2021/12/22 更新

写真a

イチノセ ユウキ
一瀬 勇規
ICHINOSE Yuki
所属
環境生命科学学域 教授
職名
教授
外部リンク

学位

  • 農学修士 ( 岡山大学 )

  • 理学博士 ( 大阪大学 )

研究キーワード

  • Plant Molecular Biology

  • 植物病理学

  • Plant Pathology

  • Plant Genetic Engineering

研究分野

  • 環境・農学 / 植物保護科学

  • 環境・農学 / 植物保護科学

学歴

  • 大阪大学   Graduate School, Division of Natural Science  

    - 1988年

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  • 大阪大学    

    - 1988年

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    国名: 日本国

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  • 岡山大学   Faculty of Agriculture  

    - 1983年

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    国名: 日本国

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  • 岡山大学   Faculty of Agriculture  

    - 1983年

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経歴

  • 国立大学法人岡山大学   環境生命科学研究科   教授

    2018年4月 - 現在

  • 国立大学法人岡山大学   グローバル人材育成院   教授

    2017年4月 - 2018年3月

  • 国立大学法人岡山大学   環境生命科学研究科   教授

    2007年4月 - 2017年3月

  • 国立大学法人岡山大学   自然科学研究科   教授

    2005年4月 - 2007年3月

  • - Okayama University, Professor

    2001年

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  • - 岡山大学 教授

    2001年

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  • 岡山大学   自然科学研究科   助教授

    1999年4月 - 2001年3月

  • 岡山大学

    1989年 - 2001年

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  • 岡山大学

    1989年 - 2001年

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  • 大阪大学

    1989年

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  • Osaka University, Research Assistant

    1989年

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▼全件表示

所属学協会

  • 日本植物病理学会

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  • 植物微生物研究会

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  • American Society for Microbiology

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  • International Society for Molecular Plant-Microbe Interactions

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委員歴

  • 日本植物病理学会   理事  

    2020年1月 - 2021年12月   

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    団体区分:学協会

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  • 日本植物病理学会   関西部会部会長  

    2019年4月 - 2020年3月   

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    団体区分:学協会

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  • Plant Cell Physiology編集委員(2005-2007)  

    2005年 - 2007年   

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論文

  • Role of trehalose synthesis in Ralstonia syzygii subsp. indonesiensis PW1001 in inducing hypersensitive response on eggplant (Solanum melongena cv. Senryo-nigou) 査読

    37 ( 6 )   566 - 579   2021年12月

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    担当区分:最終著者, 責任著者  

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  • Identification of chemoreceptor proteins for amino acids involved in host plant infection in Pseudomonas syringae pv. tabaci 6605 査読

    Stephany Angelia Tumewu, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

    Microbiological Research   253   126869 - 126869   2021年12月

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    担当区分:最終著者, 責任著者   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 BV  

    DOI: 10.1016/j.micres.2021.126869

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  • Vfr targets promoter of genes encoding methyl-accepting chemotaxis protein in Pseudomonas syringae pv. tabaci 6605 査読

    Keisuke Ogura, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Fumiko Taguchi, Yuki Ichinose

    Biochemistry and Biophysics Reports   26   100944 - 100944   2021年7月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    DOI: 10.1016/j.bbrep.2021.100944

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  • Complete Genome Sequence of Pseudomonas amygdali pv. tabaci Strain 6605, a Causal Agent of Tobacco Wildfire Disease 査読

    Hidenori Matsui, Takafumi Nishimura, Shuta Asai, Sachiko Masuda, Ken Shirasu, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

    MICROBIOLOGY RESOURCE ANNOUNCEMENTS   10 ( 28 )   2021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:AMER SOC MICROBIOLOGY  

    Pseudomonas amygdali pv. tabaci strain 6605 is the bacterial pathogen causing tobacco wildfire disease that has been used as a model for elucidating virulence mechanisms. Here, we present the complete genome sequence of P. amygdali pv. tabaci 6605 as a circular chromosome from reads using a PacBio sequencer.

    DOI: 10.1128/MRA.00405-21

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  • Cluster II che genes of Pseudomonas syringae pv. tabaci 6605, orthologs of cluster I in Pseudomonas aeruginosa, are required for chemotaxis and virulence 査読

    Stephany Angelia Tumewu, Yujiro Ogawa, Takumi Okamoto, Yuka Sugihara, Hajime Yamada, Fumiko Taguchi, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

    Molecular Genetics and Genomics   2021年1月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    DOI: 10.1007/s00438-020-01745-y

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    その他リンク: http://link.springer.com/article/10.1007/s00438-020-01745-y/fulltext.html

  • HopH1 effectors of Pseudomonas syringae pv. tomato DC3000 and pv. syringae B728a induce HR cell death in nonhost eggplant Solanum torvum 査読

    Kamrun Nahar, Takafumi Mukaihara, Fumiko Taguchi, Hidenori Matsui, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshiteru Noutoshi, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   87 ( 1 )   24 - 29   2021年1月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Springer Science and Business Media LLC  

    DOI: 10.1007/s10327-020-00961-z

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    その他リンク: http://link.springer.com/article/10.1007/s10327-020-00961-z/fulltext.html

  • Identification of effector candidate genes of Rhizoctonia solani AG-1 IA expressed during infection in Brachypodium distachyon 査読

    Sobhy S. H. Abdelsalam, Yusuke Kouzai, Megumi Watanabe, Komaki Inoue, Hidenori Matsui, Mikihiro Yamamoto, Yuki Ichinose, Kazuhiro Toyoda, Seiji Tsuge, Keiichi Mochida, Yoshiteru Noutoshi

    Scientific Reports   10 ( 1 )   2020年12月

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    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>
    <italic>Rhizoctonia solani</italic> is a necrotrophic phytopathogen belonging to basidiomycetes. It causes rice sheath blight which inflicts serious damage in rice production. The infection strategy of this pathogen remains unclear. We previously demonstrated that salicylic acid-induced immunity could block <italic>R. solani</italic> AG-1 IA infection in both rice and <italic>Brachypodium distachyon</italic>. <italic>R. solani</italic> may undergo biotrophic process using effector proteins to suppress host immunity before necrotrophic stage. To identify pathogen genes expressed at the early infection process, here we developed an inoculation method using <italic>B. distachyon</italic> which enables to sample an increased amount of semi-synchronous infection hyphae. Sixty-one <italic>R. solani secretory effector-like protein</italic> genes (<italic>RsSEPGs</italic>) were identified using in silico approach with the publicly available gene annotation of <italic>R. solani</italic> AG-1 IA genome and our RNA-sequencing results obtained from hyphae grown on agar medium. Expression of <italic>RsSEPGs</italic> was analyzed at 6, 10, 16, 24, and 32 h after inoculation by a quantitative reverse transcription-polymerase chain reaction and 52 genes could be detected at least on a single time point tested. Their expressions showed phase-specific patterns which were classified into 6 clusters. The 23 <italic>RsSEPGs</italic> in the cluster 1–3 and 29 <italic>RsSEPGs</italic> in the cluster 4–6 are expected to be involved in biotrophic and necrotrophic interactions, respectively.

    DOI: 10.1038/s41598-020-71968-x

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    その他リンク: http://www.nature.com/articles/s41598-020-71968-x

  • Ralstonia solanacearum Type III Effector RipAC Targets SGT1 to Suppress Effector-Triggered Immunity 査読

    Masahito Nakano, Yuki Ichinose, Takafumi Mukaihara

    Plant and Cell Physiology   2020年9月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Oxford University Press (OUP)  

    <title>Abstract</title>
    Ralstonia solanacearum injects type III effectors into host cells to cause bacterial wilt in Solanaceae plants. To identify R. solanacearum effectors that suppress effector-triggered immunity (ETI) in plants, we evaluated R. solanacearum RS1000 effectors for their ability to suppress a hypersensitive response (HR) induced by the avirulence (Avr) effector RipAA in Nicotiana benthamiana. Out of the 11 effectors tested, 4 suppressed RipAA-triggered HR cell death. Among them, RipAC contains tandem repeats of the leucine-rich repeat (LRR) motif, which serves as the structural scaffold for a protein–protein interaction. We found that the LRR domain of RipAC was indispensable for the suppression of HR cell death during the recognition of RipAA and another Avr effector RipP1. By yeast two-hybrid screening, we identified N. benthamiana SGT1, an adaptor protein that forms a molecular chaperone complex with RAR1, as a host factor of the RipAC target. RipAC interacted with NbSGT1 in yeast and plant cells. Upon the formation of the molecular chaperone complex, the presence of RipAC markedly inhibits the interaction between NbSGT1 and NbRAR1. The RipAA- and RipP1-triggered HR cell deaths were not observed in NbSGT1-silenced plants. The introduction of RipAC was complementary to the reduced growth of the R. solanacearum mutant strain in N. benthamiana. These findings indicate that R. solanacearum uses RipAC to subvert the NbSGT1-mediated formation of the molecular chaperone complex and suppress ETI responses during the recognition of Avr effectors.

    DOI: 10.1093/pcp/pcaa122

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  • Role of Two Sets of RND-Type Multidrug Efflux Pump Transporter Genes, mexAB-oprM and mexEF-oprN, in Virulence of Pseudomonas syringae pv. tabaci 6605 査読

    Yuki Ichinose, Takafumi Nishimura, Minori Harada, Ryota Kashiwagi, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Fumiko Taguchi, Daigo Takemoto, Hidenori Matsui

    PLANT PATHOLOGY JOURNAL   36 ( 2 )   148 - 156   2020年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:KOREAN SOC PLANT PATHOLOGY  

    Pseudomonas syringae pv. tabaci 6605 has two multidrug resistance (MDR) efflux pump transporters, MexAB-OprM and MexEF-OprN. To understand the role of these MDR efflux pumps in virulence, we generated deletion mutants, Delta mexB, Delta mexF, and Delta mexB Delta mexF, and investigated their sensitivity to plant-derived antimicrobial compounds, antibiotics, and virulence. Growth inhibition assays with KB soft agar plate showed that growth of the wild-type (WT) was inhibited by 5 mu l of 1 M catechol and 1 M coumarin but not by other plant-derived potential antimicrobial compounds tested including phytoalexins. The sensitivity to these compounds tended to increase in Delta mexB and Delta mexB Delta mexF mutants. The Delta mexB Delta mexF mutant was also sensitive to 2 M acetovanillone. The mexAB-oprM was constitutively expressed, and activated in the Delta mexF and Delta mexB Delta mexF mutant strains. The swarming and swimming motilities were impaired in Delta mexF and Delta mexB Delta rnexF mutants. The flood inoculation test indicated that bacterial populations in all mutant strains were significantly lower than that of WT, although all mutants and WT caused similar disease symptoms. These results indicate that MexAB-OprM extrudes plant-derived catechol, acetovanillone, or coumarin, and contributes to bacterial virulence. Furthermore, MexAB-OprM and MexEF-OprN complemented each other's functions to some extent.

    DOI: 10.5423/PPJ.OA.11.2019.0273

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  • PsyR, a transcriptional regulator in quorum sensing system, binds lux box-like sequence in psyI promoter without AHL quorum sensing molecule and activates psyI transcription with AHL in Pseudomonas syringae pv. tabaci 6605 査読

    Yuki Ichinose, Yousuke Tasaka, Satoru Yamamoto, Yuko Inoue, Motohiro Takata, Yukiko Nakatsu, Fumiko Taguchi, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshiteru Noutoshi, Hidenori Matsui

    JOURNAL OF GENERAL PLANT PATHOLOGY   86 ( 2 )   124 - 133   2020年3月

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    担当区分:最終著者, 責任著者   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 JAPAN KK  

    Quorum sensing (QS) is a mechanism for bacterial cell-cell communication using QS signals. N-acyl-homoserine lactones (AHLs), QS signals in Pseudomonas syringae pv. tabaci (Pta) 6605, are synthesized by an AHL synthase (PsyI) and recognized by the cognate transcription factor PsyR. To reveal the role of PsyR in virulence, we generated a psyR mutant and complemented strains of Pta 6605 and found that the psyR mutant is remarkably reduced in AHL production and ability to cause disease and propagate in host tobacco leaves. The phenotypes of complemented strains were restored to that of the wild type (WT). Because the psyR mutant lost nearly all AHL production, we investigated the function of PsyR in the transcription of psyI and production of AHL. Electrophoretic mobility shift assays suggested that the recombinant PsyR protein binds the promoter region of psyI but not psyR without AHL. The addition of AHL did not significantly affect this binding. The binding core sequence of this region was identified as a 20-bp lux box-like sequence. To reveal the function of PsyR and AHL on psyI transcription, we constructed a psyI promoter::lacZYA chimeric reporter gene, and inserted it into the WT and psyI mutant of Pta 6605. beta-galactosidase activity increased in a bacterial density-dependent manner in the WT and also in a psyI mutant after the addition of exogenous AHL. These results indicate that the solo PsyR binds the lux box in the psyI promoter and activates transcription in the concomitant presence of AHL.

    DOI: 10.1007/s10327-019-00893-3

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  • Endogenous suppressor(s) in Arabidopsis thaliana 査読

    Thanh Luan Mai, Tatsuhiro Kawasaki, Aprilia Nur Fitrianti, Le Thi Phuong, Tsugumi Shiokawa, Hiroko Tada, Hidenori Matsui, Yoshiteru Noutoshi, Mikihiro Yamamoto, Yuki Ichinose, Tomonori Shiraishi, Kazuhiro Toyoda

    JOURNAL OF GENERAL PLANT PATHOLOGY   86 ( 2 )   100 - 106   2020年3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 JAPAN KK  

    An ethyl acetate extract of Arabidopsis thaliana plants was tested for the presence of endogenous suppressor(s) (ES), and the active fraction, which partitioned into water phase contained a molecule(s) < 3000 Da based on a rough estimate using sized membrane filters. Foliar application of the ES enabled typically nonpathogenic fungi (non-adapted pathogens) to cause disease symptoms on A. thaliana. Consistently, the ES fraction severely suppressed the oxidative burst and the expression of defense-related genes such as FRK1, NHO1, WRKY22, WRKY29, PEN2, and PEN3 in plants challenged with non-adapted fungus Colletotrichum gloeosporioides or the fungal elicitor chitin.

    DOI: 10.1007/s10327-019-00897-z

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  • Antagonism between SA- and JA-signaling conditioned by saccharin in Arabidopsis thaliana renders resistance to a specific pathogen 査読

    Le Thi Phuong, Aprilia Nur Fitrianti, Mai Thanh Luan, Hidenori Matsui, Yoshiteru Noutoshi, Mikihiro Yamamoto, Yuki Ichinose, Tomonori Shiraishi, Kazuhiro Toyoda

    JOURNAL OF GENERAL PLANT PATHOLOGY   86 ( 2 )   86 - 99   2020年3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 JAPAN KK  

    Saccharin is generated from probenazole (PBZ) in plants and acts as a plant defense activator. Our study of the mechanism underlying saccharin-induced systemic acquired resistance in Arabidopsis thaliana suggests an antagonistic interaction between salicylic acid (SA)- and jasmonic acid (JA)-signaling as revealed through gene expression analyses. In wild-type plants (Col-0) exposed to saccharin, there was a consistent increase in callose deposition and in expression of SA-marker genes, PR1 and PR2, which coincided with a decrease in expression of JA-marker genes such as VSP2, LOX2 and PDF1.2. Actually, pretreatment of Col-0 with saccharin or PBZ conferred resistance to Pseudomonas syringae pv. tomato DC3000, but not to Pectobacterium carotovorum subsp. carotovorum, Botrytis cinerea, or Colletotrichum higginsianum. Enhanced expression of SA- and JA-marker genes and the augmented deposition of callose were evident after a challenge with virulent DC3000 in saccharin-pretreated plants. Consistently, pretreatment of saccharin and PBZ with SA- and JA-defective mutants led to diminished resistance in NahG-transgenic and npr1 mutant plants, but not in jar1 mutant plants, suggesting that saccharin and PBZ induce resistance in A. thaliana against Pto DC3000 mainly via activation of SA-signaling, leading to suppression of JA/ET-signaling and vice versa. Collectively, an antagonism between SA- and JA-signaling conditioned by saccharin renders resistance to a specific pathogen in Arabidopsis.

    DOI: 10.1007/s10327-019-00899-x

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  • The plant activator saccharin induces resistance to wheat powdery mildew by activating multiple defense-related genes 査読

    Le Thi Phuong, Lei Zhao, Aprilia Nur Fitrianti, Hidenori Matsui, Yoshiteru Noutoshi, Mikihiro Yamamoto, Yuki Ichinose, Tomonori Shiraishi, Kazuhiro Toyoda

    JOURNAL OF GENERAL PLANT PATHOLOGY   86 ( 2 )   107 - 113   2020年3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 JAPAN KK  

    Saccharin and its parental compound probenazole (PBZ) are plant activators of effective defense responses to (hemi) biotrophic pathogens. Here, we demonstrate that pretreatment of wheat seedlings with saccharin or PBZ results in a significant reduction of powdery mildew caused by Blumeria graminis f. sp. tritici. Transcriptional analysis revealed the expression of 15 defense-related genes including PR genes, WCI genes, LOX, AOS, NPR1, PAL and WRKY genes in wheat seedlings exposed to either saccharin or PBZ. Moreover, the saccharin- and PBZ-enhanced expression of those genes during fungal infection further proved a close correlation with increased resistance in wheat.

    DOI: 10.1007/s10327-019-00900-7

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  • Requirement of γ-Aminobutyric Acid Chemotaxis for Virulence of Pseudomonas syringae pv. tabaci 6605. 査読

    Stephany Angelia Tumewu, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

    Microbes and environments   35 ( 4 )   2020年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    γ-Aminobutyric acid (GABA) is a widely distributed non-proteinogenic amino acid that accumulates in plants under biotic and abiotic stress conditions. Recent studies suggested that GABA also functions as an intracellular signaling molecule in plants and in signals mediating interactions between plants and phytopathogenic bacteria. However, the molecular mechanisms underlying GABA responses to bacterial pathogens remain unknown. In the present study, a GABA receptor, named McpG, was conserved in the highly motile plant-pathogenic bacteria Pseudomonas syringae pv. tabaci 6605 (Pta6605). We generated a deletion mutant of McpG to further investigate its involvement in GABA chemotaxis using quantitative capillary and qualitative plate assays. The wild-type strain of Pta6605 was attracted to GABA, while the ΔmcpG mutant abolished chemotaxis to 10‍ ‍mM GABA. However, ΔmcpG retained chemotaxis to proteinogenic amino acids and succinic semialdehyde, a structural analog of GABA. Furthermore, ΔmcpG was unable to effectively induce disease on host tobacco plants in three plant inoculation assays: flood, dip, and infiltration inoculations. These results revealed that the GABA sensing of Pta6605 is important for the interaction of Pta6605 with its host tobacco plant.

    DOI: 10.1264/jsme2.ME20114

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  • A class III peroxidase PRX34 is a component of disease resistance in Arabidopsis 査読

    Lei Zhao, Le Thi Phuong, Mai Thanh Luan, Aprilia Nur Fitrianti, Hidenori Matsui, Hirofumi Nakagami, Yoshiteru Noutoshi, Mikihiro Yamamoto, Yuki Ichinose, Tomonori Shiraishi, Kazuhiro Toyoda

    JOURNAL OF GENERAL PLANT PATHOLOGY   85 ( 6 )   405 - 412   2019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 JAPAN KK  

    PRX34 mediates the oxidative burst in Arabidopsis. Here we characterized two additional Arabidopsis prx34 null mutants (prx34-2, prx34-3), besides the well-studied prx34-1. Due to a decrease in corresponding peroxidase, the activity that generates reactive oxygen species (ROS) was significantly lower in cell wall extracts of prx34-2 and prx34-3 plants. Consistently, the prx34-2 and prx34-3 exhibited reduced accumulation both of ROS and callose in Flg22-elicitor-treated leaves, leading to enhanced susceptibility to bacterial and fungal pathogens. In contrast, ectopic expression of PRX34 in the wild type caused enhanced resistance. PRX34 is thus a component for disease resistance in Arabidopsis.

    DOI: 10.1007/s10327-019-00863-9

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  • Glycosidase and glycan polymorphism control hydrolytic release of immunogenic flagellin peptides 査読 国際誌

    Pierre Buscaill, Balakumaran Chandrasekar, Nattapong Sanguankiattichai, Jiorgos Kourelis, Farnusch Kaschani, Emma L. Thomas, Kyoko Morimoto, Markus Kaiser, Gail M. Preston, Yuki Ichinose, Renier A. L. van der Hoorn

    Science   364 ( 6436 )   2019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:American Association for the Advancement of Science (AAAS)  

    Plants and animals recognize conserved flagellin fragments as a signature of bacterial invasion. These immunogenic elicitor peptides are embedded in the flagellin polymer and require hydrolytic release before they can activate cell surface receptors. Although much of flagellin signaling is understood, little is known about the release of immunogenic fragments. We discovered that plant-secreted β-galactosidase 1 (BGAL1) of Nicotiana benthamiana promotes hydrolytic elicitor release and acts in immunity against pathogenic Pseudomonas syringae strains only when they carry a terminal modified viosamine (mVio) in the flagellin O-glycan. In counter defense, P. syringae pathovars evade host immunity by using BGAL1-resistant O-glycans or by producing a BGAL1 inhibitor. Polymorphic glycans on flagella are common to plant and animal pathogenic bacteria and represent an important determinant of host immunity to bacterial pathogens.

    DOI: 10.1126/science.aav0748

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  • Loliolide, a Carotenoid Metabolite, Is a Potential Endogenous Inducer of Herbivore Resistance. 査読 国際誌

    Mika Murata, Yusuke Nakai, Kei Kawazu, Masumi Ishizaka, Hideyuki Kajiwara, Hiroshi Abe, Kasumi Takeuchi, Yuki Ichinose, Ichiro Mitsuhara, Atsushi Mochizuki, Shigemi Seo

    Plant physiology   179 ( 4 )   1822 - 1833   2019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)  

    Jasmonic acid (JA) plays an important role in the induction of herbivore resistance in many plants. However, JA-independent herbivore resistance has been suggested. An herbivore-resistance-inducing substance was isolated from Tobacco mosaic virus-infected tobacco (Nicotiana tabacum) leaves in which a hypersensitive response (HR) was induced and identified as loliolide, which has been identified as a β-carotene metabolite. When applied to tomato (Solanum lycopersicum) leaves, loliolide decreased the survival rate of the two-spotted spider mite, Tetranychus urticae, egg deposition by the same pest, and the survival rate of larvae of the common cutworm Spodoptera litura without exhibiting toxicity against these herbivores. Endogenous loliolide levels increased not only with an infestation by Slitura larvae, but also with the exogenous application of their oral secretions in tomato. A microarray analysis identified cell-wall-associated defense genes as loliolide-responsive tomato genes, and exogenous JA application did not induce the expression of these genes. Suppressor of zeaxanthin-less (szl), an Arabidopsis (Arabidopsis thaliana) mutant with a point mutation in a key gene of the β-carotene metabolic pathway, exhibited the decreased accumulation of endogenous loliolide and increased susceptibility to infestation by the western flower thrip (Frankliniella occidentalis). A pretreatment with loliolide decreased susceptibility to thrips in the JA-insensitive Arabidopsis mutant coronatine-insensitive1 Exogenous loliolide did not restore reduced electrolyte leakage in szl in response to a HR-inducing bacterial strain. These results suggest that loliolide functions as an endogenous signal that mediates defense responses to herbivores, possibly independently of JA, at least in tomato and Arabidopsis plants.

    DOI: 10.1104/pp.18.00837

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  • Quorum-dependent expression of rsmX and rsmY, small non-coding RNAs, in Pseudomonas syringae 査読

    Nakatsu, Y, Matsui, H, Yamamoto, M, Noutoshi, Y, Toyoda, K, Ichinose, Y

    Microbiological Research   223–225   72 - 78   2019年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.micres.2019.04.004

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  • Specific growth inhibitors of Ralstonia solanacearum, Xanthomonas oryzae pv. oryzae, X. campestris pv. campestris, and Clavibacter michiganensis subsp. michiganensis (共著) 査読

    Geofrey Ombiro, Taku Sawai, Yoshiteru Noutoshi, Yuta Nisina, Hidenori Matsui, Mikihiro Yamamoto, Kazuhiro Toyoda, Yuki Ichinose

    Microbiological Research   215   29 - 35   2018年10月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.micres.2018.06.005

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  • MexEF-OprN multidrug efflux pump transporter negatively controls N-acyl-homoserine lactone accumulation in pseudomonas syringae pv. Tabaci 6605 (共著) 査読

    Takahiro Sawada, Miho Eguchi, Seiya Asaki, Ryota Kashiwagi, Kousuke Shimomura, Fumiko Taguchi, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

    Molecular Genetics and Genomics   293 ( 4 )   907 - 917   2018年8月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s00438-018-1430-9

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  • Salicylic acid-dependent immunity contributes to resistance against Rhizoctonia solani, a necrotrophic fungal agent of sheath blight, in rice and Brachypodium distachyon. 査読

    Kouzai Y, Kimura M, Watanabe M, Kusunoki K, Osaka D, Suzuki T, Matsui H, Yamamoto M, Ichinose Y, Toyoda K, Matsuura T, Mori IC, Hirayama T, Minami E, Noutoshi Y

    New Phytologist   2018年1月

  • Characterization of the suppressive effects of the biological control strain VAR03-1 of Rhizobium vitis on the virulence of tumorigenic R. vitis (共著) 査読

    Kirara Saito, Megumi Watanabe, Hidenori Matsui, Mikihiro Yamamoto, Yuki Ichinose, Kazuhiro Toyoda, Akira Kawaguchi, Yoshiteru Noutoshi

    Journal of General Plant Pathology   84 ( 1 )   58 - 64   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s10327-017-0756-1

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  • AefR transcription factor negatively regulates the virulence of Pseudomonas syringae pv. tomato DC3000 査読

    Ishiga, T, Ishiga, Y, Kiyokawa, T, Maruyama, N, Betsuyaku, S, Ichinose, Y, Nomura, N

    PHYTOPATHOLOGY   107 ( 12::S )   83 - 83   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:AMER PHYTOPATHOLOGICAL SOC  

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  • Ultrastructural and Cytological Studies on Mycosphaerella pinodes Infection of the Model Legume Medicago truncatula 査読

    Tomoko Suzuki, Aya Maeda, Masaya Hirose, Yuki Ichinose, Tomonori Shiraishi, Kazuhiro Toyoda

    FRONTIERS IN PLANT SCIENCE   8   2017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:FRONTIERS MEDIA SA  

    Ascochyta (Mycosphaerella) blight on cultivated peas is primarily caused by infection through asexual spores (pycnospores) of Mycosphaerella pinodes (Berk. et Blox.) Vestergren [recently renamed Peyronellaea pinodes (Berk. & A. Bloxam) Aveskamp, Gruyter & Verkley]. Using a model pathosystem involving Medicago truncatula and Mycosphaerella pinodes strain OMP-1, we examined the histology and ultrastructure of early infection events and fungal development including penetration by appressoria, vegetative growth of infection hyphae, and host responses. On the susceptible ecotype R108-1, pycnospores germinated and grew over the surface of the epidermis, then formed an appressoria and penetrated the cuticle. Beneath the cuticle, the infection peg expanded into a hyphae that grew within the outer wall of the epidermis. Subsequently, the hyphae penetrated down within mesophyll cells and proliferated vigorously, eventually, forming asexual fruiting bodies (pycnidia). In contrast, successful penetration and subsequent growth of infection hyphae were considerably restricted in the ecotype Caliph. Detected by its reaction with cerium chloride (CeCl3) to generate electron-dense cerium perhydroxides in transmission electron micrographs, hydrogen peroxide (H2O2) accumulated in epidermal and mesophyll cells of Caliph challenged with pycnospores of M. pinodes. This intracellular localization was confirmed by energydispersive X-ray spectroscopy. Our observations thus indicate that the oxidative burst reaction leading to the generation of reactive oxygen species is associated with a local host defense response in Caliph, since no clear H2O2 accumulation was detectable in susceptible R108-1. Indeed, aberrant hyphae such as intrahyphal hyphae and dead hyphae, probably due to a local defense elicited by the fungus, were abundant in Caliph but not in R108-1. Our results on the cellular interactions between the fungus and host cells provide additional insights to understand foliar infection by M. pinodes on cultivated peas.

    DOI: 10.3389/fpls.2017.01132

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  • Pseudomonas syringae Flood-inoculation Method in Arabidopsis 査読

    Yasuhiro Ishiga, Takako Ishiga, Yuki Ichinose, Kirankumar S. Mysore

    BIO-PROTOCOL   7 ( 2 )   2017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:BIO-PROTOCOL  

    Pseudomonas syringae pv. tomato strain DC3000 (Pto DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen because of its pathogenicity on Arabidopsis thaliana. Here, we demonstrate a rapid and reliable flood-inoculation method based on young Arabidopsis seedlings grown on one-half strength MS medium. We also describe a method to evaluate the bacterial growth in Arabidopsis.

    DOI: 10.21769/BioProtoc.2106

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  • Pseudomonas syringaeの菌体密度感知機構と多剤排出ポンプの病原力における役割 招待

    一瀬勇規, 澤田貴博, 髙田基弘, 山本悟, 藤山友里, 中津有紀子, 田阪洋昌, 下村洪祐, 田口富美子, 松井英譲, 山本幹博, 能年義輝, 豊田和弘

    植物細菌病談話会論文集   27   77 - 88   2016年8月

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    担当区分:最終著者, 責任著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • Expression profiling of marker genes responsive to the defence-associated phytohormones salicylic acid, jasmonic acid and ethylene in Brachypodium distachyon. 査読

    Kouzai Y, Kimura M, Yamanaka Y, Watanabe M, Matsui H, Yamamoto M, Ichinose Y, Toyoda K, Onda Y, Mochida K, Noutoshi Y

    BMC plant biology   16 ( 1 )   59   2016年3月

  • Pseudomonas syringae pv. tomato OxyR Is Required for Virulence in Tomato and Arabidopsis 査読

    Yasuhiro Ishiga, Yuki Ichinose

    MOLECULAR PLANT-MICROBE INTERACTIONS   29 ( 2 )   119 - 131   2016年2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:AMER PHYTOPATHOLOGICAL SOC  

    Reactive oxygen species (ROS) have been shown to have a crucial role in plant defense responses and signaling pathways. In addition, ROS also have direct toxicity against pathogens. However, the molecular mechanisms of plant ROS in the direct effects against pathogens is still unclear. To investigate the function of plant ROS in the interactions of plant and bacterial pathogens, we focused on oxyR, encoding an oxidative stress regulated transcription factor in Pseudomonas syringae pv. tomato DC3000 (DC3000), and generated an Delta oxyR mutant. The DC3000 Delta oxyR mutant showed high sensitivity to oxidative stress in comparison with wild type and the complemented line. The host plants of DC3000, including tomato and Arabidopsis inoculated with the Delta oxyR mutant, clearly showed reduced disease symptoms as well as reduced bacterial populations. Expression profiles of DC3000 genes revealed that OxyR could regulate the expression of genes encoding ROS-detoxifying enzymes, including catalases (KatB and KatG), in response to ROS. We also demonstrated that the expression of katB could be regulated by OxyR during the infection of DC3000 in Arabidopsis. These results suggest that OxyR has an important role in the virulence of DC3000 by regulating the expression of genes related to oxidative stress.

    DOI: 10.1094/MPMI-09-15-0204-R

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  • Motility-mediated regulation of virulence in Pseudomonas syringae (共著) 招待 査読

    Yuki Ichinose, Takahiro Sawada, Hidenori Matsui, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshiteru Noutoshi, Fumiko Taguchi

    Physiological and Molecular Plant Pathology   95   50 - 54   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.pmpp.2016.02.005

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  • The plant cell wall as a site for molecular contacts in fungal pathogenesis (共著) 招待 査読

    Kazuhiro Toyoda, Sachiyo Yao, Mai Takagi, Maki Uchioki, Momiji Miki, Kaori Tanaka, Tomoko Suzuki, Masashi Amano, Akinori Kiba, Toshiaki Kato, Hirotaka Takahashi, Yasuhiro Ishiga, Hidenori Matsui, Yoshiteru Noutoshi, Mikihiro Yamamoto, Yuki Ichinose, Tomonori Shiraishi

    Physiological and Molecular Plant Pathology   95   44 - 49   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.pmpp.2016.02.006

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  • Protection induced by volatile limonene against anthracnose disease in Arabidopsis thaliana. 査読

    Fujioka, K, Gotoh, H, Noumi, T, Yoshida, A, Noutoshi, Y, Inagaki, Y, Yamamoto, M, Ichinose, Y, Shiraishi, T, Toyoda K

    J Gen Plant Pathol   81 ( 6 )   415 - 419   2015年

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    記述言語:英語  

    DOI: 10.1007/s10327-015-0621-z

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  • Characterization of quorum sensing-controlled transcriptional regulator MarR and Rieske (2Fe-2S) cluster-containing protein (Orf5) that are involved in resistance to environmental stresses in Pseudomonas syringae pv. tabaci 6605. 査読

    Taguchi F, Inoue Y, Suzuki T, Inagaki Y, Yamamoto M, Yoyoda K, Noutoshi Y, Shiraishi T, Ichinose Y

    Mol. Plant Pathol.   16 ( 4 )   376 - 387   2015年

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    記述言語:英語  

    DOI: 10.1111/mpp.12187

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  • Expression of Medicago truncatula ecto-apyrase MtAPY1;1 in leaves of Nicotiana benthamiana restricts necrotic lesions induced by a virulent fungus. 査読

    Toyoda K, Kawakami E, Nagai H, Shiobara-Komatsu T, Tanaka K, Inagaki Y, Ichinose Y, Shiraishi T

    J Gen Plant Pathol   80 ( 3 )   222 - 229   2014年

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    記述言語:英語  

    DOI: 10.1007/s10327-014-0510-x

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  • Ralstonia solanacearum type III secretion system effector Rip36 induces hypersensitive response in the nonhost wild eggplant Solanum torvum. 査読

    Nahar K, Matsumoto, I, Taguchi, F, Inagaki, Y, Yamamoto, M, Yoyoda, K, Shiraishi, T, Ichinose, Y, Mukaihara, T

    Mol. Plant Pathol.   15 ( (3) )   297 - 303   2014年

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    記述言語:英語  

    DOI: 10.1111/mpp.12079

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  • Allelic variation in two distinct Pseudomonas syringae flagellin epitopes modulates the strength of plant immune responses but not bacterial motility 査読

    Christopher R. Clarke, Delphine Chinchilla, Sarah R. Hind, Fumiko Taguchi, Ryuji Miki, Yuki Ichinose, Gregory B. Martin, Scotland Leman, Georg Felix, Boris A. Vinatzer

    NEW PHYTOLOGIST   200 ( 3 )   847 - 860   2013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:WILEY-BLACKWELL  

    The bacterial flagellin (FliC) epitopes flg22 and flgII-28 are microbe-associated molecular patterns (MAMPs). Although flg22 is recognized by many plant species via the pattern recognition receptor FLS2, neither the flgII-28 receptor nor the extent of flgII-28 recognition by different plant families is known. Here, we tested the significance of flgII-28 as a MAMP and the importance of allelic diversity in flg22 and flgII-28 in plant-pathogen interactions using purified peptides and a Pseudomonas syringae fliC mutant complemented with different fliC alleles. The plant genotype and allelic diversity in flg22 and flgII-28 were found to significantly affect the plant immune response, but not bacterial motility. The recognition of flgII-28 is restricted to a number of solanaceous species. Although the flgII-28 peptide does not trigger any immune response in Arabidopsis, mutations in both flg22 and flgII-28 have FLS2-dependent effects on virulence. However, the expression of a tomato allele of FLS2 does not confer to Nicotiana benthamiana the ability to detect flgII-28, and tomato plants silenced for FLS2 are not altered in flgII-28 recognition. Therefore, MAMP diversification is an effective pathogen virulence strategy, and flgII-28 appears to be perceived by an as yet unidentified receptor in the Solanaceae, although it has an FLS2-dependent virulence effect in Arabidopsis.

    DOI: 10.1111/nph.12408

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  • Flagellin glycosylation is ubiquitous in a broad range of phytopathogenic bacteria 査読

    Yuki Ichinose, Fumiko Taguchi, Masanobu Yamamoto, Mayumi Ohnishi-Kameyama, Tatsuo Atsumi, Tatsuo Atsumi, Masako Iwaki, Hiromi Manabe, Mio Kumagai, Quan Thanh Nguyen, Chi Linh Nguyen, Yoshishige Inagaki, Hiroshi Ono, Kazuhiro Chiku, Tadashi Ishii, Mitsuru Yoshida

    Journal of General Plant Pathology   79 ( 5 )   359 - 365   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Glycosylation of flagellin is known to be involved in filament stabilization, motility, and virulence in Pseudomonas syringae. Here we investigated flagellin glycosylation in other phytopathogenic bacteria. Analyses of deduced amino acid sequences, glycostaining, and molecular masses of purified flagellins revealed that flagellins from all phytopathogenic bacteria investigated were glycosylated. Furthermore, the flagellin in a glycosylation-defective mutant of Xanthomonas campestris pv. campestris (Xcc) had a reduced molecular mass, and motility and virulence of the mutant toward host leaves decreased. These results suggest that flagellin glycosylation is ubiquitous in most phytopathogenic bacteria and that flagellin glycosylation is required for virulence in Xcc. © 2013 The Phytopathological Society of Japan and Springer Japan.

    DOI: 10.1007/s10327-013-0464-4

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  • Comparative analysis of flagellin glycans among pathovars of phytopathogenic Pseudomonas syringae. 査読

    Chiku K, Yamamoto M, Ohnishi-Kameyama M, Ishii T, Yoshida M, Taguchi F, Ichinose Y, Ono H

    Carbohydrate research   375   100 - 104   2013年6月

  • Virulence factor regulator (Vfr) controls virulence-associated phenotypes in Pseudomonas syringae pv. tabaci 6605 by a quorum sensing-independent mechanism 査読

    Fumiko Taguchi, Yuki Ichinose

    Molecular Plant Pathology   14 ( 3 )   279 - 292   2013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)  

    Virulence factor regulator (Vfr) is a member of the cyclic 3′,5′-adenosine monophosphate (cAMP) receptor proteins that regulate the expression of many important virulence genes in Pseudomonas aeruginosa. The role of Vfr in pathogenicity has not been elucidated fully in phytopathogenic bacteria. To investigate the function of Vfr in Pseudomonas syringae pv. tabaci 6605, the vfr gene was disrupted. The virulence of the vfr mutant towards host tobacco plants was attenuated significantly, and the intracellular cAMP level was decreased. The vfr mutant reduced the expression of flagella-, pili- and type III secretion system-related genes and the defence response in nonhost Arabidopsis leaves. Furthermore, the expression levels of achromobactin-related genes and the iron uptake ability were decreased, suggesting that Vfr regulates positively these virulence-related genes. In contrast, the vfr mutant showed higher tolerance to antimicrobial compounds as a result of the enhanced expression of the resistance-nodulation-division family members, the mexA, mexB and oprM genes. We further demonstrated that the mutant strains of vfr and cyaA, an adenylate cyclase gene responsible for cAMP synthesis, showed a similar phenotype, suggesting that Vfr regulates virulence factors in a cAMP-dependent manner. Because there was no significant difference in the production of acylhomoserine lactone (AHL) quorum sensing molecules in the wild-type, vfr and cyaA mutant strains, Vfr might control important virulence factors by an AHL-independent mechanism in an early stage of infection by this bacterium. © 2012 The Authors. Molecular Plant Pathology © 2012 Bspp and Blackwell Publishing Ltd.

