo受精系における卵細胞のカルシウム動態解析

    浜村有希, 西巻萌, 栗原大輔, 東山哲也

    日本植物生理学会年会要旨集  2013.3.14 

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  91. 植物の動原体局在型オーロラキナーゼは微小管プラス末端制御タンパク質EB1をリン酸化する

    松永幸大, 北原英里奈, 栗原大輔, 小牧伸一郎, 高木麻衣, 佐野行己, 長島慶宜, 中神弘史, 伊藤正樹, 杉本慶子, 橋本隆, 松永朋子, 坂本卓也

    日本分子生物学会年会プログラム・要旨集(Web)  2013 

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  92. 組織深部のライブイメージングと光細胞操作で花の内部を探る Invited

    東山哲也, 栗原大輔

    日本女子大学バイオイメージングセンター第4回公開シンポジウム  2012.10.27 

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    Language:English   Presentation type:Oral presentation (invited, special)  

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  93. 光顕微操作とライブイメージングで迫るパターン形成

    栗原大輔, 牛王啓太, 東山哲也

    日本植物学会大会研究発表記録  2012.9.17 

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    Event date: 2012.9.17

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  94. アポミクシス特異的遺伝子の機能解析―hsp::ASG‐1::GFPを遺伝子導入したシロイヌナズナ組換え体の作出―

    西村佳子, 鉄村琢哉, 吉田薫, 杉田亘, 長田龍太郎, 栗原大輔, 東山哲也, CHEN L

    育種学研究  2012.9.14 

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    Language:Japanese   Presentation type:Oral presentation (general)  

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  95. シロイヌナズナin vitro胚発生系の開発を基盤とした初期胚発生のライブセル解析

    牛王啓太, 栗原大輔, 東山哲也

    日本植物学会大会研究発表記録  2012.9.14 

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  96. オーロラキナーゼのノックダウン個体のイメージング解析

    北原英里奈, 松永朋子, 伊藤正樹, 坂本卓也, 小牧伸一郎, 石田喬志, 栗原大輔, 杉本慶子, 松永幸大

    日本植物学会大会研究発表記録  2012.9.14 

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  97. カルシウム動態に着目した重複受精機構の解析

    西巻萌, 浜村有希, 栗原大輔, 東山哲也

    日本植物生理学会年会要旨集  2012.3.9 

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  98. シロイヌナズナにおける扁平で左右相称な葉の形成の仕組み:ASYMMETRIC LEAVES2(AS2)/AS1タンパク質による制御

    町田泰則, 岩崎まゆみ, 中川彩美, 高橋広夫, 松村葉子, 石橋奈々子, 羅麗蘭, 安藤沙友里, 岩川秀和, 栗原大輔, 東山哲也, 林里香, 大林祝, 杉山宗隆, PRATIWI Prananingrum, 笹部美知子, 町田千代子

    日本植物生理学会年会要旨集  2012.3.9 

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  99. Live-Cell Imaging of Double Fertilization and Embryogenesis in Plants Invited International conference

    Daisuke Kurihara, Yuki Hamamura, Tetsuya Higashiyama

    Conference Proceedings APMC 10 / ICONN 2012 / ACMM 22  2012.2.8 

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  100. シロイヌナズナ胚発生過程のライブイメージング

    栗原大輔, 牛王啓太, 東山哲也

    日本植物学会大会研究発表記録  2011.9.18 

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  101. 発生分化過程におけるオーロラキナーゼの動態解析

    松永幸大, 栗原大輔, 大村知広, 石田喬志, 北原英里奈, 浅田拓也, 万代文子, 松永朋子, 杉本慶子, 福井希一

    日本植物学会大会研究発表記録  2011.9.10 

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  102. 植物Auroraキナーゼの分裂・分化過程における機能解析

    万代文子, 栗原大輔, 内山進, 松永幸大, 福井希一

    日本生物工学会大会講演要旨集  2011.8.25 

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  103. Live-cell imaging of embryo development in Arabidopsis thaliana

    2011.6.27 

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  104. オーロラキナーゼによる発生分化調節機構のイメージング解析

    松永幸大, 栗原大輔, 大村知広, 浅田拓也, 万代文子, 福井希一

    日本植物生理学会年会要旨集  2011.3.11 

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  105. イメージング技術を用いた植物オーロラキナーゼの機能解析

    浅田拓也, 栗原大輔, 内山進, 松永幸大, 福井希一

    日本生物工学会大会講演要旨集  2010.9.25 

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  106. 細胞分裂におけるハスピンキナーゼの解析

    栗原大輔, 松永幸大, 大村知広, 東山哲也, 福井希一

    日本植物学会大会研究発表記録  2010.9.11 

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  107. イメージングによるオーロラキナーゼ・ノックダウン個体の表現型解析

