Updated on 2024/02/02

写真a

 
SHINYA Tomonori
 
Organization
Institute of Plant Science and Resources Associate Professor
Position
Associate Professor
External link

Degree

  • 博士(工学) ( 東京農工大学 )

Research Interests

  • 生物間相互作用

  • Plant-insect interactions

  • Plant immunity

  • 分子間相互作用

  • 植食性昆虫

  • 害虫

  • イネ

Research Areas

  • Life Science / Applied molecular and cellular biology

  • Environmental Science/Agriculture Science / Plant protection science

  • Environmental Science/Agriculture Science / Environmental agriculture

Education

  • Tokyo University of Agriculture and Technology   工学研究科   生命工学専攻

    1999 - 2004

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    Country: Japan

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  • Tokyo University of Agriculture and Technology    

    1995 - 1999

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Research History

  • Okayama University   資源植物科学研究所   Associate Professor

    2019

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  • Okayama University   資源植物科学研究所   Assistant Professor

    2013 - 2018

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Papers

  • Glucosylation prevents autotoxicity of stress inducible DOPA in maize seedlings Reviewed

    Takako Aboshi, Kohei Ittou, Ivan Galis, Tomonori Shinya, Tetsuya Murayama

    Plant Growth Regulation   101 ( 1 )   159 - 167   2023.9

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    L-3,4-Dihydroxyphenylalanine (L-DOPA) is a compound with strong allelopathic effects on Brassicaceae, Asteraceae, Cucurbitaceae and Hydrophyllaceae species. Although Gramineae are less affected by L-DOPA with respect to their root growth, metabolic routes that protect them from L-DOPA toxicity are poorly understood. We identified a DOPA glucoside in maize (Zea mays L., Gramineae) roots and leaves, but DOPA aglycon was not detectable in maize. Accordingly, when maize seedlings were exposed to L-DOPA solution, DOPA glucoside concentrations increased in maize leaves, suggesting that absorbed L-DOPA is rapidly converted to glucoside conjugate. When DOPA glucoside solution was applied to lettuce seeds (Lactuca sativa; Asteraceae), lettuce radicle growth was less inhibited compared to free L-DOPA. Considering that maize radicle growth is less affected by free L-DOPA, it is likely that maize seedlings protect themselves from toxicity by L-DOPA glucosylation. Interestingly, a developmental stage dependent variation in DOPA glucoside concentration was observed with highest level of metabolite detected in L2 stage maize leaves. As DOPA glucoside also increased in maize during herbivory by the bird cherry-oat aphid (Rhopalosiphum padi L.) and the Graminae generalist armyworm (Mythimna loreyi), as well as in response to treatment with the plant hormones, we propose that DOPA glucoside might be involved in various stress responses and/or defense in maize seedlings.

    DOI: 10.1007/s10725-023-01009-w

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  • Comparative analysis of sorghum (C4) and rice (C3) plant headspace volatiles induced by artificial herbivory Reviewed

    Cyprian Osinde, Islam S. Sobhy, David Wari, Son Truong Dinh, Yuko Hojo, Dandy A. Osibe, Tomonori Shinya, Arthur K. Tugume, Anthony M. Nsubuga, Ivan Galis

    Plant Signaling Behavior   2023.8

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    Publishing type:Research paper (scientific journal)   Publisher:Informa UK Limited  

    DOI: 10.1080/15592324.2023.2243064

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  • BSR1, a Rice Receptor-like Cytoplasmic Kinase, Positively Regulates Defense Responses to Herbivory. Reviewed International journal

    Yasukazu Kanda, Tomonori Shinya, Satoru Maeda, Kadis Mujiono, Yuko Hojo, Keisuke Tomita, Kazunori Okada, Takashi Kamakura, Ivan Galis, Masaki Mori

    International journal of molecular sciences   24 ( 12 )   2023.6

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

    Crops experience herbivory by arthropods and microbial infections. In the interaction between plants and chewing herbivores, lepidopteran larval oral secretions (OS) and plant-derived damage-associated molecular patterns (DAMPs) trigger plant defense responses. However, the mechanisms underlying anti-herbivore defense, especially in monocots, have not been elucidated. The receptor-like cytoplasmic kinase Broad-Spectrum Resistance 1 (BSR1) of Oryza sativa L. (rice) mediates cytoplasmic defense signaling in response to microbial pathogens and enhances disease resistance when overexpressed. Here, we investigated whether BSR1 contributes to anti-herbivore defense responses. BSR1 knockout suppressed rice responses triggered by OS from the chewing herbivore Mythimna loreyi Duponchel (Lepidoptera: Noctuidae) and peptidic DAMPs OsPeps, including the activation of genes required for biosynthesis of diterpenoid phytoalexins (DPs). BSR1-overexpressing rice plants exhibited hyperactivation of DP accumulation and ethylene signaling after treatment with simulated herbivory and acquired enhanced resistance to larval feeding. As the biological significance of herbivory-induced accumulation of rice DPs remains unexplained, their physiological activities in M. loreyi were analyzed. The addition of momilactone B, a rice DP, to the artificial diet suppressed the growth of M. loreyi larvae. Altogether, this study revealed that BSR1 and herbivory-induced rice DPs are involved in the defense against chewing insects, in addition to pathogens.