    DOI: 10.1111/mpp.12003

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  • Defects in D-Rhamnosyl Residue Biosynthetic Genes Affect Lipopolysaccharide Structure, Motility, and Cell-Surface Hydrophobicity in Pseudomonas syringae Pathovar glycinea Race 4 査読

    Kazuhiro Chiku, Kazuhiko Tsunemi, Masanobu Yamamoto, Mayumi Ohnishi-Kameyama, Mitsuru Yoshida, Tadashi Ishii, Fumiko Taguchi, Masako Iwaki, Yuki Ichinose, Hiroshi Ono

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   77 ( 3 )   505 - 510   2013年3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:TAYLOR & FRANCIS LTD  

    D-rhamnose (D-Rha) residue is a major component of lipopolysaccharides (LPSs) in strains of the phytopathogen Pseudomonas syringae pathovar glycinea. To investigate the effects of a deficiency in GDP-D-rhamnose biosynthetic genes on LPS structure and pathogenicity, we generated three mutants defective in D-Rha biosynthetic genes, encoding proteins GDP-D-mannose 4,6-dehydratase (GMD), GDP-4-keto-6-deoxy-D-mannose reductase (RMD), and a putative alpha-D-rhamnosyltransferase (WbpZ) in P. syringae pv. glyeinea race 4. The Delta gmd, Delta rmd, and Delta wbpZ mutants had a reduced O-antigen polysaccharide consisting of D-Rha residues as compared with the wild type (WT). The swarming motility of the Delta gmd, Delta rmd, and Delta wbpZ mutant strains decreased and hydrophobicity and adhesion ability increased as compared with WT. Although the mutants had truncated O-antigen polysaccharides, and altered surface properties, they showed virulence to soybean, as WT did.

    DOI: 10.1271/bbb.120736

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  • Infection-inhibition activity of avenacin saponins against the cereal pathogens Blumeria graminis f.sp. hordei, Bipolaris oryzae, and Magnaporthe oryzae. 査読

    Inagaki Y, Noutoshi Y, Fujita K, Imaoka A, Arase S, Toyoda K, Shiraishi T, Ichinose Y

    J Gen Plant Pathol,   79 ( (1) )   69 - 73   2013年

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    記述言語:英語  

    DOI: 10.1007/s10327-012-0412-8

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  • Suppression of mRNAs for lipoxygenase (LOX), allene oxide synthase (AOS), allene oxide cyclase (AOC) and 12-oxo-phytodienoic acid reductase (OPR) in pea reduces sensitivity to the phytotoxin coronatine and disease development by Mycosphaerella pinodes. 査読

    Toyoda K, Kawanishi Y, Kawamoto Y, Kurihara C, Yamagishi N, Tamura A, Yoshikawa N, Inagaki Y, Ichinose Y, Shiraishi T

    J Gen Plant Pathol   79 ( 5 )   321 - 334   2013年

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    DOI: 10.1007/s10327-013-0460-8

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  • 岡山県の栽培圃場における植物生育促進菌の探索と同定

    山際泰夫, 豊田和弘, 稲垣善茂, 一瀬勇規, 白石友紀

    岡山大学農学部学術報告   102   1 - 6   2013年

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    記述言語:日本語   掲載種別:研究論文(大学,研究機関等紀要)  

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  • β-caryophylleneの植物に対する生育促進作用および耐病性増進作用の解析

    山際泰夫, 豊田和弘, 稲垣善茂, 一瀬勇規, 百町満朗, 白石友紀

    岡山大学農学部学術報告   102   7 - 14   2013年

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    記述言語:日本語   掲載種別:研究論文(大学,研究機関等紀要)  

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  • The Medicago truncatula-Mycoshaerella pinodes interaction: a new pathosystem for dissecting the fungal suppressor-mediated plant disease susceptibility in plants. 査読

    Toyoda K, Ikeda S, Morikawa J, Hirose M, Maeda A, Suzuki T, Inagaki Y, Ichinose Y, Shiraishi T

    J Gen Plant Pathol,   79 ( (1) )   1 - 11   2013年

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    記述言語:英語  

    DOI: 10.1007/s10327-012-0405-7

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  • Plant cell walls as suppliers of potassium and sodium ions for induced resistance of pea (Pisum sativum L.) and cowpea (Vigna ungiculata L.). 査読

    Amano M, Toyota K, Kiba A, Inagaki Y, Ichinose Y, Shiraishi T

    J Gen Plant Pathol,   79 ( (1) )   12 - 17   2013年

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    記述言語:英語  

    DOI: 10.1007/s10327-012-0418-2

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  • Type IV pilin is glycosylated in Pseudomonas syringae pv. tabaci 6605 and required for surface motility and virulence 査読

    Linh Chi Nguyen, Fumiko Taguchi, Quang Minh Tran, Kana Naito, Masanobu Yamamoto, Mayumi Ohnishi-Kameyama, Hiroshi Ono, Mitsuru Yoshida, Kazuhiro Chiku, Tadashi Ishii, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Molecular Plant Pathology   13 ( 7 )   764 - 774   2012年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Type IV pilin (PilA) is a major constituent of pilus and is required for bacterial biofilm formation, surface motility and virulence. It is known that mature PilA is produced by cleavage of the short leader sequence of the pilin precursor, followed by methylation of N-terminal phenylalanine. The molecular mass of the PilA mature protein from the tobacco bacterial pathogen Pseudomonas syringae pv. tabaci 6605 (Pta 6605) has been predicted to be 12 329 Da from its deduced amino acid sequence. Previously, we have detected PilA as an approximately 13-kDa protein by immunoblot analysis with anti-PilA-specific antibody. In addition, we found the putative oligosaccharide-transferase gene tfpO downstream of pilA. These findings suggest that PilA in Pta 6605 is glycosylated. The defective mutant of tfpO (ΔtfpO) shows reductions in pilin molecular mass, surface motility and virulence towards host tobacco plants. Thus, pilin glycan plays important roles in bacterial motility and virulence. The genetic region around pilA was compared among P. syringae pathovars. The tfpO gene exists in some strains of pathovars tabaci, syringae, lachrymans, mori, actinidiae, maculicola and P. savastanoi pv. savastanoi. However, some strains of pathovars tabaci, syringae, glycinea, tomato, aesculi and oryzae do not possess tfpO, and the existence of tfpO is independent of the classification of pathovars/strains in P. syringae. Interestingly, the PilA amino acid sequences in tfpO-possessing strains show higher homology with each other than with tfpO-nonpossessing strains. These results suggest that tfpO and pilA might co-evolve in certain specific bacterial strains.

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  • Identification of natural diterpenes that inhibit bacterial wilt disease in Tobacco, Tomato and Arabidopsis 査読

    Shigemi Seo, Kenji Gomi, Hisatoshi Kaku, Hiroshi Abe, Hideharu Seto, Shingo Nakatsu, Masahiro Neya, Michie Kobayashi, Kazuhiro Nakaho, Yuki Ichinose, Ichiro Mitsuhara, Yuko Ohashi

    Plant and Cell Physiology   53 ( 8 )   1432 - 1444   2012年8月

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    掲載種別:研究論文(学術雑誌)  

    The soil-borne bacterial pathogen Ralstonia solanacearum invades a broad range of plants through their roots, resulting in wilting of the plant, but no effective protection against this disease has been developed. Two bacterial wilt disease-inhibiting compounds were biochemically isolated from tobacco and identified as sclareol and cis-abienol, labdane-type diterpenes. When exogenously applied to their roots, sclareol and cis-abienol inhibited wilt disease in tobacco, tomato and Arabidopsis plants without exhibiting any antibacterial activity. Microarray analysis identified many sclareol-responsive genes in Arabidopsis roots, including genes encoding or with a role in ATP-binding cassette (ABC) transporters, and biosynthesis and signaling of defense-related molecules and mitogen-activated protein kinase (MAPK) cascade components. Inhibition of wilt disease by sclareol was attenuated in Arabidopsis mutants defective in the ABC transporter AtPDR12, the MAPK MPK3, and ethylene and abscisic acid signaling pathways, and also in transgenic tobacco plants with reduced expression of NtPDR1, a tobacco homolog of AtPDR12. These results suggest that multiple host factors are involved in the inhibition of bacterial wilt disease by sclareol-related compounds. © The Author 2012. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

    DOI: 10.1093/pcp/pcs085

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  • Characterization of each aefR and mexT mutant in Pseudomonas syringae pv. tabaci 6605. 査読

    Kawakita Y, Taguchi F, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y

    Mol. Genet. Genomics   287   473 - 484   2012年

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    記述言語:英語  

    DOI: 10.1007/s00438-012-0693-9

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  • H2O2 production by copper amine oxidase, a component of the ecto-apyrase (ATPase)-containing protein complex(es) in the pea cell wall, is regulated by an elicitor and a suppressor from Mycosphaerella pinodes. 査読

    Toyoda K, Yasunaga E, Niwa M, Ohwatari Y, Nakashima A, Inagaki Y, Ichinose Y, Shiraishi T

    J Gen Plant Pathol,   78 ( (5) )   311 - 315   2012年

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    記述言語:英語  

    DOI: 10.1007/s10327-012-0399-1

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  • Identification of Genes Involved in the Glycosylation of Modified Viosamine of Flagellins in Pseudomonas syringae by Mass Spectrometry 査読

    Masanobu Yamamoto, Mayumi Ohnishi-Kameyama, Chi L. Nguyen, Fumiko Taguchi, Kazuhiro Chiku, Tadashi Ishii, Hiroshi Ono, Mitsuru Yoshida, Yuki Ichinose

    GENES   2 ( 4 )   788 - 803   2011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:MDPI AG  

    Previously we revealed that flagellin proteins in Pseudomonas syringae pv. tabaci 6605 (Pta 6605) were glycosylated with a trisaccharide, modified viosamine (mVio)-rhamnose-rhamnose and that glycosylation was required for virulence. We further identified some glycosylation-related genes, including vioA, vioB, vioT, fgt1, and fgt2. In this study, we newly identified vioR and vioM in a so-called viosamine island as biosynthetic genes for glycosylation of mVio in Pta 6605 by the mass spectrometry (MS) of flagellin glycan in the respective mutants. Furthermore, characterization of the mVio-related genes and MS analyses of flagellin glycans in other pathovars of P. syringae revealed that mVio-related genes were essential for mVio biosynthesis in flagellin glycans, and that P. syringae pv. syringae B728a, which does not possess a viosamine island, has a different structure of glycan in its flagellin protein.

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  • Role of Type IV Pili in Virulence of Pseudomonas syringae pv. tabaci 6605: Correlation of Motility, Multidrug Resistance, and HR-Inducing Activity on a Nonhost Plant 査読

    Fumiko Taguchi, Yuki Ichinose

    MOLECULAR PLANT-MICROBE INTERACTIONS   24 ( 9 )   1001 - 1011   2011年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    To investigate the role of type IV pili in the virulence of phytopathogenic bacteria, four mutant strains for pilus biogenesis-related genes were generated in Pseudomonas syringae pv. tabaci 6605. PilA encodes the pilin protein as a major subunit of type IV pili, and the pilO product is reported to be required for pilus assembly. The fimU and fimT genes are predicted to produce minor pilins. Western blot analysis revealed that pilA, pilO, and fimU mutants but not the fluff mutant failed to construct type IV phi. Although the swimming motility of all mutant strains was not impaired in liquid medium, they showed remarkably reduced motilities on semisolid agar medium, suggesting that type IV pili are required for surface motilities. Virulence toward host tobacco plants and hypersensitive response-inducing ability in nonhost Arabidopsis leaves of pilA, pilO, and fimU mutant strains were reduced. These results might be a consequence of reduced expression of type Ill secretion system-related genes in the mutant strains. Further, all mutant strains showed enhanced expression of resistance-nodulation-division family members mexA, mexB, and oprM, and higher tolerance to antimicrobial compounds. These results indicate that type IV pili are an important virulence factor of this pathogen.

    DOI: 10.1094/MPMI-02-11-0026

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  • Two flagellar stators and their roles in motility and virulence in Pseudomonas syringae pv. tabaci 6605. 査読

    Kanda E, Tatsuta T, Suzuki T, Taguchi F, Naito K, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y

    Mol. Genet. Genomics   285 ( (2) )   163 - 174   2011年

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: 10.1007/s00438-010-0594-8

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  • Talaromyces wortmannii FS2 emits beta-caryophyllene, which promotes plant growth and induces resistance. 査読

    Yamagiwa Y, Inagaki Y, Ichinose Y, Toyoda K, Hyakumachi M, Shiraishi T

    J. Gen. Plant Pathol.   77 ( (6) )   336 - 341   2011年

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    記述言語:英語  

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  • Degeneration of hrpZ gene in Pseudomonas syringae pv. tabaci to evade tobacco defence: an arms race between tobacco and its bacterial pathogen. 査読

    Tsunemi K, Taguchi F, Marutani M, Watanabe-Sugimoto M, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y

    Mol Plant Pathol.   12 ( (7) )   709 - 714   2011年

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: 10.1111/j.1364-3703.2011.00705.x

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  • Defects in flagellin glycosylation affect the virulence of Pseudomonas syringae pv. tabaci 6605. 査読

    Taguchi F, Yamamoto M, Ohnishi-Kameyama M, Iwaki M, Yoshida M, Ishii T, Konishi T, Ichinose Y

    Microbiology (Reading, England)   156 ( Pt 1 )   72 - 80   2010年1月

  • Large-scale analysis of full-length cDNAs from the tomato (Solanum lycopersicum) cultivar Micro-Tom, a reference system for the Solanaceae genomics 査読

    Koh Aoki, Kentaro Yano, Ayako Suzuki, Shingo Kawamura, Nozomu Sakurai, Kunihiro Suda, Atsushi Kurabayashi, Tatsuya Suzuki, Taneaki Tsugane, Manabu Watanabe, Kazuhide Ooga, Maiko Torii, Takanori Narita, Tadasu Shin-i, Yuji Kohara, Naoki Yamamoto, Hideki Takahashi, Yuichiro Watanabe, Mayumi Egusa, Motoichiro Kodama, Yuki Ichinose, Mari Kikuchi, Sumire Fukushima, Akiko Okabe, Tsutomu Arie, Yuko Sato, Katsumi Yazawa, Shinobu Satoh, Toshikazu Omura, Hiroshi Ezura, Daisuke Shibata

    BMC Genomics   11 ( 1 )   210 - 210   2010年

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

    DOI: 10.1186/1471-2164-11-210

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  • Genetic analysis of genes involved in synthesis of modified 4-amino-4,6-dideoxyglucose in flagellin of Pseudomonas syringae pv. tabaci 査読

    Linh Chi Nguyen, Masanobu Yamamoto, Mayumi Ohnishi-Kameyama, Salamah Andi, Fumiko Taguchi, Masako Iwaki, Mitsuru Yoshida, Tadashi Ishii, Tomoyuki Konishi, Kazuhiko Tsunemi, Yuki Ichinose

    Molecular Genetics and Genomics   282   595 - 605   2009年12月

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    担当区分:最終著者, 責任著者  

    Glycosylation of flagellin contributes to swimming and swarming motilities, adhesion ability, and consequently virulence in Pseudomonas syringae pv. tabaci 6605. Glycans attached to six serine residues are located in the central region of the flagellin polypeptide. The glycan structure at position Ser 201 was recently revealed to consist of two l-rhamnoses and one modified 4-amino-4,6-dideoxyglucose (viosamine). To clarify the mechanisms for glycosylation of modified viosamine, genes encoding dTDP-viosamine aminotransferase (vioA), dTDP-viosamine acetyltransferase (vioB), and viosamine-derivative transferase (vioT) were isolated and defective mutants were generated. MALDI-TOF-MS analysis of a lysyl endopeptidase-digested peptide including all six glycosylation sites from each flagellin indicated that the molecular masses of the three flagellin mutants were reduced with highly heterogeneous patterns at regular intervals of 146 Da in the mass range from m/z 13,819 to 15,732. The data indicated that the glycopeptides obtained from mutants had glycans consisting only of deoxyhexose instead of the flagellin glycans including the viosamine derivatives determined previously. The motility and virulence on host tobacco leaves were strongly impaired in the ΔvioA mutant and were weakly reduced in the ΔvioB and ΔvioT mutant strains. These results suggest that the genes vioA, vioB, and vioT are essential for glycosylation of flagellin, and accordingly are required for bacterial virulence. © 2009 Springer-Verlag.

    DOI: 10.1007/s00438-009-0489-8

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  • Study of flagella-mediated interactions between plants and Pseudomonas syringae 査読

    Yuki Ichinose

    JOURNAL OF GENERAL PLANT PATHOLOGY   75 ( 6 )   452 - 454   2009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 TOKYO  

    DOI: 10.1007/s10327-009-0192-y

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  • Structural characterization of an O-linked tetrasaccharide from Pseudomonas syringae pv. tabaci flagellin 査読

    Tomoyuki Konishi, Fumiko Taguchi, Masako Iwaki, Mayumi Ohnishi-Kameyama, Masanobu Yamamoto, Ikuko Maeda, Yoshihiro Nishida, Yuki Ichinose, Mitsuru Yoshida, Tadashi Ishii

    CARBOHYDRATE RESEARCH   344 ( 16 )   2250 - 2254   2009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:ELSEVIER SCI LTD  

    The flagellin of Pseudomonas syringae pv. tabaci is a glycoprotein that contains O-linked oligosaccharides composed of rhamnosyl and 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methylglucosyl residues. These O-linked glycans are released by hydrazinolysis and then labeled at their reducing ends with 2-aminopyridine (PA). A PA-labeled trisaccharide and a PA-labeled tetrasaccharide are isolated by normal-phase high-performance liquid chromatography. These oligosaccharides are structurally characterized using mass spectrometry and NMR spectroscopy. Our data show that P. syringae pv. tabaci flagellin is glycosylated with a tetrasaccharide, 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methyl-Glcp-(1 -&gt; 3)-alpha-L-Rhap-(1 -&gt; 2)-alpha-L-Rhap-(1 -&gt; 2)-alpha-L-Rha-(1 -&gt;, as well a trisaccharide, 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methyl-Glcp-(1 -&gt; 3)-alpha-L-Rhap-(1 -&gt; 2)-alpha-L-Rha-(1 -&gt;,which was identified in a previous study. (C) 2009 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.carres.2009.07.004

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  • Glycosylation of flagellin from Pseudomonas syringae pv. tabaci 6605 contributes to evasion of host tobacco plant surveillance system. 査読

    Taguchi F, Suzuki T, Takeuchi K, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y

    Physiol. Mol. Plant Pathol.   74   11 - 17   2009年

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: 10.1016/j.pmpp.2009.08.001

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  • Flagellin glycans from two pathovars of Pseudomonas syringae contain rhamnose in D and L configurations in different ratios and modified 4-amino-4,6-dideoxyglucose. 査読 国際誌

    Kasumi Takeuchi, Hiroshi Ono, Mitsuru Yoshida, Tadashi Ishii, Etsuko Katoh, Fumiko Taguchi, Ryuji Miki, Katsuyoshi Murata, Hanae Kaku, Yuki Ichinose

    Journal of bacteriology   189 ( 19 )   6945 - 56   2007年10月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

    Flagellins from Pseudomonas syringae pv. glycinea race 4 and Pseudomonas syringae pv. tabaci 6605 have been found to be glycosylated. Glycosylation of flagellin is essential for bacterial virulence and is also involved in the determination of host specificity. Flagellin glycans from both pathovars were characterized, and common sites of glycosylation were identified on six serine residues (positions 143, 164, 176, 183, 193, and 201). The structure of the glycan at serine 201 (S201) of flagellin from each pathovar was determined by sugar composition analysis, mass spectrometry, and (1)H and (13)C nuclear magnetic resonance spectroscopy. These analyses showed that the S201 glycans from both pathovars were composed of a common unique trisaccharide consisting of two rhamnosyl (Rha) residues and one modified 4-amino-4,6-dideoxyglucosyl (Qui4N) residue, beta-D-Quip4N(3-hydroxy-1-oxobutyl)2Me-(1-->3)-alpha-L-Rhap-(1-->2)-alpha-L-Rhap. Furthermore, mass analysis suggests that the glycans on each of the six serine residues are composed of similar trisaccharide units. Determination of the enantiomeric ratio of Rha from the flagellin proteins showed that flagellin from P. syringae pv. tabaci 6605 consisted solely of L-Rha, whereas P. syringae pv. glycinea race 4 flagellin contained both L-Rha and D-Rha at a molar ratio of about 4:1. Taking these findings together with those from our previous study, we conclude that these flagellin glycan structures may be important for the virulence and host specificity of P. syringae.

    DOI: 10.1128/JB.00500-07

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  • [MAMP signal transduction].

    Yuki Ichinose

    Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme   52 ( 6 Suppl )   635 - 41   2007年5月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • Identification of glycosylation genes and glycosylated amino acids of flagellin in Pseudomonas syringae pv. tabaci. 査読 国際誌

    Fumiko Taguchi, Kasumi Takeuchi, Etsuko Katoh, Katsuyoshi Murata, Tomoko Suzuki, Mizuri Marutani, Takayuki Kawasaki, Minako Eguchi, Shizue Katoh, Hanae Kaku, Chihiro Yasuda, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Cellular microbiology   8 ( 6 )   923 - 38   2006年6月

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    担当区分:最終著者, 責任著者   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 PUBLISHING  

    A glycosylation island is a genetic region required for glycosylation. The glycosylation island of flagellin in Pseudomonas syringae pv. tabaci 6605 consists of three orfs: orf1, orf2 and orf3. Orf1 and orf2 encode putative glycosyltransferases, and their deletion mutants, Deltaorf1 and Deltaorf2, exhibit deficient flagellin glycosylation or produce partially glycosylated flagellin respectively. Digestion of glycosylated flagellin from wild-type bacteria and non-glycosylated flagellin from Deltaorf1 mutant using aspartic N-peptidase and subsequent HPLC analysis revealed candidate glycosylated amino acids. By generation of site-directed Ser/Ala-substituted mutants, all glycosylated amino acid residues were identified at positions 143, 164, 176, 183, 193 and 201. Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) analysis revealed that each glycan was about 540 Da. While all glycosylation-defective mutants retained swimming ability, swarming ability was reduced in the Deltaorf1, Deltaorf2 and Ser/Ala-substituted mutants. All glycosylation mutants were also found to be impaired in the ability to adhere to a polystyrene surface and in the ability to cause disease in tobacco. Based on the predicted tertiary structure of flagellin, S176 and S183 are expected to be located on most external surface of the flagellum. Thus the effect of Ala-substitution of these serines is stronger than that of other serines. These results suggest that glycosylation of flagellin in P. syringae pv. tabaci 6605 is required for bacterial virulence. It is also possible that glycosylation of flagellin may mask elicitor function of flagellin molecule.

    DOI: 10.1111/j.1462-5822.2005.0674.x

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  • Flagellin Glycosylation Island in Pseudomonas syringae pv. glycinea and Its Role in Host Specificity. 査読 国際誌

    J. Bacteriol.   185 ( 22 )   6658 - 65   2003年11月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1128/JB.22.6658-6665.2003

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  • Post-translational modification of flagellin determines the specificity of HR induction. 査読

    Fumiko Taguchi, Rena Shimizu, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Plant & cell physiology   44 ( 3 )   342 - 9   2003年3月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Flagellin, a constituent of the flagellar filament, is a potent elicitor of hypersensitive cell death in plant cells. Flagellins of Pseudomonas syringae pvs. glycinea and tomato induce hypersensitive cell death in their non-host tobacco plants, whereas those of P. syringae pv. tabaci do not remarkably induce it in its host tobacco plants. However, the deduced amino acid sequences of flagellins from pvs. tabaci and glycinea are identical, indicating that post-translational modification of flagellins plays an important role in determining hypersensitive reaction (HR)-inducibility. To investigate genetically the role of modification of flagellin in HR-induction, biological and phytopathological phenotypes of a flagella-defective Delta fliC mutant and Delta fliC mutants complemented by the introduction of the flagellin gene (fliC) from different pathovars of P. syringae were investigated. The Delta fliC mutant of pv. tabaci lost flagella, motility, the ability to induce HR cell death in non-host tomato cells and virulence toward host tobacco plants, whereas all pv. tabaci complemented by the introduction of the fliC gene of pvs. tabaci, glycinea or tomato recovered all the abilities that the Delta fliC mutant had lost. These results indicate that post-translational modification of flagellins is strongly correlated with the ability to cause HR cell death.

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  • Flagella defective mutant fliD of Pseudomonas syringae pv. tabaci that secretes monomer flagellin induces strong HR on nonhost tomato cells. 査読

    Shimizu R, Taguchi F, Marutani M, Mukaihara T, Inagaki Y, Toyoda K, Shiraishi T, Ichinose Y

    Mol.Genet. Genomics   269 ( 1 )   21 - 30   2003年

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Expression of the 12-oxophytodienoic acid 10,11-reductase gene in the compatible interaction between pea and fungal pathogen 査読

    Y Ishiga, A Funato, T Tachiki, K Toyoda, T Shiraishi, T Yamada, Y Ichinose

    PLANT AND CELL PHYSIOLOGY   43 ( 10 )   1210 - 1220   2002年10月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Suppressors produced by Mycosphaerella pinodes are glycopeptides to block pea defense responses induced by elicitors. A clone, S64, was isolated as cDNA for suppressor-inducible gene from pea epicotyls. The treatment of pea epicotyls with suppressor alone induced an increase of S64 mRNA within 1 h, and it reached a maximum level at 3 h after treatment. The induction was not affected by application of the elicitor, indicating that the suppressor has a dominant action to regulate S64 gene expression. S64 was also induced by inoculation with a virulent pathogen, M. pinodes, but not by inoculation with a non-pathogen, Ascochyta rabiei, nor by treatment with fungal elicitor. The deduced structure of S64 showed high homology to 12-oxophytodienoic acid reductase (OPR) in Arabidopsis thaliana. A recombinant protein derived from S64 had OPR activity, suggesting compatibility-specific activation of the octadecanoid pathway in plants. Treatment with jasmonic acid (JA) or methyl jasmonic acid, end products of the octadecanoid pathway, inhibited the elicitor-induced accumulation of PAL mRNA in pea. These results indicate that the suppressor-induced S64 gene expression leads to the production of JA or related compounds, which might contribute to the establishment of compatibility by inhibiting the phenylpropanoid biosynthetic pathway.

    DOI: 10.1093/pcp/pcf144

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  • cDNA cloning and characterization of tobacco ABC transporter: NtPDR1 is a novel elicitor-responsive gene. 査読 国際誌

    Michiko Sasabe, Kazuhiro Toyoda, Tomonori Shiraishi, Yoshishige Inagaki, Yuki Ichinose

    FEBS letters   518 ( 1-3 )   164 - 8   2002年5月

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    担当区分:最終著者, 責任著者   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 isolated an INF1 elicitin-inducible cDNA encoding a pleiotropic drug resistance (PDR)-type ATP-binding cassette (ABC) transporter homolog (NtPDR1) in suspension-cultured tobacco Bright Yellow-2 (BY-2) cells by application of differential display PCR. The NtPDR1 (Nicotiana tabacum PDR protein 1) gene also encodes a 162 kDa protein that includes two putative hydrophilic domains containing the ABC signature motif and two putative hydrophobic domains. Expression of the NtPDR1 gene was rapidly and strongly activated by treatment of BY-2 cells with INF1 elicitin. Further, treatment of BY-2 cells with flagellin, a bacterial proteinaceous hypersensitive reaction elicitor, or yeast extract, a general elicitor, also induced NtPDR1 gene expression. These results indicate that NtPDR1 may be involved in the general defense response in tobacco. This is the first report that microbial elicitors induce the expression of a plant ABC transporter gene.

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  • Bacteriophage P4282, a parasite of Ralstonia solanacearum, encodes a bacteriolytic protein important for lyric infection of its host 査読

    H. Ozawa, H. Tanaka, Y. Ichinose, T. Shiraishi, T. Yamada

    Molecular and General Genetics   265 ( 1 )   95 - 101   2001年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    To enhance bacterial wilt resistance in tobacco expressing a foreign protein, we isolated the bacteriolytic gene from a bacteriophage that infects Ralstonia solanacearum. The bacteriolytic protein of phage P4282 isolated in Tochigi Prefecture was purified from a lysate of R. solanacearum M4S cells infected with the phage, and its bacteriolytic activity was assayed by following the decrease in the turbidity of suspensions of R. solanacearum M4S cells. The molecular weight of the bacteriolytic protein was approximately 71 kDa, and the sequence of the N-terminal 13 amino acids was determined. We used oligonucleotide probes based on this amino acid sequence to isolate the bacteriolytic gene from phage P4282 DNA. This gene of 2061 bp encodes a product of 687 amino acids, whose calaculated molecular weight was 70.12 kDa. The bacteriolytic gene was placed under the control of an inducible promoter, and the plasmid was transformed into Escherichia coli NM522. The soluble proteins extracted from E. coli NM522 cells harboring the plasmid with the bacteriolyric gene showed obvious bacteriolytic activities against several strains of R. solanacearum isolated in various districts in Japan. DNA fragments from five phages, isolated in Niigata, Aomori, Okinawa, Fukushima and Yamaguchi Prefectures, hybridized to the bacteriolytic gene of phage P4282. These observations indicate that the bacteriolytic protein shows nonspecific activity against R. solanacearum strains, and a sequence similar to that of the bacteriolytic gene is conserved in the DNA of other bacteriophages. These results indicate that the generation of transgenic (tobacco) plants expressing the bacteriolytic gene of phage P4282 might result in enhanced resistance to bacterial wilt in tobacco.

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  • Regulation of nuclear gene expression in relation to signal molecules 査読

    T Yamada, Y Ichinose, T Shiraishi, K Toyoda, Y Imura, H Seki, P Sriprasertsak, A Funado

    DELIVERY AND PERCEPTION OF PATHOGEN SIGNALS IN PLANTS   164 - 173   2001年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • Suppressors of defense - Supprescins and plant receptor molecules 査読

    T Shiraishi, K Toyoda, T Yamada, Y Ichinose, A Kiba, M Sugimoto

    DELIVERY AND PERCEPTION OF PATHOGEN SIGNALS IN PLANTS   112 - 121   2001年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • Plant cell wall with the suppressor may play a crucial role in determining specificity 査読

    T Shiraishi, A Kiba, A Inata, T Sugiura, K Toyoda, Y Ichinose, T Yamada

    MOLECULAR GENETICS OF HOST-SPECIFIC TOXINS IN PLANT DISEASES   13   343 - 353   1998年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:SPRINGER  

    The mechanism of plant host-parasite specificity is one of the most intriguing issues of contemporary plant biology. We recently found that a fungal elicitor stimulated cell wall functions such as ATP-hydrolyzing and superoxide generating activities. Furthermore, the suppressor from Mycosphaerella pinodes inhibited these activities in vitro in a strictly species-specific manner. The cell wall-bound ATPase, which is different from that of the plasma membrane in several properties, was associated with cell wall-bound peroxidase(s). In response to the elicitor, the isolated cell walls are able to produce an infection-inhibitor that may be dependent upon the superoxide generating system. On the other baud, the suppressor inhibited such production in pea cell walls and induced it in nonhost's cell walls. Thus, the effects of both fungal signals on isolated cell walls coincide with those on plant tissues. Ln this treatise, we discuss the importance of the cell wall, which is plant-specific acid the most exterior organelle, in determining host-parasite specificity.

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  • Regulation of genes for phenylpropanoid synthesis in pea by elicitor and suppressor 査読

    T Yamada, T Shiraishi, Y Ichinose, H Kato, H Seki, Y Murakami

    MOLECULAR ASPECTS OF PATHOGENICITY AND RESISTANCE: REQUIREMENT FOR SIGNAL TRANSDUCTION   151 - 162   1996年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • Fungal signals regulate ATPase and polyphosphoinositide metabolism in pea plants 査読

    T Shiraishi, T Yamada, Y Ichinose, K Toyoda, A Kiba, T Kato

    MOLECULAR ASPECTS OF PATHOGENICITY AND RESISTANCE: REQUIREMENT FOR SIGNAL TRANSDUCTION   197 - 208   1996年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • Effect of methyl jasmonate on harpin-induced hypersensitive cell heath, generation of hydrogen peroxide and expression of PAL mRNA in tobacco suspension cultured BY-2 cells 査読

    S Andi, F Taguchi, K Toyoda, T Shiraishi, Y Ichinose

    PLANT AND CELL PHYSIOLOGY   42 ( 4 )   446 - 449   1992年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Methyl jasmonate inhibited the harpin-induced defense responses such as cell death, H2O2 generation and gene expression encoding phenylalanine ammonia-lyase in tobacco suspension cultured BY-2 cells, These results suggest that MeJA may act as an endogenous suppressor for plant defense response including hypersensitive reaction.

    DOI: 10.1093/pcp/pce056

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書籍等出版物

  • 植物病理学 第2版

    眞山, 滋志, 土佐, 幸雄( 範囲: 抗菌性物質分解酵素、細菌の病原性発現機構)

    文永堂出版  2020年3月  ( ISBN:9784830041389

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    総ページ数:xii, 347p   記述言語:日本語

    CiNii Books

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  • 植物たちの戦争 : 病原体との5億年サバイバルレース

    日本植物病理学会( 担当: 共著 ,  範囲: 3-1, 3-2)

    講談社  2019年3月  ( ISBN:9784065152164

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    総ページ数:262p   記述言語:日本語

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  • 新植物病理学概論

    白石, 友紀, 秋光, 和也, 一瀬, 勇規, 寺岡, 徹, 吉川, 信幸( 範囲: 第4章(細菌病とファイトプラズマ)第12章(植物病理学とバイオテクノロジー))

    養賢堂  2012年3月  ( ISBN:9784842504940

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    総ページ数:7, 301p, 図版 [1] p   記述言語:日本語

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  • Genome-Enabled Analysis of Plant-Pathogen Interactions

    ( 担当: 共著)

    2011年 

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  • Genome-Enabled Analysis of Plant-Pathogen Interactions

    Wolpert, T, Shiraishi, T, Collmer A, Akimitsu, K, Glazebrook, J. ed( 担当: 共著)

    2011年 

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  • Genome-Enabled Analysis of Plant-Pathogen Interactions

    2011年 

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  • 植物病理学

    眞山, 滋志, 難波, 成任( 担当: 共著 ,  範囲: 細菌の病原性発現機構(p. 186 - 193))

    文永堂出版  2010年1月  ( ISBN:483004117X

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    総ページ数:333   記述言語:日本語

    CiNii Books

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  • 遺伝暗号(一瀬勇規)

    基礎演習 産業生物科学(泉本勝利 他編)岡山大学出版会(岡山)p. 83.  2009年 

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  • ポリメラーゼ連鎖反応 (PCR) (一瀬勇規)

    基礎演習 産業生物科学(泉本勝利 他編)岡山大学出版会(岡山)p. 84.  2009年 

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  • 組換えDNA技術(一瀬勇規)

    基礎演習 産業生物科学(泉本勝利 他編)岡山大学出版会(岡山)p. 88.  2009年 

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  • 遺伝子組換え作物(一瀬勇規)

    基礎演習 産業生物科学(泉本勝利 他編)岡山大学出版会(岡山)p. 89.  2009年 

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  • 全身獲得抵抗性(一瀬勇規)

    植物ゲノム科学辞典(駒嶺 穆 総編)朝倉書店(東京)p. 196  2009年 

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  • 病原性(一瀬勇規)

    植物ゲノム科学辞典(駒嶺 穆 総編)朝倉書店(東京)p. 284.  2009年 

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  • 病原力遺伝子(一瀬勇規)

    植物ゲノム科学辞典(駒嶺 穆 総編)朝倉書店(東京)p. 284.  2009年 

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  • レース特異的抵抗性(一瀬勇規)

    植物ゲノム科学辞典(駒嶺 穆 総編)朝倉書店(東京)p. 368.  2009年 

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  • Role of flagellin glycosylation in bacterial virulence. (Ichinose, Y., Taguchi, F., Takeuchi, K., Suzuki, T., Toyoda, K. and Shiraishi, T.)

    Pseudomonas syringae Pathovars and Related Pathogens - Identification, Epidemiology and Genomics (M. Fatmi et al. eds.) Springer p. 167-174.  2008年 

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  • MAMPシグナル伝達機構(一瀬勇規).

    蛋白質 核酸 酵素 増刊号「植物における環境と生物ストレスに対する応答」(島本 功・篠崎一雄・白須 賢・篠崎和子 編) 53 (6) p. 635-641.  2007年 

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  • Pseudomonas syringaeによる植物免疫の活性化とその制御機構,微生物の病原性と植物の防御応答(上田一郎編著)p. 132-143.

    北海道大学出版会 札幌  2007年 

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  • Role of flagellin glycosylation in bacterial virulence.(Ichinose, Y., Taguchi, F., Takeuchi, K., Suzuki, T., Toyoda, K. and Shiraishi, T.)

    Pseudomonas syringae Pathovars and Related Pathogens - Identification, Epidemiology and Genomics (M. Fatmi et al. eds.) Springer p. 167-174.  2007年 

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  • Bacterial flagellins as elicitors of the defense response.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Compatible-specific expression of 12-oxophytodienoic acid 10,11-reductase gene in pea.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Compatible-specific expression of 12-oxophytodienoic acid 10,11-reductase gene in pea.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Flagellin Glycosylation Island in Pseudomonas syringae and Its Role in Host Specificity.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Defense signaling snd the plant cell wall – A new signaling pathway dependent upon inorganic phosphate.

    Genomic and Genetic Analysis of Plant Parasitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Flagellin Glycosylation Island in Pseudomonas syringae and Its Role in Host Specificity.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

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  • Defense signaling snd the plant cell wall – A new signaling pathway dependent upon inorganic phosphate.

    Genomic and Genetic Analysis of Plant Parasitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

     詳細を見る

  • :植物病原菌のシグナル分子により発現が変動する遺伝子の制御機構と機能

    植物病の探究 (高松 進ら編)  2004年 

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  • Bacterial flagellins as elicitors of the defense response.

    Genomic and Genetic Analysis of Plant Paratsitism and Defense (Tsuyumu et al. eds.) APS Press (St. Paul Minnesota, USA)  2004年 

     詳細を見る

  • Suppressors of defense-Supprescins and Plant receptor molecules.

    Delivery of Pathogen Signals to Plants (N. T. Keen ed.) APS Press (St. Paul Minnesota, USA)  2001年 

     詳細を見る

  • Regulation of nuclear gene expression in relation to signal molecules.

    Delivery of Pathogen Signals to Plants (N. T. Keen ed.) APS Press (St. Paul Minnesota, USA)  2001年 

     詳細を見る

  • Suppressors of defense-Supprescins and Plant receptor molecules.

    Delivery of Pathogen Signals to Plants (N. T. Keen ed.) APS Press (St. Paul Minnesota, USA)  2001年 

     詳細を見る

  • Regulation of nuclear gene expression in relation to signal molecules.