    松永幸大, 栗原大輔, 大村知広, 浅田拓也, 福井希一

    日本植物学会大会研究発表記録  2010.9.8 

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  108. 分裂期キナーゼが制御する染色体動態からみた細胞分裂の研究 Invited

    栗原大輔

    日本植物形態学会第22回総会・大会  2010.9.8 

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  109. 花粉菅誘引物質LUREの1分子イメージング解析に向けて

    後藤宏旭, 奥田哲弘, 栗原大輔, 佐々木成江, 東山哲也

    日本植物学会大会研究発表記録  2010.9.8 

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  110. 細胞分裂期におけるヒストンH3スレオニンキナーゼの同定および解析

    栗原大輔, 松永幸大, 大村知広, 内山進, 福井希一

    日本植物学会大会研究発表記録  2009.9.19 

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    Event date: 2009.9.19

    Language:Japanese   Presentation type:Oral presentation (general)  

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  111. ライブイメージングによるオーロラキナーゼの機能解析

    松永幸大, 栗原大輔, 池田虎三, 大村知広, 内山進, 福井希一

    日本植物学会大会研究発表記録  2009.9.17 

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  112. 染色体動態イメージングによる植物器官形成モデルの構築

    大村知広, 栗原大輔, 内山進, 松永幸大, 福井希一

    日本生物工学会大会講演要旨集  2009.8.25 

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  113. シロイヌナズナのサイクリン依存性キナーゼCDKB2のDNA損傷による発現制御

    安達澄子, 栗原大輔, 松永幸大, 梅田正明

    日本植物生理学会年会要旨集  2009.3.16 

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  114. 動原体に局在するオーロラキナーゼのイメージング解析

    松永幸大, 栗原大輔, 池田虎三, 大村知広, 内山進, 福井希一

    日本植物生理学会年会要旨集  2009.3.16 

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  115. タバコBY‐2のXMAP215ホモローグTMBP200の機能ドメインの解析

    安原裕紀, 小西麻由, 松永幸大, 栗原大輔

    日本植物学会大会研究発表記録  2008.9.25 

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  116. ヒストンメチル化酵素SDG4による生殖器官分化のエピジェネティクス制御

    松永幸大, JOYCE Cartagena, 関原明, 栗原大輔, 篠崎一雄, 内山進, 福井希一

    日本遺伝学会大会プログラム・予稿集  2008.8.8 

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  117. 植物Auroraキナーゼ阻害による体細胞分裂の染色体動態変化

    松永幸大, 栗原大輔, 内山進, 福井希一

    日本植物生理学会年会要旨集  2008.3.15 

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  118. Functional analysis of plant Aurora kinase during mitosis International conference

    Daisuke Kurihara, Susumu Uchiyama, Sachihiro Matsunaga, Kiichi Fukui

    The 3rd Asian Chromosome Colloquium  2008 

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  119. 体細胞分裂におけるAuroraキナーゼの機能解析

    栗原大輔

    特定領域研究「植物メリステム」若手ワークショップ  2008 

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    Language:English   Presentation type:Oral presentation (general)  

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  120. 体細胞分裂の染色体接着を制御する新規カスケードの発見

    松永幸大, 高田秀昭, 森本晃弘, 馬楠, 栗原大輔, 大野(真庭)理香, 内山進, 福井希一

    日本遺伝学会大会プログラム・予稿集  2007.9.7 

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  121. 植物Auroraキナーゼの染色体動態における機能解析

    松永幸大, 栗原大輔, 内山進, 福井希一

    日本植物学会大会研究発表記録  2007.9.6 

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  122. 細胞核・染色体のダイナミクス

    松永幸大, 栗原大輔, 藤本聡, 内山進, 福井希一

    日本植物生理学会年会要旨集  2007.3.15 

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  123. Functional analysis of plant Aurora kinases in chromosome segregation during mitosis International conference

    Daisuke Kurihara

    Genetisches Seminar  2007 

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    Language:English   Presentation type:Oral presentation (invited, special)  

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  124. 植物においてヒストンH3をリン酸化するAuroraキナーゼ

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    第1回日本エピジェネティクス研究会年会  2007 

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  125. Aurora kinases are required for chromosome segregation in plants International conference

    Daisuke Kurihara, Masanori Noda, Akira Kawabe, Satoru Fujimoto, Susumu Uchiyama, Sachihiro Matsunaga, Kiichi Fukui

