2021/12/07 更新

写真a

ウエキ ショウコ
植木 尚子
UEKI Shoko
所属
資源植物科学研究所 准教授
職名
准教授

学位

  • 博士(理学) ( 東京工業大学 )

研究キーワード

  • Heterosigma akashiwo

  • ecophysiology

  • cell biology

  • molecular biology

  • microbial ecology

研究分野

  • 環境・農学 / 環境農学  / 水圏生命科学

  • ライフサイエンス / 分子生物学

  • ライフサイエンス / 細胞生物学

学歴

  • 東京工業大学   Graduate School of Bioscience and Biotechnology  

    1994年4月 - 1997年3月

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  • 筑波大学   Master's Program in Environmental Sciences  

    1992年4月 - 1994年7月

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  • 名古屋大学   School of Science  

    1988年4月 - 1992年3月

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    国名: 日本国

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経歴

  • 岡山大学   資源植物科学研究所   准教授

    2017年9月 - 現在

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  • 岡山大学   資源植物科学研究所   助教

    2011年9月 - 2017年7月

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  • ニューヨーク州立大学ストーニーブルック校   生化学・細胞生物学部

    2007年8月 - 2011年8月

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    国名:アメリカ合衆国

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  • ニューヨーク州立大学ストーニーブルック校   生化学・細胞生物学部

    1998年9月 - 2007年7月

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    国名:アメリカ合衆国

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  • 日本学術振興会   東京工業大学 生命理工学部   特別研究員

    1997年4月 - 1998年8月

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所属学協会

  • 日本微生物生態学会

    2016年 - 現在

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論文

  • Suitcase Lab: new, portable, and deployable equipment for rapid detection of specific harmful algae in Chilean coastal waters 査読

    So Fujiyoshi, Kyoko Yarimizu, Yohei Miyashita, Joaquín Rilling, Jacquelinne J. Acuña, Shoko Ueki, Gonzalo Gajardo, Oscar Espinoza-González, Leonardo Guzmán, Milko A. Jorquera, Satoshi Nagai, Fumito Maruyama

    Environmental Science and Pollution Research   2   14144 - 14155   2020年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    <title>Abstract</title>Phytoplankton blooms, including harmful algal blooms (HABs), have serious impacts on ecosystems, public health, and productivity activities. Rapid detection and monitoring of marine microalgae are important in predicting and managing HABs. We developed a toolkit, the Suitcase Lab, to detect harmful algae species in the field. We demonstrated the Suitcase Lab’s capabilities for sampling, filtration, DNA extraction, and loop-mediated isothermal amplification (LAMP) detection in cultured <italic>Alexandrium catenella</italic> cells as well as Chilean coastal waters from four sites: Repollal, Isla García, Puerto Montt, and Metri. A LAMP assay using the Suitcase Lab in the field confirmed microscopic observations of <italic>A. catenella</italic> in samples from Repollal and Isla García. The Suitcase Lab allowed the rapid detection of <italic>A. catenella</italic>, within 2 h from the time of sampling, even at a single cell per milliliter concentrations, demonstrating its usefulness for quick and qualitative on-site diagnosis of target toxic algae species. This method is applicable not only to detecting harmful algae but also to other field studies that seek a rapid molecular diagnostic test.

    DOI: 10.1007/s11356-020-11567-5

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    その他リンク: http://link.springer.com/article/10.1007/s11356-020-11567-5/fulltext.html

  • Protocols for Monitoring Harmful Algal Blooms for Sustainable Aquaculture and Coastal Fisheries in Chile 査読

    Kyoko Yarimizu, So Fujiyoshi, Mikihiko Kawai, Luis Norambuena-Subiabre, Emma-Karin Cascales, Joaquin-Ignacio Rilling, Jonnathan Vilugrón, Henry Cameron, Karen Vergara, Jesus Morón-López, Jacquelinne J. Acuña, Gonzalo Gajardo, Oscar Espinoza-González, Leonardo Guzmán, Milko A. Jorquera, Satoshi Nagai, Gemita Pizarro, Carlos Riquelme, Shoko Ueki, Fumito Maruyama

    International Journal of Environmental Research and Public Health   17 ( 20 )   7642 - 7642   2020年10月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:MDPI AG  

    Harmful algae blooms (HABs) cause acute effects on marine ecosystems due to their production of endogenous toxins or their enormous biomass, leading to significant impacts on local economies and public health. Although HAB monitoring has been intensively performed at spatiotemporal scales in coastal areas of the world over the last decades, procedures have not yet been standardized. HAB monitoring procedures are complicated and consist of many methodologies, including physical, chemical, and biological water sample measurements. Each monitoring program currently uses different combinations of methodologies depending on site specific purposes, and many prior programs refer to the procedures in quotations. HAB monitoring programs in Chile have adopted the traditional microscopic and toxin analyses but not molecular biology and bacterial assemblage approaches. Here we select and optimize the HAB monitoring methodologies suitable for Chilean geography, emphasizing on metabarcoding analyses accompanied by the classical tools with considerations including cost, materials and instrument availability, and easiness and efficiency of performance. We present results from a pilot study using the standardized stepwise protocols, demonstrating feasibility and plausibility for sampling and analysis for the HAB monitoring. Such specific instructions in the standardized protocol are critical obtaining quality data under various research environments involving multiple stations, different analysts, various time-points, and long HAB monitoring duration.

    DOI: 10.3390/ijerph17207642

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  • Phylogeographic characteristics of hypervariable regions in the mitochondrial genome of a cosmopolitan, bloom-forming raphidophyte, Heterosigma akashiwo 査読

    Shoko Ueki

    J Phycology   2019年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • A unique, highly variable mitochondrial gene with coding capacity of Heterosigma akashiwo, class Raphidophyceae 査読

    Aiko Higashi, Satoshi Nagai, Paulo S. Salomon, Shoko Ueki

    JOURNAL OF APPLIED PHYCOLOGY   29 ( 6 )   2961 - 2969   2017年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    Information related to geographical distribution and local strain composition is essential to an understanding of the dynamics of harmful algae in the environment. Previously, we identified a highly variable segment on the mitochondrial genome of Heterosigma akashiwo, a bloom-forming noxious unicellular algal species. Here, we assessed the utility of the mitochondrial hypervariable region for the phylogeographic study of the alga for different distance ranges. The sequences of H. akashiwo strains obtained from different geographic origins were successfully amplified and sequenced. We found differences among the sequences of the strains obtained from high-latitude regions of Northern Pacific/Atlantic; lower latitude regions of North America West Coast; and other regions including Brazil, Japan, Singapore, and North America East Coast. On the other hand, no strong geographic patterns for the sequences among Japanese strains were observed. Therefore, the hypervariable segment may be useful to distinguish H. akashiwo strains originated from distant regions (Atlantic/Pacific, high/low latitudes), rather than regions separated by shorter distances. The sequence contains an open reading frame coding for a protein with unknown function, and the transcription of the gene was confirmed by RNA-seq analysis. Despite the sequence variations observed among H. akashiwo strains originating in different parts of the world, three domains of the protein were highly conserved among all of the strains, suggesting that they may be important to the function of the protein.