    Delivery of Pathogen Signals to Plants (N. T. Keen ed.) APS Press (St. Paul Minnesota, USA)  2001年 

     詳細を見る

  • 病原体に対する応答

    朝倉植物生理学講座第5巻環境応答  2001年 

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  • Suppressors of defense-Supprescins and Plant receptor molecules.(共著)

    Delivery of Pathogen Signals to Plants(N. T. Keen Ed. )APS Press, St. Paul Minnesota, USA  2000年 

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  • Regulation of nuclear gene expression in relation to signal molecules.(共著)

    Delivery of Pathogen Signals to Plants(N. T. Keen Ed. )APS Press, St Paul Minnesota, USA  2000年 

     詳細を見る

  • 宿主特異性決定における植物細胞壁の役割(共著)

    東北地方における植物病理学のフロントラン日本植物病理学会東北部会創立35周年記念誌刊行会  2000年 

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  • Suppressors of defense-Supprescins and Plant receptor molecules.(共著)

    Delivery of Pathogen Signals to Plants(N. T. Keen Ed. )APS Press, St. Paul Minnesota, USA  2000年 

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  • Regulation of nuclear gene expression in relation to signal molecules.(共著)

    Delivery of Pathogen Signals to Plants(N. T. Keen Ed. )APS Press, St Paul Minnesota, USA  2000年 

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  • Suppressor as a factor determining plant-pathogen Specificity.(共著)

    Plant-Microbe Interactions Vol. 4, (Stacey, G. and Keen, N. T. eds. )APS Press, St, Paul Minnesota, USA  1999年 

     詳細を見る

  • Suppressor as a factor determining plant-pathogen Specificity.(共著)

    Plant-Microbe Interactions Vol. 4, (Stacey, G. and Keen, N. T. eds. )APS Press, St, Paul Minnesota, USA  1999年 

     詳細を見る

  • The role of suppressors in determining host-parasite specificities in plant cells(共著)

    International Review of Cytology, Academic Press  1997年 

     詳細を見る

  • 宿主特異性と防御遺伝子の発現調節(共著)

    植物細胞工学シリーズ8「分子レベルからみた植物の耐病性」  1997年 

     詳細を見る

  • 目で見る植物の耐病性(共著)

    植物細胞工学シリーズ8「分子レベルからみた植物の耐病性」  1997年 

     詳細を見る

  • The role of suppressors in determining host-parasite specificities in plant cells(共著)

    International Review of Cytology, Academic Press  1997年 

     詳細を見る

  • Regulation of the gene involved in pheylpropanoid pathway in pea by eliutor and suppressor(共著)

    Molecular Aspects of Phathogenisity and Host Resistance (Requirement of Signal Transduction) APS Press  1996年 

     詳細を見る

  • 植物の病原体認識とシグナル伝達(共著)

    植物のシグナルトランスダクション,東京化学同人  1996年 

     詳細を見る

  • Fungal signals regulate ATPase and polyphosphoinositide metabolism in pea plant(共著)

    Molecular Aspects of Phathogenisity and Host Resistance (Requirement of Signal Transduction) APS Press  1996年 

     詳細を見る

  • Regulation of the gene involved in pheylpropanoid pathway in pea by eliutor and suppressor(共著)

    Molecular Aspects of Phathogenisity and Host Resistance (Requirement of Signal Transduction) APS Press  1996年 

     詳細を見る

  • Fungal signals regulate ATPase and polyphosphoinositide metabolism in pea plant(共著)

    Molecular Aspects of Phathogenisity and Host Resistance (Requirement of Signal Transduction) APS Press  1996年 

     詳細を見る

  • Regulation of ATPase and signal transduction for pea defense responses by the suppressor and elicitor from ┣DBMycosphaerella pinodes(/)-┫DB. (共著)

    Host-Specific Toxin : Biosynthesis, Receptor and Molecular Biology (eds. K. Kohmoto and O. C. Yoder). Tottori University, Tottori.  1993年 

     詳細を見る

  • 植物病原性微生物研究法 共著

    ソフトサイエンス社  1993年 

     詳細を見る

  • Regulation of ATPase and signal transduction for pea defense responses by the suppressor and elicitor from ┣DBMycosphaerella pinodes(/)-┫DB. (共著)

    Host-Specific Toxin : Biosynthesis, Receptor and Molecular Biology (eds. K. Kohmoto and O. C. Yoder). Tottori University, Tottori.  1993年 

     詳細を見る

▼全件表示

MISC

  • タバコ野火病菌の病徴進展に寄与するエフェクターの同定

    西村隆史, 樫原沙知, 能年義輝, 山本幹博, 豊田和弘, 一瀬勇規, 松井英譲

    日本植物病理学会報   87 ( 1 )   2021年

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  • メトミノストロビンの抵抗性誘導作用に関する研究(3)メトミノストロビンはシロイヌナズナのMAPキナーゼを活性化する

    伊藤千晶, 小原七海, 佐藤穂高, 市成光広, 山田晶, 櫻本和生, 白石慎, 山本隆, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 豊田和弘, 豊田和弘

    日本農薬学会大会講演要旨集   46th (CD-ROM)   2021年

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  • メトミノストロビンの抵抗性誘導作用に関する研究 6.シロイヌナズナにおけるPTI関連遺伝子の発現に対する影響

    小原七海, 伊藤千晶, 佐藤穂高, 市成光広, 山田晶, 櫻本和生, 白石慎, 山本隆, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 豊田和弘, 豊田和弘

    日本植物病理学会大会プログラム・講演要旨予稿集   2021   2021年

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  • Pseudomonas syringae pv.tabaci 6605の走化性受容体遺伝子mcpの病原力への関与

    寺元茜, 金重慎, TUMEWU S.A., 松井英譲, 山本幹博, 能年義輝, 豊田和弘, 一瀬勇規

    日本植物病理学会大会プログラム・講演要旨予稿集   2021   2021年

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  • MARK2のリン酸化はflg22処理に伴うカロース蓄積に関与する

    松井英譲, 松井英譲, 澁谷日奈, 前田和輝, 四井いずみ, 四井いずみ, HYON G-S., 野村有子, 一瀬勇規, 中神弘史, 中神弘史

    日本植物病理学会報   87 ( 1 )   2021年

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  • メトミノストロビンの抵抗性誘導作用に関する研究(4)メトミノストロビンはシロイヌナズナにおいてPTI関連遺伝子の発現を促進する

    小原七海, 伊藤千晶, 佐藤穂高, 市成光広, 山田晶, 櫻本和生, 白石慎, 山本隆, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 豊田和弘, 豊田和弘

    日本農薬学会大会講演要旨集   46th (CD-ROM)   2021年

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  • タバコ野火病抵抗性品種はタバコ野火病菌TTSSエフェクターを認識し,HRを誘導する

    樫原沙知, 西村隆史, 能年義輝, 山本幹博, 豊田和弘, 一瀬勇規, 松井英譲

    日本植物病理学会報   87 ( 1 )   2021年

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  • シロイヌナズナのMAMP誘導性オキシダティブバーストにおける細胞壁ペルオキシダーゼとNADPHオキシダーゼの関与

    木元菜々子, 高須瑞穂, APRILIA Nur Fitrianti, 松井英譲, 能年義輝, 山本幹博, 一瀬勇規, 白石友紀, 豊田和弘

    日本植物病理学会報   87 ( 1 )   2021年

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  • シロイヌナズナの細胞内/細胞外レドックス制御におけるNADPHオキシダーゼRBOHDの役割

    高須瑞穂, 木元菜々子, 松井英譲, 能年義輝, 山本幹博, 一瀬勇規, 白石友紀, 豊田和弘

    日本植物病理学会報   87 ( 1 )   2021年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2021   2021年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2020   2020年

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    日本植物病理学会報   86 ( 1 )   2020年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2020   2020年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2020   2020年

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    日本植物病理学会報   86 ( 1 )   2020年

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    日本植物病理学会報   86 ( 1 )   2020年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2020   2020年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2020   2020年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2019   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2019   2019年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2019   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会報   85 ( 1 )   2019年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2018   2018年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2018   2018年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2018   2018年

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    日本植物病理学会報   84 ( 1 )   2018年

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    日本植物病理学会報   84 ( 1 )   2018年

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    日本植物病理学会報   84 ( 1 )   2018年

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    日本植物病理学会大会プログラム・講演要旨予稿集   2018   2018年

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  • バイオコントロール細菌Rhizobium vitis VAR03-1株のブドウ根頭がんしゅ病抑制機構の解析

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    日本植物病理学会大会プログラム・講演要旨予稿集   2018   2018年

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  • Pseudomonas syringaeの菌体密度感知機構の解析(1)

    山本悟, 藤山友里, 高田基弘, 松井英譲, 山本幹博, 能年義輝, 豊田和弘, 一瀬勇規

    日本植物病理学会報   83 ( 1 )   2017年

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  • エンドウならびにシロイヌナズナにおけるPAMP誘導性細胞外オキシダティブバースト反応の分子機構

    片岡千香子, 三木紅葉, 山崎史織, 松尾実佳, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 白石友紀, 白石友紀, 豊田和弘, 豊田和弘

    日本植物病理学会大会プログラム・講演要旨予稿集   2017   2017年

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  • 非病原性Rhizobium vitis VAR03-1株によるブドウ根頭がんしゅ病の病原性関連遺伝子の発現抑制

    齊藤晶, 渡邉恵, 松井英譲, 山本幹博, 一瀬勇規, 豊田和弘, 川口章, 能年義輝

    日本植物病理学会大会プログラム・講演要旨予稿集   2017   2017年

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  • エンドウから分離した細胞壁タンパク質のPAMP応答

    三木紅葉, 山崎史織, 片岡千香子, 松尾実佳, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 白石友紀, 白石友紀, 豊田和弘, 豊田和弘

    日本植物病理学会報   83 ( 1 )   2017年

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  • 非病原性Rhizobium vitis ARK-1株によるブドウ根頭がんしゅ病の病原性関連遺伝子の発現抑制

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    日本植物病理学会報   83 ( 1 )   2017年

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  • エクト型ATPase(PsAPY1)はエンドウ葉におけるPAMP誘導性オキシダティブバースト反応を正に調節し,不適応型の病原菌に対する非宿主抵抗性に関与する

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    日本植物病理学会報   83 ( 1 )   2017年

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    日本植物病理学会報   83 ( 1 )   2017年

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    日本植物病理学会報   83 ( 1 )   2017年

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  • シロイヌナズナにおけるPAMP誘導性オキシダティブバースト反応におけるアポプラストの役割について

    片岡千香子, 山崎史織, 松尾実佳, 三木紅葉, 松井英譲, 松井英譲, 能年義輝, 能年義輝, 山本幹博, 山本幹博, 一瀬勇規, 一瀬勇規, 白石友紀, 白石友紀, 白石友紀, 豊田和弘, 豊田和弘

    日本植物病理学会報   83 ( 1 )   2017年

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  • Pseudomonas syringae pv.tabaci6605におけるAHL合成制御機構並びにMexEFOprN多剤排出ポンプの病原力における役割の解析

    澤田貴博, 山本幹博, 松井英譲, 能年義輝, 豊田和弘, 一瀬勇規

    日本植物病理学会報   83 ( 1 )   2017年

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  • エクト型ATPase(PsAPY1)はエンドウ葉におけるリポ多糖(LPS)誘導性細胞外オキシダティブバースト反応を正に調節する

    三木紅葉, 山崎史織, 矢尾幸世, 松井英譲, 能年義輝, 山本幹博, 一瀬勇規, 白石友紀, 豊田和弘, 松井英譲, 能年義輝, 山本幹博, 一瀬勇規, 白石友紀, 白石友紀, 豊田和弘

    日本植物病理学会大会プログラム・講演要旨予稿集   2016   2016年

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  • Pseudomonas syringae pv.syringae B728aの病原力制御因子Vfrの標的遺伝子のクロマチン免疫沈降法による探索(2)

    小倉敬右, 田口富美子, 山本幹博, 松井英譲, 能年義輝, 豊田和弘, 一瀬勇規

    日本植物病理学会大会プログラム・講演要旨予稿集   2016   2016年

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  • エクト型ATPase(PsAPY1)の発現を抑制したエンドウ葉におけるジャスモン酸応答性遺伝子の発現

    矢尾幸世, 田中佳織, 奥田竜太, 三木紅葉, 松井英譲, 能年義輝, 山本幹博, 一瀬勇規, 白石友紀, 白石友紀, 豊田和弘

    日本植物病理学会報   82 ( 1 )   2016年

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  • Pseudomonas syringae pv.tabaciにおけるAHL合成制御機構の解析

    澤田貴博, 山本幹博, 松井英譲, 能年義輝, 豊田和弘, 一瀬勇規

    日本植物病理学会報   82 ( 1 )   2016年

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  • ミナトカモジグサ-紋枯病菌感染系の確立と植物ホルモンによる抵抗性機構の解析

    香西雄介, 山中由利恵, 渡邉恵, 木村麻美子, 恩田義彦, 持田恵一, 山本幹博, 松井英譲, 一瀬勇規, 豊田和弘, 能年義輝

    日本植物病理学会報   82 ( 1 )   2016年

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  • 非病原性Rhizobium vitis ARK-1株によるブドウ根頭がんしゅ病の発病発現抑制機構の解析

    齊藤晶, 渡邉恵, 松井英譲, 山本幹博, 一瀬勇規, 豊田和弘, 川口章, 能年義輝

    日本植物病理学会大会プログラム・講演要旨予稿集   2016   2016年

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  • ナス科植物青枯病菌の病原力や増殖を制御する化合物の探索

    澤井拓, 一瀬勇規, 山本幹博, 松井英譲, 能年義輝, 豊田和弘, 向原隆文, 長田裕之

    日本植物病理学会大会プログラム・講演要旨予稿集   2016   2016年

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  • Pathogenicity and virulence factors of Pseudomonas syringae

    Yuki Ichinose, Fumiko Taguchi, Takafumi Mukaihara

    Journal of General Plant Pathology   79 ( 5 )   285 - 296   2013年9月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等  

    In 1994, Oku reported that plant pathogens, mainly fungal pathogens, require three essential abilities to infect plants: to enter plants, to overcome host resistance, and to evoke disease. Because the infectious process of phytopathogenic bacteria differs from that of fungal pathogens, we have attempted to characterize pathogenicity, the ability of a pathogen to cause disease, using the phytopathogenic bacterium Pseudomonas syringae as a representative pathogen. To establish infection and incite disease development, bacteria first have to enter a plant. This process requires flagella- and type IV pili-mediated motility, and active taxis is probably necessary for effective infection. After bacteria enter a plant's apoplastic spaces, they need to overcome host plant resistance. To do this, they secrete a wide variety of hypersensitive response and pathogenicity (Hrp) effector proteins into the plant cytoplasm to interfere with pathogen/microbe-associated molecular pattern- and effector-triggered immunity, produce phytohormones and/or phytotoxins to suppress plant defense responses and extracellular polysaccharides to prevent access by antibiotics and to chelate Ca2+, and activate the multidrug resistance efflux pump to extrude antimicrobial compounds for successful colonization. Furthermore, to evoke disease, bacteria produce toxins and Hrp effectors that compromise a plant's homeostasis and injure plant cells. The expression of these virulence factors depends on the infection processes and environmental conditions. Thus, the expression and function of virulence factors interact with each other, creating complex networks in the regulation of bacterial virulence-related genes. © 2013 The Phytopathological Society of Japan and Springer Japan.

    DOI: 10.1007/s10327-013-0452-8

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  • A volatile substance from Talaromyces sp promotes the plant growth and blocks the disease development on several plants

    T. Shiraishi, Y. Yamagiwa, P. T. Le, K. Maeda, Y. Inagaki, Y. Ichinose, M. Hyakumachi, K. Toyoda

    PHYTOPATHOLOGY   101 ( 6 )   S165 - S166   2011年6月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • Siderophore pyoverdine of Pseudomonas syringae pv. tabaci 6605 is an intrinsic virulence factor on host tobacco infection. 査読

    Taguchi, F, Suzuki, T, Inagaki, Y, Toyoda, K, Shiraishi, T, Ichinose, Y

    J. Bacteriol.   192 ( 1 )   117 - 126   2010年1月

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    担当区分:最終著者, 責任著者  

    DOI: 10.1128/jb.00689-09

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  • Bacterial DNA activates immunity in Arabidopsis thaliana

    Suguru Yakushiji, Yasuhiro Ishiga, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    JOURNAL OF GENERAL PLANT PATHOLOGY   75 ( 3 )   227 - 234   2009年6月

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    記述言語:英語   出版者・発行元:SPRINGER TOKYO  

    To initiate defense responses against invasion of pathogenic organisms, animals and plants must recognize microbe-associated molecular patterns (MAMPs). In this study, the elicitor activity of bacterial DNA on the model plant Arabidopsis thaliana was examined. EcoRI-digested plasmid DNA induced defense responses such as generation of reactive oxygen species and deposition of callose, whereas SmaI- and HapII-digested plasmid DNA and EcoRI-digested herring DNA did not remarkably induce these responses. Further, methylation of the CpG sequence of plasmid DNA and Escherichia coli DNA reduced the level of the defense responses. The endocytosis inhibitors wortmannin and amantadine significantly inhibited DNA-induced defense responses. These results suggest that non-methylated CpG DNA, as a MAMP, induced defense responses in Arabidopsis and that non-methylated DNA seems to be translocated into the cytoplasm by endocytosis.

    DOI: 10.1007/s10327-009-0162-4

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  • Bacterial DNA activates immunity in Arabidopsis thaliana

    Suguru Yakushiji, Yasuhiro Ishiga, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   75 ( 3 )   227 - 234   2009年6月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:Springer Science and Business Media LLC  

    To initiate defense responses against invasion of pathogenic organisms, animals and plants must recognize microbe-associated molecular patterns (MAMPs). In this study, the elicitor activity of bacterial DNA on the model plant Arabidopsis thaliana was examined. EcoRI-digested plasmid DNA induced defense responses such as generation of reactive oxygen species and deposition of callose, whereas SmaI- and HapII-digested plasmid DNA and EcoRI-digested herring DNA did not remarkably induce these responses. Further, methylation of the CpG sequence of plasmid DNA and Escherichia coli DNA reduced the level of the defense responses. The endocytosis inhibitors wortmannin and amantadine significantly inhibited DNA-induced defense responses. These results suggest that non-methylated CpG DNA, as a MAMP, induced defense responses in Arabidopsis and that non-methylated DNA seems to be translocated into the cytoplasm by endocytosis.

    DOI: 10.1007/s10327-009-0162-4

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  • Enhanced Defense Responses in Arabidopsis Induced by the Cell Wall Protein Fractions from Pythium oligandrum Require SGT1, RAR1, NPR1 and JAR1

    Yoko Kawamura, Shigehito Takenaka, Shu Hase, Mayumi Kubota, Yuki Ichinose, Yoshinori Kanayama, Kazuhiro Nakaho, Daniel F. Klessig, Hideki Takahashi

    Plant and Cell Physiology   50 ( 5 )   924 - 934   2009年5月

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    記述言語:英語   出版者・発行元:Oxford University Press (OUP)  

    The cell wall protein fraction (CWP) is purified from the non-pathogenic biocontrol agent Pythium oligandrum and is composed of two glycoproteins (POD-1 and POD-2), which are structurally similar to class III elicitins. In tomato plants treated with CWP, jasmonic acid (JA)- and ethylene (ET)-dependent signaling pathways are activated, and resistance to Ralstonia solanaceraum is enhanced. To dissect CWP-induced defense mechanisms, we investigated defense gene expression and resistance to bacterial pathogens in Arabidopsis thaliana ecotype Col-0 treated with CWP. When the leaves of Col-0 were infiltrated with CWP, neither visible necrosis nor salicylic acid (SA)-responsive gene (PR-1 and PR-5) expression was induced. In contrast, JA-responsive gene (PDF1.2 and JR2) expression was up-regulated and the resistance to R. solanaceraum and Pseudomonas syringae pv. tomato DC3000 was enhanced in response to CWP. Such CWP-induced defense responses were completely compromised in CWP-treated coi1-1 and jar1-1 mutants with an impaired JA signaling pathway. The induction of defense-related gene expression after CWP treatment was partially compromised in ET-insensitive ein2-1 mutants, but not in SA signaling mutants or nahG transgenic plants. Global gene expression analysis using cDNA array also suggested that several other JA- and ET-responsive genes, but not SA-responsive genes, were up-regulated in response to CWP. Further analysis of CWP-induced defense responses using another eight mutants with impaired defense signaling pathways indicated that, interestingly, the induction of JA-responsive gene expression and enhanced resistance to two bacterial pathogens in response to CWP were completely compromised in rar1-1, rar1-21, sgt1a-1, sgt1b (edm1) and npr1-1 mutants. Thus, the CWP-induced defense system appears to be regulated by JA-mediated and SGT1-, RAR1- and NPR1-dependent signaling pathways.

    DOI: 10.1093/pcp/pcp044

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  • Enhanced Defense Responses in Arabidopsis Induced by the Cell Wall Protein Fractions from Pythium oligandrum Require SGT1, RAR1, NPR1 and JAR1

    Yoko Kawamura, Shigehito Takenaka, Shu Hase, Mayumi Kubota, Yuki Ichinose, Yoshinori Kanayama, Kazuhiro Nakaho, Daniel F. Klessig, Hideki Takahashi

    PLANT AND CELL PHYSIOLOGY   50 ( 5 )   924 - 934   2009年5月

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

    The cell wall protein fraction (CWP) is purified from the non-pathogenic biocontrol agent Pythium oligandrum and is composed of two glycoproteins (POD-1 and POD-2), which are structurally similar to class III elicitins. In tomato plants treated with CWP, jasmonic acid (JA)- and ethylene (ET)-dependent signaling pathways are activated, and resistance to Ralstonia solanaceraum is enhanced. To dissect CWP-induced defense mechanisms, we investigated defense gene expression and resistance to bacterial pathogens in Arabidopsis thaliana ecotype Col-0 treated with CWP. When the leaves of Col-0 were infiltrated with CWP, neither visible necrosis nor salicylic acid (SA)-responsive gene (PR-1 and PR-5) expression was induced. In contrast, JA-responsive gene (PDF1.2 and JR2) expression was up-regulated and the resistance to R. solanaceraum and Pseudomonas syringae pv. tomato DC3000 was enhanced in response to CWP. Such CWP-induced defense responses were completely compromised in CWP-treated coi1-1 and jar1-1 mutants with an impaired JA signaling pathway. The induction of defense-related gene expression after CWP treatment was partially compromised in ET-insensitive ein2-1 mutants, but not in SA signaling mutants or nahG transgenic plants. Global gene expression analysis using cDNA array also suggested that several other JA- and ET-responsive genes, but not SA-responsive genes, were up-regulated in response to CWP. Further analysis of CWP-induced defense responses using another eight mutants with impaired defense signaling pathways indicated that, interestingly, the induction of JA-responsive gene expression and enhanced resistance to two bacterial pathogens in response to CWP were completely compromised in rar1-1, rar1-21, sgt1a-1, sgt1b (edm1) and npr1-1 mutants. Thus, the CWP-induced defense system appears to be regulated by JA-mediated and SGT1-, RAR1- and NPR1-dependent signaling pathways.

    DOI: 10.1093/pcp/pcp044

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  • High level expression of a virus resistance gene, RCY1, confers extreme resistance to Cucumber Mosaic Virus in Arabidopsis thaliana.

    21 ( 11 )   1398 - 1407   2008年11月

  • High Level Expression of a Virus Resistance Gene, RCY1, Confers Extreme Resistance to Cucumber mosaic virus in Arabidopsis thaliana

    Ken-Taro Sekine, Sayaka Kawakami, Shu Hase, Mayumi Kubota, Yuki Ichinose, Jyoti Shah, Hong-Gu Kang, Daniel F. Klessig, Hideki Takahashi

    MOLECULAR PLANT-MICROBE INTERACTIONS   21 ( 11 )   1398 - 1407   2008年11月

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    記述言語:英語   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    A coiled coil-nucleotide binding site-leucine rich repeat-type resistance gene, RCY1, confers resistance to a yellow strain of Cucumber mosaic virus, CMV(Y), in Arabidopsis thaliana ecotype C24. Resistance to CMV(Y) in C24 is accompanied by a hypersensitive response (HR) that is characterized by the development of necrotic local lesions at the primary infection sites. To further study the HR and resistance to CMV(Y) in ecotype Col-0, which is susceptible to CMV(Y), Col-0 were transformed with RCY1. Systemic spread of CMV(Y) was completely suppressed in RCY1-transformed Col-0 (Col::pRCY1 lines 2 to 6), whereas virulent strain CMV(B2) spread and multiplied systemically in these transgenic lines similar to that in wild-type Col-0. Interestingly, the resistant phenotype of Col:pRCY1 varied among the lines. In lines 3 and 6, in which levels of RCY1 transcript were similar to that in wild-type C24, the HR and resistance to CMV(Y) was induced. Line 4, which expresses moderately elevated levels of RCY1 transcript, exhibited moderately enhanced resistance compared with that in C24 or line 3. In contrast, lines 2 and 5, which highly overexpress the RCY1 gene, did not exhibit either visible lesions or a micro-HR on the inoculated leaves. Moreover, virus coat protein was not detected in either inoculated or noninoculated upper leaves of these two lines, suggesting that extreme resistance (ER) to CMV(Y) was induced by high levels of expression of RCY1. Furthermore, in transgenic lines expressing hemagglutinin (HA) epitope-tagged RCY1 (Col::pRCY1-HA), high levels of accumulation of RCY1-HA protein were also correlated with the ER phenotype. Global gene expression analysis in line 2, which highly overexpresses RCY1, indicated that expression of several defense-related genes were constitutively elevated compared with wild-type Col-0. Despite this, line 2 did not have enhanced resistance to other avirulent and virulent pathogens. Take together, constitutive accumulation of high levels of RCY1 protein appears to regulate the strength of RCY1-conferred resistance in a gene-for-gene manner and implies that ER and HR-associated resistance differ only in the strength of resistance.

    DOI: 10.1094/MPMI-21-11-1398

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  • Biological Control of Crown Gall of Grapevine, Rose, and Tomato by Nonpathogenic Agrobacterium vitis Strain VAR03-1

    A. Kawaguchi, K. Inoue, Y. Ichinose

    PHYTOPATHOLOGY   98 ( 11 )   1218 - 1225   2008年11月

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    担当区分:最終著者   記述言語:英語   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    A nonpathogenic strain of Agrobacterium vitis VAR03-1 was tested as a biological control agent for crown gall of grapevine (Vitis vinifera). When roots of grapevine, rose (Rose multiflora), and tomato (Lycopersicon esculentum) were soaked in a cell suspension of antagonists before planting in soil infested with tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens, respectively, treatment with VAR03-1 significantly reduced the number of plants with tumors and disease severity in the three plant species. The inhibitory effects of treatment with VAR03-1 and the nonpathogenic A. rhizogenes strain K84 on crown gall of rose and tomato were almost identical, and the inhibitory effect of VAR03-1 on grapevine was superior to that of K84. Moreover, VAR03-1 greatly controlled crown gall of grapevine due to tumorigenic A. vitis in the field. VAR03-1 established populations averaging 10(6) colony forming units (CFU)/g of root in the rhizosphere of grapevine and persisted on roots for 2 years. VAR03-1 was bacteriocinogenic, producing a halo of inhibition against those three species of Agrobacterium. This is the first report that a nonpathogenic strain, VAR03-1, can effectively control crown gall caused by tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens.

    DOI: 10.1094/PHYTO-98-11-1218

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  • Biological Control of Crown Gall of Grapevine, Rose, and Tomato by Nonpathogenic Agrobacterium vitis Strain VAR03-1

    A. Kawaguchi, K. Inoue, Y. Ichinose

    PHYTOPATHOLOGY   98 ( 11 )   1218 - 1225   2008年11月

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    記述言語:英語   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    A nonpathogenic strain of Agrobacterium vitis VAR03-1 was tested as a biological control agent for crown gall of grapevine (Vitis vinifera). When roots of grapevine, rose (Rose multiflora), and tomato (Lycopersicon esculentum) were soaked in a cell suspension of antagonists before planting in soil infested with tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens, respectively, treatment with VAR03-1 significantly reduced the number of plants with tumors and disease severity in the three plant species. The inhibitory effects of treatment with VAR03-1 and the nonpathogenic A. rhizogenes strain K84 on crown gall of rose and tomato were almost identical, and the inhibitory effect of VAR03-1 on grapevine was superior to that of K84. Moreover, VAR03-1 greatly controlled crown gall of grapevine due to tumorigenic A. vitis in the field. VAR03-1 established populations averaging 10(6) colony forming units (CFU)/g of root in the rhizosphere of grapevine and persisted on roots for 2 years. VAR03-1 was bacteriocinogenic, producing a halo of inhibition against those three species of Agrobacterium. This is the first report that a nonpathogenic strain, VAR03-1, can effectively control crown gall caused by tumorigenic A. vitis, A. rhizogenes, and A. tumefaciens.

    DOI: 10.1094/PHYTO-98-11-1218

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  • Amino Acid Sequence of Bacterial Microbe-Associated Molecular Pattern flg22 Is Required for Virulence 国際誌

    Kana Naito, Fumiko Taguchi, Tomoko Suzuki, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR PLANT-MICROBE INTERACTIONS   21 ( 9 )   1165 - 1174   2008年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    Flagellin proteins derived from Pseudomonas syringae pv. tabaci 6605 and flg22(Pa) (QRLSTGSRINSAKDDAAGLQIA), one of the microbe-associated molecular patterns (MAMP) in bacterial flagellin, induce cell death and growth inhibition in Arabidopsis thaliana. To examine the importance of aspartic acid (D) at position 43 from the N-terminus of a flagellin in its elicitor activity, D43 was replaced with valine (V) and alanine (A) in P. syringae pv. tabaci flagellin and flg22(Pta). The abilities of flagellins from P syringae pv. tabaci D43V and D43A to induce cell death and growth inhibition were reduced, whereas the abilities of flg22(Pta)D43V and flg22(Pta)D43A were abolished. These results indicate that D43 is important for elicitor activity in P. syringae pv. tabaci. When tobacco plants were inoculated with each bacterium by the spray method, both P. syringae pv. tabaci D43V and D43A mutants had remarkably reduced ability to cause disease symptoms. Both mutants had reduced or no swimming and swarming motilities and adhesion ability. In P. syringae pv. tabaci D43V, little flagellin protein was detected and few flagella were observed by electron microscopy. These results indicate that mutant flagella are unstable and that flagellar motility is impaired. Thus, the amino acid residue required for MAMP activity is important for the intrinsic flagellar function.

    DOI: 10.1094/MPMI-21-9-1165

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  • Amino Acid Sequence of Bacterial Microbe-Associated Molecular Pattern flg22 Is Required for Virulence

    Kana Naito, Fumiko Taguchi, Tomoko Suzuki, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR PLANT-MICROBE INTERACTIONS   21 ( 9 )   1165 - 1174   2008年9月

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    記述言語:英語   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

    Flagellin proteins derived from Pseudomonas syringae pv. tabaci 6605 and flg22(Pa) (QRLSTGSRINSAKDDAAGLQIA), one of the microbe-associated molecular patterns (MAMP) in bacterial flagellin, induce cell death and growth inhibition in Arabidopsis thaliana. To examine the importance of aspartic acid (D) at position 43 from the N-terminus of a flagellin in its elicitor activity, D43 was replaced with valine (V) and alanine (A) in P. syringae pv. tabaci flagellin and flg22(Pta). The abilities of flagellins from P syringae pv. tabaci D43V and D43A to induce cell death and growth inhibition were reduced, whereas the abilities of flg22(Pta)D43V and flg22(Pta)D43A were abolished. These results indicate that D43 is important for elicitor activity in P. syringae pv. tabaci. When tobacco plants were inoculated with each bacterium by the spray method, both P. syringae pv. tabaci D43V and D43A mutants had remarkably reduced ability to cause disease symptoms. Both mutants had reduced or no swimming and swarming motilities and adhesion ability. In P. syringae pv. tabaci D43V, little flagellin protein was detected and few flagella were observed by electron microscopy. These results indicate that mutant flagella are unstable and that flagellar motility is impaired. Thus, the amino acid residue required for MAMP activity is important for the intrinsic flagellar function.

    DOI: 10.1094/MPMI-21-9-1165

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  • (129) サポニン生合成に関与するイネOxidosqualene Cyclase遺伝子の分子解析(平成20年度日本植物病理学会大会講演要旨)

    稲垣 善茂, 今岡 敦子, 藤田 景子, 荒瀬 栄, 豊田 和弘, 白石 友紀, 一瀬 勇規, Osbourn Anne

    日本植物病理學會報   74 ( 3 )   201 - 201   2008年8月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • (308) Pseudomonas syringaeのフラジェリン糖鎖修飾構造とその合成に関わる遺伝子の機能解析 I(平成20年度日本植物病理学会大会講演要旨)

    常見 和彦, 田口 富美子, 竹内 香純, 石井 忠, 吉田 充, 小野 裕嗣, 亀山 眞由美, 一瀬 勇規

    日本植物病理學會報   74 ( 3 )   2008年8月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • (127) Medicago truncatula アピラーゼMtAPY1;1の機能解析(平成20年度日本植物病理学会大会講演要旨)

    塩原 泰樹, 藤田 景子, 稲垣 善茂, 一瀬 勇規, 豊田 和弘, 白石 友紀

    日本植物病理學會報   74 ( 3 )   200 - 200   2008年8月

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  • Phylogenetic and serological analyses reveal genetic diversity of Agrobacterium vitis strains in Japan

    A. Kawaguchi, H. Sawada, Y. Ichinose

    PLANT PATHOLOGY   57 ( 4 )   747 - 753   2008年8月

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    担当区分:最終著者   記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    Twenty-eight strains of Agrobacterium vitis, including both tumorigenic and nonpathogenic phenotypes involving 26 isolated in Japan and strains NCPPB 3554(T) and NCPPB 2562 isolated in Australia and Greece, respectively, were characterized by means of a slide agglutination test (SAT) using antisera prepared against somatic antigens. Phylogenetic analyses were also carried out, using the results of repetitive sequence-based polymerase chain reaction and the partial nucleotide sequences of pyrG, recA and rpoD. The A. vitis strains separated into four serogroups (A to D) based on the SAT reactivity. The phylogenetic analyses showed A. vitis strains separated into four tumorigenic groups (A to D) and one nonpathogenic group (E). Serogroups A to C corresponded exactly to genetic groups A to C, respectively, whereas serogroup D further divided into two distinct genetic groups, D and E. Genetic group E was isolated in Okayama Prefecture, Japan, and all strains of it have been found to coexist with tumorigenic strains belonging to the other groups within the same galled grapevine tissues. These results suggest that A. vitis strains are genetically heterogeneous and can be separated into several genetic groups. The differences between the nucleotide sequences of pyrG, recA and rpoD of the genetic groups will enable the development of a simple and convenient monitoring method, which will increase understanding of the dynamics of each genetic group of A. vitis in the natural environment.

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  • Phylogenetic and serological analyses reveal genetic diversity of Agrobacterium vitis strains in Japan

    A. Kawaguchi, H. Sawada, Y. Ichinose

    PLANT PATHOLOGY   57 ( 4 )   747 - 753   2008年8月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    Twenty-eight strains of Agrobacterium vitis, including both tumorigenic and nonpathogenic phenotypes involving 26 isolated in Japan and strains NCPPB 3554(T) and NCPPB 2562 isolated in Australia and Greece, respectively, were characterized by means of a slide agglutination test (SAT) using antisera prepared against somatic antigens. Phylogenetic analyses were also carried out, using the results of repetitive sequence-based polymerase chain reaction and the partial nucleotide sequences of pyrG, recA and rpoD. The A. vitis strains separated into four serogroups (A to D) based on the SAT reactivity. The phylogenetic analyses showed A. vitis strains separated into four tumorigenic groups (A to D) and one nonpathogenic group (E). Serogroups A to C corresponded exactly to genetic groups A to C, respectively, whereas serogroup D further divided into two distinct genetic groups, D and E. Genetic group E was isolated in Okayama Prefecture, Japan, and all strains of it have been found to coexist with tumorigenic strains belonging to the other groups within the same galled grapevine tissues. These results suggest that A. vitis strains are genetically heterogeneous and can be separated into several genetic groups. The differences between the nucleotide sequences of pyrG, recA and rpoD of the genetic groups will enable the development of a simple and convenient monitoring method, which will increase understanding of the dynamics of each genetic group of A. vitis in the natural environment.

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  • Polyphosphate kinase is essential for swarming motility, tolerance to environmental stresses, and virulence in Pseudomonas syringae pv. tabaci 6605

    Md. Mijan Hossain, Chiharu Tani, Tomoko Suzuki, Fumiko Taguchi, Tatsuhiro Ezawa, Yuki Ichinose

    PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY   72 ( 4-6 )   122 - 127   2008年7月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD  

    Polyphosphate kinase (PPK), encoded by the ppk gene, is a principal enzyme responsible for synthesis of inorganic polyphosphate (poly P) from ATP in many Gram-negative bacteria. In order to elucidate the functions of poly P in Pseudomonas syringae pv. tabaci 6605, an in-frame deletion mutant of the ppk gene (Delta ppk) was constructed. The Delta ppk mutant did not accumulate poly P, whereas the wild-type strain accumulated a large quantity. The mutant had reduced swarming motility, even though it retains swimming motility like the parental strain. The mutant exhibited increased sensitivity to prolonged incubation and environmental stresses, such as heat shock and oxidative stress and reduced exopolysaccharide (EPS) production compared to the wild-type. Northern blot analysis revealed that expression of the rpoS gene, encoding the stationary phase sigma factor RpoS, was reduced in Delta ppk in the logarithmic phase, indicating that rpoS is regulated by the ppk gene. The poly P deficient mutant had significantly reduced ability to cause disease in its host tobacco plant and in planta growth of the mutant was also significantly reduced in host tobacco leaves as compared to the wild-type strain. Thus, our results suggest that poly P plays an important role in the virulence of P. syringae pv. tabaci 6605. (C) 2008 Elsevier Ltd. All rights reserved.

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  • Polyphosphate kinase is essential for swarming motility, tolerance to environmental stresses, and virulence in Pseudomonas syringae pv. tabaci 6605

    Md. Mijan Hossain, Chiharu Tani, Tomoko Suzuki, Fumiko Taguchi, Tatsuhiro Ezawa, Yuki Ichinose

    PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY   72 ( 4-6 )   122 - 127   2008年7月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD  

    Polyphosphate kinase (PPK), encoded by the ppk gene, is a principal enzyme responsible for synthesis of inorganic polyphosphate (poly P) from ATP in many Gram-negative bacteria. In order to elucidate the functions of poly P in Pseudomonas syringae pv. tabaci 6605, an in-frame deletion mutant of the ppk gene (Delta ppk) was constructed. The Delta ppk mutant did not accumulate poly P, whereas the wild-type strain accumulated a large quantity. The mutant had reduced swarming motility, even though it retains swimming motility like the parental strain. The mutant exhibited increased sensitivity to prolonged incubation and environmental stresses, such as heat shock and oxidative stress and reduced exopolysaccharide (EPS) production compared to the wild-type. Northern blot analysis revealed that expression of the rpoS gene, encoding the stationary phase sigma factor RpoS, was reduced in Delta ppk in the logarithmic phase, indicating that rpoS is regulated by the ppk gene. The poly P deficient mutant had significantly reduced ability to cause disease in its host tobacco plant and in planta growth of the mutant was also significantly reduced in host tobacco leaves as compared to the wild-type strain. Thus, our results suggest that poly P plays an important role in the virulence of P. syringae pv. tabaci 6605. (C) 2008 Elsevier Ltd. All rights reserved.

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  • Gac two-component system in Pseudomonas syringae pv. tabaci is required for virulence but not for hypersensitive reaction

    Mizuri Marutani, Fumiko Taguchi, Yujiro Ogawa, Md. Mijan Hossain, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR GENETICS AND GENOMICS   279 ( 4 )   313 - 322   2008年4月

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    記述言語:英語   出版者・発行元:SPRINGER  

    Pseudomonas syringae pv. tabaci 6605 causes wildfire disease on host tobacco plants. To investigate the regulatory mechanism of the expression of virulence, Gac two-Component system-defective mutants, Delta gacA and Delta gacS, and a double mutant, Delta gacA Delta gacS, were generated. These mutants produced smaller amounts of N-acyl homoserine lactones required for quorum sensing, had lost swarming motility, and had reduced expression of virulence-related hrp genes and the algT gene required for exopolysaccharide production. The ability of the mutants to cause disease symptoms in their host tobacco plant was remarkably reduced, while they retained the ability to induce hypersensitive reaction (HR) in the nonhost plants. These results indicated that the Gac two-component system of P. syringae pv. tabaci 6605 is indispensable for virulence on the host plant, but not for HR induction in the nonhost plants.