    DOI: 10.3390/ijms241210395

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  • The rice wound-inducible transcription factor RERJ1 sharing same signal transduction pathway with OsMYC2 is necessary for defense response to herbivory and bacterial blight. Reviewed International journal

    Ioana Valea, Atsushi Motegi, Naoko Kawamura, Koichi Kawamoto, Akio Miyao, Rika Ozawa, Junji Takabayashi, Kenji Gomi, Keiichirou Nemoto, Akira Nozawa, Tatsuya Sawasaki, Tomonori Shinya, Ivan Galis, Koji Miyamoto, Hideaki Nojiri, Kazunori Okada

    Plant Molecular Biology   109 ( 4-5 )   651 - 666   2022.7

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    KEY MESSAGE: This study describes biological functions of the bHLH transcription factor RERJ1 involved in the jasmonate response and the related defense-associated metabolic pathways in rice, with particular focus on deciphering the regulatory mechanisms underlying stress-induced volatile emission and herbivory resistance. RERJ1 is rapidly and drastically induced by wounding and jasmonate treatment but its biological function remains unknown as yet. Here we provide evidence of the biological function of RERJ1 in plant defense, specifically in response to herbivory and pathogen attack, and offer insights into the RERJ1-mediated regulation of metabolic pathways of specialized defense compounds, such as monoterpene linalool, in possible collaboration with OsMYC2-a well-known master regulator in jasmonate signaling. In rice (Oryza sativa L.), the basic helix-loop-helix (bHLH) family transcription factor RERJ1 is induced under environmental stresses, such as wounding and drought, which are closely linked to jasmonate (JA) accumulation. Here, we investigated the biological function of RERJ1 in response to biotic stresses, such as herbivory and pathogen infection, using an RERJ1-defective mutant. Transcriptome analysis of the rerj1-Tos17 mutant revealed that RERJ1 regulated the expression of a typical family of conserved JA-responsive genes (e.g., terpene synthases, proteinase inhibitors, and jasmonate ZIM domain proteins). Upon exposure to armyworm attack, the rerj1-Tos17 mutant exhibited more severe damage than the wildtype, and significant weight gain of the larvae fed on the mutant was observed. Upon Xanthomonas oryzae infection, the rerj1-Tos17 mutant developed more severe symptoms than the wildtype. Among RERJ1-regulated terpene synthases, linalool synthase expression was markedly disrupted and linalool emission after wounding was significantly decreased in the rerj1-Tos17 mutant. RERJ1 appears to interact with OsMYC2-a master regulator of JA signaling-and many OsJAZ proteins, although no obvious epistatic interaction was detected between them at the transcriptional level. These results indicate that RERJ1 is involved in the transcriptional induction of JA-mediated stress-responsive genes via physical association with OsMYC2 and mediates defense against herbivory and bacterial infection through JA signaling.

    DOI: 10.1007/s11103-021-01186-0

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  • Chitooligosaccharide elicitor and oxylipins synergistically elevate phytoalexin production in rice Reviewed International journal

    Tomonori Shinya, Koji Miyamoto, Kenichi Uchida, Yuko Hojo, Emi Yumoto, Kazunori Okada, Hisakazu Yamane, Ivan Galis