    20th IUBMB Congress  2006 

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    Event date: 2006

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  126. 染色体分離を制御する植物Auroraキナーゼ

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    染色体学会2006年度(第57回)年会 

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  127. 細胞分裂期において植物Auroraキナーゼは染色体分離を制御する

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    日本植物形態学会第18回総会・大会  2006 

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  128. 細胞分裂期における植物Auroraキナーゼの機能解析

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    「植物の生殖過程におけるゲノム障壁」ワークショップ 

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    Language:English   Presentation type:Oral presentation (general)  

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  129. Auroraキナーゼ阻害剤と植物のヒストンH3リン酸化を抑制し細胞周期進行を阻害する

    栗原大輔, 野田勝紀, 河辺昭, 藤本聡, 内山進, 松永幸大, 福井希一

    日本分子生物学会年会講演要旨集 

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    Event date: 2005.11.25

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  130. 植物における分裂期特異的なヒストンH3のリン酸化

    栗原大輔, 松永幸大, 藤本聡, 内山進, 福井希一

    イネ・シロイヌナズナ合同ワークショップ 

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    Event date: 2005.3

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  131. 細胞分裂を制御する植物Auroraキナーゼの動態解析

    松永幸大, 河辺昭, 中川勝之, 栗原大輔, 米田新, 馳沢盛一郎, 内山進, 福井希一

    日本植物生理学会年会要旨集 

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    Event date: 2005.3

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  132. Auroraキナーゼ阻害剤は植物のヒストンH3リン酸化を抑制し細胞周期進行を阻害する

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    日本分子生物学会第28回年会 

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    Event date: 2005

    Language:English   Presentation type:Oral presentation (general)  

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  133. アラビドプシスAuroraキナーゼの体細胞分裂過程における動態解析

    松永幸大, 河辺昭, 栗原大輔, 中川勝之, 内山進, 福井希一

    日本分子生物学会年会プログラム・講演要旨集 

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    Event date: 2004.11.25

    Language:English   Presentation type:Oral presentation (general)  

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  134. In vitro における植物Aurora キナーゼとヒストンH3との相互作用

    栗原大輔, 河邊昭, 中川勝之, 内山進, 松永幸大, 福井希一

    第56回日本生物工学会大会  

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    Event date: 2004.8.25

    Language:English   Presentation type:Oral presentation (general)  

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  135. Interaction of plant Aurora kinases with histone H3 in vitro

    Daisuke Kurihara, Akira Kawabe, Katsuyuki Nakagawa, Masanori Noda, Hiroaki Nakano, Susumu Uchiyama, Sachihiro Matsunaga, Kiichi Fukui

    The 2nd Asian Chromosome Colloquium 

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    Event date: 2004

    Language:English   Presentation type:Oral presentation (general)  

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  136. 動物培養細胞への導入による動植物受精因子のライブ解析

    中島耕大, Clari Valansi, 栗原大輔, 佐々木成江, Benjamin Podbilewicz, 東山哲也

    日本植物形態学会第31回大会  2019.9.14 

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  137. 二光子イメージングによる一対一受精機構の解明

    水多陽子, 栗原大輔, 東山哲也

    日本植物形態学会第31回大会  2019.9.14 

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  138. ライブイメージングで迫るシロイヌナズナ受精卵の極性化機構 Invited

    植田美那子, 木全祐資, 田中小百合, 檜垣匠, 栗原大輔, 東山哲也

    日本植物学会第83回大会  2019.9.15 

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  139. シロイヌナズナ新規核膜融合因子Gex1の細胞内動態の解析

    鈴木 千晴, 矢部 あやか, 栗原 大輔, 佐藤 良勝, 東山 哲也, 西川 周一

    日本植物学会第83回大会  2019.9.17 

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  140. シロイヌナズナ初期胚の細胞運命制御機構に関わる因子の探索

    栗原大輔, 大谷悠登, 石田喬志, 澤進一郎, 東山哲也

    日本植物形態学会第31回大会  2019.9.14 

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  141. AS2 と協調的に働く核小体局在タンパク質 RNA HELICASE10 の AS2 body の局在における機能の解析

    安藤沙友里, 岩井雅斗, 小島晶子, 栗原大輔, 東山哲也, 町田泰則, 町田千代子

    第61回日本植物生理学会年会  2020.3.21 

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  142. 動物培養細胞を用いた動植物の受精因子群の機能解析