    DOI: 10.1007/s10811-017-1142-2

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  • A hypervariable mitochondrial protein coding sequence associated with geographical origin in a cosmopolitan bloom-forming alga, Heterosigma akashiwo 査読

    Aiko Higashi, Satoshi Nagai, Sergio Seone, Shoko Ueki

    BIOLOGY LETTERS   13 ( 4 )   2017年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ROYAL SOC  

    Geographical distributions of phytoplankton species can be defined by events on both evolutionary time and shorter scales, e.g. recent climate changes. Additionally, modern industrial activity, including the transport of live fish and spat for aquaculture and aquatic microorganisms in ship ballast water, may aid the spread of phytoplankton. Obtaining a reliable marker is key to gaining insight into the phylogeographic history of a species. Here, we report a hypervariable mitochondrial gene in the cosmopolitan bloom forming alga, Heterosigma akashiwo. We compared the entire mitochondrial genome sequences of seven H.. akashiwo strains from Japanese and North American coastal waters and identified a hypervariable segment. The region codes for a hypothetical protein with no defined function, and its variations between Japanese and North American isolates were prominent, while the sequences were more conserved among Japanese strains and North American isolates. Comparison of the sequence in isolates obtained from different geographical points in the Northern Hemisphere revealed that the sequence variations largely correlated with latitude and longitude (i.e. Pacific/Atlantic oceans). Our results demonstrate the usefulness of the sequence in determining the phylogeographic history of H. akashiwo.

    DOI: 10.1098/rsbl.2016.0976

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  • Chloroplast genome sequences of seven strains of the bloom-forming raphidophyte Heterosigma akashiwo 査読

    Sergio Seoane, Kiwamu Hyodo, Shoko Ueki

    Genome Announcements   5 ( 41 )   2017年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:American Society for Microbiology  

    We report here the complete chloroplast genome sequences of seven strains of the bloom-forming raphidophyte Heterosigma akashiwo. These ~160-kb sequences contain 124 protein-, 6 rRNA-, and 34 tRNA-coding sequences. Notable sequence variations were observed among these seven sequenced and two previously characterized strains.

    DOI: 10.1128/genomeA.01030-17

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  • Selective growth promotion of bloom-forming raphidophyte Heterosigma akashiwo by a marine bacterial strain 査読

    Aiko Higashi, Yoshiko Fujitani, Natsuko Nakayama, Akio Tani, Shoko Ueki

    HARMFUL ALGAE   60   150 - 156   2016年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Algal bloom is typically caused by aberrant propagation of a single species, resulting in its predomination in the local population. While environmental factors including temperature and eutrophication are linked to bloom, the precise mechanism of its formation process is still obscure. Here, we isolated a bacterial strain that promotes growth of Heterosigma akashiwo, a Raphidophyceae that causes harmful algal blooms. Based on 16S rRNA gene sequence, the strain was identified as Altererythrobacter ishigakiensis, a member of the class Alphaproteobacteria. When added to culture, this strain facilitated growth of H. akashiwo and increased its cell culture yield significantly. Importantly, this strain did not affect the growth of other raphidophytes, Chattonella ovate and C. antiqua, indicating that it promotes growth of H. akashiwo in a species-specific manner. We also found that, in co-culture, H. akashiwo suppressed the growth of C. ovate. When A. ishigakiensis was added to the mixed culture, H. akashiwo growth was facilitated while C ovate propagation was markedly suppressed, indicating that the presence of the bacterium enhances the dominance of H. akashiwo over C ovate. This is the first example of selective growth promotion of H. akashiwo by a marine bacterium, and may exemplify importance of symbiotic bacterium on algal bloom forming process in general. (C) 2016 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.hal.2016.11.009

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  • Evolution and Phylogeny of Large DNA Viruses, Mimiviridae and Phycodnaviridae Including Newly Characterized Heterosigma akashiwo Virus 査読

    Fumito Maruyama, Shoko Ueki

    FRONTIERS IN MICROBIOLOGY   7   1942   2016年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:FRONTIERS MEDIA SA  

    Nucleocytoplasmic DNA viruses are a large group of viruses that harbor double stranded DNA genomes with sizes of several 100 kbp, challenging the traditional concept of viruses as small, simple 'organisms at the edge of life.' The most intriguing questions about them may be their origin and evolution, which have yielded the variety we see today. Specifically, the phyletic relationship between two giant dsDNA virus families that are presumed to be close, Mimiviridae, which infect Acantharnoeba, and Phycodnaviridae, which infect algae, is still obscure and needs to be clarified by indepth analysis. Here, we studied Mimiviridae Phycodnaviridae phylogeny including the newly identified Heterosigma akashiwo virus strain HaV53. Gene-to-gene comparison of HaV53 with other giant dsDNA viruses showed that only a small proportion of HaV53 genes show similarities with the others, revealing its uniqueness among Phycodnaviridae. Phylogenetic/genomic analysis of Phycodnaviridae including HaV53 revealed that the family can be classified into four distinctive subfamilies, namely, Megaviridae (Mimivirus-like), Chlorovirus-type, and Coccolitho/Phaeovirus-type groups, and HaV53 independent of the other three groups. Several orthologs found in specific subfamilies while absent from the others were identified, providing potential family marker genes. Finally, reconstruction of the evolutionary history of Phycodnaviridae and Mimiviridae revealed that these viruses are descended from a common ancestor with a small set of genes and reached their current diversity by differentially acquiring gene sets during the course of evolution. Our study illustrates the phylogeny and evolution of Mimiviridac Phycodnaviridae and proposes classifications that better represent phyletic relationships among the family members.

    DOI: 10.3389/micb.2016.01942

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  • Complete genome sequence of a phycodnavirus, Heterosigma akashiwo virus strain 53 査読

    Yoshitoshi Ogura, Tetsuya Hayashi, Shoko Ueki

    Genome Announcements   4 ( 6 )   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:American Society for Microbiology  

    We report the complete genome sequence of Heterosigma akashiwo virus strain 53. The virus is a member of the Phycodnaviridae, one of the families regarded as giant double-stranded DNA viruses. The 274,793-bp genome contained 246 protein-coding and 3 tRNA-coding sequences.

    DOI: 10.1128/genomeA.01279-16

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  • Mitochondrial genome sequences of four strains of the bloom-forming raphidophyte Heterosigma akashiwo 査読

    Yoshitoshi Ogura, Natsuko Nakayama, Tetsuya Hayashi, Shoko Ueki

    Genome Announcements   4 ( 6 )   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:American Society for Microbiology  

    We report here the complete mitochondrial genome sequences of four strains of bloom-forming raphidophytes from Heterosigma akashiwo. These 39-kb sequences contain 42 protein-, two rRNA-, and 26 tRNA-coding sequences. Notable sequence variations were observed among these four newly sequenced and three previously characterized strains, suggesting their potential usage as strain-specific markers.