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  • Gac two-component system in Pseudomonas syringae pv. tabaci is required for virulence but not for hypersensitive reaction

    Mizuri Marutani, Fumiko Taguchi, Yujiro Ogawa, Md. Mijan Hossain, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR GENETICS AND GENOMICS   279 ( 4 )   313 - 322   2008年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:SPRINGER  

    Pseudomonas syringae pv. tabaci 6605 causes wildfire disease on host tobacco plants. To investigate the regulatory mechanism of the expression of virulence, Gac two-Component system-defective mutants, Delta gacA and Delta gacS, and a double mutant, Delta gacA Delta gacS, were generated. These mutants produced smaller amounts of N-acyl homoserine lactones required for quorum sensing, had lost swarming motility, and had reduced expression of virulence-related hrp genes and the algT gene required for exopolysaccharide production. The ability of the mutants to cause disease symptoms in their host tobacco plant was remarkably reduced, while they retained the ability to induce hypersensitive reaction (HR) in the nonhost plants. These results indicated that the Gac two-component system of P. syringae pv. tabaci 6605 is indispensable for virulence on the host plant, but not for HR induction in the nonhost plants.

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    その他リンク: http://link.springer.com/article/10.1007/s00438-007-0309-y/fulltext.html

  • Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana

    Kuniaki Higashi, Yasuhiro Ishiga, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR GENETICS AND GENOMICS   279 ( 3 )   303 - 312   2008年3月

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    記述言語:英語   出版者・発行元:SPRINGER  

    Flagellin, a component of the flagellar filament of Pseudomonas syringae pv. tabaci 6605 (Pta), induces hypersensitive reaction in its non-host Arabidopsis thaliana. We identified the WRKY41 gene, which belongs to a multigene family encoding WRKY plant-specific transcription factors, as one of the flagellin-inducible genes in A. thaliana. Expression of WRKY41 is induced by inoculation with the incompatible pathogen P. syringae pv. tomato DC3000 (Pto) possessing AvrRpt2 and the non-host pathogens Pta within 6-h after inoculation, but not by inoculation with the compatible Pto. Expression of WRKY41 was also induced by inoculation of A. thaliana with an hrp-type three secretion system (T3SS)-defective mutant of Pto, indicating that effectors produced by T3SS in the Pto wild-type suppress the activation of WRKY41. Arabidopsis overexpressing WRKY41 showed enhanced resistance to the Pto wild-type but increased susceptibility to Erwinia carotovora EC1. WRKY41-overexpressing Arabidopsis constitutively expresses the PR5 gene, but suppresses the methyl jasmonate-induced PDF1.2 gene expression. These results demonstrate that WRKY41 may be a key regulator in the cross talk of salicylic acid and jasmonic acid pathways.

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  • Modulation of defense signal transduction by flagellin-induced WRKY41 transcription factor in Arabidopsis thaliana 国際誌

    Kuniaki Higashi, Yasuhiro Ishiga, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    MOLECULAR GENETICS AND GENOMICS   279 ( 3 )   303 - 312   2008年3月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:SPRINGER  

    Flagellin, a component of the flagellar filament of Pseudomonas syringae pv. tabaci 6605 (Pta), induces hypersensitive reaction in its non-host Arabidopsis thaliana. We identified the WRKY41 gene, which belongs to a multigene family encoding WRKY plant-specific transcription factors, as one of the flagellin-inducible genes in A. thaliana. Expression of WRKY41 is induced by inoculation with the incompatible pathogen P. syringae pv. tomato DC3000 (Pto) possessing AvrRpt2 and the non-host pathogens Pta within 6-h after inoculation, but not by inoculation with the compatible Pto. Expression of WRKY41 was also induced by inoculation of A. thaliana with an hrp-type three secretion system (T3SS)-defective mutant of Pto, indicating that effectors produced by T3SS in the Pto wild-type suppress the activation of WRKY41. Arabidopsis overexpressing WRKY41 showed enhanced resistance to the Pto wild-type but increased susceptibility to Erwinia carotovora EC1. WRKY41-overexpressing Arabidopsis constitutively expresses the PR5 gene, but suppresses the methyl jasmonate-induced PDF1.2 gene expression. These results demonstrate that WRKY41 may be a key regulator in the cross talk of salicylic acid and jasmonic acid pathways.

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  • (72)タバコ野火病菌べん毛タンパク質フラジェリン糖鎖のべん毛構造・運動能における役割(関西部会講演要旨,平成19年度地域部会講演要旨)

    田口 富美子, 柴田 敏史, 鈴木 智子, 小川 裕二郎, 相沢 慎一, 一瀬 勇規

    日本植物病理學會報   74 ( 1 )   75 - 75   2008年2月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • Effects of glycosylation on swimming ability and flagellar polymorphic transformation in Pseudomonas syringae pv. tabaci 6605 国際誌

    Fumiko Taguchi, Satoshi Shibata, Tomoko Suzuki, Yujiro Ogawa, Shin-Ichi Aizawa, Kasumi Takeuchi, Yuki Ichinose

    JOURNAL OF BACTERIOLOGY   190 ( 2 )   764 - 768   2008年1月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    The role of flagellin glycosylation on motility was investigated in Pseudomonas syringae pv. tabaci. The swimming activity of glycosylation-defective mutants was prominently decreased in a highly viscous medium. The mutants showed differences in polymorphic transitions and in the bundle formation of flagella, indicating that glycosylation stabilizes the filament structure and lubricates the rotation of the bundle.

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  • Effects of glycosylation on swimming ability and flagellar polymorphic transformation in Pseudomonas syringae pv. tabaci 6605

    Fumiko Taguchi, Satoshi Shibata, Tomoko Suzuki, Yujiro Ogawa, Shin-Ichi Aizawa, Kasumi Takeuchi, Yuki Ichinose

    JOURNAL OF BACTERIOLOGY   190 ( 2 )   764 - 768   2008年1月

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    記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    The role of flagellin glycosylation on motility was investigated in Pseudomonas syringae pv. tabaci. The swimming activity of glycosylation-defective mutants was prominently decreased in a highly viscous medium. The mutants showed differences in polymorphic transitions and in the bundle formation of flagella, indicating that glycosylation stabilizes the filament structure and lubricates the rotation of the bundle.

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  • Molecular analysis for genome structure and gene expression of Oxidosqualene Cyclase genes in rice seedling

    Yoshishige Inagaki, Yukako Mutsukado, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose, Anne Osbourn

    GENES & GENETIC SYSTEMS   82 ( 6 )   521 - 521   2007年12月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:GENETICS SOC JAPAN  

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  • Flagellin Glycans from two pathovars of Pseudomonas syringae contain rhamnose in D and L configurations in different ratios and modified 4-amino-4,6-dideoxyglucose

    Kasumi Takeuchi, Hiroshi Ono, Mitsuru Yoshida, Tadashi Ishii, Etsuko Katoh, Fumiko Taguchi, Ryuji Miki, Katsuyoshi Murata, Hanae Kaku, Yuki Ichinose

    JOURNAL OF BACTERIOLOGY   189 ( 19 )   6945 - 6956   2007年10月

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    記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    Flagellins from Pseudomonas syringae pv. glycinea race 4 and Pseudomonas syringae pv. tabaci 6605 have been found to be glycosylated. Glycosylation of flagellin is essential for bacterial virulence and is also involved in the determination of host specificity. Flagellin glycans from both pathovars were characterized, and common sites of glycosylation were identified on six serine residues (positions 143, 164, 176, 183, 193, and 201). The structure of the glycan at serine 201 (S201) of flagellin from each pathovar was determined by sugar composition analysis, mass spectrometry, and H-1 and C-13 nuclear magnetic resonance spectroscopy. These analyses showed that the S201 glycans from both pathovars were composed of a common unique trisaccharide consisting of two rhamnosyl (Rha) residues and one modified 4-amino-4,6-dideoxyglucosyl (Qui4N) residue, beta-D-Quip4N(3-hydroxy-1-oxobutyl)2Me-(1 -&gt; 3)-alpha-L-Rhap-(1 -&gt; 2)-alpha-L-Rhap. Furthermore, mass analysis suggests that the glycans on each of the six serine residues are composed of similar trisaccharide units. Determination of the enantiomeric ratio of Rha from the flagellin proteins showed that flagellin from P. syringae pv. tabaci 6605 consisted solely Of L-Rha, whereas P. syringae pv. glycinea race 4 flagellin contained both L-Rha and D-Rha at a molar ratio of about 4:1. Taking these findings together with those from our previous study, we conclude that these flagellin glycan structures may be important for the virulence and host specificity of P. syringae.

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  • N-terminal domain including conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana

    Kana Naito, Yasuhiro Ishiga, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   73 ( 4 )   281 - 285   2007年8月

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    記述言語:英語  

    Flagellin in Pseudomonas syringae is a potent elicitor of defense responses including hypersensitive cell death in dicot plants. The oligopeptides flg22 consisting of 22 conserved amino acids near the N-terminus of flagellins is reported to induce plant defense responses. Because glycosylation of the central domain of flagellin affects its elicitor activity, we investigated whether any peptide sequence in addition to flg22 is required for flagellin-induced hypersensitive reaction. A study of recombinant flagellin polypeptides indicated that the N-terminal domain including the conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana. © 2007 The Phytopathological Society of Japan and Springer.

    DOI: 10.1007/s10327-007-0017-9

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  • N-terminal domain including conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana

    Kana Naito, Yasuhiro Ishiga, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   73 ( 4 )   281 - 285   2007年8月

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    記述言語:英語  

    Flagellin in Pseudomonas syringae is a potent elicitor of defense responses including hypersensitive cell death in dicot plants. The oligopeptides flg22 consisting of 22 conserved amino acids near the N-terminus of flagellins is reported to induce plant defense responses. Because glycosylation of the central domain of flagellin affects its elicitor activity, we investigated whether any peptide sequence in addition to flg22 is required for flagellin-induced hypersensitive reaction. A study of recombinant flagellin polypeptides indicated that the N-terminal domain including the conserved flg22 is required for flagellin-induced hypersensitive cell death in Arabidopsis thaliana. © 2007 The Phytopathological Society of Japan and Springer.

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  • Suppression of Cdc27B expression induces plant defence responses

    Chikako Kudo, Tomoko Suzuki, Sumie Fukuoka, Shuta Asai, Hiroko Suenaga, Michiko Sasabe, Yoshitaka Takano, Tetsuro Okuno, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose, Yoshi-Shige Inagaki

    MOLECULAR PLANT PATHOLOGY   8 ( 4 )   365 - 373   2007年7月

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    記述言語:英語   出版者・発行元:BLACKWELL PUBLISHING  

    Non-host resistance is the most general form of disease resistance in plants because it is effective against most phytopathogens. The importance of hypersensitive responses (HRs) in non-host resistance of Nicotiana species to the oomycete Phytophthora is clear. INF1 elicitin, an elicitor obtained from the late-blight pathogen Phytophthora infestans, is sufficient to induce a typical HR in Nicotiana species. The molecular mechanisms that underlie the non-host resistance component of plant defence responses have been investigated using differential-display polymerase chain reaction (PCR) in a model HR system between INF1 elicitin and tobacco BY-2 cells. Differential-display PCR has revealed that Cdc27B is down-regulated in tobacco BY- 2 cells after treatment with INF1 elicitin. Cdc27B is one of 13 essential components of the anaphase- promoting complex or cyclosome ( APC/ C)-type E3 ubiquitin ligase complex in yeast. This APC/C-type E3 ubiquitin ligase complex regulates G2-to-M phase transition of the cell cycle by proteolytic degradation. In this study, we investigated the roles of this gene, NbCdc27B, in plant defence responses using virus-induced gene silencing. Suppression of NbCdc27B in Nicotiana benthamiana plants induced defence responses and a gain of resistance to Colletotrichum lagenarium fungus. Elicitin-induced hypersensitive cell death (HCD) was inhibited mildly in plants silenced with tobacco rattle virus:: Cdc27B. Cdc27B could manage the signalling pathways of plant defence responses as a negative regulator without HCD.

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  • Suppression of Cdc27B expression induces plant defence responses

    Chikako Kudo, Tomoko Suzuki, Sumie Fukuoka, Shuta Asai, Hiroko Suenaga, Michiko Sasabe, Yoshitaka Takano, Tetsuro Okuno, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose, Yoshi-Shige Inagaki

    MOLECULAR PLANT PATHOLOGY   8 ( 4 )   365 - 373   2007年7月

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    記述言語:英語   出版者・発行元:BLACKWELL PUBLISHING  

    Non-host resistance is the most general form of disease resistance in plants because it is effective against most phytopathogens. The importance of hypersensitive responses (HRs) in non-host resistance of Nicotiana species to the oomycete Phytophthora is clear. INF1 elicitin, an elicitor obtained from the late-blight pathogen Phytophthora infestans, is sufficient to induce a typical HR in Nicotiana species. The molecular mechanisms that underlie the non-host resistance component of plant defence responses have been investigated using differential-display polymerase chain reaction (PCR) in a model HR system between INF1 elicitin and tobacco BY-2 cells. Differential-display PCR has revealed that Cdc27B is down-regulated in tobacco BY- 2 cells after treatment with INF1 elicitin. Cdc27B is one of 13 essential components of the anaphase- promoting complex or cyclosome ( APC/ C)-type E3 ubiquitin ligase complex in yeast. This APC/C-type E3 ubiquitin ligase complex regulates G2-to-M phase transition of the cell cycle by proteolytic degradation. In this study, we investigated the roles of this gene, NbCdc27B, in plant defence responses using virus-induced gene silencing. Suppression of NbCdc27B in Nicotiana benthamiana plants induced defence responses and a gain of resistance to Colletotrichum lagenarium fungus. Elicitin-induced hypersensitive cell death (HCD) was inhibited mildly in plants silenced with tobacco rattle virus:: Cdc27B. Cdc27B could manage the signalling pathways of plant defence responses as a negative regulator without HCD.

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  • Ultrastructural features of Mycosphaerella pinodes Infection and differenced in host susceptibility responses among Medicago truncatula Ecotypes.

    Suzuki, T, Maeda, A, Hirose, M, Ichinose, Y, Toyoda, K, Shiraishi, T

    J. Electr. Microsc. Technol. Med. Biol.   20 ( 2 )   167 - 168   2007年

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  • Ultrastructural features of Mycosphaerella pinodes Infection and differenced in host susceptibility responses among Medicago truncatula Ecotypes.

    Suzuki, T, Maeda, A, Hirose, M, Ichinose, Y, Toyoda, K, Shiraishi, T

    J. Electr. Microsc. Technol. Med. Biol.   20 ( 2 )   167 - 168   2007年

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  • Elicitin-responsive lectin-like receptor kinase genes in BY-2 cells

    Michiko Sasabe, Kana Naito, Hiroko Suenaga, Takako Ikeda, Kazuhiro Toyodak, Yoshishige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    DNA SEQUENCE   18 ( 2 )   152 - 159   2007年

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    記述言語:英語   出版者・発行元:INFORMA HEALTHCARE-TAYLOR & FRANCIS  

    The inhibition of elicitor-induced plant defense responses by the protein kinase inhibitors K252a and staurosporine indicates that defense responses require protein phosphorylation. We isolated a cDNA clone encoding Nicotiana tabacum lectin-like receptor protein kinase 1 ( NtlecRK1), an elicitor-responsive gene; in tobacco bright yellow ( BY-2) cells by a differential display method. NtlecRK forms a gene family with at least three members in tobacco. All three NtlecRK genes potentially encode the N-terminal legume lectin domain, transmembrane domain and C-terminal Ser/Thr-type protein kinase domain. Green fluorescent protein ( GFP) fusion showed that the NtlecRK1 protein was located on the plasma membrane. In addition, NtlecRK1 and 3 were responsive to INF1 elicitin and the bacterial elicitor harpin. These results indicate that NtlecRKs are membrane-located protein kinases that are induced during defense responses in BY-2 cells.

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  • Elicitin-responsive lectin-like receptor kinase genes in BY-2 cells

    Michiko Sasabe, Kana Naito, Hiroko Suenaga, Takako Ikeda, Kazuhiro Toyodak, Yoshishige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    DNA SEQUENCE   18 ( 2 )   152 - 159   2007年

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    記述言語:英語   出版者・発行元:INFORMA HEALTHCARE-TAYLOR & FRANCIS  

    The inhibition of elicitor-induced plant defense responses by the protein kinase inhibitors K252a and staurosporine indicates that defense responses require protein phosphorylation. We isolated a cDNA clone encoding Nicotiana tabacum lectin-like receptor protein kinase 1 ( NtlecRK1), an elicitor-responsive gene; in tobacco bright yellow ( BY-2) cells by a differential display method. NtlecRK forms a gene family with at least three members in tobacco. All three NtlecRK genes potentially encode the N-terminal legume lectin domain, transmembrane domain and C-terminal Ser/Thr-type protein kinase domain. Green fluorescent protein ( GFP) fusion showed that the NtlecRK1 protein was located on the plasma membrane. In addition, NtlecRK1 and 3 were responsive to INF1 elicitin and the bacterial elicitor harpin. These results indicate that NtlecRKs are membrane-located protein kinases that are induced during defense responses in BY-2 cells.

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  • A homologue of the 3-oxoacyl-(acyl carrier protein) synthase III gene located in the glycosylation island of Pseudomonas syringae pv. tabaci regulates virulence factors via N-acyl homoserine lactone and fatty acid synthesis 国際誌

    Fumiko Taguchi, Yujiro Ogawa, Kasumi Takeuchi, Tomoko Suzuki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    JOURNAL OF BACTERIOLOGY   188 ( 24 )   8376 - 8384   2006年12月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    Pseudomonas syringae pv. tabaci 6605 possesses a genetic region involved in flagellin glycosylation. This region is composed of three open reading frames: orf1, orf2, and orf3. Our previous study revealed that orf1 and orf2 encode glycosyltransferases; on the other hand, orf3 has no role in posttranslational modification of flagellin. Although the function of Orf3 remained unclear, an orf3 deletion mutant (Delta orf3 mutant) had reduced virulence on tobacco plants. Orf3 shows significant homology to a 3-oxoacyl-(acyl carrier protein) synthase III in the fatty acid elongation cycle. The Delta orf3 mutant had a significantly reduced ability to form acyl homoserine lactones (AHLs), which are quorum-sensing molecules, suggesting that Orf3 is required for AHL synthesis. In comparison with the wild-type strain, swarming motility, biosurfactant production, and tolerance to H2O2 and antibiotics were enhanced in the Delta orf3 mutant. A scanning electron micrograph of inoculated bacteria on the tobacco leaf surface revealed that there is little extracellular polymeric substance matrix surrounding the cells in the Delta orf3 mutant. The phenotypes of the Delta orf3 mutant and an AHL synthesis (Delta psyI) mutant were similar, although the mutant-specific characteristics were more extreme in the Delta orf3 mutant. The swarming motility of the Delta orf3 mutant was greater than that of the Delta psyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the Delta orf3 mutant. Furthermore, the amounts of iron and biosurfactant seem to be involved in biofilm development under quorum-sensing regulation in P. syringae pv. tabaci 6605.

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  • A homologue of the 3-oxoacyl-(acyl carrier protein) synthase III gene located in the glycosylation island of Pseudomonas syringae pv. tabaci regulates virulence factors via N-acyl homoserine lactone and fatty acid synthesis

    Fumiko Taguchi, Yujiro Ogawa, Kasumi Takeuchi, Tomoko Suzuki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    JOURNAL OF BACTERIOLOGY   188 ( 24 )   8376 - 8384   2006年12月

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    記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    Pseudomonas syringae pv. tabaci 6605 possesses a genetic region involved in flagellin glycosylation. This region is composed of three open reading frames: orf1, orf2, and orf3. Our previous study revealed that orf1 and orf2 encode glycosyltransferases; on the other hand, orf3 has no role in posttranslational modification of flagellin. Although the function of Orf3 remained unclear, an orf3 deletion mutant (Delta orf3 mutant) had reduced virulence on tobacco plants. Orf3 shows significant homology to a 3-oxoacyl-(acyl carrier protein) synthase III in the fatty acid elongation cycle. The Delta orf3 mutant had a significantly reduced ability to form acyl homoserine lactones (AHLs), which are quorum-sensing molecules, suggesting that Orf3 is required for AHL synthesis. In comparison with the wild-type strain, swarming motility, biosurfactant production, and tolerance to H2O2 and antibiotics were enhanced in the Delta orf3 mutant. A scanning electron micrograph of inoculated bacteria on the tobacco leaf surface revealed that there is little extracellular polymeric substance matrix surrounding the cells in the Delta orf3 mutant. The phenotypes of the Delta orf3 mutant and an AHL synthesis (Delta psyI) mutant were similar, although the mutant-specific characteristics were more extreme in the Delta orf3 mutant. The swarming motility of the Delta orf3 mutant was greater than that of the Delta psyI mutant. This was attributed to the synergistic effects of the overproduction of biosurfactants and/or alternative fatty acid metabolism in the Delta orf3 mutant. Furthermore, the amounts of iron and biosurfactant seem to be involved in biofilm development under quorum-sensing regulation in P. syringae pv. tabaci 6605.

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  • ダイズ斑点細菌病菌のフラジェリン糖鎖解析

    竹内香純, 田口富美子, 加藤悦子, 三木隆二, 村田勝義, 賀来華江, 一瀬勇規

    日本植物病理学会報   72 ( 4 )   310   2006年11月

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    記述言語:日本語  

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  • Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

    Tomonari Kasai, Tomoko Suzuki, Kozue Ono, Ken'Ichi Ogawa, Yoshishige Inagaki, Yuki Ichinose, Kazuhiro Toyoda, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 5 )   265 - 272   2006年10月

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    記述言語:英語  

    We previously reported that the release of O2 - from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. © 2006 The Phytopathological Society of Japan and Springer-Verlag.

    DOI: 10.1007/s10327-006-0283-y

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  • Pea extracellular Cu/Zn-superoxide dismutase responsive to signal molecules from a fungal pathogen

    Tomonari Kasai, Tomoko Suzuki, Kozue Ono, Ken'Ichi Ogawa, Yoshishige Inagaki, Yuki Ichinose, Kazuhiro Toyoda, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 5 )   265 - 272   2006年10月

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    記述言語:英語  

    We previously reported that the release of O2 - from isolated pea cell walls was enhanced by a 70-kDa glycoprotein elicitor but was suppressed by mucin-type glycopeptide suppressors (supprescins A and B) prepared from pycnospore germination fluid of Mycosphaerella pinodes, causal agent of Mycosphaerella blight of pea. Here, we show that superoxide dismutase (SOD) in the apoplast fluid/cell wall of pea seedlings responds to the fungal elicitor and suppressor molecules. In a pharmacological study and with internal amino acid sequencing, the apoplastic SOD in a pea cultivar Midoriusui was found to be a Cu/Zn type SOD. We cloned a full-length cDNA of the Cu/Zn-SOD and designated it as PsCu/Zn-SOD1. An increase in PsCu/Zn-SOD1 mRNA and the PsCu/Zn-SOD1 protein was induced by treatment with the elicitor more intensively than by wounding. Such induction by the elicitor or wounding, however, was inhibited by the concomitant presence of supprescins. The SOD activity of recombinant PsCu/Zn-SOD1 was regulated directly by these signal molecules in a manner similar to their effect on the SOD activity in the apoplastic fluid and in the cell wall-bound proteins. Based on these findings, we discuss a role for PsCu/Zn-SOD1 in the pea defense response. © 2006 The Phytopathological Society of Japan and Springer-Verlag.

    DOI: 10.1007/s10327-006-0283-y

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  • A binding protein for fungal signal molecules in the cell wall of Pisum sativum

    Akinori Kiba, Takako Ohgawara, Kazuhiro Toyoda, Miho Inoue-Ozaki, Tadahiro Takeda, Uppalapati Srinivasa Rao, Toshiaki Kato, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 4 )   228 - 237   2006年8月

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    記述言語:英語  

    In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules. © The Phytopathological Society of Japan and Springer-Verlag 2006.

    DOI: 10.1007/s10327-006-0278-8

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  • A pea NTPase, PsAPY1, recognizes signal molecules from microorganisms

    Akinori Kiba, Kazuhiro Toyoda, Kazuaki Yoshioka, Kami Tsujimura, Hirotaka Takahashi, Yuki Ichinose, Tadahiro Takeda, Toshiaki Kato, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 4 )   238 - 246   2006年8月

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    記述言語:英語  

    Apyrases (E.C.3.6.1.5
    NTP-NDPases) are distributed in the cytosol, nuclei, cytoskeleton, and on the surface of plant cells. Some may play an important role in signal transduction from exogenous stimuli. We previously found a protein of ca. 55-kDa (CWP-55) in an ATPase-rich fraction from the pea cell wall bound to the elicitor and supprescins (suppressors of defense) from pea pathogen Mycosphaerella pinodes. We cloned the cDNA of CWP-55 that coincided with PsAPY1, one of two NTPase clones in a pea cDNA library. An analysis with a green fluorescent protein fusion protein indicated that PsAPY1 was distributed in the cell wall, nucleus, and cytoplasm. The recombinant PsAPY1 expressed in Escherichia coli had ATP-hydrolyzing activity responsive not only to the elicitor and supprescins from the pea pathogen but also to other elicitors such as a bacterial harpin, a yeast extract, and a synthetic glycopeptide. Biotinylated fungal signal molecules were bound to the recombinant PsAPY1 specifically. Resonant mirror detection confirmed such binding characteristics of PsAPY1. Based on these results, we discuss the role of cell-wall-bound NTPases in recognizing and responding to microorganisms on the cell wall surface. © The Phytopathological Society of Japan and Springer-Verlag 2006.

    DOI: 10.1007/s10327-006-0279-7

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  • A pea NTPase, PsAPY1, recognizes signal molecules from microorganisms

    Akinori Kiba, Kazuhiro Toyoda, Kazuaki Yoshioka, Kami Tsujimura, Hirotaka Takahashi, Yuki Ichinose, Tadahiro Takeda, Toshiaki Kato, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 4 )   238 - 246   2006年8月

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    記述言語:英語  

    Apyrases (E.C.3.6.1.5
    NTP-NDPases) are distributed in the cytosol, nuclei, cytoskeleton, and on the surface of plant cells. Some may play an important role in signal transduction from exogenous stimuli. We previously found a protein of ca. 55-kDa (CWP-55) in an ATPase-rich fraction from the pea cell wall bound to the elicitor and supprescins (suppressors of defense) from pea pathogen Mycosphaerella pinodes. We cloned the cDNA of CWP-55 that coincided with PsAPY1, one of two NTPase clones in a pea cDNA library. An analysis with a green fluorescent protein fusion protein indicated that PsAPY1 was distributed in the cell wall, nucleus, and cytoplasm. The recombinant PsAPY1 expressed in Escherichia coli had ATP-hydrolyzing activity responsive not only to the elicitor and supprescins from the pea pathogen but also to other elicitors such as a bacterial harpin, a yeast extract, and a synthetic glycopeptide. Biotinylated fungal signal molecules were bound to the recombinant PsAPY1 specifically. Resonant mirror detection confirmed such binding characteristics of PsAPY1. Based on these results, we discuss the role of cell-wall-bound NTPases in recognizing and responding to microorganisms on the cell wall surface. © The Phytopathological Society of Japan and Springer-Verlag 2006.

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  • A binding protein for fungal signal molecules in the cell wall of Pisum sativum

    Akinori Kiba, Takako Ohgawara, Kazuhiro Toyoda, Miho Inoue-Ozaki, Tadahiro Takeda, Uppalapati Srinivasa Rao, Toshiaki Kato, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 4 )   228 - 237   2006年8月

     詳細を見る

    記述言語:英語  

    In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules. © The Phytopathological Society of Japan and Springer-Verlag 2006.

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  • Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

    Hirotaka Takahashi, Kazuhiro Toyoda, Yuzo Hirakawa, Kunihiko Morishita, Toshiaki Kato, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 3 )   143 - 151   2006年6月

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    記述言語:英語  

    Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. © The Phytopathological Society of Japan and Springer-Verlag 2006.

    DOI: 10.1007/s10327-005-0267-3

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  • Identification of glycosylation genes and glycosylated amino acids of flagellin in Pseudomonas syringae pv. tabaci

    F Taguchi, K Takeuchi, E Katoh, K Murata, T Suzuki, M Marutani, T Kawasaki, M Eguchi, S Katoh, H Kaku, C Yasuda, Y Inagaki, K Toyoda, T Shiraishi, Y Ichinose

    CELLULAR MICROBIOLOGY   8 ( 6 )   923 - 938   2006年6月

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    記述言語:英語   出版者・発行元:BLACKWELL PUBLISHING  

    A glycosylation island is a genetic region required for glycosylation. The glycosylation island of flagellin in Pseudomonas syringae pv. tabaci 6605 consists of three orfs: orf1, orf2 and orf3. Orf1 and orf2 encode putative glycosyltransferases, and their deletion mutants, Delta orf1 and Delta orf2, exhibit deficient flagellin glycosylation or produce partially glycosylated flagellin respectively. Digestion of glycosylated flagellin from wild-type bacteria and non-glycosylated flagellin from Delta orf1 mutant using aspartic N-peptidase and subsequent HPLC analysis revealed candidate glycosylated amino acids. By generation of site-directed Ser/Ala-substituted mutants, all glycosylated amino acid residues were identified at positions 143, 164, 176, 183, 193 and 201. Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) analysis revealed that each glycan was about 540 Da. While all glycosylation-defective mutants retained swimming ability, swarming ability was reduced in the Delta orf1, Delta orf2 and Ser/Ala-substituted mutants. All glycosylation mutants were also found to be impaired in the ability to adhere to a polystyrene surface and in the ability to cause disease in tobacco. Based on the predicted tertiary structure of flagellin, S176 and S183 are expected to be located on most external surface of the flagellum. Thus the effect of Ala-substitution of these serines is stronger than that of other serines. These results suggest that glycosylation of flagellin in P. syringae pv. tabaci 6605 is required for bacterial virulence. It is also possible that glycosylation of flagellin may mask elicitor function of flagellin molecule.

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  • Induction of defense responses in pea tissues by inorganic phosphate

    Tomoharu Kawahara, Hiroko Namba, Kazuhiro Toyoda, Tomonari Kasai, Megumi Sugimoto, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 3 )   129 - 136   2006年6月

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    記述言語:英語  

    When inorganic phosphate, a common and essential element for organisms, was applied endogenously, a rejection reaction and superoxide generation were induced in pea tissues but phytoalexin production was not. Phosphate-induced superoxide generation was sensitive to cycloheximide (CHX) and salicylhydroxamic acid (SHAM), indicating that part of the generation was dependent upon the expression of peroxidase gene(s). Peroxidases (POXs) are well known not only to scavenge hydrogen peroxide with phenolics but also to generate superoxide via NADH oxidation in the presence of p-coumaric acid and manganese ion. We cloned five pea POX cDNAs that are predicted to be located outside of the cells. The accumulation of five POX mRNAs, NTPase mRNA, and phenylalanine ammonia-lyase mRNA was measured by semiquantitative reverse transcription-polymerase chain reaction. The expression of the five POX genes was induced by a fungal elicitor. On the other hand, inorganic phosphate induced the accumulation of POX11, POX14, and POX21 mRNAs but not of POX13, POX29, and PsPAL1 mRNAs within 1-3∈h after treatment of pea seedlings. In view of these findings, we discuss inorganic phosphate as a signal transmitter inducing part of the plant defense responses. © The Phytopathological Society of Japan and Springer-Verlag 2006.

    DOI: 10.1007/s10327-005-0261-9

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  • Induction of defense responses in pea tissues by inorganic phosphate

    Tomoharu Kawahara, Hiroko Namba, Kazuhiro Toyoda, Tomonari Kasai, Megumi Sugimoto, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 3 )   129 - 136   2006年6月

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    記述言語:英語  

    When inorganic phosphate, a common and essential element for organisms, was applied endogenously, a rejection reaction and superoxide generation were induced in pea tissues but phytoalexin production was not. Phosphate-induced superoxide generation was sensitive to cycloheximide (CHX) and salicylhydroxamic acid (SHAM), indicating that part of the generation was dependent upon the expression of peroxidase gene(s). Peroxidases (POXs) are well known not only to scavenge hydrogen peroxide with phenolics but also to generate superoxide via NADH oxidation in the presence of p-coumaric acid and manganese ion. We cloned five pea POX cDNAs that are predicted to be located outside of the cells. The accumulation of five POX mRNAs, NTPase mRNA, and phenylalanine ammonia-lyase mRNA was measured by semiquantitative reverse transcription-polymerase chain reaction. The expression of the five POX genes was induced by a fungal elicitor. On the other hand, inorganic phosphate induced the accumulation of POX11, POX14, and POX21 mRNAs but not of POX13, POX29, and PsPAL1 mRNAs within 1-3∈h after treatment of pea seedlings. In view of these findings, we discuss inorganic phosphate as a signal transmitter inducing part of the plant defense responses. © The Phytopathological Society of Japan and Springer-Verlag 2006.

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  • Localization and responsiveness of a cowpea apyrase VsNTPase1 to phytopathogenic microorganisms

    Hirotaka Takahashi, Kazuhiro Toyoda, Yuzo Hirakawa, Kunihiko Morishita, Toshiaki Kato, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   72 ( 3 )   143 - 151   2006年6月

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    記述言語:英語  

    Apyrases (NTPases) are associated with both compatible and incompatible interactions between plants and microorganisms. Previously we reported that the ATPase activities of cell-wall-bound apyrases of several leguminous plants, such as pea, cowpea, soybean, and kidney bean, were enhanced by a glycoprotein elicitor and were inhibited in a species-specific manner by mucin-type glycopeptide suppressors secreted from a pea pathogenic fungus, Mycosphaerella pinodes. In this study, we isolated two apyrase genes, VsNTPase1 and VsNTPase2, from a cDNA library of Vigna sinensis Endl. cv. Sanjakusasage. Based on phylogenetic analysis, VsNTPase1 may belong to a group that responds to environmental stimuli. In a transient assay using DNA bombardment, a fusion protein of green fluorescent protein (GFP) and the N-terminal putative signal sequence of VsNTPase1 was distributed in the nucleus, cytoplasm (cytoskeletal structure), and cell wall. On the other hand, a fusion protein of GFP and the N-terminal putative VsNTPase2-signal sequence was localized in the cytoplasm, especially in small particles (perhaps mitochondria). A recombinant VsNTPase1 expressed in Spodoptera frugiperda 21 cells responded directly to signal molecules from several phytopathogenic microorganisms. Here, we discuss the role of apyrases in recognizing and responding to exogenous signals. © The Phytopathological Society of Japan and Springer-Verlag 2006.

    DOI: 10.1007/s10327-005-0267-3

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  • Functional analysis of Gac two-component system in P syringae pv. tabaci

    M. Marutani, Y. Ogawa, F. Taguchi, Y. Inagaki, K. Toyoda, T. Shiraishi, Y. Ichinose

    PHYTOPATHOLOGY   96 ( 6 )   S74 - S74   2006年6月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER PHYTOPATHOLOGICAL SOC  

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  • 植物の非宿主抵抗性

    一瀬勇規

    化学と生物   44 ( 8 )   556 - 562   2006年

  • AtOPR3高発現シロイヌナズナを用いた植物病原菌相互作用におけるOPR subgroup 2の機能解析

    松井英譲, 豊田和弘, 稲垣善茂, 白石友紀, 一瀬勇規

    日本植物病理学会報   72 ( 4 )   2006年

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  • (18) 病原性相互作用で発現が変動する植物遺伝子の機能解析2 : 小胞体-ゴルジ間の小胞輸送に関わるRer1遺伝子ホモログの病原菌応答(平成17年度日本植物病理学会大会講演要旨)

    吉廣 美由貴, 廣瀬 昌也, 藤田 景子, 稲垣 善茂, 一瀬 勇規, 豊田 和弘, 白石 友紀

    日本植物病理學會報   71 ( 3 )   190 - 190   2005年8月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • Flagellin from Pseudomonas syringae pv. tabaci induced hrp-independent HR in tomato

    Mizuri Marutani, Fumiko Taguchi, Rena Shimizu, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   71 ( 4 )   289 - 295   2005年8月

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    記述言語:英語  

    We have previously shown that flagellin of Pseudomonas syringae pv. tabaci is an elicitor that induces a hypersensitive reaction (HR) in nonhost tomato cells. Flagellin is the major HR elicitor produced by this pathogen, as shown by the inability of a flagellin-defective mutant, ΔfliC, to induce HR. Also, a ΔfliD mutant that secretes large amounts of monomer flagellins induces a strong HR in tomato. In this study, the possible involvement of an Hrp type III secretion system (TTSS) in flagellin-induced HR was investigated using flagella-defective mutants or Hrp TTSS-defective mutants. The hrcC gene encodes HrcC protein, which is required for Hrp pilus formation in the outer membrane. An hrcC mutation, introduced into the wild-type, ΔfliC, and ΔfliD mutants of P. syringae pv. tabaci did not affect swimming motility or flagellin secretion, whereas all ΔhrcC, ΔfliC, and ΔfliD mutants lost the ability to cause disease on host tobacco leaves. However, the ΔhrcC mutant and the ΔfliD/ΔhrcC double mutant were still able to induce HR cell death, expression of one of the defense-related genes hsr203J, and the generation of hydrogen peroxide in nonhost tomato cells. Thus, flagellin is required for both pathogenicity in host tobacco and HR in nonhost tomato. On the other hand, hrp TTSS is necessary for pathogenicity on host tobacco but is not indispensable to induce HR in nonhost tomato. These results clearly show that flagellin-induced HR is hrp-independent in tomato. © The Phytopathological Society of Japan and Springer-Verlag 2005.

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  • Defense responses of Arabidopsis thaliana inoculated with Pseudomonas syringae pv. tabaci wild type and defective mutants for flagellin (ΔfliC) and flagellin-glycosylation (Δorf1)

    Yasuhiro Ishiga, Kasumi Takeuchi, Fumiko Taguchi, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   71 ( 4 )   302 - 307   2005年8月

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    記述言語:英語  

    Flagellin, an essential component of the bacterial flagellar filament, is capable of inducing a hypersensitive response (HR), including cell death, in a nonhost plant. A flagellin-defective mutant (ΔfliC) of Pseudomonas syringae pv. tabaci lacks both the flagellar filament and motility, whereas a flagellin-glycosylation-defective mutant (Δorf1) retains the flagellar filament but lacks the glycosyl modification of flagellin protein. To investigate the role of flagellin protein and its glycosylation in the interaction with its nonhost Arabidopsis thaliana, we analyzed plant responses after inoculation with these bacteria. Inoculation with wild-type P. syringae pv. tabaci induced HR, with the generation of reactive oxygen species and cell death. In contrast, inoculation with either ΔfliC or Δorf1 mutant induced a low level of HR, and inoculated leaves developed a disease-like yellowing. These mutant bacteria multiplied better than the wild-type bacteria in A. thaliana. These results indicate that A. thaliana expresses a defense reaction in response to the bacterial flagellin with its glycosyl structure. © The Phytopathological Society of Japan and Springer-Verlag 2005.

    DOI: 10.1007/s10327-005-0201-8

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  • Defense responses of Arabidopsis thaliana inoculated with Pseudomonas syringae pv. tabaci wild type and defective mutants for flagellin (∆fliC) and flagellin-glycosylation (∆orf1).