    中島耕大, Clari Valansi, 栗原大輔, 佐々木 成江, Benjamin Podbilewicz, 東山哲也

    日本植物学会第83回大会  2019.9.16 

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  143. 深部イメージングで探る植物生殖の謎

    水多陽子, 栗原大輔, 東山哲也

    日本植物学会第83回大会  2019.9.16 

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  144. 化合物スクリーニングにより見出された新規細胞分裂阻害剤 Invited

    木全祐資, 佐藤綾人, 桑田啓子, 鈴木孝征, 山田萌恵, 栗原大輔, 山田朋美, 東山哲也, 植田美那子

    日本植物学会第83回大会  2019.9.15 

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  145. 植物において染色体分離を制御するAuroraキナーゼ

    栗原大輔, 野田勝紀, 河邊昭, 藤本聡, 内山進, 松永幸大, 福井希一

    日本遺伝学会大会プログラム・予稿集  2007.9.7 

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  146. シロイヌナズナの細胞分裂に関わるヒストンH3スレオニンキナーゼの解析

    栗原大輔, 松永幸大, 大村知広, 東山哲也, 福井希一

    日本植物形態学会第22回総会・大会  2010 

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  147. シロイヌナズナの細胞分裂に関わるヒストンH3スレオニンキナーゼの解析

    栗原大輔, 松永幸大, 大村知広, 東山哲也, 福井希一

    Plant Morphol  2011.4 

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Research Project for Joint Research, Competitive Funding, etc. 1

  1. 膜融合による植物への長鎖DNA導入技術の開発

    Grant number:JPMJPR18K4  2018.10 - 2022.03

    さきがけ 

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    Grant type:Competitive

KAKENHI (Grants-in-Aid for Scientific Research) 7

  1. 植物雌性配偶体をモデルとした細胞運命制御機構の解明

    2022.04

    科学技術振興機構 (JST)  創発的研究支援事業 

    栗原大輔

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    Authorship:Principal investigator 

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  2. 植物胚発生における胚性再獲得と全能性消失機構の解明

    Grant number:20H05358  2020.04 - 2022.03

    科学研究費補助金  新学術領域研究(研究領域提案型)

    栗原 大輔

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    Authorship:Principal investigator 

    Grant amount:\7800000 ( Direct Cost: \6000000 、 Indirect Cost:\1800000 )

    本研究の目的は、我々が確立したin vitro胚発生系を利用し、植物受精胚の「全能性プログラム」制御に関わる因子を明らかにすることである。本研究では、[1]植物ホルモンであるオーキシンが全能性獲得に必要であるか、[2]全能性消失はいつ、どのように起こるのかという二点に着目して研究を行い、計画研究班の哺乳動物研究との連携により、生命の根幹をなす仕組みに潜む普遍的なメカニズムの発見に繋げていきたい。

  3. Investigation of the relationship between positional information and cell identity by controlling the nuclear dynamics using optical manipulation

    Grant number:18K19331  2018.06 - 2020.03

    Grant-in-Aid for Scientific Research  Grant-in-Aid for Challenging Research (Exploratory)

    Kurihara Daisuke

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    Authorship:Principal investigator 

    Grant amount:\6370000 ( Direct Cost: \4900000 、 Indirect Cost:\1470000 )

    In this study, we aimed to clarify the relationship between nuclear positional information and cell fate specification to elucidate the molecular mechanisms of cell fate specification in plants using a simple female gametophyte formation process with a small number of cells. We established the in vitro female gametophyte development system of Arabidopsis and monitored the dynamics of female gametophyte development in real-time. Analysis of the spatiotemporal information of nuclear dynamics and the expression of cell fate marker during the female gametophyte development suggested that the cell fate specification initiated earlier than previously thought. We also clarified the problem to be overcome in the intracellular organelle manipulation technology in the plant cells.

  4. 植物胚発生における細胞間コミュニケーションによる細胞運命制御機構の解明

    Grant number:17H03697  2017.04 - 2020.03

    科学研究費補助金  基盤研究(B)

  5. Analysis of the cell division patterning during plant early embryogenesis by developing the optical manipulation techniques

    Grant number:23870014  2011 - 2012

    Grant-in-Aid for Scientific Research 

    KURIHARA Daisuke

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    Authorship:Principal investigator 

    Grant amount:\3250000 ( Direct Cost: \2500000 、 Indirect Cost:\750000 )

    Multicellular animals and plants develop from the single-celled zygote to form the mature embryo. During embryogenesis, the zygote follows a pattern of cell division to form the body plan of the embryo. In this study, I performed live-cell imaging using in vitroovule culture and the fluorescent marker lines in Arabidopsis thaliana. The data of live-cell imaging indicated that the timings of cell division are not strictly synchronized even in 4-cell embryo stage. Moreover, I established the gene induction system in a single-cell level in Arabidopsis embryo using the optical manipulation technique.