    DOI: 10.1128/genomeA.01288-16

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  • Plasmodesmata-associated proteins: Can we see the whole elephant? 査読

    Shoko Ueki, Vitaly Citovsky

    Plant Signaling and Behavior   9 ( 2 )   e27899   2014年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Landes Bioscience  

    Encased in rigid cell walls, plant cells have evolved unique channel structures, plasmodesma (Pd), to create a pathway for molecular exchange between adjacent cells. Pd are basically cytoplasmic channels through the cell wall, which are lined by plasma membrane, and contain a modified strand of ER that spans them. These structures provide cytoplasmic and membrane continuity between connected cells, and that continuity is utilized for short and long distance molecular trafficking. Pd sphincters, made from constricting the Pd openings by outer layers of callose, together with the ER strand that occludes the Pd lumen set the upper limit for the size of molecules that can freely diffuse through the cytoplasmic component of the Pd channel. This limit, called the size exclusion limit (SEL), is a major factor that restricts macromolecular transport through Pd. © 2014 Landes Bioscience.

    DOI: 10.4161/psb.27899

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  • Identification of a movement protein of Mirafiori lettuce big-vein ophiovirus 査読

    Akihiro Hiraguri, Shoko Ueki, Hideki Kondo, Koji Nomiyama, Takumi Shimizu, Tamaki Ichiki-Uehara, Toshihiro Omura, Nobumitsu Sasaki, Hiroshi Nyunoya, Takahide Sasaya

    Journal of General Virology   94 ( 5 )   1145 - 1150   2013年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Mirafiori lettuce big-vein virus (MiLBVV) is a member of the genus Ophiovirus, which is a segmented negative-stranded RNA virus. In microprojectile bombardment experiments to identify a movement protein (MP) gene of ophioviruses that can trans-complement intercellular movement of an MP-deficient heterologous virus, a plasmid containing an infectious clone of a tomato mosaic virus (ToMV) derivative expressing the GFP was co-bombarded with plasmids containing one of three genes from MiLBVV RNAs 1, 2 and 4 onto Nicotiana benthamiana. Intercellular movement of the movement-defective ToMV was restored by co-expression of the 55 kDa protein gene, but not with the two other genes. Transient expression in epidermal cells of N. benthamiana and onion showed that the 55 kDa protein with GFP was localized on the plasmodesmata. The 55 kDa protein encoded in the MiLBVV RNA2 can function as an MP of the virus. This report is the first to describe an ophiovirus MP. © 2013 SGM.

    DOI: 10.1099/vir.0.050005-0

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  • Transient gene expression in epidermal cells of plant leaves by biolistic DNA delivery 査読

    Shoko Ueki, Shimpei Magori, Benoît Lacroix, Vitaly Citovsky

    Methods in Molecular Biology   940   17 - 26   2013年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Transient gene expression is a useful approach for studying the functions of gene products. In the case of plants, Agrobacterium infiltration is a method of choice for transient introduction of genes for many species. However, this technique does not work efficiently in some species, such as Arabidopsis thaliana. Moreover, the infection of Agrobacterium is known to induce dynamic changes in gene expression patterns in the host plants, possibly affecting the function and localization of the proteins to be tested. These problems can be circumvented by biolistic delivery of the genes of interest. Here, we present an optimized protocol for biolistic delivery of plasmid DNA into epidermal cells of plant leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol allows efficient and reproducible transient expression of diverse genes in Arabidopsis, Nicotiana benthamiana and N. tabacum, and is suitable for studies of the biological function and subcellular localization of the gene products directly in planta. The protocol also can be easily adapted to other species by optimizing the delivery gas pressure. © 2013 Springer Science+Business Media, LLC.

    DOI: 10.1007/978-1-62703-110-3_2

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  • To Gate, or Not to Gate: Regulatory Mechanisms for Intercellular Protein Transport and Virus Movement in Plants 査読

    Shoko Ueki, Vitaly Citovsky

    MOLECULAR PLANT   4 ( 5 )   782 - 793   2011年9月

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    記述言語:英語   出版者・発行元:OXFORD UNIV PRESS  

    Cell-to-cell signal transduction is vital for orchestrating the whole-body physiology of multi-cellular organisms, and many endogenous macromolecules, proteins, and nucleic acids function as such transported signals. In plants, many of these molecules are transported through plasmodesmata (Pd), the cell wall-spanning channel structures that interconnect plant cells. Furthermore, Pd also act as conduits for cell-to-cell movement of most plant viruses that have evolved to pirate these channels to spread the infection. Pd transport is presumed to be highly selective, and only a limited repertoire of molecules is transported through these channels. Recent studies have begun to unravel mechanisms that actively regulate the opening of the Pd channel to allow traffic. This macromolecular transport between cells comprises two consecutive steps: intracellular targeting to Pd and translocation through the channel to the adjacent cell. Here, we review the current knowledge of molecular species that are transported though Pd and the mechanisms that control this traffic. Generally, Pd traffic can occur by passive diffusion through the trans-Pd cytoplasm or through the membrane/lumen of the trans-Pd ER, or by active transport that includes protein-protein interactions. It is this latter mode of Pd transport that is involved in intercellular traffic of most signal molecules and is regulated by distinct and sometimes interdependent mechanisms, which represent the focus of this article.

    DOI: 10.1093/mp/ssr060

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  • Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro 査読

    Shoko Ueki, Benoit Lacroix, Vitaly Citovsky

    JOVE-JOURNAL OF VISUALIZED EXPERIMENTS   ( 54 )   2011年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOURNAL OF VISUALIZED EXPERIMENTS  

    Validating interactions between different proteins is vital for investigation of their biological functions on the molecular level. There are several methods, both in vitro and in vivo, to evaluate protein binding, and at least two methods that complement the shortcomings of each other should be conducted to obtain reliable insights.
    For an in vivo assay, the bimolecular fluorescence complementation (BiFC) assay represents the most popular and least invasive approach that enables to detect protein-protein interaction within living cells, as well as identify the intracellular localization of the interacting proteins 1,2. In this assay, non-fluorescent N-and C-terminal halves of GFP or its variants are fused to tested proteins, and when the two fusion proteins are brought together due to the tested proteins' interactions, the fluorescent signal is reconstituted3-6. Because its signal is readily detectable by epifluorescence or confocal microscopy, BiFC has emerged as a powerful tool of choice among cell biologists for studying about proteinprotein interactions in living cells 3. This assay, however, can sometimes produce false positive results. For example, the fluorescent signal can be reconstituted by two GFP fragments arranged as far as 7 nm from each other due to close packing in a small subcellular compartment, rather that due to specific interactions7.
    Due to these limitations, the results obtained from live cell imaging technologies should be confirmed by an independent approach based on a different principle for detecting protein interactions. Co-immunoprecipitation (Co-IP) or glutathione transferase (GST) pull-down assays represent such alternative methods that are commonly used to analyze protein-protein interactions in vitro. However, iIn these assays, however, the tested proteins must be readily soluble in the buffer that supportsused for the binding reaction. Therefore, specific interactions involving an insoluble protein cannot be assessed by these techniques.
    Here, we illustrate the protocol for the protein membrane overlay binding assay, which circumvents this difficulty. In this technique, interaction between soluble and insoluble proteins can be reliably tested because one of the proteins is immobilized on a membrane matrix. This method, in combination with in vivo experiments, such as BiFC, provides a reliable approach to investigate and characterize interactions faithfully between soluble and insoluble proteins. In this article, binding between Tobacco mosaic virus (TMV) movement protein (MP), which exerts multiple functions during viral cell-to-cell transport8-14, and a recently identified plant cellular interactor, tobacco ankyrin repeat-containing protein (ANK) 15, is demonstrated using this technique.