    J. Gen. Plant Pathol.   71 ( 4 )   302 - 307   2005年8月

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  • Regulation of elicitin-induced ethylene production in suspension-cultured tobacco BY-2 cells

    Dirk Schenke, Kana Naito, Kazuhiro Toyoda, Yoshishige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   71 ( 4 )   273 - 279   2005年8月

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    記述言語:英語  

    INF1 elicitin, a proteinaceous elicitor produced by Phytophthora infestans, induces a hypersensitive response in tobacco BY-2 cells. In response to elicitin, tobacco cells produce both reactive oxygen species (ROS) and ethylene (ET). To investigate the regulation of elicitin-induced ET production, we pharmacologically analyzed the effects of several chemicals on ET production. Inhibitors of ROS generation or ROS chelators efficiently inhibited ET production, whereas simultaneous treatment of a superoxide anion-generating system with salicylhydroxamic acid recovered ET production. In an in vitro experiment, superoxide anion was necessary and sufficient for conversion of 1-aminocyclopropane-1-carboxylate (ACC) to ET because ET was produced from ACC solely in the presence of the superoxide-generating chemical KO2. ET production was also inhibited by lipoxygenase (LOX) inhibitors, indicating a possible involvement of LOX-mediated generation of superoxide anion and ET production itself. Furthermore, elicitin-induced ET production was completely inhibited by the protein synthesis inhibitor cycloheximide but recovered after exogenous application of ACC, indicating that de novo protein synthesis is required for ACC accumulation, leading to ET production. We also investigated the effects of several phytohormones on elicitor-induced ET production and discuss their role in the defense response. © The Phytopathological Society of Japan and Springer-Verlag 2005.

    DOI: 10.1007/s10327-005-0197-0

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  • Regulation of elicitin-induced ethylene production in suspension-cultured tobacco BY-2 cells

    Dirk Schenke, Kana Naito, Kazuhiro Toyoda, Yoshishige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   71 ( 4 )   273 - 279   2005年8月

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    記述言語:英語  

    INF1 elicitin, a proteinaceous elicitor produced by Phytophthora infestans, induces a hypersensitive response in tobacco BY-2 cells. In response to elicitin, tobacco cells produce both reactive oxygen species (ROS) and ethylene (ET). To investigate the regulation of elicitin-induced ET production, we pharmacologically analyzed the effects of several chemicals on ET production. Inhibitors of ROS generation or ROS chelators efficiently inhibited ET production, whereas simultaneous treatment of a superoxide anion-generating system with salicylhydroxamic acid recovered ET production. In an in vitro experiment, superoxide anion was necessary and sufficient for conversion of 1-aminocyclopropane-1-carboxylate (ACC) to ET because ET was produced from ACC solely in the presence of the superoxide-generating chemical KO2. ET production was also inhibited by lipoxygenase (LOX) inhibitors, indicating a possible involvement of LOX-mediated generation of superoxide anion and ET production itself. Furthermore, elicitin-induced ET production was completely inhibited by the protein synthesis inhibitor cycloheximide but recovered after exogenous application of ACC, indicating that de novo protein synthesis is required for ACC accumulation, leading to ET production. We also investigated the effects of several phytohormones on elicitor-induced ET production and discuss their role in the defense response. © The Phytopathological Society of Japan and Springer-Verlag 2005.

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  • Flagellin from Pseudomonas syringae pv. tabaci induced hrp-independent HR in tomato

    Mizuri Marutani, Fumiko Taguchi, Rena Shimizu, Yoshishige Inagaki, Kazuhiro Toyoda, Tomonori Shiraishi, Yuki Ichinose

    Journal of General Plant Pathology   71 ( 4 )   289 - 295   2005年8月

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    記述言語:英語  

    We have previously shown that flagellin of Pseudomonas syringae pv. tabaci is an elicitor that induces a hypersensitive reaction (HR) in nonhost tomato cells. Flagellin is the major HR elicitor produced by this pathogen, as shown by the inability of a flagellin-defective mutant, ΔfliC, to induce HR. Also, a ΔfliD mutant that secretes large amounts of monomer flagellins induces a strong HR in tomato. In this study, the possible involvement of an Hrp type III secretion system (TTSS) in flagellin-induced HR was investigated using flagella-defective mutants or Hrp TTSS-defective mutants. The hrcC gene encodes HrcC protein, which is required for Hrp pilus formation in the outer membrane. An hrcC mutation, introduced into the wild-type, ΔfliC, and ΔfliD mutants of P. syringae pv. tabaci did not affect swimming motility or flagellin secretion, whereas all ΔhrcC, ΔfliC, and ΔfliD mutants lost the ability to cause disease on host tobacco leaves. However, the ΔhrcC mutant and the ΔfliD/ΔhrcC double mutant were still able to induce HR cell death, expression of one of the defense-related genes hsr203J, and the generation of hydrogen peroxide in nonhost tomato cells. Thus, flagellin is required for both pathogenicity in host tobacco and HR in nonhost tomato. On the other hand, hrp TTSS is necessary for pathogenicity on host tobacco but is not indispensable to induce HR in nonhost tomato. These results clearly show that flagellin-induced HR is hrp-independent in tomato. © The Phytopathological Society of Japan and Springer-Verlag 2005.

    DOI: 10.1007/s10327-005-0200-9

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  • Identification of genes expressed during spore germination of Mycosphaerella pinodes

    Hiroyuki Takahara, Kazuhiro Toyoda, Gento Tsuji, Yasuyuki Kubo, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   71 ( 3 )   190 - 195   2005年6月

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    記述言語:英語  

    Mycosphaerella blight, caused by Mycosphaerella pinodes, is one of the major diseases of cultivated pea (Pisum sativum L.). To isolate the genes that are up- and down-regulated during spore germination, suppression subtraction hybridization (SSH) was performed between ungerminated and germinated spores. The 232 and 128 clones from forward and reverse libraries, respectively, were collected, sequenced, and analyzed with a BLASTX homology search. About 95% of the 32 selected clones were expressed during spore germination on a paper sheet and during infection of pea leaves. We discuss the applicability of the SSH libraries for analyzing M. pinodes genes involved in the early stage of infection. © The Phytopathological Society of Japan and Springer-Verlag 2005.

    DOI: 10.1007/s10327-005-0185-4

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  • 植物病原細菌Pseudomonas syringaeのべん毛タンパク質フラジェリンの糖鎖修飾と植物相互作用

    竹内香純, 田口富美子, 三木隆二, 安田千裕, 村田勝義, 加藤悦子, 加藤静恵, 賀来華江, 稲垣善茂, 豊田和弘, 白石友紀, 一瀬勇規

    植物微生物研究会研究交流会講演要旨集   14th   (JA)120,(EN)121   2005年3月

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    記述言語:日本語  

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  • Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens

    Hideki Takahashi, Ayano Shimizu, Tsutomu Arie, Syofi Rosmalawati, Sumire Fukushima, Mari Kikuchi, Yasufumi Hikichi, Ayami Kanda, Akiko Takahashi, Akinori Kiba, Kohei Ohnishi, Yuki Ichinose, Fumiko Taguchi, Chihiro Yasuda, Motoichiro Kodama, Mayumi Egusa, Chikara Masuta, Hiroyuki Sawada, Daisuke Shibata, Koichi Hori, Yuichiro Watanabe

    Journal of General Plant Pathology   71 ( 1 )   8 - 22   2005年2月

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    記述言語:英語  

    Lycopersicon esculentum cultivar Micro-Tom is a miniature tomato with many advantages for studies of the molecular biology and physiology of plants. To evaluate the suitability of Micro-Tom as a host plant for the study of pathogenesis, Micro-Tom plants were inoculated with 16 well-known fungal, bacterial, and viral pathogens of tomato. Athelia rolfsii, Botryotinia fuckeliana, Oidium sp., Phytophthora infestans, and Sclerotinia sclerotiorum caused typical symptoms and sporulated abundantly on Micro-Tom. Micro-Tom was resistant to Alternaria alternata, Corynespora cassiicola, and Fusarium oxysporum. When Micro-Tom was inoculated with 17 isolates of Ralstonia solanacearum, many isolates induced wilt symptoms. Agrobacterium tumefaciens also was pathogenic, causing crown galls on stem tissue after needle prick inoculation. In Micro-Tom sprayed with Pseudomonas syringae pv. tomato, P. s. pv. tabaci, or P. s. pv. glycinea, bacterial populations did not increase, and yellow lesions appeared only on leaves sprayed with P. s. pv. tomato. Tomato mosaic virus, Tomato aspermy virus, and Cucumber mosaic virus systemically infected Micro-Tom, which developed symptoms characteristic of other cultivars of tomato after infection with the respective virus. These results indicated that Micro-Tom was generally susceptible to most of the important tomato pathogens and developed typical symptoms, whereas certain pathogens were restricted by either hypersensitive resistance or nonhost resistance on Micro-Tom. Therefore, an assortment of Micro-Tom-pathogen systems should provide excellent models for studying the mechanism of susceptible and resistant interactions between plants and pathogens. © The Phytopathological Society of Japan and Springer-Verlag 2005.

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  • Catalog of Micro-Tom tomato responses to common fungal, bacterial, and viral pathogens

    Hideki Takahashi, Ayano Shimizu, Tsutomu Arie, Syofi Rosmalawati, Sumire Fukushima, Mari Kikuchi, Yasufumi Hikichi, Ayami Kanda, Akiko Takahashi, Akinori Kiba, Kohei Ohnishi, Yuki Ichinose, Fumiko Taguchi, Chihiro Yasuda, Motoichiro Kodama, Mayumi Egusa, Chikara Masuta, Hiroyuki Sawada, Daisuke Shibata, Koichi Hori, Yuichiro Watanabe

    Journal of General Plant Pathology   71 ( 1 )   8 - 22   2005年2月

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    記述言語:英語  

    Lycopersicon esculentum cultivar Micro-Tom is a miniature tomato with many advantages for studies of the molecular biology and physiology of plants. To evaluate the suitability of Micro-Tom as a host plant for the study of pathogenesis, Micro-Tom plants were inoculated with 16 well-known fungal, bacterial, and viral pathogens of tomato. Athelia rolfsii, Botryotinia fuckeliana, Oidium sp., Phytophthora infestans, and Sclerotinia sclerotiorum caused typical symptoms and sporulated abundantly on Micro-Tom. Micro-Tom was resistant to Alternaria alternata, Corynespora cassiicola, and Fusarium oxysporum. When Micro-Tom was inoculated with 17 isolates of Ralstonia solanacearum, many isolates induced wilt symptoms. Agrobacterium tumefaciens also was pathogenic, causing crown galls on stem tissue after needle prick inoculation. In Micro-Tom sprayed with Pseudomonas syringae pv. tomato, P. s. pv. tabaci, or P. s. pv. glycinea, bacterial populations did not increase, and yellow lesions appeared only on leaves sprayed with P. s. pv. tomato. Tomato mosaic virus, Tomato aspermy virus, and Cucumber mosaic virus systemically infected Micro-Tom, which developed symptoms characteristic of other cultivars of tomato after infection with the respective virus. These results indicated that Micro-Tom was generally susceptible to most of the important tomato pathogens and developed typical symptoms, whereas certain pathogens were restricted by either hypersensitive resistance or nonhost resistance on Micro-Tom. Therefore, an assortment of Micro-Tom-pathogen systems should provide excellent models for studying the mechanism of susceptible and resistant interactions between plants and pathogens. © The Phytopathological Society of Japan and Springer-Verlag 2005.

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  • Identification of the genes expressed during spore germination of Mycosphaerella pinodes.

    J. Gen. Plant Pathol.   71 (3), 190-195.   2005年

  • Mechanism of flagellin-induced hypersensitive cell death in Arabidopsis thaliana

    Y Ishiga, YC Lin, M Marutani, F Taguchi, Y Inagaki, K Toyoda, T Shiraishi, Y Ichinose

    PLANT AND CELL PHYSIOLOGY   46   S94 - S94   2005年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • Pea apoplastic Cu/Zn-SOD responsive to pathogenic signal molecules

    T Kasai, K Ono, T Suzuki, K Toyoda, Y Inagaki, Y Ichinose, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   46   S94 - S94   2005年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • Pseudomonas syringaeのべん毛を介した植物相互作用

    一瀬勇規, 田口富美子, 竹内香純, 石賀康博, 村田勝義, 加藤悦子, 丸谷瑞理, 三木隆二, 安田千裕

    日本植物病理学会植物感染生理談話会論文集   ( 40 )   13 - 24   2004年8月

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    記述言語:日本語  

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  • Agrobacterium tumefaciens-mediated transformation as a tool for random mutagenesis of Colletotrichum trifolii

    Hiroyuki Takahara, Gento Tsuji, Yasuyuki Kubo, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   70 ( 2 )   93 - 96   2004年4月

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    記述言語:英語  

    We transformed Colletotrichum trifolii, the causal agent of alfalfa anthracnose, using Agrobacterium tumefaciens as a new tool for random insertional mutagenesis. Fungal spores of C. trifolii were transformed with T-DNA including the hygromycin phosphotransferase gene (hph). Southern analysis showed that every randomly selected transformant had a unique hybridization pattern of T-DNA, suggesting that the T-DNA was randomly integrated into the fungal genome. More significantly, about 75% of transformants had a single copy of the T-DNA. The results demonstrate that insertional mutagenesis via A. tumefaciens is a useful tool for studying the function of C. trifolii genes.

    DOI: 10.1007/s10327-003-0099-y

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  • Agrobacterium tumefaciens-mediated transformation as a tool for random mutagenesis of Colletotrichum trifolii

    Hiroyuki Takahara, Gento Tsuji, Yasuyuki Kubo, Mikihiro Yamamoto, Kazuhiro Toyoda, Yoshishige Inagaki, Yuki Ichinose, Tomonori Shiraishi

    Journal of General Plant Pathology   70 ( 2 )   93 - 96   2004年4月

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    記述言語:英語  

    We transformed Colletotrichum trifolii, the causal agent of alfalfa anthracnose, using Agrobacterium tumefaciens as a new tool for random insertional mutagenesis. Fungal spores of C. trifolii were transformed with T-DNA including the hygromycin phosphotransferase gene (hph). Southern analysis showed that every randomly selected transformant had a unique hybridization pattern of T-DNA, suggesting that the T-DNA was randomly integrated into the fungal genome. More significantly, about 75% of transformants had a single copy of the T-DNA. The results demonstrate that insertional mutagenesis via A. tumefaciens is a useful tool for studying the function of C. trifolii genes.

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  • Structure and expression of 12-oxophytodienoate reductase (subgroup I) genes in pea, and characterization of the oxidoreductase activities of their recombinant products

    H Matsui, G Nakamura, Y Ishiga, H Toshima, Y Inagaki, K Toyoda, T Shiraishi, Y Ichinose

    MOLECULAR GENETICS AND GENOMICS   271 ( 1 )   1 - 10   2004年2月

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    担当区分:最終著者, 責任著者   記述言語:英語   出版者・発行元:SPRINGER-VERLAG  

    Recently, we observed that expression of a pea gene (S64) encoding an oxophytodienoic acid reductase (OPR) was induced by a suppressor of pea defense responses, secreted by the pea pathogen Mycosphaerella pinodes. Because it is known that OPRs are usually encoded by families of homologous genes, we screened for genomic and cDNA clones encoding members of this putative OPR family in pea. We isolated five members of the OPR gene family from a pea genomic DNA library, and amplified six cDNA clones, including S64, by RT-PCR (reverse transcriptase-PCR). Sequencing analysis revealed that S64 corresponds to PsOPR2, and the amino acid sequences of the predicted products of the six OPR-like genes shared more than 80% identity with each other. Based on their sequence similarity, all these OPR-like genes code for OPRs of subgroup I, i.e., enzymes which are not required for jasmonic acid biosynthesis. However, the genes varied in their exon/intron organization and in their promoter sequences. To investigate the expression of each individual OPR-like gene, RT-PCR was performed using gene-specific primers. The results indicated that the OPR-like gene most strongly induced by the inoculation of pea plants with a compatible pathogen and by treatment with the suppressor from M. pinodes was PsOPR2. Furthermore, the ability of the six recombinant OPR-like proteins to reduce a model substrate, 2-cyclohexen-1-one (2-CyHE), was investigated. The results indicated that PsOPR1, 4 and 6 display robust activity, and PsOPR2 has a most remarkable ability to reduce 2-CyHE, whereas PsOPR3 has little and PsOPR5 does not reduce this compound. Thus, the six OPR-like proteins can be classified into four types. Interestingly, the gene structures, expression profiles, and enzymatic activities used to classify each member of the pea OPR-like gene family are clearly correlated, indicating that each member of this OPR-like family has a distinct function.

    DOI: 10.1007/s00438-003-0948-6

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  • Differential regulation of MBP kinases by a glycoproptein elicitor and a polypeptide suppressor from Mycosphaerella pinodes in pea

    Uppalapati, SR, K Toyoda, Yasuhiro, I, Y Ichinose, T Shiraishi

    PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY   64 ( 1 )   17 - 25   2004年1月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    A polypeptide fungal suppressor from a pea pathogen Mycosphaerella pinodes plays a key role in pathogenesis by suppressing elicitor-induced defense response(s) in pea (Pisum sativum L). In this study, we show that treatment of pea tissues with the polysaccharide elicitor secreted by M. pinodes results in rapid increased activation of two myelin basic protein (MBP)-dependent kinases p44 (approximate to44 kDa) and p48 (approximate to48 kDa) within 15-30 min upon elicitation. Interestingly, the suppressor inhibited the elicitor-induced activation of only p44 kinase. While the defense-inducing signalling molecules, chitosan and salicylic acid (SA) activated the p44 and p48 kinases, methyl jasmonate (MeJA) did not. The abiotic stress signals, abscisic acid (ABA), NaCl and wounding activated the p48 kinase alone. These results demonstrate that MAPKs are differentially activated in response to pathogen invasion and abiotic stress in pea. Furthermore, specific inhibition of elicitor-induced p44 kinase activation by a MAPKK inhibitor, PD098059 and protein kinase inhibitor, K252a correlated with the suppression of elicitor-induced phenylalanine ammonia lyase (PAL) gene expression, supporting a role for p44 in the elicitor-induced defense response(s) in pea. Inhibition of p44 by the phosphoinositide (PI) turnover inhibitor, neomycin (a fungal suppressor mimic), and potentiation of p44 by the diacylglycerol (DAG) kinase inhibitor, R59022 indicated that p44 may be acting downstream of (PI) metabolism. Taken together, our results indicate that suppressor of defense elicitation from M. pinodes acts through inhibition of a MAPK (p44), possibly through a PI signaling pathway, facilitating the establishment of basic compatibility during infection of pea. (C) 2004 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.pmpp.2004.05.003

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  • Differential regulation of MBP kinases by a glycoproptein elicitor and a polypeptide suppressor from Mycosphaerella pinodes in pea

    Uppalapati, SR, K Toyoda, Yasuhiro, I, Y Ichinose, T Shiraishi

    PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY   64 ( 1 )   17 - 25   2004年1月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD  

    A polypeptide fungal suppressor from a pea pathogen Mycosphaerella pinodes plays a key role in pathogenesis by suppressing elicitor-induced defense response(s) in pea (Pisum sativum L). In this study, we show that treatment of pea tissues with the polysaccharide elicitor secreted by M. pinodes results in rapid increased activation of two myelin basic protein (MBP)-dependent kinases p44 (approximate to44 kDa) and p48 (approximate to48 kDa) within 15-30 min upon elicitation. Interestingly, the suppressor inhibited the elicitor-induced activation of only p44 kinase. While the defense-inducing signalling molecules, chitosan and salicylic acid (SA) activated the p44 and p48 kinases, methyl jasmonate (MeJA) did not. The abiotic stress signals, abscisic acid (ABA), NaCl and wounding activated the p48 kinase alone. These results demonstrate that MAPKs are differentially activated in response to pathogen invasion and abiotic stress in pea. Furthermore, specific inhibition of elicitor-induced p44 kinase activation by a MAPKK inhibitor, PD098059 and protein kinase inhibitor, K252a correlated with the suppression of elicitor-induced phenylalanine ammonia lyase (PAL) gene expression, supporting a role for p44 in the elicitor-induced defense response(s) in pea. Inhibition of p44 by the phosphoinositide (PI) turnover inhibitor, neomycin (a fungal suppressor mimic), and potentiation of p44 by the diacylglycerol (DAG) kinase inhibitor, R59022 indicated that p44 may be acting downstream of (PI) metabolism. Taken together, our results indicate that suppressor of defense elicitation from M. pinodes acts through inhibition of a MAPK (p44), possibly through a PI signaling pathway, facilitating the establishment of basic compatibility during infection of pea. (C) 2004 Elsevier Ltd. All rights reserved.

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  • 耐病性のシグナリングモデル,新

    分子レベルからみた植物の耐病性 (細胞工学別冊),(島本 功ら編) 秀潤社 (東京)   pp.14-17.   2004年

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  • 植物の病原菌に体する受容性と抵抗性

    新版 分子レベルからみた植物の耐病性 (細胞工学別冊),(島本 功ら編) 秀潤社   pp.74-81.   2004年

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  • PDR-type ABC transporter and tobacco defense response against pathogen

    Y Ichinose, D Schenke, M Sasabe, Y Inagaki, K Toyoda, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   45   S1 - S1   2004年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • Genomic structure of the NtPDR1 gene, harboring the two miniature inverted-repeat transposable elements, NtToya1 and NtStowaway101.

    Genes Genet. Syst.   78 ( 6 )   409 - 418   2003年12月

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    担当区分:最終著者, 責任著者   記述言語:英語  

    DOI: 10.1266/ggs.78.409

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  • Differential effects of flagellins from Pseudomonas syringae pv. tabaci, tomato and glycinea on plant defense response

    F Taguchi, R Shimizu, R Nakajima, K Toyoda, T Shiraishi, Y Ichinose

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   41 ( 2 )   165 - 174   2003年2月

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    記述言語:英語   出版者・発行元:EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER  

    To investigate the factor that determines incompatible interactions between Pseudomonas syringae pv. tabaci and non-host plants, an elicitor of hypersensitive reaction (HR) was partially purified from the supernatant of a nutrient-poor medium of bacterial culture by DEAE column chromatography. The major protein in the elicitor-active fractions was identified as a flagellin which is a component of flagellar filaments. The flagellins purified from P. syringae pv. tomato and glycinea, incompatible pathogens of tobacco plants, induced fragmentation of chromosomal DNA and oxidative burst accompanied by programmed cell death in tobacco (Nicotiana tabacum) Bright Yellow (BY-2) cells, but the flagellin from pv. tabaci, a compatible pathogen, did not. However, the amino acid sequences of flagellins deduced from fliC genes showed a high homology among these P. syringae pathovars. In particular, the amino acid sequences of pv. tabaci and glycinea are completely identical. However, both recombinant flagellins produced in Escherichia coli possess HR-inducing activity in BY-2 cells. These results indicate that the post-translational modification of flagellins has an important role for HR-inducing ability in tobacco cells. Furthermore, we discuss the cause of a different elicitor activity among flagellins on tobacco cells and the role of flagellins in the determining specificity. (C) 2002 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

    DOI: 10.1016/S0981-9428(02)00018-9

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  • Need for flagella for complete virulence of Pseudomonas syringae pv. tabaci: genetic analysis with flagella-defective mutants, DfliC and DfliD, in host tobacco plants.

    J. Gen. Plant Pathol.   69 (4) 244-249.   2003年

  • Need for flagella for complete virulence of Pseudomonas syringae pv. tabaci: genetic analysis with flagella-defective mutants, DfliC and DfliD, in host tobacco plants.

    J. Gen. Plant Pathol.   69 (4) 244-249.   2003年

  • Role of flagella and flagellin in plant-Pseudomonas syrignae interactions. In Japan/ Taiwan Symposium on Molecular Biology of Functional Regulation in Plant and Microbe (N. Sako, H. Yaegashi and M.-K. Yang eds.)

    Showado Co. (Saga, Japan)   pp. 158-166.   2003年

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  • Phylogenetic Classification of Dof type Transcription Factors in Pea (Pisum sativum)

    Naoki Nakamura, Mizuri Marutani, Shiroh Sanematsu, Kazuhiro Toyoda, Yoshi-shige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    Plant Biotechnology   20 ( 3 )   247 - 252   2003年

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    記述言語:英語  

    The Dof protein family is a group of plant transcription factors carrying highly conserved 52 residues referred to as the Dof domain, and belongs to the C4 zinc-finger transcription factors. We isolated various PsDof genes by PCR using the Dof-domain nucleotide sequence of the PsDofl gene from a cDNA library of elicitor-treated pea epicotyls, and these isolated PsDof genes were then classified phylogenetically. Since the obtained genes (PsDofl to PsDofT) were scattered over various positions of the phylogenetic tree, they were expected to perform various functions as Dof-type transcription factors. From their positions in the tree, it is expected that the PsDofZ and PsDof5 genes are defense related, as is the PsDofl gene. © 2003, Japanese Society for Plant Cell and Molecular Biology. All rights reserved.

    DOI: 10.5511/plantbiotechnology.20.247

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  • Phylogenetic Classification of Dof type Transcription Factors in Pea (Pisum sativum)

    Naoki Nakamura, Mizuri Marutani, Shiroh Sanematsu, Kazuhiro Toyoda, Yoshi-shige Inagaki, Tomonori Shiraishi, Yuki Ichinose

    Plant Biotechnology   20 ( 3 )   247 - 252   2003年

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    記述言語:英語  

    The Dof protein family is a group of plant transcription factors carrying highly conserved 52 residues referred to as the Dof domain, and belongs to the C4 zinc-finger transcription factors. We isolated various PsDof genes by PCR using the Dof-domain nucleotide sequence of the PsDofl gene from a cDNA library of elicitor-treated pea epicotyls, and these isolated PsDof genes were then classified phylogenetically. Since the obtained genes (PsDofl to PsDofT) were scattered over various positions of the phylogenetic tree, they were expected to perform various functions as Dof-type transcription factors. From their positions in the tree, it is expected that the PsDofZ and PsDof5 genes are defense related, as is the PsDofl gene. © 2003, Japanese Society for Plant Cell and Molecular Biology. All rights reserved.

    DOI: 10.5511/plantbiotechnology.20.247

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  • Expression of allene oxide synthase and allene oxide cyclase in the interactions between pea and fungal pathogens.

    J. Gen. Plant Pathol.   69 (6) In press   2003年

  • Expression of allene oxide synthase and allene oxide cyclase in the interactions between pea and fungal pathogens.

    J. Gen. Plant Pathol.   69 (6) In press   2003年

  • Role of Flagella and Flagellin in Plant - Pseudomonas syringae Interactions

    Pseudomonas syringae and Related Pathogens (N. S. Iacobellis ed.) Kluwer AcademicPress (Dordrecht, The Netherland)   pp. 311-318.   311 - 318   2003年

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    担当区分:筆頭著者, 責任著者   記述言語:英語  

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  • Post-translational modification of flagellin determines the specificity of HR induction.

    Plant Cell Physiol.   44 (3) 342-349.   2003年

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  • 植物病原細菌のべん毛を介した植物相互作用.

    化学と生物   41 (8) 511-516   2003年

  • Possible Involvement of AAAG Motif and PsDof1 in Elicitor-Induced Gene Expression in Pea.

    The Scientific Reports of The Faculty of Agriculture, Okayama University.   92, in press   2003年

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  • Cloning and characterization of pea apyrases: involvement of PsAPY1 in response to signal molecules from the pea pathogen Mycosphaerella pinodes.

    J. Gen. Plant Pathol.   69: 33-38.   2003年

  • Cloning and characterization of pea apyrases: involvement of PsAPY1 in response to signal molecules from the pea pathogen Mycosphaerella pinodes.

    J. Gen. Plant Pathol.   69: 33-38.   2003年

  • The DfliD mutant of Pseudomonas syringae pv. tabaci, which secretes flagellin monomers, induces a strong hypersensitive reaction (HR) in non-host tomato cells.

    Mol. Genet. Genomics   269: 21-30.   2003年

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  • Role of flagella and flagellin in plant-Pseudomonas syrignae interactions. In Japan/ Taiwan Symposium on Molecular Biology of Functional Regulation in Plant and Microbe (N. Sako, H. Yaegashi and M.-K. Yang eds.)

    Showado Co. (Saga, Japan)   pp. 158-166.   2003年

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  • Possible Involvement of AAAG Motif and PsDof1 in Elicitor-Induced Gene Expression in Pea.

    The Scientific Reports of The Faculty of Agriculture, Okayama University.   92, in press   2003年

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  • (321)Pseudomonas syringaeの生産する過敏感反応誘導因子(5)P.syringaeの生産するflagellinのタバコに対する過敏感反応誘導特異性(平成14年度 日本植物病理学会大会講演要旨)

    田口 富美子, 清水 鈴菜, 竹内 香純, 池田 陽子, 稲垣 善茂, 白石 友紀, 一瀬 勇規

    日本植物病理學會報   68 ( 2 )   242 - 243   2002年8月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • (322)Pseudomonas syringaeの生産する過敏感反応誘導因子(6)P.syringaeのflagellinの糖鎖修飾に関わる推定遺伝子領域の解析(平成14年度 日本植物病理学会大会講演要旨)

    竹内 香純, 川崎 敬之, 田口 富美子, 清水 鈴菜, 池田 陽子, 稲垣 善茂, 白石 友紀, 一瀬 勇規

    日本植物病理學會報   68 ( 2 )   2002年8月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • Pseudomonas syringaeの生産する過敏感反応誘導因子(4)べん毛変異体を用いたflagellinの機能解析(関西部会講演要旨)

    清水 鈴菜, 田口 富美子, 池田 陽子, 豊田 和弘, 白石 友紀, 稲垣 善茂, 一瀬 勇規

    日本植物病理學會報   68 ( 1 )   2002年4月

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    記述言語:日本語   出版者・発行元:日本植物病理学会  

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  • Pseudomonas syringae pv. tabaciの単量体Flagellinは,非宿主植物トマト細胞にHR誘導因子として働く

    清水鈴菜, 田口富美子, 池田陽子, 豊田和弘, 白石友紀, 稲垣善茂, 一瀬勇規

    日本植物生理学会年会要旨集   42nd   238   2002年3月

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  • HR誘導性細菌エリシターflagellinのHR誘導及び病原性における翻訳後修飾の重要性

    田口富美子, 池田陽子, 清水鈴菜, 竹内香純, 稲垣善茂, 白石友紀, 一瀬勇規

    日本植物生理学会年会要旨集   42nd   238   2002年3月

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  • cDNA cloning and characterization of tobacco ABC transporter: NtPDR1 is a novel elicitor-responsive gene.

    FEBS Letters   518, 164-168   2002年

  • Molecular cloning of cDNA for a novel pea Dof protein, PsDof1, and its DNA binding activity to the promoter of PsDof1 gene.

    Plant Biotechnol.   19 ( 4 )   251 - 260   2002年

  • Molecular cloning of cDNA for a novel pea Dof protein, PsDof1, and its DNA binding activity to the promoter of PsDof1 gene.

    SEKI H, NAKAMURA N, MARUTANI M, OKABE T, SANEMATSU S, INAGAKI Y, TOYODA K, SHIRAISHI T, ICHINOSE Y

    Plant Biotechnol.   19 ( 4 )   251 - 260   2002年

  • Flagellin, a constituent of a flagella filament in Pseudomonas syringae pv. tabaci, is an intense HR elicitor as a monomer molecule on nonhost tomato cells

    R Shimizu, F Taguchi, Y Ikeda, K Toyoda, T Shiraishi, Y Inagaki, Y Ichinose

    PLANT AND CELL PHYSIOLOGY   43   S201 - S201   2002年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • Cell wall-plasma membrane continuum is involved in transduction of pathogen signals

    K Toyoda, AM Kiba, T Kawahara, M Sugimoto, RS Uppalapati, Y Inagaki, Y Ichinose, M Yamamoto, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   43   S10 - S10   2002年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • The importance of post-translational modification of bacterial HR-elicitor, flagellin on HR-inducing ability and pathogenicity

    F Taguchi, Y Ikeda, R Shimizu, K Takeuchi, Y Inagaki, T Shiraishi, Y Ichinose

    PLANT AND CELL PHYSIOLOGY   43   S201 - S201   2002年

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:OXFORD UNIV PRESS  

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  • Biochemical and molecular biological studies on infection (Ascochyta rabiei)-induced thaumatin-like proteins from chickpea plants (Cicer arietinum L.)

    T Hanselle, Y Ichinose, W Barz

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   56 ( 11-12 )   1095 - 1107   2001年11月

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    記述言語:英語   出版者・発行元:VERLAG Z NATURFORSCH  

    A pathogenesis-related protein induced by infection with Ascochyta rabiei was purified from intercellular washing fluid of chickpea (Cicer arietinum L.) leaves. Amino-terminal sequencing identified the protein, named PR-5a, as a thaumatin-like protein. The isoelectric point was determined with 6.5 and the molecular mass is 16 kDa. Therefore., chickpea PR-5a is the first dicot member of a TLP subgroup containing small TLPs with a molecular weight between 15 and 18 kDa. PR-5a shows no antifungal activity towards A. rabiei. Screening of a chickpea cDNA library led to the isolation of a cDNA clone (p5a-241) for this protein. A second cDNA clone (ELR112) encoding a TLP was isolated using differential hybridisation of cDNA libraries obtained from elicited and water treated cell suspension cultures of chickpea. The deduced protein (PR-5b) has a molecular mass of 22 kDa. PR-5b is postulated to be located in the vacuole due to the presence of a respective N-terminal signal peptide and a carboxy-terminal extension. Southern blot analyses showed that ELR112 and p5a-241 represent single copy genes. During fungal infection of chickpea plants expression of both genes proceeds much faster in an A. rabiei resistant cultivar than in a susceptible one.

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  • Biochemical and molecular biological studies on infection (Ascochyta rabiei)-induced thaumatin-like proteins from chickpea plants (Cicer arietinum L.)

    T Hanselle, Y Ichinose, W Barz

    ZEITSCHRIFT FUR NATURFORSCHUNG C-A JOURNAL OF BIOSCIENCES   56 ( 11-12 )   1095 - 1107   2001年11月

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    記述言語:英語   出版者・発行元:VERLAG Z NATURFORSCH  

    A pathogenesis-related protein induced by infection with Ascochyta rabiei was purified from intercellular washing fluid of chickpea (Cicer arietinum L.) leaves. Amino-terminal sequencing identified the protein, named PR-5a, as a thaumatin-like protein. The isoelectric point was determined with 6.5 and the molecular mass is 16 kDa. Therefore., chickpea PR-5a is the first dicot member of a TLP subgroup containing small TLPs with a molecular weight between 15 and 18 kDa. PR-5a shows no antifungal activity towards A. rabiei. Screening of a chickpea cDNA library led to the isolation of a cDNA clone (p5a-241) for this protein. A second cDNA clone (ELR112) encoding a TLP was isolated using differential hybridisation of cDNA libraries obtained from elicited and water treated cell suspension cultures of chickpea. The deduced protein (PR-5b) has a molecular mass of 22 kDa. PR-5b is postulated to be located in the vacuole due to the presence of a respective N-terminal signal peptide and a carboxy-terminal extension. Southern blot analyses showed that ELR112 and p5a-241 represent single copy genes. During fungal infection of chickpea plants expression of both genes proceeds much faster in an A. rabiei resistant cultivar than in a susceptible one.

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  • Generation of hydrogen peroxide is not required for harpin-induced apoptotic cell death in tobacco BY-2 cell suspension culture

    Y Ichinose, S Andi, R Doi, R Tanaka, F Taguchi, M Sasabe, K Toyoda, T Shiraishi, T Yamada

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   39 ( 9 )   771 - 776   2001年9月

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    担当区分:筆頭著者   記述言語:英語   出版者・発行元:GAUTHIER-VILLARS/EDITIONS ELSEVIER  

    To characterize molecular and biochemical mechanisms of hypersensitive reaction (HR) in plants, a tobacco suspension culture of BY-2 was treated with the proteinaceous HR elicitor harpin from several pathovars of Pseudomonas syringae. Tobacco BY-2 cells are sensitive to harpins derived from non-pathogenic pathovars of P. syringae, such as pvs. pisi, tomato and glycinea. These three harpins induce apoptotic cell death accompanied by DNA fragmentation in BY-2. Because the cell death was also accompanied by rapid generation of hydrogen peroxide (H2O2), one of the active oxygen species, we investigated the role of H2O2 in harpin-induced cell death. Although treatment with diphenylene iodium chloride (DPI) reduced, and catalase completely abolished, the harpin-induced generation of H2O2, the frequency of cell death was not affected at all. Treatment with superoxide dismutase (SOD) enhanced the generation of H2O2, but the cell death was unaffected. These results indicate that harpin-induced apoptotic cell death does not require oxidative burst. (C) 2001 Editions scientifiques et medicales Elsevier SAS

    DOI: 10.1016/S0981-9428(01)01300-6

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  • Contrary operations of Box-I element of pea phenylalanine ammonia-lyase gene 1 promoter for organ-specific expression

    Y Imura, H Seki, K Toyoda, Y Ichinose, T Shiraishi, T Yamada

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   39 ( 5 )   355 - 362   2001年5月

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    記述言語:英語   出版者・発行元:GAUTHIER-VILLARS/EDITIONS ELSEVIER  

    Expression of genes (PSPAL) encoding phenylalanine ammonia-lyase from pea (Pisum sativum L.) is regulated in response to various environmental stimuli and during plant development. We examined the cis-regulatory elements in PSPAL1 promoter for organ-specific expression by determining the sequences specifically associated with nuclear proteins on its promoter in each pea organ. In vivo dimethyl sulfate (DMS) footprinting analysis showed putative protein bindings on AC-rich sequences including Box-I in particular in roots and stems in which PSPAL mRNA were highly accumulated. The potential role of the AC-rich element was investigated by fusing PSPAL1 promoter with Box-I deleted to the reporter gene P-glucuronidase (GUS) and transforming the construct into tobacco plants (Nicotiana tabacum). GUS activity controlled under this Box-I deletion promoter was significantly reduced in roots and leaves, whereas drastically increased in stems as compared with the activity of wild-type PSPALI promoter. Histochemical GUS staining indicated the activity was elevated in vascular tissues of stem by deleting Box-I. These results suggest that Box-I in PSPALI contributes to a positive regulation in root and leaf, but might also function as a negative regulator in stem, especially in xylem tissue. (C) 2001 Editions scientifiques et medicales Elsevier SAS.

    DOI: 10.1016/S0981-9428(01)01248-7

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  • Contrary operations of Box-I element of pea phenylalanine ammonia-lyase gene 1 promoter for organ-specific expression

    Y Imura, H Seki, K Toyoda, Y Ichinose, T Shiraishi, T Yamada

    PLANT PHYSIOLOGY AND BIOCHEMISTRY   39 ( 5 )   355 - 362   2001年5月

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    記述言語:英語   出版者・発行元:GAUTHIER-VILLARS/EDITIONS ELSEVIER  

    Expression of genes (PSPAL) encoding phenylalanine ammonia-lyase from pea (Pisum sativum L.) is regulated in response to various environmental stimuli and during plant development. We examined the cis-regulatory elements in PSPAL1 promoter for organ-specific expression by determining the sequences specifically associated with nuclear proteins on its promoter in each pea organ. In vivo dimethyl sulfate (DMS) footprinting analysis showed putative protein bindings on AC-rich sequences including Box-I in particular in roots and stems in which PSPAL mRNA were highly accumulated. The potential role of the AC-rich element was investigated by fusing PSPAL1 promoter with Box-I deleted to the reporter gene P-glucuronidase (GUS) and transforming the construct into tobacco plants (Nicotiana tabacum). GUS activity controlled under this Box-I deletion promoter was significantly reduced in roots and leaves, whereas drastically increased in stems as compared with the activity of wild-type PSPALI promoter. Histochemical GUS staining indicated the activity was elevated in vascular tissues of stem by deleting Box-I. These results suggest that Box-I in PSPALI contributes to a positive regulation in root and leaf, but might also function as a negative regulator in stem, especially in xylem tissue. (C) 2001 Editions scientifiques et medicales Elsevier SAS.