  6. 光操作技術の開発による植物胚発生におけるヒストンH3バリアントの機能解析

    Grant number:09J05807  2009.04 - 2010.12

    科学研究費補助金 

  7. ヒストンH3リン酸化の可視化による染色体構造構築メカニズムの解明

    Grant number:06J09144  2006.04 - 2009.03

    科学研究費補助金 

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Industrial property rights 2

  1. 植物組織透明化剤

    栗原大輔

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    Application no:特願2015-245977  Date applied:2015.12.17

    Announcement no:特開2017-108684  Date announced:2017.6.22

    Country of applicant:1  

  2. 植物細胞分裂抑制剤

    植田美那子, 南保正和, 栗原大輔, 桑田啓子, 大川妙子

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    Application no:特願2016-028537  Date applied:2016.2.18

    Announcement no:特開2017-145218  Date announced:2017.8.24

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Teaching Experience (Off-campus) 1

  1. Seminar in Bioscience B

    2020.8 Chubu University)

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Social Contribution 3

  1. 植物のからだを覗いてみよう

    Lecturer

    豊橋市自然史博物館  名古屋大学出前授業in豊橋2016  2016.12.4

  2. 植物を透明にする薬

    Appearance

    CBCラジオ  多田しげおの気分爽快!!朝からP・O・N  2015.11.9

  3. 特別企画「最新の顕微鏡技術で花の中をのぞいてみよう!」

    Demonstrator

    JST・ERATO東山ライブホロニクスプロジェクト  公開講座「花の神秘:最先端技術でみる光と映像の世界」  2015.11.7

Media Coverage 26

  1. LOOK DEEP INSIDE

    2016.2.23

  2. 植物を丸ごと透明化する試薬 名大研究チームが開発

    朝日新聞  2015.12.31

  3. The Breakthrough of the “See through” Plant

    2015.12.24

  4. これから解剖は必要ない?植物の丸ごと透明化を達成

    化学 2016年1月号   2015.12.18

  5. 内部構造が丸見え! 植物を透明にする新技術を開発

    子供の科学 2016年1月号  2015.12.10

  6. 植物を透明化し、内部を観る新技術"ClearSee"

    JST サイエンスポータル  2015.12.4

  7. 植物が透明に!そのままで観察 試薬開発で実現 名大が新技術

    科学新聞  2015.11.20

  8. 植物を透明化、切らずに内部観察 名大が試薬開発

    日本経済新聞  2015.11.19

  9. 植物を丸ごと透明化 解剖せずに内部観察

    産経フォト  2015.11.18

  10. 植物を丸ごと透明化する新技術 細胞レベルを観察 名古屋大

    ハザードラボ  2015.11.2

  11. 植物 透明化、内部見えた

    中日新聞朝刊  2015.10.30

  12. 名大,植物の透明化技術を開発

    Optronics Online  2015.10.30

  13. 葉緑素を取り除き透明化する薬品開発

    NHK 東海ニュース  2015.10.29

  14. 植物を透明化する試薬「ClearSee」を開発 内部構造の観察が可能に

    バイオの杜  2015.10.29

  15. 植物 透明化する薬

    読売新聞朝刊  2015.10.29

  16. 植物の丸ごと透明化に成功

    Yahoo! ニュース  2015.10.28

  17. 植物を丸ごと透明化 解剖不要で内部の観察可能に 名古屋大が成功

    ITmediaニュース  2015.10.28

  18. 植物受精卵 成長を観察

    読売新聞朝刊  2015.9.6

  19. 名大、植物受精卵を行きたまま観察するシステムを開発

    財経新聞  2015.7.15

  20. 植物受精卵 ライブで観察

    毎日新聞夕刊  2015.7.11

  21. 植物の受精卵 生きたまま観察

    Yahoo! ニュース  2015.7.11

  22. 細胞分裂する植物受精卵撮影

    CBCテレビ ニュース  2015.7.10

  23. 名大、植物の受精卵が分裂する様子を生きたまま観察できる手法開発ー「胚」形成を観察

    日刊工業新聞  2015.7.10

  24. 植物の受精卵分裂を初観察

    中日新聞朝刊  2015.7.10

  25. 植物受精卵の分裂 生きたまま観察

    朝日新聞朝刊  2015.7.10

  26. 植物の受精卵分裂を可視化=生きたまま観察、世界初ー名大

    時事通信  2015.7.10

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