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  • Biology of callose (β-1,3-glucan) turnover at plasmodesmata. 査読

    Zavaliev R, Ueki S, Epel BL, Citovsky V

    Protoplasma   248 ( 1 )   117 - 130   2011年1月

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  • ANK, a Host Cytoplasmic Receptor for the Tobacco mosaic virus Cell-to-Cell Movement Protein, Facilitates Intercellular Transport through Plasmodesmata 査読

    Shoko Ueki, Roman Spektor, Danielle M. Natale, Vitaly Citovsky

    PLOS PATHOGENS   6 ( 11 )   e1001201   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Plasmodesma (PD) is a channel structure that spans the cell wall and provides symplastic connection between adjacent cells. Various macromolecules are known to be transported through PD in a highly regulated manner, and plant viruses utilize their movement proteins (MPs) to gate the PD to spread cell-to-cell. The mechanism by which MP modifies PD to enable intercelluar traffic remains obscure, due to the lack of knowledge about the host factors that mediate the process. Here, we describe the functional interaction between Tobacco mosaic virus (TMV) MP and a plant factor, an ankyrin repeat containing protein (ANK), during the viral cell-to-cell movement. We utilized a reverse genetics approach to gain insight into the possible involvement of ANK in viral movement. To this end, ANK overexpressor and suppressor lines were generated, and the movement of MP was tested. MP movement was facilitated in the ANK-overexpressing plants, and reduced in the ANK-suppressing plants, demonstrating that ANK is a host factor that facilitates MP cell-to-cell movement. Also, the TMV local infection was largely delayed in the ANK-suppressing lines, while enhanced in the ANK-overexpressing lines, showing that ANK is crucially involved in the infection process. Importantly, MP interacted with ANK at PD. Finally, simultaneous expression of MP and ANK markedly decreased the PD levels of callose, beta-1,3-glucan, which is known to act as a molecular sphincter for PD. Thus, the MP-ANK interaction results in the downregulation of callose and increased cell-to-cell movement of the viral protein. These findings suggest that ANK represents a host cellular receptor exploited by MP to aid viral movement by gating PD through relaxation of their callose sphincters.

    DOI: 10.1371/journal.ppat.1001201

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  • A cell-to-cell macromolecular transport assay in Planta utilizing biolistic bombardment. 査読

    Ueki S, Meyers BL, Yasmin F, Citovsky V

    Journal of visualized experiments : JoVE   ( 42 )   2010年8月

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  • Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment 査読

    Shoko Ueki, Benoit Lacroix, Alexander Krichevsky, Sondra G. Lazarowitz, Vitaly Citovsky

    NATURE PROTOCOLS   4 ( 1 )   71 - 77   2009年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Transient gene expression is an indispensable tool for studying functions of gene products. In the case of plants, transient introduction of genes by Agrobacterium infiltration is a method of choice for many species. However, this technique does not work efficiently in Arabidopsis leaf tissue, the most widely used model system for basic plant biology research. Here we present an optimized protocol for biolistic delivery of plasmid DNA into the epidermis of Arabidopsis leaves, which can be easily performed using the Bio-Rad Helios gene gun system. This protocol yields efficient and reproducible transient expression of diverse genes and is exemplified here for use in a functional assay of a transcription repressor and for the subcellular localization and cell-to-cell movement of plant viral movement protein. This protocol is suitable for studies of biological function and subcellular localization of the gene product of interest directly in planta by utilizing different types of activity-based assays. Using this procedure, the data are obtained after 2-4 d of work.

    DOI: 10.1038/nprot.2008.217

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  • Identification of an interactor of cadmium ion-induced glycine-rich protein involved in regulation of callose levels in plant vasculature 査読

    S Ueki, Citovsky, V

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   102 ( 34 )   12089 - 12094   2005年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATL ACAD SCIENCES  

    Cadmium-induced glycine-rich protein (cdiGRP) is a cell wall-associated factor that increases callose levels in plant vasculature. To better understand the cdiGRP/callose regulation system, we identified a tobacco protein, GrIP (cdiGRP-interacting protein, GrIP), that associates with cdiGRP and localizes at the plant cell wall. Constitutive overexpression of GrIP enhanced the accumulation of the cdiGRP protein and callose in vasculature-associated cells with or without treatment with cadmium ions. That GrIP gene expression was not affected by cadmium ions indicated that GrIP does not directly modulate the callose levels induced by the treatment. Instead, GrIP most likely functions by further elevating the accumulated amount of cdiGRP, the expression of which is up-regulated by the cadmium ions. Interestingly, the levels of cdiGRP mRNA were not affected by constitutive expression of GrIP, demonstrating that the enhancement in cdiGRP protein accumulation by GrIP overexpression occurs posttranslationally. Collectively, these observations suggest that GrIP interacts with cdiGRP and increases its level of accumulation; in turn, the elevated amounts of cdiGRP induce callose deposits in the plant cell walls. Therefore, GrIP and cdiGRP represent sequentially acting factors in a biochemical pathway that regulates callose accumulation in the plant vasculature.

    DOI: 10.1073/pnas.0505927102

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  • Control improves with age: Intercellular transport in plant embryos and adults 査読

    S Ueki, Citovsky, V

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   102 ( 6 )   1817 - 1818   2005年2月

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    記述言語:英語   出版者・発行元:NATL ACAD SCIENCES  

    DOI: 10.1073/pnas.0409785102

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  • The systemic movement of a tobamovirus is inhibited by a cadmium-ion-induced glycine-rich protein 査読

    S Ueki, Citovsky, V

    NATURE CELL BIOLOGY   4 ( 7 )   478 - 485   2002年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Systemic movement is central to plant viral infection. Exposure of tobacco plants to low levels of cadmium ions blocks the systemic spread of turnip vein-clearing tobamovirus (TVCV). We identified a tobacco glycine-rich protein, cdiGRP, specifically induced by low concentrations of cadmium and expressed in the cell walls of plant vascular tissues. Constitutive cdiGRP expression inhibited systemic transport of TVCV, whereas suppression of cdiGRP production allowed TVCV movement in the presence of cadmium. cdiGRP exerted its inhibitory effect on TVCV transport by enhancing callose deposits in the vasculature. So cdiGRP may function to control plant viral systemic movement.