    DOI: 10.1016/S0981-9428(01)01248-7

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  • Molecular cloning and functional analysis of pea cDNA E86 encoding homologous protein to hypersensitivity-related hsr203J

    Y Ichinose, Y Hisayasu, S Sanematsu, Y Ishiga, H Seki, K Toyoda, T Shiraishi, T Yamada

    PLANT SCIENCE   160 ( 5 )   997 - 1006   2001年4月

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    記述言語:英語   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    Clone E86 was isolated as cDNA for elicitor-inducible gene From pea epicotyls by differential screening. The deduced amino acid sequence of E86 showed high homology to hypersensitivity-related protein hsr203J in tobacco and also showed significant homologies to the Ser-active hydrolases, such as mammalian hormone-sensitive lipases, bacterial lipases and esterases. E86 polypeptide possesses consensus amino acid sequence motifs (His-Gly) and (Gly-X-Ser-X-Gly) conserved in lipases and esterases and showed esterase degradation of p-nitrophenyl butyrate. Northern blot analysis revealed that the E86-transcript is abundant in roots and stems and was induced by fungal elicitor in pea epicotyls. However, elicitor-induced accumulation of E86 mRNA was significantly inhibited by the fungal suppressor. Furthermore the expression of the genes encoding E86 and phenylalanine ammonia-lyase was induced within 1 h after the inoculation of a nonpathogen. but it was delayed for 5 h by the inoculation of a compatible pathogen. These results suggest that the elicitor-induced Ser-active hydrolase derived from E86 gene might be related to the plant defense responses. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

    DOI: 10.1016/S0168-9452(01)00343-0

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  • Molecular cloning and functional analysis of pea cDNA E86 encoding homologous protein to hypersensitivity-related hsr203J

    Y Ichinose, Y Hisayasu, S Sanematsu, Y Ishiga, H Seki, K Toyoda, T Shiraishi, T Yamada

    PLANT SCIENCE   160 ( 5 )   997 - 1006   2001年4月

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    記述言語:英語   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    Clone E86 was isolated as cDNA for elicitor-inducible gene From pea epicotyls by differential screening. The deduced amino acid sequence of E86 showed high homology to hypersensitivity-related protein hsr203J in tobacco and also showed significant homologies to the Ser-active hydrolases, such as mammalian hormone-sensitive lipases, bacterial lipases and esterases. E86 polypeptide possesses consensus amino acid sequence motifs (His-Gly) and (Gly-X-Ser-X-Gly) conserved in lipases and esterases and showed esterase degradation of p-nitrophenyl butyrate. Northern blot analysis revealed that the E86-transcript is abundant in roots and stems and was induced by fungal elicitor in pea epicotyls. However, elicitor-induced accumulation of E86 mRNA was significantly inhibited by the fungal suppressor. Furthermore the expression of the genes encoding E86 and phenylalanine ammonia-lyase was induced within 1 h after the inoculation of a nonpathogen. but it was delayed for 5 h by the inoculation of a compatible pathogen. These results suggest that the elicitor-induced Ser-active hydrolase derived from E86 gene might be related to the plant defense responses. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

    DOI: 10.1016/S0168-9452(01)00343-0

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  • Bacteriophage P4282, a parasite of ┣DB Ralstonia solanacearum (/)-┫DB, encodes a bacteriolytic protein important for lytic infection of its host.

    OZAWA H, TANAKA H, ICHINOSE Y, SHIRAISHI T, YAMADA T

    Mol. Gen. Genet.   265 ( 1 )   95 - 101   2001年3月

  • Bacteriophage P4282, a parasite of Ralstonia solanacearum, encodes a bacteriolytic protein important for lytic infection of its host

    H Ozawa, H Tanaka, Y Ichinose, T Shiraishi, T Yamada

    MOLECULAR GENETICS AND GENOMICS   265 ( 1 )   95 - 101   2001年3月

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    記述言語:英語   出版者・発行元:SPRINGER-VERLAG  

    To enhance bacterial wilt resistance in tobacco expressing a foreign protein, we isolated the bacteriolytic gene from a bacteriophage that infects Ralstonia solanacearum The bacteriolytic protein of phage P4282 isolated in Tochigi Prefecture was purified from a lysate of R. solanacearum M4S cells infected with the phage, and its bacteriolytic activity was assayed by following the decrease in the turbidity of suspensions of R. solanacearum M4S cells. The molecular weight of the bacteriolytic protein was approximately 71 kDa, and the sequence of the N-terminal 13 amino acids was determined. We used oligonucleotide probes based on this amino acid sequence to isolate the bacteriolytic gene from phage P4282 DNA. This gene of 2061 bp encodes a product of 687 amino acids, whose calaculated molecular weight was 70.12 kDa. The bacteriolytic gene was placed under the control of an inducible promoter, and the plasmid was transformed into Escherichia coli NM522. The soluble proteins extracted from E. coli NM522 cells harboring the plasmid with the bacteriolytic gene showed obvious bacteriolytic activities against several strains of R. solanacearum isolated in various districts in Japan. DNA fragments from five phages, isolated in Niigata. Aomori, Okinawa, Fukushima and Yamaguchi Prefectures, hybridized to the bacteriolytic gene of phage P4282. These observations indicate that the bacteriolytic protein shows nonspecific activity against R. solanacearum strains, and a sequence similar to that of the bacteriolytic gene is conserved in the DNA of other bacteriophages. These results indicate that the generation of transgenic (tobacco) plants expressing the bacteriolytic gene of phage P4282 might result in enhanced resistance to bacterial wilt in tobacco.

    DOI: 10.1007/s004380000389

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  • HarpinPsta from ┣DB Pseudomonas syringae (/)-┫DB pv.┣DB tabaci (/)-┫DB is defective and deficient in its gene expression and HR-inducing activity.

    J. Gen. Plant Pathol.   67 ( 2 )   116 - 123   2001年

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  • Effect of harpin from four pathovars of ┣DB Pseudomonas syringae (/)-┫DB on pea defense responses.

    J. Gen. Plant Pathol.   67 ( 2 )   148 - 151   2001年

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  • Effect of harpin from four pathovars of ┣DB Pseudomonas syringae (/)-┫DB on pea defense responses.

    TANAKA R, TAGUCHI F, ICHINOSE Y, TOYODA K, SHIRAISHI T, YAMADA T

    J. Gen. Plant Pathol.   67 ( 2 )   148 - 151   2001年

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  • 初期・表層シグナル伝達系と防御システム

    化学と生物   39 ( 10 )   531 - 537   2001年

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  • Methyl jasmonate inhibits Harpin-induced hypersensitive cell death, generation of hydrogen peroxide and expression of defense genes in tobacco suspension cultured BY-2 cells.

    Plant Cell Physiol.   42 ( 4 )   446 - 449   2001年

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  • HarpinPsta from ┣DB Pseudomonas syringae (/)-┫DB pv.┣DB tabaci (/)-┫DB is defective and deficient in its gene expression and HR-inducing activity.

    J. Gen. Plant Pathol.   67 ( 2 )   116 - 123   2001年

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  • Gramine increase associated with rapid and transient systemic resistance in barley seedlings induced by mechanical and biological stresses.

    Matsuo, H, Taniguchi, K, Hiramoto, T, Yamada, T, Ichinose, Y, Toyoda, K, Takeda, K, Shiraishi, T

    Plant Cell Physiol.   42 ( 10 )   1103 - 1111   2001年

  • Potentiation of phytoalexin accumulation in elicitor-treated epicotyls of pea (Pisum sativum) by a diacylglycerol kinase inhibitor

    K Toyoda, T Kawahara, Y Ichinose, T Yamada, T Shiraishi

    JOURNAL OF PHYTOPATHOLOGY   148 ( 11-12 )   633 - 636   2000年12月

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    記述言語:英語   出版者・発行元:WILEY-BLACKWELL  

    When epicotyl tissues of pea were treated with a diacylglycerol (DAG) kinase inhibitor (R59022), enhanced induction of the phytoalexin accumulation occurred which was induced by fungal elicitor. The marked induction was associated with a sustained accumulation of phenylalanine ammonia-lyase (PAL)-mRNA and the consequent increase in PAL activity. These results suggest that inhibition of DAG breakdown leads to increased induction of phytoalexin accumulation and support the hypothesis that DAG kinase negatively regulates the signal transduction.

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  • Phytoalexin accumulation in elicitor-treated epicotyis of pea(Pisum sativum L.)by a diacylglycerol kinase inhibitor.(共著)

    Journal of Phytopathology   148 ( 11-12 )   633 - 636   2000年12月

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    記述言語:英語  

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  • Independent pathways leading to apoptotic cell death, oxidative burst and defense gene expression in response to elicitin in tobacco cell suspension culture

    M Sasabe, K Takeuchi, S Kamoun, Y Ichinose, F Govers, K Toyoda, T Shiraishi, T Yamada

    EUROPEAN JOURNAL OF BIOCHEMISTRY   267 ( 16 )   5005 - 5013   2000年8月

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    記述言語:英語   出版者・発行元:BLACKWELL SCIENCE LTD  

    We characterized pharmacologically the hypersensitive cell death of tobacco BY-2 cells that followed treatments with Escherichia coli preparations of INF1, the major secreted elicitin of the late blight pathogen Phytophthora infestans. INF1 elicitin treatments resulted in fragmentation and 180 bp laddering of tobacco DNA as early as 3 h post-treatment. INF1 elicitin also induced rapid accumulation of H2O2 typical of oxidative burst, and the expression of defense genes such as phenylalanine ammonia-lyase (PAL) gene at 1 h and 3 h after elicitin treatment, respectively. To investigate the involvement of the oxidative burst and/or the expression of defense genes in the signal transduction pathways leading to hypersensitive cell death, we analyzed the effect of several chemical inhibitors of signal transduction pathways on the various responses. The results indicated that (a) the cell death required serine proteases, Ca2+ and protein kinases, (b) the oxidative burst was involved in Ca2+ and protein kinase mediated pathways, but elicitin-induced AOS was neither necessary nor sufficient for cell death and PAL gene expression, and (c) the signaling pathway of PAL gene expression required protein kinases. These results suggest that the three signal transduction pathways leading to cell death, oxidative burst and expression of defense genes branch in the early stages that follow elicitin recognition by tobacco cells.

    DOI: 10.1046/j.1432-1327.2000.01553.x

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  • Characterization of cDNAs encoding two glycine-rich proteins in chickpea (Cicer arietinum L.) : Accumulation in response to fungal infection and other stress factors.(共著)

    CORNELS H, ICHINOSE Y, BARZ W

    Plant Science   154 ( 1 )   83 - 88   2000年5月

  • Cytochalasin A inhibits the binding of phenylalanine ammonia-lyase mRNA to ribosomes during induction of phytoalexin in pea seedlings

    M Sugimoto, K Toyoda, Y Ichinose, T Yamada, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   41 ( 2 )   234 - 238   2000年2月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Cytochalasin A (CA) blocked the accumulation of phytoalexin and phenylalanine ammonia-lyase (PAL)-protein in pea tissues treated with a fungal elicitor but scarcely affected the PAL-mRNA content. Further analysis showed that CA decreased the PAL-mRNA bound to ribosomes. These results indicate that actin filaments are tightly associated with the translational process of the PAL gene.

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  • Genes expressed in Ascochyta rabiei-inoculated chickpea plants and elicited cell cultures as detected by differential cDNA-hybridization.(共著)

    Zeitschrift fuer Naturforschung   55 ( 1-2 )   44 - 54   2000年1月

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    記述言語:英語  

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  • lmportance of AC-rich Element on Pea Phenylalanine Ammonia-Lyase Gene 1 Promoter for Expression lnduced by Nonpathogen Attack.(共著)

    IMURA Y, IGUCHI S, TOYODA K, ICHINOSE Y, SHIRAISHI T, YAMADA T

    Journal of General Plant Pathol.ogy   66 ( 2 )   123 - 127   2000年

  • Potentiation of phytoalexin accumulation in elicitor-treated epicotyls of pea (Pisum sativum) by a diacylglycerol kinase inhibitor

    K. Toyoda, T. Kawahara, Y. Ichinose, T. Yamada, T. Shiraishi

    Journal of Phytopathology   148 ( 11-12 )   633 - 636   2000年

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    記述言語:英語  

    When epicotyl tissues of pea were treated with a diacylglycerol (DAG) kinase inhibitor (R59022), enhanced induction of the phytoalexin accumulation occurred which was induced by fungal elicitor. The marked induction was associated with a sustained accumulation of phenylalanine ammonia-lyase (PAL)-mRNA and the consequent increase in PAL activity. These results suggest that inhibition of DAG breakdown leads to increased induction of phytoalexin accumulation and support the hypothesis that DAG kinase negatively regulates the signal transduction.

    DOI: 10.1046/j.1439-0434.2000.00568.x

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  • Potentiation of phytoalexin accumulation in elicitor-treated epicotyls of pea (┣DB Pisum sativum (/)-┫DB L.) by a diacylglycerol kinase inhibitor.

    J. Phytopathol.   148 ( 11-12 )   633 - 636   2000年

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  • lndependent pathways leading to apoptotic cell death,oxidative burst and defense gene expression in response to elicitin in tobacco cell suspension culture.(共著)

    SASABE M, TAKEUCHI K, ICHINOSE Y, TOYODA K, SHIRAISHI T, YAMADA T, KAMOUN S, GOVERS F

    European Journal of Biochemistry   267 ( 16 )   5005 - 5013   2000年

  • Cytochalasin A inhibits the binding of phenylalanine ammonia-lyass mRNA to ribosomes during induction of phytoalexin in pea seedlings.(共著)

    Plant Cell Physiology   41   234 - 238   2000年

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  • lmportance of AC-rich Element on Pea Phenylalanine Ammonia-Lyase Gene 1 Promoter for Expression lnduced by Nonpathogen Attack.(共著)

    Journal of General Plant Pathol.ogy   66 ( 2 )   123 - 127   2000年

  • Induction of defense responses by synthetic glycopeptides that have a partial structure of the elicitor in the spore germination fluid of Mycosphaerella pinodes

    A Kiba, T Takeda, T Kanemitsu, K Toyoda, Y Ichinose, T Yamada, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   40 ( 9 )   978 - 985   1999年9月

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

    A high molecular weight elicitor (&gt;70 kDa) from spore germination fluid of a pea pathogen, Mycosphaerella pinodes, has a partial structure of beta-D-Glc-(1--&gt;6)-alpha-D-Man-(1--&gt;6)-D-Man, which is O-glycosidically attached to serine in the protein moiety. To elucidate the minimum structure for the elicitor activity to pea plants, the effects of nine glycopeptides including beta-D-Glc-(1--&gt;6)-alpha-D-Man-(1--&gt;6)-D-Man-O-Ser (No. 1) to [beta-D-Glc-(1--&gt;6)-alpha-D-Man-(1--&gt;6)-D-Man](3)-O-Ser(3)-Pro(3) (No. 9) on the infection by M. pinodes, superoxide generation and ATPase activity were measured. The glycopeptides [beta-D-Glc-(1--&gt;6)-alpha-D-Man-(1--&gt;6)-D-Man]-O-Ser(2)-Pro(2) (No. 3) to No. 9 induced rejection reaction of pea tissue against M. pinodes. The glycopeptides No. 3 to No. 9 also induced superoxide generation on uninjured pea leaves. Moreover, the glycopeptides No. 3 to No. 9 induced in vitro the activation of cell wall-bound ATPase and superoxide generation system in the protein fraction solubilized from pea cell wall. The results indicate that the synthetic glycopeptides, No. 3 to No. 9, are available to analyze the signal transduction cascade leading to defense responses and the receptor for the elicitor.

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  • Functional analysis of retrotransposons in pea

    H Kato, P Sriprasertsak, H Seki, Y Ichinose, T Shiraishi, T Yamada

    PLANT AND CELL PHYSIOLOGY   40 ( 9 )   933 - 941   1999年9月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    The 5'-upstream regions of the plant active defense genes in pea (PSPAL2 and PSCHS1) exhibit significant nucleotide sequence identity to part of a copia-type retrotransposon. To characterize the retrotransposon in pea putative reverse transcriptase sequences (Psr) were amplified by PCR from cDNA prepared from protoplasts derived from pea suspension cultured cells. Psr genes can be classified into at least eight subgroups (A-H) according to their nucleotide sequence similarities. A subgroup PsrC was induced by the treatment of etiolated pea epicotyls with fungal elicitor within 30 min but was hardly induced by protoplasting of pea suspension cultured cells, whereas subgroups of PsrA and PsrB were highly induced by protoplasting. Interestingly, the retrotransposon sequence present at the 5'-upstream region of PSCHS1 which was induced by the treatment with fungal elicitor belonged to subgroup PsrC. The retrotransposon might play an important role in reorganization of the genes and optimization of the gene expression in response to various environmental stimuli such as an attack by phytopathogens or protoplasting.

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  • Changes in in vivo DNA-protein interactions in pea phenylalanine ammonia-lyase and chalcone synthase gene promoter induced by fungal signal molecules

    H Seki, Y Nagasugi, Y Ichinose, T Shiraishi, T Yamada

    PLANT AND CELL PHYSIOLOGY   40 ( 1 )   88 - 95   1999年1月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Elicitor-induced protein bindings were detected on the several particular sequence motifs in two members of the pea PAL and CHS gene promoters by in vivo footprinting analyses. However, elicitor-induced changes rapidly reverted back to the uninduced pattern after the subsequent application of suppressor from M. pinodes.

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  • Changes in in vivo DNA-potein interactions in pea phenylalanine ammonia-lyase and chalcone synthase gene promoter induced by fungal signal molecules.(共著)

    Plant Cell Physiology   40 ( 1 )   88 - 95   1999年1月

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  • Molecular cloning of a cDNA encoding a putative DNA-binding zinc-finger protein in pea.(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   88   25 - 30   1999年

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  • Expression of the chimeric pea PSPAL2 promoter in transgenic tobacco in response to fungal ingress and injury.(共著)

    Permpong SRIPRASERTSAK, Andi SALAMAH, Yoshiyuki IMURA, Yuki ICHINOSE, Tomonori SHIRAISHI, Tetsuji YAMADA

    Annals of Phytopathological Society of Japan   65 ( 2 )   123 - 130   1999年

  • Expression of the chimeric pea PSPAL2 promoter in transgenic tobacco in response to fungal ingress and injury.(共著)

    井村 善之, 一瀬 勇規, 白石 友紀, 山田 哲治

    Annals of Phytopathological Society of Japan   65 ( 2 )   123 - 130   1999年

  • Cloning and Structural Characterization of hrp Locus of Pseudomonas syringae pv. pisi.(共著)

    Hideki NAKADA, Aiko TAHARA, Masaharu HAYASHI, Rena SHIMIZU, Rui TANAKA, Yuki ICHINOSE, Tomonori SHIRAISHI, Tetsuji YAMADA

    Annals of Phytopathological Society of Japan   65 ( 2 )   147 - 152   1999年

  • Tranformation of the endophyte Neotyphodium With the iaaM gene(共著)

    Ahmad YUNUS, Shinji KAWAMATA, Tadayuki SHIMANUKI, Yasuhiro MURAKAMI, Yuki ICHINOSE, Tomonori SHIRAISHI, Tetsuji YAMADA

    Annals of Phytopathological Society of Japan   65 ( 2 )   192 - 196   1999年

  • Tranformation of the endophyte Neotyphodium With the iaaM gene(共著)

    川又 伸治, 島貫 忠幸, 村上 泰弘, 一瀬 勇規, 白石 友紀, 山田 哲治

    Annals of Phytopathological Society of Japan   65 ( 2 )   192 - 196   1999年

  • Functional Analysis of retrotransposons in pea.(共著)

    Plant Cell Phyiology   40   933 - 941   1999年

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  • Cloning and Structural Characterization of hrp Locus of Pseudomonas syringae pv. pisi.(共著)

    中田 秀毅, 田原 愛子, 林 昌治, 清水 鈴菜, 田中 塁, 一瀬 勇規, 白石 友紀, 山田 哲治

    Annals of Phytopathological Society of Japan   65 ( 2 )   147 - 152   1999年

  • cNDA cloning and gene expression of three small GTP-binding proteins in defense response of chickpea.(共著) 査読

    Biochimica et Biophysica Acta   1489 ( 2-3 )   462 - 466   1999年

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:速報,短報,研究ノート等(学術雑誌)  

    DOI: 10.1016/S0167-4781(99)00201-8

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  • 植物病原菌と防御機構(共著)

    蛋白質・核酸・酵素   44   2308 - 2314   1999年

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  • Molecular cloning of a cDNA encoding a putative DNA-binding zinc-finger protein in pea.(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   88   25 - 30   1999年

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  • Co-purification of plasma membrane ATPase and phosphatidylinositol kinase from pea plasma membrane.(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   88   31 - 38   1999年

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  • Co-purification of plasma membrane ATPase and phosphatidylinositol kinase from pea plasma membrane.(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   88   31 - 38   1999年

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  • Inductions of defense responses by synthetic glycopeptides that have a partial structure of the elicitor in the spore germination fluid of Mycosphaerella pinodes.(共著)

    Plant Cell Phyiology   40   978 - 985   1999年

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  • Structural analysis of a putative hypovirulent plasmid, pJTPS1, found in a spontaneous avirulent mutant of Ralstonia solanacearum.(共著)

    Rena SHIMIZU, Korehiro AKAISHI, Hideaki NEGISHI, Hiroshi TANAKA, Yuki ICHINOSE, Tomonori SHIRAISHI, Tetsuji YAMADA

    Annals of Phytopathological Society of Japan   65 ( 2 )   184 - 188   1999年

  • Structural analysis of a putative hypovirulent plasmid, pJTPS1, found in a spontaneous avirulent mutant of Ralstonia solanacearum.(共著)

    清水 鈴菜, 赤石 維衆, 根岸 秀明, 田中 博, 一瀬 勇規, 白石 友紀, 山田 哲治

    Annals of Phytopathological Society of Japan   65 ( 2 )   184 - 188   1999年

  • Interaction between cell wall and plasma membrane via RGD motif is implicated in plant defense response.(共著)

    Plant Cell Physiology   39 ( 11 )   1245 - 1249   1998年11月

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    記述言語:英語  

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  • Interaction between cell wall and plasma membrane via RGD motif is implicated in plant defense responses

    A Kiba, M Sugimoto, K Toyoda, Y Ichinose, T Yamada, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   39 ( 11 )   1245 - 1249   1998年11月

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

    Integrin- and vitronectin-like proteins were found to exist in the pea plasma membrane and cell wall, respectively. The hexapeptide GRGDSP but not GRGESP inhibited either the binding of cell wall protein(s) to plasma membrane protein(s) or the defense response. A possible role of linkage via RGD motif in plant defenses is discussed.

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  • SPECIFIC REGULATION OF CELL WALL-BOUND ATPASE BY THE SUPPRESSOR FROM MYCOSPHAERELLA PINODES(3) -IDENTIFICATION OF ELICITOR- AND SUPPRESSOR-BINDING PROTEINS IN CELL WALL-BOUND ATPASE FRACTION FROM PEA PLANT-

    KIBA Akinori, SUGIMOTO Megumi, TOYODA Kazuhiro, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   39   S144 - S144   1998年5月

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  • ISOLATION AND CHARACTERIZATION OF ELICITOR-RESPONSIVE GENES IN PEA

    ICHINOSE Yuki, SANEMATSU Shiroh, ENDOH Ai, OKABE Takeharu, HISAYASU Yumiko, SEKI Hikaru, SHIRAISHI Tomonori, TOYODA Kazuhiro, YAMADA Tetsuji

    Plant and cell physiology   39   S143 - S143   1998年5月

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  • In vivo FOOTPRINTING ANALYSIS OF cis-REGULATORY ELEMENTS INVOLVED IN THE ELICITOR-MEDIATED ACTIVATION AND SUPPRESSOR-MEDIATED DEACTIVATION OF PEA PAL AND CHS

    SEKI Hikaru, ICHINOSE Yuki, NAGASUGI Yumi, TOYODA Kazuhiro, SHIRAISHI Tomonori, YAMADA Tetsuji

    Plant and cell physiology   39   S43 - S143   1998年5月

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  • RECOGNITION OF FUNGAL SUPPRESSOR BY PEA PLANTS II

    SUGIURA Tetsuya, KIBA Akinori, AOYAGI Masaaki, TOYODA Kazuhiro, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   39   S144 - S144   1998年5月

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  • PURIFICATION AND ANALYSIS OF ELICITOR BINDING-PROTEIN IN CELL WALL FRACTION FROM PEA

    SUGIMOTO Megumi, KIBA Akinori, TOYODA Kazuhiro, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   39   S143 - S143   1998年5月

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  • Cis-regulatory elements and trans-acting factors involved in the activation of a member of elicitor-responsive pea chalcone synthase gene family, PSCHS2(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   91 - 97   1998年

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  • Phosphorylation of phosphatidylinositols and production of lysophospholipid in pea plasma membrane are coordinately regulated by elicitor-and suppressor from Mycosphaerella pinodes(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   109 - 116   1998年

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  • Cis-regulatory elements and trans-acting factors involved in the activation of a member of elicitor-responsive pea chalcone synthase gene family, PSCHS2(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   91 - 97   1998年

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  • Transformation of mutualistic fungal Acremonium endophyte(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   99 - 107   1998年

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  • Phosphorylation of phosphatidylinositols and production of lysophospholipid in pea plasma membrane are coordinately regulated by elicitor-and suppressor from Mycosphaerella pinodes(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   109 - 116   1998年

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  • Transformation of mutualistic fungal Acremonium endophyte(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   87   99 - 107   1998年

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  • Elevation of diacylglycerol during the early stage of elictitor-signal transduction in pea(Pisum sativum).(共著)

    Kazuhiro TOYODA, Tomoharu KAWAHARA, Rumi MIZUKOSHI, Masashi KOYAMA, Yuki ICHINOSE, Tetsuji YAMADA, Tomonori SHIRAISHI

    Annals of Phytopathological Society of Japan   64 ( 5 )   485 - 487   1998年

  • Elevation of diacylglycerol during the early stage of elictitor-signal transduction in pea(Pisum sativum).(共著)

    豊田 和弘, 河原 智治, 水越 留美, 小山 昌史, 一瀬 勇規, 山田 哲治, 白石 友紀

    Annals of Phytopathological Society of Japan   64 ( 5 )   485 - 487   1998年

  • Plant cell wall my recognize pathogenic microorganisms

    Tomonori SHIRAISHI, Akinori KIBA, Kazuhiro TOYODA, Yuki ICHINOSE, Tetsuji YAMADA

    KASEAA   36 ( 4 )   226 - 233   1998年

  • Functional Analysis of the Putative cis-elements Involved in Activation by Elicitor and Deactivation by Suppressor in the Promoter of a Pea Gene for Phenylalanine Ammonia-1yase(PSPAL1).(共著)

    Plant Cell Physiology   38 ( 12 )   1403 - 1408   1997年12月

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    記述言語:英語  

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  • Superoxide generation in extracts from isolated plant cell walls is regulated by fungal signal molecules.(共著)

    KIBA A, MIYAKE C, TOYODA K, ICHINOSE Y, YAMADA T, SHIRAISHI T

    Phytopathology   87 ( 8 )   846 - 852   1997年8月

  • Temporal and spatial pattern of expression of the pea phenylalanine ammonia-lyase gene1 promoter in transgenic tobacco

    S Kawamata, K Shimoharai, Y Imura, M Ozaki, Y Ichinose, T Shiraishi, H Kunoh, T Yamada

    PLANT AND CELL PHYSIOLOGY   38 ( 7 )   792 - 803   1997年7月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Genes encoding phenylalanine ammonia-lyase (PAL) form a small multigene family with at least three members in pea. Tissue-specific expression of the promoter of a member of PAL gene family (PSPAL1) was investigated in the transgenic tobacco transformants carrying the different modes of chimeric fusion between the PSPAL1 promoter and a bacterial beta-glucuronidase (GUS) gene. In stems, at least, strict correlation was found between steady-state levels of Gus-mRNA and enzyme activity. Significantly high level of GUS activity was observed in roots, particularly in meristematic tissues and the pigmented region of petals of transgenic tobacco carrying the translational fusion type B (-1,394 to +140 of PSPAL1 connected to Gus), followed by moderately high level of GUS activity carrying the translational fusion type A(-1,394 to +117). GUS expression in tissues of mature leaves, however, was very low in these constructs. Extremely low GUS activity was observed in the transformants of transcriptional fusion type (-1,394 to +5), whilst no activity was detected carrying non-transcription fusion type (-1,394 to -27). Furthermore, the pattern of the PSPAL1 expression was characterized in response to pathogen ingress and woundings in transgenic tobacco carrying the translational fusion type B. Woundings itself triggered marked expression of PSPAL1-driven GUS expression at the wounded sites. Inoculation of nonpathogens, Phytophthora capsici, P. boehmeriae and Erisiphe graminis f. sp. hordei, both caused rapid and very clear GUS expression zone along with the development of hypersensitive cell death area where callose was accumulated, however, the inoculation of a pathogen, P. nicotiana caused slow and hazy GUS expression zone along with the lesion development. These results suggest that the expression of pea PSPAL1 promoter is regulated in a similar fashion, at least in a part, in pea and transgenic tobacco, under the plant development and various environmental cues.

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  • Putative cis-regulatory elements for elicitor-mediated transcriptional activation and molecular evolution of the pea chalcone synthase genes.

    Y Ichinose, H Seki, M Ito, T Shiraishi, T Yamada

    PLANT PHYSIOLOGY   114 ( 3 )   1175 - 1175   1997年7月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC PLANT PHYSIOLOGISTS  

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  • Specific regulation of cell wall-bound ATPase and peroxidase by the suppressor for defense response.

    A Kiba, A Inata, C Miyake, K Toyoda, Y Ichinose, T Yamada, T Shiraishi

    PLANT PHYSIOLOGY   114 ( 3 )   1176 - 1176   1997年7月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC PLANT PHYSIOLOGISTS  

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  • Con A-binding protein in plasma membranes may participate in signal-transduction cascades that lead to defense responses in pea.

    K Toyoda, M Sugimoto, T Kawahara, Y Ichinose, T Yamada, T Shiraishi

    PLANT PHYSIOLOGY   114 ( 3 )   1502 - 1502   1997年7月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:AMER SOC PLANT PHYSIOLOGISTS  

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  • Association between ion fluxes and defense responses in pea and cowpea tissues

    M Amano, K Toyoda, Y Ichinose, T Yamada, T Shiraishi

    PLANT AND CELL PHYSIOLOGY   38 ( 6 )   698 - 706   1997年6月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    The glycopeptide elicitor from a pea pathogen, Mycosphaerella pinodes, induced rapid alkalinization and increases in levels of Na+ and K+ ions in the extracellular solution upon contact with pea and cowpea tissues, The presence of monensin, nigericin, lidocaine, quinidine or phenytoin together with the elicitor markedly inhibited these changes, whereas the presence of valinomycin, gramicidin D, tetraethylammonium, CsCl and aminopyridine did not. The production of phytoalexins in pea and cowpea tissues was also strongly inhibited by the simultaneous presence of the former reagents but not of the latter reagents. Inhibitory effects on the production of phytoalexins were diminished when monensin, nigericin or a Na+-channel blocker was applied 3 h after the start of treatment with elicitor, Furthermore, orthovanadate and neomycin, which suppress defense responses in both tissues, also inhibited the above mentioned changes. By contrast, the species-specific suppressor from M. pinodes inhibited the elicitor-induced release of Na+ and K+ ions from pea tissues, but, conversely, by itself it elicited either the defense response or the release of Na+ and K+ ions from cowpea tissues. The results indicate that these ion-related changes, in particular the efflux of Na+ and K+ ions, might be closely associated with the signal transduction system for defense responses at the tissue level.

    DOI: 10.1093/oxfordjournals.pcp.a029223

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  • Molecular evolution and functional relevance of the chalcone synthase genes of pea

    M Ito, Y Ichinose, H Kato, T Shiraishi, T Yamada

    MOLECULAR & GENERAL GENETICS   255 ( 1 )   28 - 37   1997年6月

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    記述言語:英語   出版者・発行元:SPRINGER VERLAG  

    We have isolated seven genomic chalcone synthase (CHS) genes and six classes of CNS cDNA from elicitor-treated pea tissues. Comparison of the nucleotide sequences of the coding regions revealed the existence of eight members of the CHS gene family in pea. These can essentially be divided into three groups (PSCHS1, 2 and 8; PSCHS3, 4 and 5; and PSCHS6 and 7) on the basis of nucleotide and/or amino acid sequence comparisons of the coding regions, introns and promoter regions. We previously reported that the accumulation of CHS mRNAs is induced by elicitor treatment. Accumulation of CHS mRNA was observed mainly in roots and very little was found in floral organs. To specifically detect expression of each CHS gene in various types of pea cells, S1 nuclease protection assays were performed. Interestingly, the classification of the eight members of the CHS gene family based on the sequence identity was found to reflect their expression patterns as determined by the S1 nuclease protection assay. The first group of CHS genes, PSCHS1, 2 and 8, was strongly induced not only by elicitor treatment and UV irradiation but is also constitutively expressed in root and flower tissues. The second group, PSCHS3, 4 and 5, was also strongly induced by elicitor treatment and UV irradiation but is constitutively expressed only in root. Expression of the third group, PSCHS6 and 7 was barely detectable in any of the organs tested and was not influenced by environmental stimuli such as elicitor or UV. Furthermore, sequence analysis of the promoter region of each member of the CHS gene family revealed that putative cis-regulatory elements, such as Box-I, Box-II and G-Box, were conserved only in PSCHS1, 2, 3, 4 and 5. From these results we propose that an ancestral CHS gene might have given rise to defense response-related (UV irradiation- and elicitor-responsive) and -unrelated (unresponsive) genes at an early stage of evolution, followed by divergence within these subclasses based upon the developmental program in pea.

    DOI: 10.1007/s004380050471

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  • Molecular evolution and functional relevance of the chalcone synthase genes of pea

    M Ito, Y Ichinose, H Kato, T Shiraishi, T Yamada

    MOLECULAR & GENERAL GENETICS   255 ( 1 )   28 - 37   1997年6月

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    記述言語:英語   出版者・発行元:SPRINGER VERLAG  

    We have isolated seven genomic chalcone synthase (CHS) genes and six classes of CNS cDNA from elicitor-treated pea tissues. Comparison of the nucleotide sequences of the coding regions revealed the existence of eight members of the CHS gene family in pea. These can essentially be divided into three groups (PSCHS1, 2 and 8; PSCHS3, 4 and 5; and PSCHS6 and 7) on the basis of nucleotide and/or amino acid sequence comparisons of the coding regions, introns and promoter regions. We previously reported that the accumulation of CHS mRNAs is induced by elicitor treatment. Accumulation of CHS mRNA was observed mainly in roots and very little was found in floral organs. To specifically detect expression of each CHS gene in various types of pea cells, S1 nuclease protection assays were performed. Interestingly, the classification of the eight members of the CHS gene family based on the sequence identity was found to reflect their expression patterns as determined by the S1 nuclease protection assay. The first group of CHS genes, PSCHS1, 2 and 8, was strongly induced not only by elicitor treatment and UV irradiation but is also constitutively expressed in root and flower tissues. The second group, PSCHS3, 4 and 5, was also strongly induced by elicitor treatment and UV irradiation but is constitutively expressed only in root. Expression of the third group, PSCHS6 and 7 was barely detectable in any of the organs tested and was not influenced by environmental stimuli such as elicitor or UV. Furthermore, sequence analysis of the promoter region of each member of the CHS gene family revealed that putative cis-regulatory elements, such as Box-I, Box-II and G-Box, were conserved only in PSCHS1, 2, 3, 4 and 5. From these results we propose that an ancestral CHS gene might have given rise to defense response-related (UV irradiation- and elicitor-responsive) and -unrelated (unresponsive) genes at an early stage of evolution, followed by divergence within these subclasses based upon the developmental program in pea.

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  • dad-1, a putative programmed cell death suppressor gene in rice

    Y Tanaka, T Makishima, M Sasabe, Y Ichinose, T Shiraishi, T Nishimoto, T Yamada

    PLANT AND CELL PHYSIOLOGY   38 ( 3 )   379 - 383   1997年3月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    The human dad-1 cDNA homolog was isolated from rice plants. The amino acid sequence of the predicted protein product is well conserved in both animals and plants. This rice dad-1 homolog can rescue the temperature-sensitive dad-1 mutants of hamster cells from apoptotic death, suggesting that the rice dad-1 homolog also functions as a suppressor for programmed cell death.

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  • RECOGNITION OF FUNGAL SUPPRESSOR BY PEA PLANTS -SIGNIFICANCE OF CELL WALL-

    SUGIURA Tetsuya, KIBA Akinori, AOYAGI Masaaki, KATO Toshiaki, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   38   s58   1997年3月

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  • THE SIGNIFICANCE OF ACTIVE OXYGEN SPEACIES IN ELICITATION OF DEFENSE RESPONSES IN PEA PLANTS

    INATA Atsuko, FUJII Masahiro, KIBA Akinori, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   38   s92   1997年3月

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  • EFFECTS OF SYNTHETIC GLYCOPEPTIDE-ELICITORS ON DEFENSE RESPONSES OF PEA PLANTS

    KIBA Akinori, INATA Atsuko, TAKEDA Tadahiro, KANEMITSU Takuya, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   38   s93   1997年3月

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  • PLANT CELL WALL - THE PRIMARY APPARATUS RECOGNIZING AND RESPONDING TO FUNGAL SIGNALS

    SHIRAISHI Tomonori, YAMADA Tetsuji, ICHINOSE Yuki, KIBA Akinori

    Plant and cell physiology   38   s11   1997年3月

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  • Combined effects of multiple cis-acting elements in elicitor-mediated activation of PSCHS1 gene

    H Seki, Y Ichinose, M Ito, T Shiraishi, T Yamada

    PLANT AND CELL PHYSIOLOGY   38 ( 1 )   96 - 100   1997年1月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Promoter of a gene encoding chalcone synthase 1 (PSCHS1), a member of the defense-related genes in pea, was analyzed by transient transfection assay. The results demonstrated that in addition to the previously identified AT-rich sequences, at least four distinct cis-acting elements were required for maximal fungal elicitor-mediated activation of PSCHS1.

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  • Combined effects of multiple cis-acting elements in elicitor-mediated activation of PSCHS1 gene

    H Seki, Y Ichinose, M Ito, T Shiraishi, T Yamada

    PLANT AND CELL PHYSIOLOGY   38 ( 1 )   96 - 100   1997年1月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Promoter of a gene encoding chalcone synthase 1 (PSCHS1), a member of the defense-related genes in pea, was analyzed by transient transfection assay. The results demonstrated that in addition to the previously identified AT-rich sequences, at least four distinct cis-acting elements were required for maximal fungal elicitor-mediated activation of PSCHS1.