    Plant Molecular Biology   109 ( 4-5 )   595 - 609   2022.7

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    KEY MESSAGE: We show that in rice, the amino acid-conjugates of JA precursor, OPDA, may function as a non-canonical signal for the production of phytoalexins in coordination with the innate chitin signaling. The core oxylipins, jasmonic acid (JA) and JA-Ile, are well-known as potent regulators of plant defense against necrotrophic pathogens and/or herbivores. However, recent studies also suggest that other oxylipins, including 12-oxo-phytodienoic acid (OPDA), may contribute to plant defense. Here, we used a previously characterized metabolic defense marker, p-coumaroylputrescine (CoP), and fungal elicitor, chitooligosaccharide, to specifically test defense role of various oxylipins in rice (Oryza sativa). While fungal elicitor triggered a rapid production of JA, JA-Ile, and their precursor OPDA, rice cells exogenously treated with the compounds revealed that OPDA, rather than JA-Ile, can stimulate the CoP production. Next, reverse genetic approach and oxylipin-deficient rice mutant (hebiba) were used to uncouple oxylipins from other elicitor-triggered signals. It appeared that, without oxylipins, residual elicitor signaling had only a minimal effect but, in synergy with OPDA, exerted a strong stimulatory activity towards CoP production. Furthermore, as CoP levels were compromised in the OPDA-treated Osjar1 mutant cells impaired in the oxylipin-amino acid conjugation, putative OPDA-amino acid conjugates emerged as hypothetical regulators of CoP biosynthesis. Accordingly, we found several OPDA-amino acid conjugates in rice cells treated with exogenous OPDA, and OPDA-Asp was detected, although in small amounts, in the chitooligosaccharide-treated rice. However, as synthetic OPDA-Asp and OPDA-Ile, so far, failed to induce CoP in cells, it suggests that yet another presumed OPDA-amino acid form(s) could be acting as novel regulator(s) of phytoalexins in rice.

    DOI: 10.1007/s11103-021-01217-w

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    Other Link: https://link.springer.com/article/10.1007/s11103-021-01217-w/fulltext.html

  • Integrated view of plant metabolic defense with particular focus on chewing herbivores Reviewed

    David Wari, Takako Aboshi, Tomonori Shinya, Ivan Galis

    Journal of Integrative Plant Biology   64 ( 2 )   449 - 475   2022.2

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

    Success of plants largely depends on their ability to defend against herbivores. Since emergence of the first voracious consumers, plants maintained adapting their structures and chemistry to escape from extinction. The constant pressure was further accelerated by adaptation of herbivores to plant defenses, which all together sparked the rise of a chemical empire comprised of thousands of specialized metabolites currently found in plants. Metabolic diversity in the plant kingdom is truly amazing, and although many plant metabolites have already been identified, a large number of potentially useful chemicals remain unexplored in plant bio-resources. Similarly, biosynthetic routes for plant metabolites involve many enzymes, some of which still wait for identification and biochemical characterization. Moreover, regulatory mechanisms that control gene expression and enzyme activities in specialized metabolism of plants are scarcely known. Finally, understanding of how plant defense chemicals exert their toxicity and/or repellency against herbivores remains limited to typical examples, such as proteinase inhibitors, cyanogenic compounds and nicotine. In this review, we attempt summarizing the current status quo in metabolic defense of plants that is predominantly based on the survey of ubiquitous examples of plant interactions with chewing herbivores.

    DOI: 10.1111/jipb.13204

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  • Phytohormone‐dependent plant defense signaling orchestrated by oral bacteria of the herbivore Spodoptera litura Reviewed

    Yukiyo Yamasaki, Hiroka Sumioka, Mayu Takiguchi, Takuya Uemura, Yuka Kihara, Tomonori Shinya, Ivan Galis, Gen‐ichiro Arimura

    New Phytologist   231 ( 5 )   2029 - 2038   2021.9

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    DOI: 10.1111/nph.17444

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/nph.17444

  • Herbivore-induced and constitutive volatiles are controlled by different oxylipin-dependent mechanisms in rice. Reviewed International journal

    Kadis Mujiono, Tilisa Tohi, Islam S Sobhy, Yuko Hojo, Tomonori Shinya, Ivan Galis

    Plant, cell & environment   44 ( 8 )   2687 - 2699   2021.8

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    Despite the importance of volatile organic compounds (VOCs) for plants, control mechanisms for their basal and stress-induced biosynthesis and release remain unclear. We sampled and characterized headspace and internal leaf volatile pools in rice (Oryza sativa), after a simulated herbivory treatment, which triggers an endogenous jasmonate burst. Certain volatiles, such as linalool, were strongly upregulated by simulated herbivory stress. In contrast, other volatiles, such as β-caryophyllene, were constitutively emitted and fluctuated according to time of day. Transcripts of the linalool synthase gene transiently increased 1-3 h after exposure of rice to simulated herbivory, whereas transcripts of caryophyllene synthase peaked independently at dawn. Unexpectedly, although emission and accumulation patterns of rice inducible and constitutive VOCs were substantially different, both groups of volatiles were compromised in jasmonate-deficient hebiba mutants, which lack the allene oxide cyclase (AOC) gene. This suggests that rice employs at least two distinct oxylipin-dependent mechanisms downstream of AOC to control production of constitutive and herbivore-induced volatiles. Levels of the JA precursor, 12-oxo-phytodienoic acid (OPDA), were correlated with constitutive volatile levels suggesting that OPDA or its derivatives could be involved in control of volatile emission in rice.