    DOI: 10.1038/ncb806

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  • RNA commutes to work: regulation of plant gene expression by systemically transported RNA molecules 査読

    S Ueki, Citovsky, V

    BIOESSAYS   23 ( 12 )   1087 - 1090   2001年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Although long-distance movement of endogenous mRNAs in plants is well established, the functional contributions of these transported RNA molecules has remained unclear. In a recent report, Kim et al.((1)) showed that systemically transported mRNA is capable of causing phenotypic change In developing tissue. Here, this finding and its significance are reviewed and discussed In detail. In addition, in order to give proper perspective, long-distance transport of other types of RNAs, e.g., RNA elicitors of post-transcriptional gene silencing and RNA genomes of plant viruses, and its possible regulation are discussed. (C) 2001 John Wiley & Sons, Inc.

    DOI: 10.1002/bies.10027

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  • Inhibition of systemic onset of post-transcriptional gene silencing by non-toxic concentrations of cadmium 査読

    S Ueki, Citovsky, V

    PLANT JOURNAL   28 ( 3 )   283 - 291   2001年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL SCIENCE LTD  

    Post-transcriptional gene silencing (PTGS) is an important mechanism for regulation of plant gene expression and virus-plant interactions. To better understand this process, the heavy metal cadmium was identified as a specific inhibitor in two different PTGS systems, constitutive and inducible. The pattern of cadmium-induced inhibition of PTGS allowed several insights into PTGS development. First, cadmium treatment prevented only systemic but not local onset of PTGS, uncoupling between these two modes of PTGS. Second, non-toxic, but not toxic, levels of cadmium inhibited PTGS, suggesting induction of a pathway that interferes with PTGS. Third, cadmium effects on PTGS closely paralleled those on the movement of tobamoviruses, suggesting that both processes may share common steps in their systemic transport pathways. Interestingly, these effects of cadmium do not represent a general property of toxic metal ions because two other such elements, that is zinc and aluminum, did not interfere with PTGS and viral systemic movement.

    DOI: 10.1046/j.1365-313X.2001.01145.x

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  • Activity-independent cell adhesion to tissue-type transglutaminase is mediated by alpha 4 beta 1 integrin 査読

    T Isobe, H Takahashi, S Ueki, J Takagi, Y Saito

    EUROPEAN JOURNAL OF CELL BIOLOGY   78 ( 12 )   876 - 883   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:URBAN & FISCHER VERLAG  

    Transglutaminases (TGases) are enzymes which catalyze cross-link formation between glutamine residues and lysine residues in substrate proteins. We have previously reported that one of the TGases, blood coagulation factor XIIIa (FXIIIa), is capable of mediating adhesion of various cells. In this paper, we report for the first time that tissue-type transglutaminase (TGc) also has cell adhesion activity TGc-coated plastic surface promoted adhesion and spreading of cells in a TGc concentration-dependent manner. However, there are some obvious differences between cell adhesion mediated by TGc and FXIIIa, As was reported previously, the adhesion to FXIIIa is dependent on its TGase activity In contrast, the TGc-mediated cell adhesion is independent of its TGase activity: 1) The modification of the active center cysteine with iodoacetamide blocked the enzyme activity without any effect on cell adhesion; 2) the addition of Mg2+ did not induce the enzyme activity, but it was as effective as Ca2+ for cell adhesion; 3) the addition of NH4+ inhibited the enzyme activity but did not affect the cell adhesion significantly. The integrins involved in these cell adhesions are quite different. In the case of FXIIIa, av()3 and alpha 5 beta 1 integrins are involved and consequently the RGD peptide substantially inhibited the adhesion. On the other hand, the cell adhesion to TGc is mediated by alpha 4 beta 1 integrin but not alpha 5 beta 1; a CS-1 peptide, which represents the binding site of fibronectin to alpha 4 beta 1 integrin, completely inhibited the cell adhesion to TGc, It is possible that TGc and FXIIIa mag mediate cell adhesion under different physiological and pathological situations.

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  • 12-hydroxy-5Z,8Z,10E,14Z, eicosatetraenoic acid (12-HETE) stimulates cAMP production in normal human fibroblasts 査読

    S Ueki, J Takagi, Y Kobayashi, F Sato, Y Saito

    JOURNAL OF CELLULAR PHYSIOLOGY   178 ( 1 )   63 - 68   1999年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    We report here that the 12-lipoxygenase metabolite of arachidonic acid, 12-hydroxy-5Z, 8Z, 10E, 14Z, eicosatetraenoic acid (12-HETE), stimulates cAMP production in human fibroblasts among various cultured cell lines tested. Although 12-HETE seemed to stimulate the phospholipase C (PLC)-protein kinase C (PKC) system, inhibitors against PLC and PKC did not reduce the cAMP production induced by 12-HETE, indicating that the activation of PLC-PKC system is not positively coupled with the stimulation of cAMP production. On the other hand, the cAMP production induced by 12-HETE was dependent on the Ca2+/calmodulin system in the cells. The results suggest that 12-HETE specifically stimulates Ca2+/calmodulin-dependent adenylyl cyclase to increase cAMP level in the fibroblasts. (C) 1999 Wiley-Liss, Inc.

    DOI: 10.1002/(SICI)1097-4652(199901)178:1<63::AID-JCP8>3.0.CO;2-J

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  • The 'ligand-induced conformational change' of alpha 5 beta 1 integrin - Relocation of alpha 5 subunit to uncover the beta 1 stalk region 査読

    J Tsuchida, S Ueki, Y Takada, Y Saito, J Takagi

    JOURNAL OF CELL SCIENCE   111   1759 - 1766   1998年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Integrin heterodimers undergo a conformational change upon the binding of ligand to their extracellular domains. An anti-beta 1 integrin monoclonal antibody AG89 can detect such a conformational change since it recognizes a ligand-inducible epitope in the stalk-like region of beta 1 subunits, The binding of a I-125-labeled AG89 Fab fragment to alpha 5 beta 1 integrins on K562 cells was assessed and analyzed by the Scatchard method. High affinity binding sites for AG89 are present on cells treated with ligand peptide. In addition, results revealed that cells treated with EDTA also express AG89 binding sites with the same affinity although the number of binding sites is 4-fold lower. AG89 immunoprecipitated alpha 5 beta 1 complexes from surface-labeled K562 cells treated with ligand peptide. By contrast, it immunoprecipitated only beta 1 chains when the ligand peptide was absent, suggesting that high affinity binding sites on EDTA-treated cells are associated with nonfunctional beta 1 monomer. Additional studies show that the epitope for AG89 is constitutively exposed on mutant pi that cannot complex with alpha 5. These data suggest that the AG89 epitope is masked by the alpha 5 subunit. Ligand binding and integrin activation may uncover the beta 1 stalk region by triggering a conformational shift of alpha 5 relative to beta 1.