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  • Orthovanadate induces phytoalexiu production in pea suspension-cultured cells(共著)

    Scientific Reprots of the Fawlty of Agriculture OKAYAMA UNIVERSITY   86   33 - 41   1997年

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  • Mechanisms of the plant disease resistance-Focusing on the suppressor-(共著)

    32 ( 1 )   49 - 59   1997年

  • Association between ion fluxes and defense responses in pea and cowpea tissues(共著)

    Plant Cell Physiology   38 ( 6 )   698 - 706   1997年

  • Temporal and spatial pattern of expression of the pea phenylalanine ammoria-lyase gene 1 promoter in transgenic tobacco(共著)

    Plant Cell Physiology   38 ( 7 )   792 - 803   1997年

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  • The Effects of Expression of Procaryotic-type iaa Genes in Agrobacterium tumefaciems on Plant Tumorigenicity.(共著)

    Yasuhiro MURAKAMI, Tomoki NISHINO, Tsuguyoshi TAIRA, Ahmad YUNUS, Yuki ICHINOSE, Tomonori SHIRAISHI, Tetsuji YAMADA

    Annals of the Phytopathological Society of Japan   63 ( 5 )   377 - 380   1997年

  • The Effects of Expression of Procaryotic-type iaa Genes in Agrobacterium tumefaciems on Plant Tumorigenicity.(共著)

    村上 泰弘, 西野 友規, 平嗣 良, 一瀬 勇規, 白石 友紀, 山田 哲治

    Annals of the Phytopathological Society of Japan   63 ( 5 )   377 - 380   1997年

  • Orthovanadate induces phytoalexiu production in pea suspension-cultured cells(共著)

    Scientific Reprots of the Fawlty of Agriculture OKAYAMA UNIVERSITY   86   33 - 41   1997年

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  • dad-1, a putative programmed cell death suppressor gene in rice(共著)

    Plant Cell Physiology   38 ( 3 )   379 - 383   1997年

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  • Mechanisms of the plant disease resistance-Focusing on the suppressor-(共著)

    32 ( 1 )   49 - 59   1997年

  • 植物の病害抵抗性機構-特にサプレッサーについて-

    植物の化学調節   32 ( 1 )   49 - 59   1997年

  • The role of suppressors in determining host-parasite specificities in plant cells

    T Shiraishi, T Yamada, Y Ichinose, A Kiba, K Toyoda

    INTERNATIONAL REVIEW OF CYTOLOGY - A SURVEY OF CELL BIOLOGY, VOL 172   172   55 - 93   1997年

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:ELSEVIER ACADEMIC PRESS INC  

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  • Analysis of cis-regulatory elements involved in the activation of a member of chalcone synthase gene family (PsChs1) in pea

    H Seki, Y Ichinose, H Kato, T Shiraishi, T Yamada

    PLANT MOLECULAR BIOLOGY   31 ( 3 )   479 - 491   1996年6月

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    記述言語:英語   出版者・発行元:KLUWER ACADEMIC PUBL  

    Cis-regulatory elements involved in the activation of the plant defense-related gene encoding chalcone synthase 1 (PsChs1) in pea (Pisum sativum L.) were examined by transient transfection, gel mobility shift assay and in vitro DNase I-footprinting analysis. Transient transfection assay revealed that a 61 bp DNA fragment spanning from -242 to -182 of PsChs1 was required for the maximal promoter activity and possibly involved in the enhancement of elicitor-mediated activation. Nuclear isolate from elicitor-treated pea epicotyl tissues contained some factor(s) that specifically bound to this DNA fragment to form a complex with low mobility (LMC, low mobility complex) in gel mobility shift assay. DNase I-footprinting analysis of LMC revealed that among three protected regions detected in a 61 bp DNA fragment, two regions contained identical AT-rich sequence, TAAAATACT. Site directed mutation in either or both identical sequences, T&lt;(AA)under bar&gt;AATACT to T&lt;(GG)under bar&gt;AATACT, resulted in the reduction or loss in the ability to form LMC. Detailed analysis of 61 bp DNA fragment demonstrated that the region from -242 to -226 containing promoter-distal TAAAATACT motif was imperative for the maximal elicitor-mediated activation of PsChs1.

    DOI: 10.1007/BF00042222

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  • Specific response of partially purified cell wall-bound ATPases to fungal suppressor(共著)

    Plant Cell Physiology   37 ( 2 )   207 - 214   1996年3月

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    記述言語:英語  

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  • Suppression of ATPase activity in pea cells during infecfion by a compatible race of Pseudomouas syriugae pv. pisi(共著)

    Annals of Phytopathological Society of Japan   1996年

  • Species-specific suppression of superoxide-anion generation of surfaces of pea leaves by the suppressor from Mycosphaerella pinodes(共著)

    Annals of the Phytopathological Society of JAPAN   62 ( 5 )   508 - 512   1996年

  • Species-specific suppression of superoxide-anion generation of surfaces of pea leaves by the suppressor from Mycosphaerella pinodes(共著)

    KIBA A, TOYODA K, ICHINOSE Y, YAMADA T, SHIRAISHI T

    Annals of the Phytopathological Society of JAPAN   62 ( 5 )   508 - 512   1996年

  • Analysis of cis-regulatory elements involued in the activation of a member of chalcoue synthase gene family (PsCHS1) in pea(共著)

    Plant Molecular Biology   1996年

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  • Suppression of ATPase activity in pea cells during infecfion by a compatible race of Pseudomouas syriugae pv. pisi(共著)

    Annals of Phytopathological Society of Japan   1996年

  • EXPRESSION OF HETEROLOGOUS PAL GENE IN MUSKMELON (CUCUMIS-MELO L CV BIRDIE)

    NM SALEH, SH TAN, Y ICHINOSE, T YAMADA

    ASIA-PACIFIC JOURNAL OF MOLECULAR BIOLOGY AND BIOTECHNOLOGY   3 ( 3 )   206 - 214   1995年9月

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    記述言語:英語   出版者・発行元:MALAYSIAN SOC MOLECULAR BIOLOGY BIOTECHNOLOGY  

    A system to develop PAL gene expression using particle gun was established in Cucumis melo L. These techniques enable us to study the regulation of pea phenylalanine ammonia-lyase (PAL) promoters in a heterologous plant system. The effect of UV irradiation on the regulation of the PAL promoters under different regime of UV irradiation were tested on the mesophyll protoplasts of melon. Maximum induction of GUS expression were observed in muskmelon leaves electroporated with two members of pea PAL promoters, PSPAL1 and PSPAL2 at 120 sec and 80 sec, respectively. Further confirmation of the GUS expression in the tissue of muskmelon were carried out using particle bombardment. Similar pattern of induction of GUS expression by UV irradiation have been observed, which suggests that common mechanism of PAL gene expression may exist in both muskmelon plant.

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  • Expression of heterologous pal gene in Muskmelon(Cucumis melo L. Birdie)(共著)

    Asia pacific Journal of Molecular Biology and Biotechndogy   3 ( 3 )   206 - 214   1995年9月

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    記述言語:英語  

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  • PLANT-LECTINS INDUCE THE PRODUCTION OF A PHYTOALEXIN IN PISUM-SATIVUM

    K TOYODA, K MIKI, Y ICHINOSE, T YAMADA, T SHIRAISHI

    PLANT AND CELL PHYSIOLOGY   36 ( 5 )   799 - 807   1995年7月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    The effects of several plant lectins on the production of a pea phytoalexin, pisatin, were examined. Con A, PHA, PNA and PSA each induced the production of pisatin in pea epicotyl tissues, demonstrating that plant lectins can act as elicitors. The production of pisatin in response to PHA, PNA or PSA was not affected by the simultaneous presence of the respective hapten sugars, whereas haptens specific for Con A, such as alpha-D-mannose and methyl-alpha-D-mannoside, abolished the induction of pisatin by Con A. These results indicate that the elicitor effect of Con A is attributable to its ability to bind to specific carbohydrates in pea cells. Induction of the production of pisatin by Con A was markedly inhibited by the suppressor derived from a pea pathogen. Mycosphaerella pinodes, and by several inhibitors related to signal-transduction pathways. It is suggested, therefore, that the Con A-induced production of pisatin in pea tissues might be associated with activation of a signal-transduction pathway. An additive effect on the accumulation of pisatin was observed when Con A was present with a polysaccharide elicitor from M. pinodes, suggesting that exogenous Con A does not compete with the recognition site(s) for the fungal elicitor in pea cells. The present data also indicate that Con A may be useful for characterization of the signal-transduction system that leads to the synthesis of phytoalexin in pea epicotyl tissues.

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  • SPECIFIC-INHIBITION OF CELL WALL-BOUND ATPASES BY FUNGAL SUPPRESSOR FROM MYCOSPHAERELLA-PINODES

    A KIBA, K TOYODA, Y ICHINOSE, T YAMADA, T SHIRAISHI

    PLANT AND CELL PHYSIOLOGY   36 ( 5 )   809 - 817   1995年7月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Activities of phosphatases were found in the fractions which were solubilized from cell walls of both pea and cowpea seedlings with 0.5 M NaCl. These phosphatases hydrolyzed triphosphonucleotides in the order: UTP = CTP &gt; GTP &gt; ATP; and UTP = GTP &gt; CTP = ATP, respectively. The activities of a pyrophosphatase and a p-nitrophenylphosphatase were also detected in these fractions. The suppressor in the spore germination fluid of a pea pathogen, Mycosphaerella pinodes, inhibited all of these phosphatase activities in the fraction solubilized from pea cell walls, but it rather enhanced only the activity of the ATPase among those phosphatases from the cowpea cell wall. Hydrolysis of ATP by a cell wall fraction of pea was also markedly inhibited by the suppressor, while hydrolysis of ATP by similar fractions from cowpea, kidney bean and soybean were rather enhanced by the suppressor, as well as by the elicitor. Thus, the cell wall-bound ATPases responded to the suppressor species-specifically. These cell wall-bound ATPases seemed to be different from the plasma membrane ATPases in several respects. The results suggest that plants recognize the fungal signals not only on their plasma membranes but also on their cell walls and, moreover that putative receptors for the fungal signals might be located close to cell wall-bound ATPases or might even be these ATPases themselves.

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  • Plant lectins induce the production of a phytolexin in Pisum sativum(共著)

    Plant Cell Physiology   36 ( 5 )   799 - 807   1995年7月

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    記述言語:英語  

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  • Specific inhibition of cell wall-bound ATPase by fungal suppressor from Mycosphaerella pinodes(共著)

    Plant Cell Physiology   36 ( 5 )   809 - 817   1995年7月

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    記述言語:英語  

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  • Establishment of a system for the transient expression of GUS gene in electroporated muskmelon (Cucumis melo L) protoplasts. (共著)

    Asia Pacific Journal of Molecular Biology and Biotechnology   3 ( 2 )   156 - 160   1995年6月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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  • A SUPPRESCIN FROM A PHYTOPATHOGENIC FUNGUS DEACTIVATES TRANSCRIPTION OF A PLANT DEFENSE GENE ENCODING PHENYLALANINE AMMONIA-LYASE

    M WADA, H KATO, K MALIK, P SRIPRASERTSAK, Y ICHINOSE, T SHIRAISHI, T YAMADA

    JOURNAL OF MOLECULAR BIOLOGY   249 ( 3 )   513 - 519   1995年6月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)   出版者・発行元:ACADEMIC PRESS (LONDON) LTD  

    Both elicitor and supprescin (suppressor) are present in the pycnospore germination fluid of a pea pathogen Mycospharella pinodes. A nuclear run-on assay revealed that supprescin rapidly deactivated elicitor-triggered transcription of the gene encoding phenylalanine ammonia-lyase in pea epicotyl tissues. The mechanism underlying the deactivation of the plant defense gene by signal molecules secreted from the fungal pathogen was investigated. Cis-acting sequences and trans-acting factors responsive to supprescin in a TATA-proximal region of a member of the phenylalanine ammonia-lyase gene family in pea were examined in vitro. Gel mobility-shift assays and DNase I footprinting analysis revealed that the promoter region of PSPAL2 was modified by the binding of nuclear factors at multiple sites that were possibly involved in supprescin-mediated deactivation. The prominent changes by supprescin were observed at boxes 2 and 4 and near exonic sequences.

    DOI: 10.1006/jmbi.1995.0313

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  • CHARACTERIZATION OF NUCLEAR FACTORS FOR ELICITOR-MEDIATED ACTIVATION OF THE PROMOTER OF THE PEA PHENYLALANINE AMMONIA-LYASE GENE

    H KATO, M WADA, K MURAYA, K MALIK, T SHIRAISHI, Y ICHINOSE, T YAMADA

    PLANT PHYSIOLOGY   108 ( 1 )   129 - 139   1995年5月

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    記述言語:英語   出版者・発行元:AMER SOC PLANT PHYSIOLOGISTS  

    The nuclear factors presumably associated with the activation of the gene encoding phenylalanine ammonia-lyase by a fungal elicitor were characterized in pea (Pisum sativum L.) epicotyls. The TATA-proximal region was dissected and putative cis-regulatory elements in the promoter of pea phenylalanine ammonia-lyase gene 1 were examined by gel-mobility shift and in vitro footprinting analyses. Specific binding of the nuclear factors to the promoter-proximal regions of pea phenylalanine ammonia-lyase gene 1 associated with elicitor-mediated activation was detected at a region containing consensus sequence motifs of boxes 2 and 4 and other AT-rich sequences. The analyses of DNA fragments containing the deleted promoter regions suggested that a residue from -183 to -173 (ATTAGTAAGTGAT) was essential for a maximal activity of forming low-mobility complex (LMC) in the gel-mobility shift assay, and synthetic oligonucleotides confirmed the presence of at least one nuclear component associated with the formation of an active LMC. Competition experiments and treatment with Hoechst 33258 provided direct evidence that the formation of LMC with the promoter fragments from genes encoding phenylalanine ammonia-lyase and chalcone synthase in pea contained one or more of the same proteins that recognize AT-rich sequence motifs for binding. It also suggests that common high-mobility group-like proteins might be involved in the regulation of elicitor-inducible genes in pea.

    DOI: 10.1104/pp.108.1.129

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  • RELATIONSHIP BETWEEN DEFENSE RESPONSESAND ION FLUX

    AMANO Masashi, TOYODA Kazuhiro, ICHINOSE Yuki, YAMADA Tetsuji, SHIRAISHI Tomonori

    Plant and cell physiology   36   S119   1995年3月

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  • TRANSCRIPTIONAL REGULATION OF THE GENEENCODING CHALCONE SYNTHASE IN PEA BY FUNGAL ELICITOR

    SEKI Hikaru, ICHINOSE Yuki, SHIRAISHI Tomonori, YAMADA Tetsuji

    Plant and cell physiology   36   S120   1995年3月

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  • REGULATION OF PLANT DEFENSE RESPONSES BY SIGNALS FROM FUNGAL PATHOGENS

    T SHIRAISHI, T YAMADA, Y ICHINOSE

    NIPPON NOGEIKAGAKU KAISHI-JOURNAL OF THE JAPAN SOCIETY FOR BIOSCIENCE BIOTECHNOLOGY AND AGROCHEMISTRY   69 ( 2 )   174 - 177   1995年2月

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    記述言語:日本語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:JAPAN SOC BIOSCI BIOTECHN AGROCHEM  

    DOI: 10.1271/nogeikagaku1924.69.174

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  • THE RELATIONSHIP BETWEEN SYSTEMIC RESISTANCE INDUCED BY PRUNING AND ACCUMULATION OF ANTIFUNGAL SUBSTANCES IN BARLEY SEEDLINGS

    T HIRAMOTO, N ABE, R TOBIMATSU, T SHIRAISHI, H OKU, T YAMADA, Y ICHINOSE

    JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT   143 ( 1 )   43 - 46   1995年1月

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    記述言語:英語   出版者・発行元:BLACKWELL WISSENSCHAFTS VERLAG GMBH  

    Infection by a compatible race of Erysiphe graminis f. sp. hordei on barley secondary leaves was significantly suppressed upon pruning of the primary leaves when E. graminis hordei was inoculated 3-12 h after the pruning, but it was rather enhanced during 15-21 h. The accumulation of antifungal substances was detected in hot ethanol extracts of barley seedlings from 15-27 h after pruning the primary leaves. Taking the time of the infection process of a challenger (E. graminis hordei) into consideration, timing of systemic resistance induced upon pruning coincided with the accumulation of antifungal substances.

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  • SIGNAL MOLECULES INDUCING SYSTEMIC RESISTANCE AND SUSCEPTIBILITY IN BARLEY SEEDLINGS

    T HIRAMOTO, R TOBIMATSU, N ABE, T SHIRAISHI, H OKU, T YAMADA, Y ICHINOSE

    JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT   143 ( 1 )   47 - 51   1995年1月

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    記述言語:英語   出版者・発行元:BLACKWELL WISSENSCHAFTS VERLAG GMBH  

    Exudate collected from the cut end of barley seedlings exhibited both activities that induced systemic resistance and susceptibility against Erysiphe graminis f. sp. hordei race Hh4 depending on the time after pruning. Exudates collected between 3-6 h after pruning showed maximum activity that induced systemic resistance, whereas those during 9-12 h conversely induced susceptibility in barley seedlings. The accumulation of antifungal substances in barley leaves correlates to the timing of induced resistance. The antifuntingal substances were water-soluble and severely affected the infection of E. graminis f. sp. hordei.

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  • Characterization of nuclear factors for elicitor-mediated activation of the promoter of the pea phenylalanine ammonia-lyase gene 1. (共著)

    KATO H, WADA M, MURAYA K, MALIK K, SHIRAISHI T, ICHINOSE Y, YAMADA T

    Plant Physiology   108 ( 1 )   129 - 139   1995年

  • Regulation of plant defense responses by signals from fangal pathogeus

    Tomonori Shiraishi, Tetsuji Yamada, Yuki Ichinose

    Nippon N(]E87C7[)geikagaku Kaishi   69 ( 2 )   174 - 177   1995年

  • 病原菌シグナルと植物の防御応答 共著

    白石 友紀, 山田 哲治, 一瀬 勇規

    日本農芸化学会誌   69 ( 2 )   174 - 177   1995年

  • H+ - translocating activity in proteoliposomes reconstituted with pea plasma membrane ATPase and its inhibition by fungal suppnessor from Mycosphaerella pinodes(共著)

    Annals of Phytopathological Society of Japan   61   134 - 136   1995年

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  • Supprescin from a phytopathogenic fungus deactivates transcription of a plant defense gene encodig phenylalanine ammonia-lyase. (共著)

    Journal of Molecular Biology   1995年

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  • Signal molecules inducing systemic resistance and susceptibility in barley seedling. (共著)

    Journal of Phytopathology   143   1995年

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  • The relationship between systemic resistance induced by pruning and accumulation of anitifungal substances in barley seedling. (共著) 査読

    Journal of Phytopathology   143   47 - 51   1995年

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    担当区分:最終著者  

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  • Suppression of the activation of chitinase and β-1, 3-glucanase in pea epicotyls by endogenous suppressor from ┣DBPisum sativum(/)-┫DB. (共著)

    Annals of the Phytopathological Society of Japan   1995年

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  • H+ - translocating activity in proteoliposomes reconstituted with pea plasma membrane ATPase and its inhibition by fungal suppnessor from Mycosphaerella pinodes(共著)

    Annals of Phytopathological Society of Japan   61   134 - 136   1995年

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  • Suppression of the activation of chitinase and β-1, 3-glucanase in pea epicotyls by endogenous suppressor from ┣DBPisum sativum(/)-┫DB. (共著)

    Annals of the Phytopathological Society of Japan   1995年

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  • Supperssors : Determinants of specificity produced by plant pathogens. (共著)

    Plant Cell Phisiology   35 ( 8 )   1107 - 1119   1994年12月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等  

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  • Suppressor produced by a pea pathogen, ┣DBM(/)-┫DB. pinodes. (共著)

    Grain Legumes   4   22 - 23   1994年

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  • Suppressor produced by a pea pathogen, ┣DBM(/)-┫DB. pinodes. (共著)

    Grain Legumes   4   22 - 23   1994年

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  • Rapid Changes in Polyphosphoinositide Metabolism in Pea in Response to Fungal Signals 共著

    Plant Cell Physiology   34 ( 5 )   729 - 735   1993年7月

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    記述言語:英語  

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  • ORGANIZATION OF THE GENES ENCODING CHALCONE SYNTHASE IN PISUM-SATIVUM

    C AN, Y ICHINOSE, T YAMADA, Y TANAKA, T SHIRAISHI, H OKU

    PLANT MOLECULAR BIOLOGY   21 ( 5 )   789 - 803   1993年3月

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    記述言語:英語   出版者・発行元:KLUWER ACADEMIC PUBL  

    To analyze the regulation of defense-related genes by signal molecules produced by phytopathogens, we isolated genes that encode chalcone synthase (CHS) in Pisum sativum. We have obtained seven independent genomic clones that contain at least seven classes of CHS genes, identified by the hybridization analysis to CHS cDNA and by the restriction mapping analysis. Two of the genomic clones (clone 5 and 6) each contain two CHS genes in a tandem repeat. The nucleotide sequence analysis of CHS genomic clone 5 revealed that PsCHS1 and PsCHS2 were corresponding genes of the CHS cDNA clones, pCC6 and pCC2, respectively, as reported earlier. Both genes are interrupted by a single intron of 88 nucleotides with identical sequences, although exonic sequences and 5'-flanking sequences are divergent. Nucleotide sequences of the introns in five other classes of CHS genes showed that three classes had an intron of 87 nt with a striking homology to each other, but that the intron of the other two classes of CHS genes showed heterogeneity both in size and nucleotide sequence. 5'-upstream regions of PsCHS1 and PsCHS2 did not show sequence homology except the 31 bp identical sequence that contains the CCTACC motif resembling the box-1 sequence. Both PsCHS1 and PsCHS2 genes are shown to be induced by fungal elicitor by a primer extension analysis and a transient transformation analysis using pea protoplasts prepared from suspension cultured-cells.

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  • Organization of the genes encoding chalcone synthase in Pisum sativum 共著

    Plant Molecular Biology   21 ( 5 )   789 - 803   1993年3月

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    記述言語:英語  

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  • MOLECULAR-CLONING OF PHENYLALANINE AMMONIA-LYASE CDNA FROM PISUM-SATIVUM

    S KAWAMATA, T YAMADA, Y TANAKA, P SRIPRASERTSAK, H KATO, Y ICHINOSE, H KATO, T SHIRAISHI, H OKU

    PLANT MOLECULAR BIOLOGY   20 ( 1 )   167 - 170   1992年10月

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    記述言語:英語   出版者・発行元:KLUWER ACADEMIC PUBL  

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  • Phenylalanine ammonia-lyase genes from Pisum sativum : Structure, organ-specific expression and regulation by fungal elicitor and suppressor(共著)

    Plant Cell Physiology   33 ( 6 )   715 - 725   1992年9月

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    記述言語:英語  

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  • AN ENDOGENOUS SUPPRESSOR OF THE DEFENSE RESPONSE IN PISUM-SATIVUM

    K NASU, T SHIRAISHI, H YOSHIOKA, N HORI, Y ICHINOSE, T YAMADA, H OKU

    PLANT AND CELL PHYSIOLOGY   33 ( 5 )   617 - 626   1992年7月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Healthy pea plants contain a substance, tentatively called "endogenous suppressor", which specifically suppresses the accumulation of pisatin in pea plants that is induced by treatment with CuCl2 or an elicitor from Mycosphaerella pinodes. This suppressor elicits the accumulation of phytoalexins in other legumes, such as kidney bean, soybean and cowpea. The endogenous suppressor functions to delay the accumulation of pisatin, the activation of phenylalanine ammonialyase (PAL) and the accumulation of mRNAs for PAL and chalcone synthase induced by the elicitor from M. pinodes. The substance specifically induces susceptibility to nonpathogens, such as Mycosphaerella ligulicola and M. melonis, in pea out of four species of legume tested, but the effect is not cultivar-specific. Thus, the endogenous suppressor in healthy pea plants suppresses a series of self-defense reactions and induces susceptibility in pea plants in a species-specific manner, being similar to the exogenous fungal suppressor from the pea pathogen, M. pinodes.

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  • 2-SUPPRESSORS, SUPPRESCINS-A AND SUPPRESCINS-B, SECRETED BY A PEA PATHOGEN, MYCOSPHAERELLA-PINODES

    T SHIRAISHI, K SAITOH, HM KIM, T KATO, M TAHARA, H OKU, T YAMADA, Y ICHINOSE

    PLANT AND CELL PHYSIOLOGY   33 ( 5 )   663 - 667   1992年7月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    Two mucin-type glycopeptides that suppressed the production of pisatin, a phytoalexin of pea, were purified from a pea pathogen, Mycosphaerella pinodes. The structures of Supprescin A (Mr, 452) and Supprescin B (Mr, 959) were determined by an analysis of amino acid sequences and C-13- and H-1-NMR.

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  • Regulation of phosphoinositide metabolism in pea plasma membranes by elicitor and suppressor from a pea pathogen, Mycosphaerella pinodes(共著)

    Plant Cell Physiology   33 ( 4 )   445 - 452   1992年6月

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    記述言語:英語  

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  • ENDOGENOUS ELICITOR PRESENT IN BARLEY-SEEDS .2. MECHANISMS OF THE INDUCTION OF THE RESISTANCE IN BARLEY LEAVES TO ERYSIPHE-GRAMINIS

    T HIRAMOTO, R TOBIMATSU, T SHIRAISHI, T YAMADA, Y ICHINOSE, H OKU

    JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT   135 ( 2 )   167 - 176   1992年6月

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    記述言語:英語   出版者・発行元:BLACKWELL WISSENSCHAFTS-VERLAG GMBH  

    Endogenous elicitor(s) present in barley seeds induce the accumulation of antifungal substances effective against powdery mildew fungi. Antifungal substances induced by the treatment with barley seeds extract coincides with the one induced by the inoculation with compatible or incompatible races of powdery mildew fungi on thin-layer chromatography analysis. The resistance induced by the treatment with barley seeds extract correlates to the accumulation of the antifungal substances in barley leaves. Furthermore, the activity of phenylalanine ammonia-lyase was also induced by the treatment with barley seeds extract.

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  • TRANSIENT EXPRESSION IN ELECTROPORATED PEA PROTOPLASTS - ELICITOR RESPONSIVENESS OF A PHENYLALANINE AMMONIA-LYASE PROMOTER

    T HASHIMOTO, T YAMADA, A TADA, S KAWAMATA, Y TANAKA, P SRIPRASERTSAK, Y ICHINOSE, H KATO, S IZUTSU, T SHIRAISHI, H OKU, Y OHTSUKI

    PLANT CELL REPORTS   11 ( 4 )   183 - 187   1992年5月

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    記述言語:英語   出版者・発行元:SPRINGER VERLAG  

    High yields of viable pea protoplasts were produced from suspension cultured cells and the conditions for the optimum transient expression of the chloramphenicol acetyltransferase (CAT) gene fused to the CaMV 35S promoter after electroporation were investigated. Conditions for elicitor induction of a member of the phenylalanine ammonia-lyase (PAL) gene family in pea was also investigated using a chimeric gene carrying 480 bp of the putative promoter region of gPAL1 connected to bacterial cat gene and nos terminator. CAT activity was considerably induced by the treatment with fungal elicitor (&gt; 100-mu-g/ml glucose equivalent) isolated from Mycosphaerella pinodes, a pea pathogen.

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  • MOLECULAR-CLONING OF CHALCONE SYNTHASE CDNAS FROM PISUM-SATIVUM

    Y ICHINOSE, S KAWAMATA, T YAMADA, CC AN, T KAJIWARA, T SHIRAISHI, H OKU

    PLANT MOLECULAR BIOLOGY   18 ( 5 )   1009 - 1012   1992年3月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)   出版者・発行元:KLUWER ACADEMIC PUBL  

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  • Orthovanadate suppresses on accumulation of phenylalanine ammonia-lyase mRNA and chalcone synthase mRNA in pea epicotyls that is induced by an elicitor from Mycosphaerella pinodes(共著)

    Plant Cell Physiology   33 ( 2 )   201 - 204   1992年3月

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    記述言語:英語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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  • Molecular cloning of phenylalanine ammonia-lyase cDNA from Pisum sativum 共著

    Plant Molecular Biology   20,167-170   1992年

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  • Suppression of the activation of chitinase and β-1,3-glucanase in pea epicotyls by orthovanadate and a suppressor from Mycosphae-rella pinodes 共著

    Annals of the Phytopathological Society of JAPAN   58,405-410   1992年

  • Suppression of the activation of chitinase and β-1,3-glucanase in pea epicotyls by orthovanadate and a suppressor from Mycosphae-rella pinodes 共著

    Annals of the Phytopathological Society of JAPAN   58,405-410   1992年

  • An endogenous suppressor of the defense response in Pisum sativum 共著

    Plant Cell Physiology   33,617-626   1992年

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  • Endogenous elicitor present in barley seeds II. Mechanisms of the induction of the resistance in barley leaves to Erysiphe graminis 共著

    Journal of Phytopathology   135,167-176   1992年

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  • A reporter gene expression under the control of a pea phenylalanine ammonia-lyase gene promoter(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   80,51-60   1992年

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  • TWO suppressors, suppresciws A and B secreted by a pea pathogen, Mycosphaerella pinodes 共著

    Plant Cell Physiology   33,663-667   1992年

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  • TWO suppressors, suppresciws A and B secreted by a pea pathogen, Mycosphaerella pinodes 共著

    Plant Cell Physiology   33,663-667   1992年

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  • Molecular cloning of chalcone synthase cDNA frow Pisum sativum 共著

    Plant Molecular Biology   18,1009-1012   1992年

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  • A reporter gene expression under the control of a pea phenylalanine ammonia-lyase gene promoter(共著)

    Scientific Reports of the Faculty of Agriculture Okayama University   80,51-60   1992年

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  • Transient expression in electroporated pea protoplasts : Elicitor responsiveness of a phenylalanine ammonia-lyase promoter 共著

    Plant Cell Reports   11,183-187   1992年

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  • Inhibition of ATPase in pea plasma membranes in situ by a suppressor from a pea pathogen, Mycospharella pinodes 共著

    Plant Cell Physiology   32 ( 7 )   1067 - 1075   1991年10月

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    記述言語:英語  

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  • エンドウ動的遺伝子の発現機構. 招待

    山田哲治, 白石友紀, 一瀬勇規, 奥 八郎, 川又伸治, 田中良和, Spirasertsak, P, 安 成才, 吉岡博文, 橋本忠明

    生物相互間の認識機構   27   56-65   1991年

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    記述言語:日本語   掲載種別:記事・総説・解説・論説等(学術雑誌)  

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  • SUPPRESSION OF PISATIN PRODUCTION AND ATPASE ACTIVITY IN PEA PLASMA-MEMBRANES BY ORTHOVANADATE, VERAPAMIL AND A SUPPRESSOR FROM MYCOSPHAERELLA-PINODES

    H YOSHIOKA, T SHIRAISHI, T YAMADA, Y ICHINOSE, H OKU

    PLANT AND CELL PHYSIOLOGY   31 ( 8 )   1139 - 1146   1990年12月

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    記述言語:英語   出版者・発行元:JAPANESE SOC PLANT PHYSIOLOGISTS  

    A pea pathogen, Mycosphaerella pinodes, secretes both an elicitor and a suppressor for the accumulation of pisatin, a major phytoalexin of pea, into the spore germination fluid. The effects of elicitor and the suppressor on the ATPase activity in pea plasma membranes was examined. The ATPase was sensitive to orthovanadate and dicyclohexylcarbodiimide but insensitive to nitrate and azide; it was unaffected by the elicitor but was markedly inhibited by the suppressor (50-mu-g.ml-1, bovine serum albumin equivalents) or verapamil (100-mu-M). The accumulation of pisatin induced by the elicitor was delayed for 3 to 6 h in the presence of orthovanadate or verapamil to an extent similar to that in the presence of the suppressor. The relationship between the inhibition of plasma membrane ATPase activity and the suppression of the active defense reaction that involves the production of pisatin in the pea plant is discussed.

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  • Identification of mouse endodermal cytoskeletal protein C (endo C) with cytokeratin no. 19 共著

    Biochemical and Biophysical Research Communications   167 ( 2 )   644 - 647   1990年3月

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    担当区分:筆頭著者   記述言語:英語  

    DOI: 10.1016/0006-291X(90)92073-9

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  • Suppression of pisatin production and ATPase activity in pea plasma membranes by orthovanadate, verapamil and a suppressor from Mycosphaerella pinodes 共著

    Plant Cell Physiology   31,1139-1146   1990年

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  • MOLECULAR-CLONING AND CHARACTERIZATION OF CDNA-ENCODING MOUSE CYTOKERATIN-NO-19

    Y ICHINOSE, K HASHIDO, H MIYAMOTO, T NAGATA, M NOZAKI, T MORITA, A MATSUSHIRO

    GENE   80 ( 2 )   315 - 323   1989年8月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/0378-1119(89)90295-3

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  • Molecular cloning and characerization of cDNA encoding for mouse cytokeratin no. 19 共著

    Gene   80,315-323   1989年

  • NUCLEOTIDE-SEQUENCE AND STRUCTURE OF THE MOUSE CYTOKERATIN ENDOB GENE

    Y ICHINOSE, T MORITA, FY ZHANG, S SRIMAHASONGCRAM, MLC TONDELLA, M MATSUMOTO, M NOZAKI, A MATSUSHIRO

    GENE   70 ( 1 )   85 - 95   1988年10月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/0378-1119(88)90107-2

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  • Nucleotide sequence of mouse endoA cytokeratin cDNA reveals a poly-peptide characterized to the type II keratin subfamily 共著

    Gene   68 ( 1 )   109 - 117   1988年8月

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  • Nucleotide sequence and structure of a mouse cytokeratin endoB gene 共著

    Gene   70,85-95   1988年

  • Regulation of pisatin biosynthesis in pea leaves by elicitor and suppressor produced by Mycosphaerella pinodes 共著

    Annals of the Phytopathological Society of JAPAN   52,53-58   1986年

  • Regulation of pisatin biosynthesis in pea leaves by elicitor and suppressor produced by Mycosphaerella pinodes 共著

    Annals of the Phytopathological Society of JAPAN   52,53-58   1986年

▼全件表示

Works(作品等)

  • Flagellar motility and structure in phytopathogenic bacteria

    2004年

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  • Glycosyl structure of flagellin in phytopathogenic bacteria

    2001年

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  • 植物病原細菌のべん毛糖鎖構造の解析

    2001年

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  • Molecular study of plant defense genes

    1996年

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  • 植物の生体防御遺伝子の分子生物学的解析

    1996年

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受賞

  • 平成21年度日本植物病理学会 学会賞受賞

    2009年  

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    受賞国:日本国

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  • 日本植物生理学会論文賞

    1994年  

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    受賞国:日本国

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共同研究・競争的資金等の研究

  • 植物病原細菌のGac/Rsm菌体密度感知機構による病原性関連遺伝子の発現制御機構

    研究課題/領域番号:19H02956  2019年04月 - 2022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)

    一瀬 勇規, 松井 英譲

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

    Pseudomonas syringaeなど多くの植物病原細菌では菌体密度感知分子としてN-アシルホモセリンラクトン(AHL)を生産し、AHLの濃度により細菌密度を検知し、病原力関連遺伝子を発現すると言われているが、本研究代表者は、P. syringae pv. tomato DC3000 (PtoDC3000) など多くのP. syringae分離株のAHL合成酵素遺伝子やAHL結合性転写因子遺伝子が変異しておりAHLを生産しないことから、AHLを介しない新たな菌体密度感知システムが存在していると推察した。これまでに高菌体密度のPtoDC3000はタンパク質をコードしないsmall RNAであるrsmXやrsmYの発現を高めること、これらの発現はGacS/GacA二成分制御系の支配下にあることを明らかにした。PtoDC3000のrsmX1, rsmX2, rsmX3, rsmX4, rsmX5, rsmY, rsmZのそれぞれを欠損したシングル変異株を作出して運動能、病原力などを比較解析した。その結果、rsmX3, rsmX4, rsmX5の各変異株ではswimming運動能、バイオフィルム形性能、GABA利用能、病原力について若干低下する傾向が見られた。また、AHLを生産するPtoDC3000を作出したが、シロイヌナズナに対する病原力の低下は観察されなかった。また、GacS/GacA二成分制御系の制御に関わることが知られているretS, ladSの欠損変異株を作出したので、その表現型を解析しているところである。rsmX2のプロモーターにレポーター遺伝子を連結させたトランスポゾンを転移させてrsmX2の発現が低下する変異株のスクリーニングを行なったが、これまでのところ明確に低下した変異株は得られていない。

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  • 植物病原細菌多剤排出ポンプの病原力としての役割

    研究課題/領域番号:16K14861  2016年04月 - 2019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

    一瀬 勇規, 松井 英譲

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    配分額:3770000円 ( 直接経費:2900000円 、 間接経費:870000円 )

    Pseudomonas syringae pv. tabaci (Pta) 6605の主要な多剤排出ポンプトランスポーターMexAB-OprMとMexEF-OprNの病原力としての役割を解析した。mexB変異株,mexF変異株,mexBmexFの2重変異株はアセトバニロン,カテコール,クマリンにより増殖が抑制され,タバコに対する病原力が低下した。これらの結果は,MexAB-OprMとMexEF-OprNがアセトバニロン,カテコール,クマリンを排出し,Pta6605の病原力発現において重要な役割を担っていることを示している。

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  • 菌体密度感知機構によるT3SS遺伝子発現制御機構の解析

    研究課題/領域番号:15H04458  2015年04月 - 2018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)

    一瀬 勇規

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    配分額:16640000円 ( 直接経費:12800000円 、 間接経費:3840000円 )

    タバコ野火病菌(Pta)の菌体密度感知機構転写因子(PsyR)の組換えタンパク質はアシルホモセリンラクトン(AHL)不在下でAHL合成酵素遺伝子プロモーターに結合し、AHL存在下で転写を増高させる正の転写制御因子であることを明らかにした。また、PsyRはタイプIII 分泌機構の転写制御遺伝子であるhrpL遺伝子プロモーターに存在する非完全逆方向反復配列に結合することを明らかにした。Pta及びAHL を生産しないトマト斑葉細菌病菌では、高菌体密度でnoncoding small RNAであるrsmX, rsmYが高発現し、Gac/Rsmシグナル経路の重要性が推察された。

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  • 植物病原細菌の化学物質による感染行動制御機構の解析

    研究課題/領域番号:26660035  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

    一瀬 勇規

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

    植物病原細菌Pseudmonas syringaeには45前後の走化性センサータンパク質(mcp)遺伝子が存在しており、多くの化合物に対して走化性応答を示す可能性が高い。まず、高い運動能を有するP. s. pv. tabaci 6605がカイネチンなど10の化合物に対し正の走化性を示すことを明らかにした。次に植物病原性Pseudomonadに特有な27mcp遺伝子について、各欠損変異株を作出したところ、mcp4, mcp15b, mcp24の欠損変異株ではカイネチンに対する応答性が低下した。これらの遺伝子のコードするMCPはカイネチンに対するセンサータンパク質である可能性が示唆された。

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  • 植物病原細菌における病原性遺伝子発現制御機構のグローバルネットワークの解明

    研究課題/領域番号:24380028  2012年04月 - 2015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)

    一瀬 勇規

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    配分額:19240000円 ( 直接経費:14800000円 、 間接経費:4440000円 )

    Pseudomonas syringaeの運動能を担うべん毛とタイプIV線毛はそれぞれ菌体密度感知分子であるアシルホモセリンラクトン(AHL)とcAMPの合成に必要であり、AHLの合成には転写因子AefRとPsyRが、cAMPの合成には転写因子Vfrを必要とした。前者の欠損は多剤排出ポンプ遺伝子mexEF/oprNの、後者の欠損はmexAB/oprMの発現を増高させ、薬剤耐性能が高まった。PsyRとVfrはhrp遺伝子の発現制御にも関わることが判明した。

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  • 青枯病菌のタンパク質分解酵素エフェクターによる過敏感反応誘導と罹病化機構の解明

    研究課題/領域番号:24658042  2012年04月 - 2014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

    一瀬 勇規, 向原 隆文

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    配分額:4160000円 ( 直接経費:3200000円 、 間接経費:960000円 )

    ナス科植物青枯病菌が分泌する病原性因子エフェクターの中から,本菌の非宿主で台木ナスのトルバムビガーに細胞死を伴う激しい防御応答(HR)を誘導するRip36を同定した.Rip36はZnプロテアーゼモチーフを有するが,このモチーフに変異を導入するとHR誘導能を失ったことから,Rip36はトルバムビガーの標的タンパク質を分解することでHRを誘導したと考えられた.一方,rip36欠損変異株の宿主ナスにおける増殖能は野生株と変わらず,ナスに対する病原性に必須の因子ではないことが判明した.