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  • Classification of 'activation' antibodies against integrin beta 1 chain 査読

    J Tsuchida, S Ueki, Y Saito, J Takagi

    FEBS LETTERS   416 ( 2 )   212 - 216   1997年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    We compared the effects of two anti-beta 1 integrin activating antibodies, TS2/16 and AG89, on K562 cell adhesion to fibronectin. Though both antibodies effectively induced cell adhesion, the EC50 for AG89 was more than 200-fold higher than that for TS2/16, Scatchard analysis of the data from [I-125]Fab fragment binding to the cells revealed that the TS2/16 epitope is exposed constitutively on all the beta 1 integrin molecules, while only 3% of the beta 1 integrins on resting K562 cells bear the AG89 epitope. Calculation of the actual number of each antibody bound to the cell during the cell adhesion assay revealed that induction of cell adhesion can be accomplished by binding much fewer AG89 molecules compared to TS2/16. Thus, AG89 and TS2/16 represent distinct classes of anti-integrin activating antibodies that show completely different binding characteristics as well as different activation effects on the integrin molecule upon binding. (C) 1997 Federation of European Biochemical Societies.

    DOI: 10.1016/S0014-5793(97)01206-4

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  • Dual functions of transglutaminase in novel cell adhesion 査読

    S Ueki, J Takagi, Y Saito

    JOURNAL OF CELL SCIENCE   109   2727 - 2735   1996年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    Transglutaminases (TGases) are enzymes which catalyze cross-link formation between glutamine residues and lysine residues in substrate proteins, In the present study, we report for the first time that a representative enzyme, blood coagulation factor XIIIa (FXIIIa), is capable of mediating adhesion of various cells, When coated on plastic surfaces FXIIIa promoted adhesion and spreading of various cells of both normal and tumor origin, in a concentration-dependent manner, The adhesion was not inhibited by antibodies against possible contaminants in the enzyme preparation such as fibronectin and vitronectin, but was completely inhibited by a polyclonal antibody against the enzyme, Therefore, if there were any contaminating cell adhesive substrates in the enzyme preparation, they cannot account for the observed cell adhesion to the enzyme; FXIIIa itself mediates the cell adhesion, Furthermore, phosphorylation of tyrosine residues in 120 kDa and 70 kDa proteins was clearly shown in human fibroblasts adhering to the enzyme, Formation of actin stress fibers was also unambiguously observed in the adhering cells, These biochemical reactions, which are also observed when cells adhere to a typical cell adhesion protein, fibronectin, are believed to be of importance in the process of cell adhesion, This adhesion activity of FXIIIa was dependent on its TGase activity, because both a modification of the active center cysteine with iodoacetamide and the addition of ammonium ion abolished the cell adhesion activity along with the enzyme activity, The cell adhesion to fibronectin, however, was not affected by these treatments, The effects of various anti-integrin antibodies suggested that both alpha v beta 3 and beta 1 family integrins participated in the cell adhesion to FXIIIa, Taken together, these data demonstrate for the first time that there is a unique TGase activity-mediated cell adhesion, This novel function of the enzyme may be of physiological importance.

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講演・口頭発表等

  • Characterization of Marine Bacteria That Support Growth of Harmful Algae Under Nutritionally Limiting Conditions. 招待

    Shoko Ueki

    Annual Meeting of Society of Microbiology in Chile  2021年11月30日 

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    開催年月日: 2021年11月29日 - 2021年11月30日

    記述言語:英語   会議種別:口頭発表(招待・特別)  

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  • リン欠乏条件下においてHeterosigma akashiwoがリン源とするポリリン酸合成細菌の発見.

    宇佐美文子, 小原静香, 隠塚俊満, 近藤健, 中嶋昌紀, 小池一彦, 植木尚子

    微生物生態学会年会  2021年10月30日 

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    開催年月日: 2021年10月30日 - 2021年11月2日

    記述言語:日本語   会議種別:ポスター発表  

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  • Gene Manipulation Techniques in Heterosigma Akashiwo

    Nanami Nakayama , Shoko Ueki , Ayano Satoh

    日本分子生物学会年会  日本分子生物学会

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    開催年月日: 2020年12月2日 - 2020年12月4日

    記述言語:英語   会議種別:口頭発表(一般)  

    開催地:online  

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  • 赤潮原因種Heterosigma akashiwoの遺伝子操作技術

    中山七海,植木尚子,佐藤あやの

    日本生物工学会西日本支部大会(第5回講演会)  日本生物工学会

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    開催年月日: 2020年11月14日

    記述言語:日本語   会議種別:口頭発表(一般)  

    開催地:岡山理科大学  

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  • 赤潮原因藻ヘテロシグマの系統地理学的マーカーの確立を目指した研究

    妹尾美紀, 平松諒也, Anette Engesmo, Brian D. Bill, Vera L. Trainer, 長井敏, 門田有希, 植木尚子

    日本育種学会・第 11 回中国地域育種談話会  岡山大学

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    開催年月日: 2019年12月21日 - 2019年12月22日

    記述言語:日本語   会議種別:ポスター発表  

    開催地:岡山大学津島キャンパス自然科学研究科棟 大講義室  

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  • 赤潮原因藻ヘテロシグマのバイオテクノロジー的利用をめざした遺伝子導入法の検討

    佐藤あやの、楠本恭平、植木尚子

    赤潮原因藻ヘテロシグマのバイオテクノロジー的利用をめざした遺伝子導入法の検討  日本分子生物学会

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    開催年月日: 2019年12月3日 - 2019年12月6日

    記述言語:日本語   会議種別:ポスター発表  

    開催地:福岡国際会議場  

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  • Toward understanding ecophysiology of a bloom-forming alga, Heterosigma akashiwo 招待

    Shoko Ueki

    International Symposium: “Advances in Environmental Microbiology and Microbial Biotechnology”  N?cleo Cient?fico Tecnol?gico de la Universidad de La Frontera BIOREN-UFRO

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    開催年月日: 2018年12月6日 - 2018年12月8日

    記述言語:英語   会議種別:口頭発表(一般)  

    開催地:Pucon, Chile  

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  • A hypervariable mitochondrial gene associated with geographical origin in a cosmopolitan bloom-forming alga, Heterosigma akashiwo

    Shoko Ueki

    International Conference for Harmful Algae  International Society for the Study of Harmful Algae

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    開催年月日: 2018年10月21日 - 2018年10月26日

    記述言語:英語   会議種別:ポスター発表  

    開催地:Nantes, France  

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  • 赤潮原因藻ヘテロシグマのミトコンドリア・ゲノム上に存在する水域特異的な超可変領域配列についての研究

    植木 尚子

    2017環境微生物学系学会合同年会  東北大学

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    開催年月日: 2017年8月29日 - 2017年8月31日

    記述言語:日本語   会議種別:口頭発表(一般)  

    開催地:東北大学  

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  • Whole Genome sequence of Heterosigma akashiwo virus 53

    Shoko Ueki

    8th Aquatic Virus workshop  Plymouth University

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    開催年月日: 2016年7月10日 - 2016年7月12日

    記述言語:英語   会議種別:口頭発表(一般)  

    開催地:Plymouth, Devon, UK  

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  • 大型二本鎖DNAウイルス:多因子・多層制御による宿主感染機序の理解を目指して

    植木尚子

    新学術領域研究『ゲノム支援』拡大班会議  新学術領域研究『ゲノム支援』遺伝研事務局

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    開催年月日: 2015年8月27日 - 2015年8月28日

    記述言語:日本語   会議種別:口頭発表(一般)  

    開催地:京都国際会議場  

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  • 大型二本鎖DNAウイルス:多因子・多層制御による宿主感染機序の理解を目指して

    植木尚子

    ウイルス感染現象における宿主細胞コンピテンシーの分子基盤 第6回領域会議  ウイルス感染現象における宿主細胞コンピテンシーの分子基盤 領域事務局

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    開催年月日: 2015年5月24日 - 2015年5月25日

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • Growth promotion of Heterosigma akashiwo by marine microorganisms; implication of marine bacterium in bloom formation

    Shoko Ueki

    16th International Conference of Harmful Algae 

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    開催年月日: 2014年10月26日 - 2014年10月31日

    記述言語:英語   会議種別:口頭発表(一般)  

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  • ウイルス・宿主共存機構:宿主個体群構造ダイナミクスの生理生態学的・数理学的解析

    植木 尚子

    新学術領域研究『ゲノム支援』拡大班会議 

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    開催年月日: 2014年8月20日 - 2014年8月21日

    記述言語:日本語   会議種別:ポスター発表  

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共同研究・競争的資金等の研究

  • アスタキサンチン産生海洋細菌と赤潮原因藻の共生による赤潮の新規発生機構の解明

    研究課題/領域番号:21K19148  2021年07月 - 2024年03月

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

    植木 尚子

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    配分額:6240000円 ( 直接経費:4800000円 、 間接経費:1440000円 )

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  • 赤潮発生に必要な栄養基盤形成機構に海洋環境微生物の物質代謝が果たす役割の解明

    研究課題/領域番号:21H02285  2021年04月 - 2025年03月

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

    植木 尚子, 隠塚 俊満, 小原 静夏, 近藤 健

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    配分額:17160000円 ( 直接経費:13200000円 、 間接経費:3960000円 )

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  • 人為起源化学物質が沿岸域の基礎生産に及ぼす影響:低栄養や強光との複合影響の解明

    研究課題/領域番号:20H03067  2020年04月 - 2023年03月

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

    隠塚 俊満, 小池 一彦, 植木 尚子, 小原 静夏

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    配分額:17550000円 ( 直接経費:13500000円 、 間接経費:4050000円 )

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  • 赤潮形成を促進する海洋細菌の単離同定と、赤潮動態予測法の開発

    2019年04月 - 2021年03月

    特定非営利活動法人 瀬戸内海研究会議  大阪湾圏域の海域環境再生・創造に関する助成制度 

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    担当区分:研究代表者 

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  • チリにおける持続可能な沿岸漁業及び養殖に資する赤潮早期予測システムの構築と運用

    2018年04月 - 2023年03月

    日本科学技術振興機構  地球規模課題対応国際科学技術協力プログラム  生物資源

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    担当区分:研究代表者 

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  • 赤潮発生の謎を解く 〜赤潮原因藻の増殖速度制御機構の解明〜

    2018年10月 - 2019年09月

    積水化学工業株式会社  積水化学 自然に学ぶものづくり助成 

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    担当区分:研究代表者 

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  • 実用的物質生産系構築にむけたゲノム情報に基づく新規生合成システムのリデザイン

    研究課題/領域番号:16H06449  2016年06月 - 2021年03月

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

    渡辺 賢二, 植木 尚子

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    配分額:72020000円 ( 直接経費:55400000円 、 間接経費:16620000円 )

    担子菌ウシグソヒトヨタケCoprinopsis cinereaにおける内因性遺伝子の強制発現
    C. cinereaを宿主とした異種遺伝子発現を行う準備段階として、まずはC. cinerea自身の遺伝子を強制発現させることを試みた。既に我々は本遺伝子についてcDNAを取得した後にクローニングし、出芽酵母を用いて異種発現させることでその産物がオルセリン酸 (1)であることを報告している。そこでCC1G_05377の上流に強制発現が可能と考えられるb-tublin、EF3、DED19の各プロモーターを配置したプラスミドを構築し、これをC. cinerea 326株に導入した。PCRによって遺伝子導入が確認された変異体をMYG培地にて振盪培養し、その代謝産物をLC-MSにて解析したところ、それぞれ最大で約10倍(pb-tublin)、約100倍(pEF3)、約250倍(pDED1)と野生株よりも生産性の向上したクローンを得ることに成功した。
    続いてcop6遺伝子(CC1G_03563)に着目した。本遺伝子はセスキテルペン合成酵素Cop6をコードしており、FPPよりa-cupreneneを生合成することが報告されている。C. cinerea由来のa-cuprenene関連化合物としてはlagopodin類が知られており、加えてlagopodin類がさらに代謝されて生成したと考えられるhytoyol類の構造がつい最近、報告された。しかし、lagopodinおよびhytoyol生合成におけるcop6遺伝子の関与について直接的な証拠はこれまでに得られていなかった。そこでcop6遺伝子についてDED1プロモーターによる強制発現を試みた。得られた形質転換体は、野生株と比較してlagopodin類が約2-3倍、hytoyol類の生産性が約15倍に増加したクローンを見出すことに成功した。

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  • コレラ菌から地球規模での水の衛生微生物学的安全性を保証する

    研究課題/領域番号:16H05830  2016年04月 - 2020年03月

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

    丸山 史人, 竹村 太地郎, 中川 一路, 野中 里佐, 植木 尚子, 村瀬 一典

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    配分額:17420000円 ( 直接経費:13400000円 、 間接経費:4020000円 )

    本年度も特にベトナム、日本のコレラの解析を進めている。ベトナムでは多くの途上国と同様抗菌薬の使用量が非常に多く、臨床的な使用とともに農水産物の生産段階での多量の使用が大きな問題となっている。多くのビブリオ属細菌はエビ等の病原体として知られており、養殖池でビブリオ病予防のために抗菌薬が長期間使用されることも多い。ヒトにおける薬剤耐性菌感染に関する解析は十分とは言えないが多数報告されているのに比較し、環境中における薬剤耐性菌の調査研究は非常に報告が少ない。本研究ではエビ養殖池、農村部での農業・生活用水、河川水の解析を行い、ヒトにおける薬剤耐性菌侵淫とそのゲノム情報との比較解析を行うことで、耐性遺伝子の伝達特性、拡散・消長の過程を微生物生態学的見地から明らかにすることを目的としている。本年度はベトナム北部ナムディン省60検体、タイビン省40検体、ハイフォン市近郊46検体の環境水(河川、農業生活用水)および養殖池の検体解析を行った。選択培養法によりV. choleraeを分離した。現在分離株のうち20検体の全ゲノム解析を進めるとともに、2016年度に収集した株の全ゲノム配列を用いて薬剤耐性遺伝子の検索を進めている。また愛媛大学沿岸環境科学研究センター共同利用研究として「Vibrio属細菌が利用する遺伝子伝達機構の多様性解明と日本沿岸からのVibrio choleraの分離」にも採択され、愛媛県河川・沿岸域および台湾養殖場からのV. choleraeの全ゲノム配列解析中であり、遺伝的多様性解析および薬剤耐性プロファイルを明らかにしているので、これを継続していく。