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  • MAMP認識に関わるシロイヌナズナ受容体様キナーゼの網羅的解析

    研究課題/領域番号:21380030  2009年 - 2011年

    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)

    一瀬 勇規, 稲垣 善茂

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    配分額:18850000円 ( 直接経費:14500000円 、 間接経費:4350000円 )

    微生物(あるいは病原体)由来特有分子パターンとして、細菌の非メチル化DNAを同定した。病原体由来分子パターンとしてはハーピンの認識機構に取り組み、タバコ野火病菌は、タバコが獲得した高度なハーピン応答性防御機構を回避すべく、ハーピン遺伝子に内部欠損を加え、タバコの認識と防御応答を避けることにより病原性を発揮することを逆遺伝学的に明らかにしたが、シロイヌナズナにおける細菌DNAやハーピンの受容体分子の同定には至らなかった。

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  • 植物病原細菌の病原性糖タンパク質糖鎖の構造解析と病害防除への利用

    2007年 - 2011年

    新技術・新分野創出のための基礎研究推進事業 

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    資金種別:競争的資金

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  • 植物の葉における斑入りと環境適応

    研究課題/領域番号:19657017  2007年 - 2008年

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    坂本 亘, 一瀬 勇規

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    配分額:3500000円 ( 直接経費:3500000円 )

    本研究では、植物における斑入り突然変異が個体レベルでの光合成としては負に働くか、環境適応に有利に働く場合があることを実証する実験をモデル植物で試みた。特に、環境適応の1つとして斑入りが「病害抵抗性」を示す可能性について萌芽的研究を行った。これまでの研究で、代表者らが解析を進めているシロイヌナズナの斑入り変異var2では光化学系IIの維持に必要な葉緑体プロテアーゼが欠損しており、その結果として葉緑体に高レベルの活性酸素を蓄積することを明らかにしている。昨年度は、これらの活性酸素が病原細菌への抵抗性に関与するかどうかをP. syringae pv tomato DC3000を用いて調べ、野生型に比べ、病原細菌の増殖が抑制されることが明らかとなった。今年度は、これらの抵抗性の原因として斑入り変異では自然免疫の活性化が起きている可能性、葉緑体の活性酸素自身がDC3000への抵抗性に寄与する可能性について調べた。自然免疫活性化の指標となるPR1遣伝子の発現上昇がvar2の葉で観察されず、またこれらの活性化に関わるシグナル因子であるサリチル酸の上昇も見られなかった。他の遺伝子発現の変化もマイクロアレイ等で解析したところ、やはり自然免疫活性化に関する遺伝子発現上昇を支持する結果は得られなかったが、var2の斑入り葉では活性酸素消去系に関わる遺伝子発現が上昇していることが明らかとなった。特に斑入りの白色組織でSODなどの消去系酵素が働いており、抵抗性に寄与する可能性も示唆された。一方で、斑入りを示さない光化学系II酸素発生系複合体に関する突然変異体でも活性酸素が蓄積し、var2よりは弱いがDC3000に抵抗性を示す結果が得られたことから、葉緑体の活性酸素が自然免疫系を活性化するよりもむしろ、葉緑体で蓄積する活性酸素が直接病原細菌に毒性を示す可能性が示唆された。

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  • 植物病原細菌のフラジェリンによる植物免疫の活性化と病原菌によるその回避機構

    研究課題/領域番号:18380035  2006年 - 2008年

    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)

    一瀬 勇規

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    配分額:17920000円 ( 直接経費:15100000円 、 間接経費:2820000円 )

    植物病原細菌Pseudomonas syringae pv. tabaci(Pta)の鞭毛構成タンパク質フラジェリンは非宿主植物に対し激しい防御応答(HR)を誘導するが、宿主タバコには顕著には誘導しない。本研究ではフラジェリン分子中のHR誘導ドメインがアミノ末端側の保存配列(flg22)に存在することを見出した。また、フラジェリンに結合するラムノースや修飾ビオサミンなどの糖鎖は、鞭毛の安定性を高めた。従って、糖鎖にはflg22の露出を防ぐことで防御応答を回避する機能が推測された

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  • 植物オルガネラ間相互作用による異物認識機構に関する分子解析

    研究課題/領域番号:15108001  2003年 - 2007年

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

    白石 友紀, 一瀬 勇規, 稲垣 善茂, 豊田 和弘

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    配分額:109720000円 ( 直接経費:84400000円 、 間接経費:25320000円 )

    感染サイクルの大半を細胞外(植物表面や細胞間隙)で営む病原糸状菌や細菌をモデルとして、分子パターン認識と病原性エフェクターの作用機構について解析してきた(H15〜18年度)。今年度は、糸状菌由来エフェクター(サプレッサー)の分子標的である宿主植物の細胞壁アピラーゼの相互作用分子の解析を進め、TOF/MS解析から複数の情報伝達や酸化還元関連分子の存在を示した。また、アピラーゼで生成するリン酸は細胞壁に構成的に存在するペルオキシダーゼ(POX)依存性の活性酸素生成を亢進し、さらに一群の細胞外POX遺伝子を転写レベルから活性化することを示した。一方、表層でのパターン認識に続く過敏感細胞死の分子機構について、エリシチンをモデルに解析した結果、細胞周期(M期)制御系の構成因子NbCdc27Bは過敏感細胞死には直接関与しないが、抵抗性機能発現にそのC末端領域が必要である可能性が示唆された。また、細菌由来分子パターン(フラジェリン)による下流の情報伝達・遺伝子応答についてマイクロアレーで解析し、植物固有の転写因子WRKY41を同定した。WRKY41はフラジェリン処理で急速に発現が誘導されるが、エフェクターにより発現が抑えられる。さらに、WRKY41高発現体ではPseudomonas syringaeに対する抵抗性が増高したが、逆にErwinia carotovoraに対する感受性は高まった。この結果は、WRKY41はSA系に対して正に作用していることを示す。しかし、アピラーゼ高発現体の解析から、SA経路やJA経路とは異なる情報伝達系の存在も示されている。以上から、細胞壁には植物独自の異物認識機構が存在し、細胞壁始発のシグナルは感染防御を担う様々な細胞小器官へ情報伝達され、固有の細胞内因子の活性化あるいはフィードバック機構(増幅)を介して、最終応答(抵抗性発現)が制御されているものと考察した。

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  • 植物の罹病時特異的発現遺伝子の制御機構の解析とその利用に関わる基礎研究

    研究課題/領域番号:14656019  2002年 - 2003年

    日本学術振興会  科学研究費助成事業 萌芽研究  萌芽研究

    一瀬 勇規, 稲垣 善茂

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    配分額:3200000円 ( 直接経費:3200000円 )

    エンドウ褐紋病菌の生産するサプレッサーは、エリシターによって誘導されるエンドウの防御応答を特異的に抑制あるいは遅延させる。本病原性因子サプレッサーの親和性確立における機能を解析するために、エンドウにおけるサプレッサー応答性遺伝子のcDNAの単離を行い、クローンS64が既に同定されている。S64の遺伝子産物はジャスモン酸合成経路の12-oxophytodienoic acid reductase(OPR)と高い相同性を示す。そこでジャスモン酸合成系の他の酵素であるallene oxide synthase(AOS)とallene oxide cyclase(AOC)をコードする遺伝子のcDNAを単離し、ノーザン法、RT-PCR法によりエリシター・サプレッサー応答、病原菌・非病原菌に対する応答を解析した。その結果、AOSの遺伝子発現はサプレッサー処理や病原菌接種により誘導されることが明らかとなった。一方、S64の遺伝子を単離するためにエンドウゲノムDNAライブラリーよりハイブリダイゼーションによりスクリーニングを行うと、S64以外にOPRと推定される遺伝子が計5個単離され、OPRは遺伝子ファミリーを形成していることが明らかにされた。更に対応するcDNAを単離する試みにおいて新たに6個目のOPR cDNAを同定した。合計6種のエンドウOPR遺伝子について、個々の発現解析を行うとともに、個々の組換えタンパク質を生産し、モデル基質である2-cyclohexen-1-one(CyHE)に対する還元活性を解析した。その結果、S64に相当するPsOPR2遺伝子の発現が最も強くサプレッサー処理・病原菌接種に応答し、その組換えタンパク質が最も高いCyHE還元活性を示した。これらのことより6種のエンドウOPRはそれぞれ異なる基質・発現特性を有していると推測された。現在PsORP2の高発現タバコ、シロイヌナズナ並びにPsOPR2のプロモーターをレポーター遺伝子に繋いだ形質転換植物体を作出している。

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  • 植物の感染応答に伴うプログラム細胞死の制御機構

    研究課題/領域番号:12052215  2000年 - 2005年

    日本学術振興会  科学研究費助成事業 特定領域研究  特定領域研究

    眞山 滋志, 朴 杓允, 一瀬 勇規, 秋光 和也

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    配分額:68200000円 ( 直接経費:68200000円 )

    本年度の研究成果の主要なものは下記の通りである。
    1)セルフリー系を用いた植物アポトーシスの分子機構解析
    単離核を用いたセルフリー系によるエンバクのアポトーシス様細胞死の分子機構の解析を行い、エンバクのDNAラダー化に関与すると考えられるDNaseII様のヌクレアーゼP28を同定した。また、これに関与するプロテアーゼは、カスペースおよびグランザイム特異的な阻害剤によって活性が阻害されない、システインプロテアーゼであることが示された。また、エンバクのアポトーシス様細胞死の過程で、細胞質、ミトコンドリア、葉緑体由来の各種rRNAが分解されることを植物細胞で初めて明らかとした。
    2)過敏感反応のシグナル伝達系の解析
    植物に過敏感細胞死を誘導する複数種の病原菌由来タンパク質性エリシターを用い、過敏感反応シグナル伝達系を解析した。また、新規過敏感反応誘導性エリシターとしてP.syringaeのべん毛繊維タンパク質であるフラジェリンを同定した。更に、植物の防御応答の抑制に関わることが予想される遺伝子としてオクタデカノイド代謝系のOPR遺伝子を単離し、その機能を解析した
    3)宿主特異的毒素,ACR毒素の作用機序の解明
    本研究者らは、宿主特異的ACR毒素の第一次作用点は宿主細胞ミトコンドリアにあり、この毒素のレセプター探索を進め、ドーパミン、グルタミン酸、黄体形成ホルモンレセプター等と部分的配列類似性を示すミトコンドリアゲノム遺伝子ACRSを単離した。さらに、ACRS遺伝子を介した宿主特異性決定機構の解析を進展させ、毒素への感受性/抵抗性は本遺伝子転写物へのプロセッシングの有無により制御されることを明らかにした。

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  • 植物防御応答遺伝子の発現を誘導する転写因子の解析

    研究課題/領域番号:12460023  2000年 - 2002年

    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)

    一瀬 勇規, 豊田 和弘

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    配分額:13200000円 ( 直接経費:13200000円 )

    本研究ではエリシター処理エンドウから単離された新規転写因子E84(ERDP)について解析を進めた。E84の遺伝子産物はアミノ末端付近に植物特有のDof DNA結合タンパク質ドメインを有する転写因子である。大腸菌で発現させたE84組換えタンパク質はAAAG配列を有するDNA断片に結合することがランダム結合選別法により明らかにされた。また、E84結合配列の一つBS4/37(TCATTTAAAGTGTTTT)をモデルとしてAAAG前後の配列に変異を導入して結合実駿を行ってみるとAAAGの3'側のTGT配列も高い結合親和性のために必要な配列であることが明らかとなった。また、エリシター応答性のPsCHS1遺伝子のプロモーターとの結合実験を行ってみたところ、AAAG配列を有する断片に結合した。一方、E84結合配列BS4/37を4つ直列に連結し、CAMV 35Sプロモーターの最小単位に連結させ、エンドウプロトプラストを用いたトランジェントトランスフェクションアッセイを行った。その結果、エリシター処理によりレポーターの活性が高まり、AAAG配列の直列繰り返し配列がエリシターによる転写活性化に関与することが示された。また、E84タンパク質はE84遺伝子自身のプロモーター配列にも結合することより、正のオートレギュレーションを行っている可能性も示唆された。以上のことはE84タンパク質及びその結合配列がエリシターによる転写活性化に貢献している可能性を示している。また、E84以外にDofドメインをアミノ末端付近に有する新規Dof遺伝子を6種単離し、エンドウでDof遺伝子がファミリーを形成していることが示された。これらの知見は植物防御応答時における様々な防御遺伝子の転写活性化の解明に繋がるものと考えている。

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  • 植物病原菌由来のシグナル物質により発現が変動する宿主遺伝子の解析

    研究課題/領域番号:09660051  1997年 - 1999年

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

    一瀬 勇規, 白石 友紀

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    配分額:3300000円 ( 直接経費:3300000円 )

    本研究では、植物病原菌由来のシグナル物質(エリシター・サプレッサー)によって発現が変動する植物遺伝子を数多く単離し、発現のカイネティクスを解析するとともに、遺伝子産物の宿主-病原菌相互作用における機能を解明することを目的とした。
    その結果、エリシター応答性遺伝子として、既知のPRタンパク質遺伝子や細胞壁の強化に関わる遺伝子、ファイトアレキシン合成系酵素遺伝子のcDNAの他、転写因子のcDNAを始め、ポリアミン合成系酵素のcDNA、HRに関連すると報告されているタンパク質のcDNAなど多数の新規遺伝子が単離された。これらのうちいくつかの遺伝子については更に遺伝子産物の機能解析を進めている。特にエリシター応答性の転写因子についてはそのDNA結合配列を明らかにし、本結合配列が既知の防御遺伝子プロモーターに存在することを見い出した。本転写因子はエリシターによりその発現が誘導されることから、病原菌シグナルの受容から防御遺伝子発現に至るカスケードにおいて極めて重要な分子であると考えている。
    また、サプレッサーによって発現が誘導される遺伝子としては、細胞壁の分解酵素遺伝子、根粒形成初期に発現されるノジュリン遺伝子、ジャスモン酸合成系酵素のホモログなどが単離された。これら遺伝子の中でジャスモン酸合成系酵素のホモログは実際に病原菌接種の場においても特異的に発現が誘導されることが明らかにされた。植物が罹病化していく過程で発現が誘導される遺伝子の同定は、植物罹病化の分子機構解明に繋がるもの重要な成果と考えている。

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  • リゾチーム遺伝子を導入した青枯れ病抵抗性トランスジェニック・ナス科作物の育種

    研究課題/領域番号:09356002  1997年 - 1999年

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

    山田 哲治, 一瀬 勇規, 田原 誠, 小澤 英徳, 田中 博

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    配分額:17600000円 ( 直接経費:17600000円 )

    ナス科植物の青枯れ病・立枯病の防除には、化学的な方法、耕種的な方法および抵抗性品種の導入などが用いられてきたが、被害を皆無にするような防除方法は確立されていない。そこで本研究では、タバコ立枯病細菌感染ファージP4282からリゾチーム遺伝子を同定・単離し、さらに申請者らが長年に渡って研究してきた根組織特異的に発現するエンドウ・PAL(フェニールアラニンアンモニアリアーゼ)遺伝子のプロモータを利用し、ファージ・リゾチーム遺伝子とPALプロモータを連結したキメラ遺伝子をトマト・タバコ植物体に導入して、青枯れ・立枯れ病抵抗性トランスジェニック植物体を作出することを目的としたものである。
    本研究の成果として特に以下の内容は意義が大きいものと考えている。
    1.タバコ立枯病細菌に感染するファージP4282からリゾチームタンパク質を単離精製し、対応する遺伝子の単離、構造解析を行った。
    2.エンドウのPAL1プロモーター中の発現制御シスエレメント(Box I,Box II,Box IV)をタンデムにリピートさせた高発現型強力プロモーターを開発した。本プロモーターは特に根にでCaMV35Sプロモーターの6.5倍の高い発現を示した。
    3.恒常的に高発現するCaMVの35Sプロモーター、エンドウPAL1プロモーター及び上記リピートプロモーターとリゾチーム遺伝子を融合し、タバコに導入して形質転換体を作出した。
    4.上記のリゾチーム遺伝子を発現させた形質転換タバコでは高い溶菌活性を確認した。
    5.PAL1プロモーターとGUS遺伝子の融合遺伝子を組込んだ形質転換タバコに、LUX遺伝子を導入した青枯れ病菌を接種することにより病原菌の挙動と植物の防御応答を同時に解析できるモニタリングシステムを確立した。

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  • ストレス応答遺伝子とアポトーシス関連遺伝子の器官特異的発現と環境シグナル応答

    研究課題/領域番号:09251211  1997年

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    山田 哲治, 一ノ瀬 勇規

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    配分額:2000000円 ( 直接経費:2000000円 )

    色素生産やストレス応答において、特に重要な役割を担うフェニールプロパイド二次代謝合成系の鍵酵素をコードするPALおよびCHS遺伝子をモデルとして、多重遺伝子ファミリーの各メンバーの植物各器官・組織特異的発現に関わる制御領域を同定するとともに、鍵となる制御タンパク質をコードする遺伝子を単離して、フェニールプロパイド二次代謝経路を支配する遺伝的制御プログラムを分子生物学的に解析すること、また、植物の病原体攻撃に対する植物の典型的な防御機構に、過敏感細胞死(Hypersensitive Cell Death:HR)において物のアポトーシス現象を正および負に調節する遺伝子を探索し、これらの調節因子が、植物の器官形成にどのような役割を果たしているのかを明らかにすることを狙いとしている。
    環境シグナル及び器官特異的発現に関与するシス領域のLM-PCRによる解析
    病原菌エリシター,あるいはサプレッサーによる遺伝子発現誘導応答に必要と考えられる重要なシス因子の候補に関して、PSPAL-1,-2およびPSCHS-1,-2を中心にLM(Ligation mediated)-PCR footprinting解析を行ったところ、エリシター処理によるこれらの遺伝子のプロモータの活性化にはACに富むシ-クネンスボックス(Box l)が関与していることを示す顕著なfootprintが確認された。現在、器官特異的に発現に関与する特異的なfootprintおよびサプレッサー処理による遺伝子発現抑制に関与するfootprint解析を行っている。
    アポトーシス関連遺伝子のクローニング
    タバコBY-2細胞を用いて、Activation taggingを行い、エリシター処理に対して過敏感細胞死を遅延したり、起こさなくなった変異体細胞を得た。これらの細胞からplasmid rescueを行い、タバコ・ゲノミックDNAの挿入を確認したところ、ひとつのカルス集団には約5種類のタバコ・ゲノミックDNAが挿入されていた。そこでDNA断片を再度、組換えプラスミドに挿入し、タバコBY-2細胞に遺伝子挿入したところ、一つのラインでエリシター処理しても細胞死を起こさない変異体が出現した。現在本DNAの塩基配列決定を行っている。

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  • フェニールプロパノイド合成系鍵酵素遺伝子の器官特異的発現と環境シグナル応答

    研究課題/領域番号:08262215  1996年

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    山田 哲治, 一瀬 勇規

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    配分額:2500000円 ( 直接経費:2500000円 )

    1.遺伝子改変プロモータを用いたLoss of functionの実験結果から、PSPAL1遺伝子5′-プロモータ領域に含まれるエリシターよる発現誘導応答に必要なシス因子として、box 1(L-box)およびbox 2(P-box)があげられる.このことはbox 2とbox4を4-8ユニット繰り返し配列で繋いだキメラ・プロモータを用いたGain function実験からも更に裏付けられた.トランジェントアッセイの解析に加え、box 1(L-box),box 2(P-box),box 4(AT-box)各ボックスシークエンスを除去した遺伝子改変プロモータをGUSレポーター遺伝子に連結したpBI系遺伝子組換えプラスミドをタバコに導入し、トランスジェニックタバコにおけるin planta loss of function実験を行った.その結果、これらのボックスシークエンスは、エリシターやUV照射による発現誘導応答に必要であるばかりでなく、basalなPSPAL1のプロモータの発現にも影響を及ぼすことが明らかとなった.これらのシス因子は単独ではプロモータの活性化に及ぼす効果が少なく、これらのボックスに結合する調節タンパク質が協調的に働いていると考えられる.
    2.エンドウカルコン合成酵素(CHS)遺伝子は少なくとも8つのメンバーからなる遺伝子ファミリーを形成していることが明らかとなり、各々のメンバーの組織特異的・環境刺激特異的発現パターンを調べたところ、CHS遺伝子ファミリーの8つのメンバーは少なくとも3つのサブファミリーにグループ分けすることができることが明らかとなった。これらのうち2つのエリシター応答性グループのプロモータにはBox-1,G-Box,Box-2配列が保存されており、これらの配列がエリシター応答に重要であることが、塩基置換を導入したプロモータのトランジェント・トランスフェクション・アッセイによっても確認された。また、エリシター等のシグナルによる急激なCHS活性の増加は転写の活性化のみならず、遺伝子ファミリーの各メンバーの協調的な活性化、並びに転写後の調節機構にも基因することが推察された。

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  • フェニールプロパノイド合成系鍵酵素遺伝子の器官特異的発現と環境シグナル応答

    研究課題/領域番号:07270216  1995年

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    山田 哲治, 一瀬 勇規

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    配分額:2700000円 ( 直接経費:2700000円 )

    植物のフェニールプロパノイド二次代謝経路はアントシアニン色素、リグニンを中心とする細胞外皮の構築、紫外線毒の吸収、あるいは病原菌の攻撃に対するファイトアレキシン合成など、多岐にわたる機能を果たしている。エンドウ・フェニールプロパノイド合成系の重要な鍵酵素、フェニールアラニンアンモニア・リアーゼ(PAL)およびカルコン合成酵素(CHS)をコードする遺伝子は多重遺伝子ファミリーを形成しているが、それぞれのメンバーの植物各器官および組織特異的発現、あるいは外的環境要因による遺伝子発現調節機構に関しては未知の部分が多い。本研究では、色素生産やストレス応答において特に重要な役割を担うフェニールプロパノイド二次代謝合成系の鍵酵素をコードするPAL遺伝子、およびCHS遺伝子をモデルとして、多重遺伝子ファミリーの各メンバーの植物各器官・組織特異的発現に関わる制御領域を同定するとともに、鍵となる制御タンパク質をコードする遺伝子を単離して、フェニールプロパノイド二次代謝経路を支配する遺伝的制御プログラムを分子生物学的に解析することを目的としている。以下に成果の概要を列記する。
    1. 遺伝子改変プロモータを用いたLoss of functionの実験結果から、PSPAL1遺伝子5′-プロモータ領域に含まれるエリシターによる発現誘導応答に必要なシス因子として、box1 (L-box) ,box2 (P-box) ,box4 (AT-box)があげられた。このことはbox2とbox4を2-4ユニット繰り返し配列で繋いだキメラ・プロモータを用いたGain of function実験からも更に裏付けられた。これらのシス因子は単独ではプロモータ活性化に及ぼす効果が少なく、協調的に働いていると予想された.
    2. 上記トランジエントアッセイの解析に加え、box1 (L-box) ,box2 (P-box) ,box4 (AT-box)各ボックスシークエンスを除去した遺伝子改変プロモータをGUSレポータ遺伝子に連結したpBI系遺伝子組換えプラスミドをタバコに導入し、トランスジェニックタバコにおけるin planta loss of function実験を行った.その結果、これらのボックスシークエンスは、エリシターやUV照射による発現誘導応答に必要であるばかりでなく、basalなPSPAL1,PSPAL2プロモータの発現にも影響を及ぼすことが明らかとなった.現在、これらのボックスシークエンスの器官・組織特異的発現に果たす役割を検討している。

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  • 病原菌シグナル伝達と植物防御遺伝子応答におけるブラックボックスの分子生物学的解明

    研究課題/領域番号:06404008  1994年 - 1996年

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

    白石 友紀, 一瀬 勇規, 山田 哲治

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    配分額:29300000円 ( 直接経費:29300000円 )

    エンドウ褐紋病菌の生産するエリシターとサプレッサーは防御応答のスイッチのon-offを行うシグナル分子である。これらの受容から遺伝子応答に至る流れの解明を目指し研究を行った。原形質膜における情報伝達解析と並行して、細胞壁画分から、エリシター・サプレッサー感応性の酵素群を検索した結果、ATPase(NTPase)、パーオキシデ-ス(POX)、アスコルビン酸オキシデ-スがエリシターによって活性が増高し、サプレッサーによって種特異的に阻害を受けた。分離細胞壁におけるスーパーオキサイドと感染阻害物質の生産には連携が認められ、病原菌シグナルで制御される事も明らかとなった。さらに、細胞壁にはヴィトロネクチン様蛋白分子が存在し、原形質膜蛋白質とRGD配列を介して連携しており、ファイトアレキシン生産のシグナル伝達を担うことが判ってきた。このように植物細胞壁は病原菌シグナルの認識・受容と変換・増幅の場であると同時に防御応答の最前線であり、細胞壁が種特異性決定に重要な役割を果たす可能性が示唆された。
    病原菌シグナル因子による植物防御遺伝子の発現調節に必要なシス・トランス因子の割り出しを進めた。PAL,CHS遺伝子プロモータにGUSを連結した遺伝子を導入したトランスジェニック植物やトランジェントアッセイにより以下の結果を得た。1)PSPAL1,PSPAL2各遺伝子5′-プロモータ領域のエリシターやUV応答に必要なシス因子の候補はbox1(L-box),box2(P-box),box4(AT-box)である。2)PSPAL1,2、およびPSCHS1,2のすべてのプロモータ領域のAT-rich配列はDNA/タンパク質複合体を形成し、結合には核タンパク質のリン酸化が必要である。3)エリシター応答のカルコン合成酵素遺伝子のプロモータにはBox-1,G-Box,Box-2配列が重要である。

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  • 植物の病原菌に対する防御反応応答と遺伝子発現制御に関する研究

    研究課題/領域番号:06454063  1994年 - 1995年

    日本学術振興会  科学研究費助成事業 一般研究(B)  一般研究(B)

    山田 哲治, 一瀬 勇規

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    配分額:7200000円 ( 直接経費:7200000円 )

    植物と微生物の総合作用における初期認識には、植物の防御反応を誘導する物質、エリシターと、エリシターに拮抗的に働くサブレッサーなどの、菌由来シグナル因子が、病原性・非病原性を決定する要因として非常に重要な役割を担っている。本研究では、これらの病原菌由来シグナル因子による植物防御応答遺伝子の同調した発現調節に必要なシス・トランス因子の割り出し行うため、生化学的・分子遺伝学的手法を用いて解析したものである.以下に実験結果を列記する。
    1.PSPAL1. 2、および PSCHS1, 2のプロモータ領域に存在するATリッチシークエンスはアクテイベータ-として機能し、本ATリッチシークエンスに結合する共通のタンパク質はHMG-I/Yに類似するDNAペンデイングに関与するタンパク質である可能性が大きい。
    2.PSPAL1. PSPAL2遺伝子5'-プロモータ領域に含まれるエリシターによる発現誘導応答に必要なシス因子の候補として特に、box 2 (P-box), box 4 (AT box)が必要であり、さらにbox 1との共存下でその効果が増幅されることが明らかとなった。
    3.トランスジェニックタバコにおけるキメラ遺伝子の発現様式からPSPAL1, PSPAL2プロモータはともに、根・下位茎における組織特異的発現に関与していることが示唆された。また、PSPAL1プロモータは、さらに花弁の色素沈着部における特異的発現に関与していることが明らかとなった。GUSレポーター遺伝子との連結で、非転写融合型(-1,394から-27まで)、転写融合型(-1,394から+5まで、並びに二つのタイプ(type A, -1,394から+115 ; type B, -1,394から+140)の翻訳融合型を構築しGUSの発現レベルを調べたところ、翻訳融合型タイプBで最も発現が高く、続いて翻訳融合型タイプBであったが、転写融合型、非転写融合型では、活性はほとんど認められなかったことから、エキソニックな領域にPSPAL1プロモータの活性化因子が存在するものと考えられる.

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  • エンドウカルコン合成酵素遺伝子の病原菌シグナルによる発現制御機構

    研究課題/領域番号:06760050  1994年

    日本学術振興会  科学研究費助成事業 奨励研究(A)  奨励研究(A)

    一瀬 勇規

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    配分額:1000000円 ( 直接経費:1000000円 )

    エリシターによりエンドウCHS遺伝子の転写誘導に必要なDNAシス領域を同定するために、まず、様々な長さのPSCHS1、PSCHS2遺伝子5´上流域をレポーター遺伝子であるクロラムフェニコールアセチルトランスフェラーゼ(CAT)遺伝子に結合させてキメラ遺伝子を構築した。次に、これらのキメラ遺伝子をエンドウ培養細胞由来のプロトプラストにエレクトロポレーション法で導入して、一時的なCATの活性を測定した。この時エレクトロポレーション後にエリシター処理を行う区と行わない区を設けてエリシター処理による影響も解析した。その結果、PSCHS1では少なくとも-242から下流に、PSCHS2では-377から下流にエリシター応答性転写活性領域が存在することが明らかとなった。更にCaMVの35Sミニマルプロモーターを用いたgain of function実験からPSCHS1の-242から-181までの61塩基だけでエリシターに応答してレポーター遺伝子の発現を増高させることが明らかになった。次に、両遺伝子の転写制御に関わる核内タンパク質因子について、ゲルシフトアッセイ法により解析した。その結果、PSCHS1の-242から-181の61塩基の断片やPSCHS2の-347から-158までの189塩基の断片はエリシター処理エンドウ組織から調製された核タンパク質と強く結合して移動度の遅い複合体(LMC)を形成することが明らかとなった。LMCの形成は核タンパク質を予めアルカリフォスファターゼで前処理することによって阻害されたことから、LMCの形成にはリン酸化が必要であると考えられる。また、エリシターで転写が誘導されるエンドウのPSPAL1とPSPAL2のプロモーターにも同様なLMCを形成する領域が存在しており、それぞれのLMCの形成は互いにLMCを形成するDNA断片により競合することが明らかにされた。以上の結果から、エリシターによる防御遺伝子の転写誘導にはLMCを形成する共通因子が関与している可能性が示唆された。

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  • フェニールプロパノイド合成系鍵酵素遺伝子の器官特異的発現と環境シグナル応答

    研究課題/領域番号:06278214  1994年

    日本学術振興会  科学研究費助成事業 重点領域研究  重点領域研究

    山田 哲治, 一瀬 勇規

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    配分額:2400000円 ( 直接経費:2400000円 )

    植物のフェニールプロパノイド二次代謝経路はアントシアニン色素、リグニンを中心とする細胞外皮の構築、紫外線毒の吸収、あるいは病原菌の攻撃に対するファイトアレキシン合成など、多岐にわたる機能を果たしている。エンドウ・フェニールプロパノイド合成系の重要な鍵酵素、フェニールアラニンアンモニア・リアーゼ(PAL)およびカルコン合成酵素(CHS)をコードする遺伝子は多重遺伝子ファミリーを形成しているが、それぞれのメンバーの植物各器官および組織特異的発現、あるいは外的環境要因による遺伝子発現調節機構に関しては未知の部分が多い。本研究では、色素生産やストレス応答において特に重要な役割を担うフェニールプロパノイド二次代謝合成系の鍵酵素をコードするPAL遺伝子、およびCHS遺伝子をモデルとして、多重遺伝子ファミリーの各メンバーの植物各器官・組織特異的発現に関わる制御領域を同定するとともに、鍵となる制御タンパク質をコードする遺伝子を単離して、フェニールプロパノイド二次代謝経路を支配する遺伝的制御プログラムを分子生物学的に解析することを目的としている。以下に成果の概要を列記する。
    1.Loss of function実験からPSPAL1,PSPAL2各遺伝子5′-プロモータ領域に含まれるエリシターあるいはUV照射による発現誘導応答に必要なシス因子の候補としてbox1(L-box),box2(P-box),box4(AT-box)があげられた。このことはゲル・モビリテイー・シフトアッセイやDNase I-,OP-Cu footprinting analysisによっても確かめられた。
    2.トランスジェニックタバコにおけるキメラ遺伝子の発現様式からPSPAL1,PSPAL2プロモータはともに、根.下位茎における組織特異的発現に関与していることが示唆された。また、PSPAL1プロモータは、さらに花弁の色素沈着部における特異的発現に関与していることが明らかとなった。
    3.PSPAL1,2、およびPSCHS1,2のすべてのプロモータ領域のATリッチシークエンスに結合する共通のタンパク質はHMG-I/Yに類似するDNAベンデイングに関与するタンパク質である可能性が大きい。

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  • 植物低抗性遺伝子の発現制御

    研究課題/領域番号:03044100  1991年

    日本学術振興会  科学研究費助成事業 国際学術研究  国際学術研究

    山田 哲治, KADO Clarenc, 一瀬 勇規, 白石 友紀, 奥 八郎

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    配分額:3000000円 ( 直接経費:3000000円 )

    植物の病害低抗性の中でもファイトアレキシン合成経路の鍵酵素として特に重要な役割を担うフェニルアラニンアンモニアリア-ゼ(PAL)をコ-ドする遺伝子とカルコン合成酵素(CHS)をコ-ドする遺伝子について構造解析を行うとともに、各々の遺伝子にレポ-タ-遺伝子を融合させたキメラ遺伝子を作成し、植物細胞に導入した後、植物病原菌由来のエリシタ-を処理してレポ-タ-遺伝子発現に対する影響を解析した。更にGUSをレポ-タ-遺伝子としたキメラ遺伝子をタバコあるいはエンドウ細胞に導入したトランスジェニック植物体における遺伝子発現様式を調べた。
    1.PAL遺伝子に関する研究
    PAL遺伝子のプロモ-タ-配列にレポ-タ-遺伝子として発光遺伝子(Lux)、βーグルクロニダ-ゼ(GUS)遺伝子、クロラムフェニコ-ルアセチルトランスフェラ-ゼ(CAT)遺伝子を融合したキメラ遺伝子の作成を行った。このうちpBI101をベ-スとしたPALーGUSのキメラ遺伝子はタバコ及びエンドウにバイナリ-ベクタ-法を用いて導入し、形質転換されたトランスジェニックタバコ及びトランスジェニックエンドウカルスを得たが、一部のトランスジェニック植物体でエンドウ褐紋病菌由来エリシタ-や病原菌接種によってGUS活性の増高が見られた。現在顕著なGUS活性を示した個体におけるキメラ遺伝子の染色体DNAへの挿入部位を調べている。一方、カリフォルニア大学デ-ヴィス校においてデュポン社製パ-ティクルガンを用い、各種のキメラ遺伝子を導入し、トランジェットなLux、あるいはGUSの発現を組織レベルで観察したところ、コントロ-ル実験での35SCaMVプロモ-タ-を有するキメラ遺伝子では顕著な活性が一部の細胞で見られたが、PALプロモ-タ-を連絡したキメラ遺伝子の導入ではエリシタ-処理によっても顕著な発現が観察されなかった。また、CAT遺伝子をレポ-タ-遺伝子とした系においては、キメラ遺伝子を主にエンドウの培養細胞由来のプロトプラストにエレクトロポレ-ション法を用いて導入し、トランジェントなCATの発現を測定することによってgPALー1遺伝子の転写調節に必要なシスエレメントの検索を行った。その結果、-340から-140までの上流領域にエリシタ-によって転写誘導を受け、エリシタ-による低抗性の誘導を阻害するオルトバナジン酸(原形質膜ATPア-ゼの阻害剤)により抑制されるエンドウgPALー1遺伝子の発現調節領域が存在することが明らかとなった。
    2.CHS遺伝子に関する解析
    エンドウのCHS遺伝子についてはエリシタ-処理を施したcDNAライブラリ-から、多数のcDNAのクロ-ニングを行い構造の解析によって3種の主要な2種のCHS遺伝子がゲノム上にタンデムに並んで存在していることを明かにし、上述のCAT遺伝子を用いたトランジェントアッセイによってこの2つのCHS遺伝子(PsCHS1,PsCHS2)はエリシタ-の処理を施さなくてもCATの活性を増高させるシスエレメントを各々の上流(PsCHS1:-1493〜+80;PsCHS2:-1889〜+83)にもつことが明らかにされた。
    3.植物病原菌が生産するシグナル物質の単離調製法と情報伝達機構の解析手法の研究調査
    平成3年12月にカリフォルニア大学カド教授を招へいし、岡山大学において長年進められてきた植物病原菌が生産するシグナル物質に関する研究を中心に調査を行った。カド教授は植物病原糸状菌の生産するエリシタ-・サプレッサ-の単離調製を行った。更にエリシタ-・サプレッサ-による植物細胞の初期情報伝達系に対する影響を解析するために岡山大学で研究されているATPア-ゼを始めとする種々の情報伝達系に関与する酵素活性の測定法や解析法について視察並びに討議を行った。

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  • 宿主ー病原菌の特異性と動的抵抗性遺伝子群の発現調節

    研究課題/領域番号:02404012  1990年 - 1991年

    日本学術振興会  科学研究費助成事業 一般研究(A)  一般研究(A)

    奥 八郎, 一瀬 勇規, 山田 哲治, 白石 友紀

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    配分額:19100000円 ( 直接経費:19100000円 )

    エンドウ褐紋病菌の生産するエリシタ-及びサプレッサ-の宿主エンドウの防御応答におけるシグナル伝達とファイトアレキシン合成経路の鍵酵素PAL並びにCHS遺伝子の発現制御に対する影響ついて解析を進めたところ、以下の概要を得た。
    1 原形質膜ATPaseに対する作用 サプレッサ-はエンドウを含めた非宿主植物から調製した原形質膜ATPaseを阻害するが、組織化学的に調べるとエンドウのATPase活性だけを特異的に阻害した。また、バナジン酸(非特異的P型ATPase阻害剤)はサプレッサ-と同様、エリシタ-によるPAL、CHSの発現誘導を遅延した。
    2 原形質膜におけるポリホスホイノシチド(PI)代謝に及ぼす影響 エリシタ-処理区ではリピド・キナ-ゼの活性化によりPIP,PIP2がリン酸化されるが、サプレッサ-共存下ではこのリン酸化が顕著に抑制された。
    3 PAL遺伝子の構造と発現調節 エンドウPAL遺伝子マルチジ-ンファミリ-のうち、二つのメンバ-のゲノム遺伝子の構造決定を行ない、それぞれの遺伝子PSPAL1,PSPAL2の組織特異的な発現量を調べたところ、根と下部茎での発現が非常に高く、他の器官での発現は低かったことから、根特異的発現に関与する調節領域が含まれることが示唆された。また、エンドウ・プロトプラストへ、種々の長さのプロモ-タ-領域を挿入したレポ-タ-遺伝子発現用ベクタ-を導入し、エリシタ-による発現誘導はバナジン酸による発現抑制を調べたところ、PSPAL1の _ー340^〜 _ー100の領域の発現制御に必要なシス因子が存在すると考えられる。
    4 カルコン合成酵素(CHS)遺伝子の構造 エンドウCHS遺伝子(PCHS1,PCHS2)はゲノミックロ-ン上にスペ-サ-領域を挟んでタンデムにクラスタ-を形成し、TATAボックスの上流には31塩基からなる同一配列が存在していた。

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