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  • 大型二本鎖DNAウイルス:多因子・多層制御による宿主感染機序の理解を目指して

    研究課題/領域番号:15H01263  2015年04月 - 2017年03月

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

    植木 尚子

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    配分額:5850000円 ( 直接経費:4500000円 、 間接経費:1350000円 )

    本研究は、赤潮原因藻ヘテロシグマ(学名 Heterosigma akashiwo)に感染する大型二本鎖DNAウイルス( 以下GDNAVと略)であるHeterosigma akashiwo virus(HaV)の特性を明らかにし、また、その感染戦略を理解するために行った。まず、HaVの1系統であるHaV53の全長配列を解読し、遺伝子予測を行ったところ、HaV53は247遺伝子を持ち、また、3つのtRNAをコードすることが明らかになった。その遺伝子配列を、他のGDNAVゲノムと比較・精査したところ、HaV53は、他のGDNAVとは異なる特徴を持ったウイルスであることが明らかとなった。
    GDNAは、Nucleocytoplasmic large dsDNA virusとも呼ばれ、その増幅・転写は、多くは宿主核周辺で、その機能を利用して起こると理解されてきた。例えば、ウイルスゲノムが増殖する際に見られる「ウイルス工場」は、核内、あるいは核に隣接して観察され、また、これまで発表されたケースでは、感染過程で検出されるRNAはポリアデニル化されたものであることが知られてきた。しかし、HaVが感染した宿主のRNAをpolyAセレクション後にRNAseqした場合には、ウイルス由来の転写物はほとんど見出されなかった。一方、total RNAからribosomal RNAを除去したものをRNAseqした場合には、ウイルス由来転写物が検出された。つまり、HaV53感染時に検出されるウイルス遺伝子由来のRNAは、HaV53感染時の遺伝子転写は、核内RNAポリメラーゼ以外の機構を利用していることを示唆している。このような例は、GDNAVでは初めての報告であり、HaVの独自性を示すものであり、また、GDNAVの多様な感染戦略の一端を明らかにしたものと言える。

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  • ウイルス・宿主共存機構:宿主個体群構造ダイナミクスの生理生態学的・数理学的解析

    研究課題/領域番号:26291092  2014年04月 - 2017年03月

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

    植木 尚子, 佐藤 昌直, 中山 奈津子, 東 藍子, 小橋 理絵子

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    配分額:15990000円 ( 直接経費:12300000円 、 間接経費:3690000円 )

    赤潮原因藻ヘテロシグマと大型二本鎖DNAウイルスHeterosigma akashiwo virus (HaV)をモデルとし、HaVに対して異なる感染応答を示す宿主混合個体群へのウイルス感染進行の過程を解析を目的として研究を行った。本研究により、HaV全長遺伝子配列を解読し、他の大型二本鎖DNAウイルスとの比較解析により、進化的・分類学的知見を得た。また、ヘテロシグマのウイルス感染への応答は、宿主遺伝子型のみではなく、宿主に随伴する細菌の種類に依存することが明らかとなった。本研究は、予定とは異なる方向に進展したが、今後のヘテロシグマおよびHaV研究の進展の基盤となる成果を上げたと考えている。

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  • 赤潮原因藻ヘテロシグマの環境における増殖ダイナミクスの分子細胞生物学的研究

    研究課題/領域番号:26660153  2014年04月 - 2016年03月

    日本学術振興会  科学研究費助成事業 挑戦的萌芽研究  挑戦的萌芽研究

    植木 尚子, 中山 奈津子

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    配分額:3900000円 ( 直接経費:3000000円 、 間接経費:900000円 )

    赤潮は赤潮原因藻の異常増殖が原因である。通常赤潮は、一種の原因藻の異常増殖によるが、この限定された増殖促進のメカニズムはこれまでに明らかになっていない。私たちは、赤潮原因藻の一種であるヘテロシグマに着目して、ヘテロシグマ増殖を促進するバクテリアを数種単離した。その一種YF1株は、ヘテロシグマ増殖は促進するが、別の赤潮原因藻シャトネラの増殖には影響しない。また、ヘテロシグマとシャトネラが共存する場合、シャトネラ増殖がヘテロシグマにより抑制されることが知られてきたが、YF1は、この抑制効果をさらに増大させた。このような種特異的なバクテリアによる藻増殖促進は、赤潮の一種優占に貢献すると考えられる。

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  • 赤潮発生メカニズムの環境微生物学的理解を目指した研究

    2013年11月 - 2014年10月

    住友財団  住友財団環境研究助成 

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    担当区分:研究代表者 

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  • 赤潮原因害藻ヘテロシグマの産業利用を可能にする基盤技術の確立

    2012年12月 - 2013年11月

    カシオ科学振興研究助成金  カシオ科学振興研究助成金 

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    担当区分:研究代表者 

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  • 赤潮プランクトンの驚異的な増殖能力を有用物質生産に利用するための基礎的研究

    2012年04月 - 2013年03月

    長瀬科学技術振興財団  長瀬科学技術振興財団研究助成金 

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    担当区分:研究代表者 

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  • トランスグルタミナーゼによる細胞接着反応に関する基礎的研究

    研究課題/領域番号:97J04169  1998年 - 1999年

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

    植木 尚子

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    配分額:2400000円 ( 直接経費:2400000円 )

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その他研究活動

  • ラボHP

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    https://www.rib.okayama-u.ac.jp/inv/index-j.html

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担当授業科目

  • 植物微生物相互作用特論 (2021年度) 前期  - その他

  • 植物環境微生物学演習 (2021年度) 前期  - その他

  • 植物環境微生物学演習 (2021年度) 後期  - その他

  • 植物環境微生物学演習 (2021年度) 後期  - その他

  • 植物環境微生物学演習 (2021年度) 前期  - その他

  • 植物生理学1 (2021年度) 第3学期  - 金1,金2

  • 植物-ウイルス/細菌相互作用学 (2021年度) 前期  - 木9~12

  • 生物資源科学特別研究 (2021年度) 通年  - その他

  • 植物環境微生物学演習 (2020年度) 前期  - その他

  • 植物環境微生物学演習 (2020年度) 後期  - その他

  • 植物-ウイルス/細菌相互作用 (2020年度) 前期  - その他

  • 生物資源科学特別研究 (2020年度) 通年  - その他

▼全件表示