    DOI: 10.1111/pce.14126

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  • Deciphering OPDA Signaling Components in the Momilactone-Producing Moss Calohypnum plumiforme. Reviewed International journal

    Hideo Inagaki, Koji Miyamoto, Noriko Ando, Kohei Murakami, Koki Sugisawa, Shion Morita, Emi Yumoto, Miyu Teruya, Kenichi Uchida, Nobuki Kato, Takuya Kaji, Yousuke Takaoka, Yuko Hojo, Tomonori Shinya, Ivan Galis, Akira Nozawa, Tatsuya Sawasaki, Hideaki Nojiri, Minoru Ueda, Kazunori Okada

    Frontiers in plant science   12   688565 - 688565   2021

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    Jasmonic acid (JA) and its biologically active form jasmonoyl-L-isoleucine (JA-Ile) regulate defense responses to various environmental stresses and developmental processes in plants. JA and JA-Ile are synthesized from α-linolenic acids derived from membrane lipids via 12-oxo-phytodienoic acid (OPDA). In the presence of JA-Ile, the COI1 receptor physically interacts with JAZ repressors, leading to their degradation, resulting in the transcription of JA-responsive genes by MYC transcription factors. Although the biosynthesis of JA-Ile is conserved in vascular plants, it is not recognized by COI1 in bryophytes and is not biologically active. In the liverwort Marchantia polymorpha, dinor-OPDA (dn-OPDA), a homolog of OPDA with two fewer carbons, and its isomer dn-iso-OPDA accumulate after wounding and are recognized by COI1 to activate downstream signaling. The moss Calohypnum plumiforme produces the antimicrobial-specialized metabolites, momilactones. It has been reported that JA and JA-Ile are not detected in C. plumiforme and that OPDA, but not JA, can induce momilactone accumulation and the expression of these biosynthetic genes, suggesting that OPDA or its derivative is a biologically active molecule in C. plumiforme that induces chemical defense. In the present study, we investigated the biological functions of OPDA and its derivatives in C. plumiforme. Searching for the components potentially involving oxylipin signaling from transcriptomic and genomic data revealed that two COI1, three JAZ, and two MYC genes were present. Quantification analyses revealed that OPDA and its isomer iso-OPDA accumulated in larger amounts than dn-OPDA and dn-iso-OPDA after wounding. Moreover, exogenously applied OPDA, dn-OPDA, or dn-iso-OPDA induced the transcription of JAZ genes. These results imply that OPDA, dn-OPDA, and/or their isomers potentially act as biologically active molecules to induce the signaling downstream of COI1-JAZ. Furthermore, co-immunoprecipitation analysis showed the physical interaction between JAZs and MYCs, indicating the functional conservation of JAZs in C. plumiforme with other plants. These results suggest that COI1-JAZ-MYC mediated signaling is conserved and functional in C. plumiforme.

    DOI: 10.3389/fpls.2021.688565

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  • Phytohormone elicitation in maize by oral secretions of specialist Mythimna separata and generalist Spodoptera litura Reviewed

    Morita M, Yamasaki Y, Shinya T, Galis I, Arimura GI

    J Plant Interact   16 ( 1 )   587 - 590   2021

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    DOI: 10.1080/17429145.2021.2006334

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  • Ethylene functions as a suppressor of volatile production in rice. Reviewed International journal

    Kadis Mujiono, Tilisa Tohi, Islam S Sobhy, Yuko Hojo, Nhan Thanh Ho, Tomonori Shinya, Ivan Galis

    Journal of Experimental Botany   71 ( 20 )   6491 - 6511   2020.10

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    We examined the role of ethylene in the production of rice (Oryza sativa) volatile organic compounds (VOCs), which act as indirect defense signals against herbivores in tritrophic interactions. Rice plants were exposed to exogenous ethylene (1 ppm) after simulated herbivory, which consisted of mechanical wounding supplemented with oral secretions (WOS) from the generalist herbivore larva Mythimna loreyi. Ethylene treatment highly suppressed VOCs in WOS-treated rice leaves, which was further corroborated by the reduced transcript levels of major VOC biosynthesis genes in ethylene-treated rice. In contrast, the accumulation of jasmonates (JA), known to control VOCs in higher plants, and transcript levels of primary JA response genes, including OsMYC2, were not largely affected by ethylene application. At the functional level, flooding is known to promote internode elongation in young rice via ethylene signaling. Consistent with the negative role of ethylene on VOC genes, the accumulation of VOCs in water-submerged rice leaves was suppressed. Furthermore, in mature rice plants, which naturally produce less volatiles, VOCs could be rescued by the application of the ethylene perception inhibitor 1-methylcyclopropene. Our data suggest that ethylene acts as an endogenous suppressor of VOCs in rice plants during development and under stress.

    DOI: 10.1093/jxb/eraa341

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  • Silencing of phosphoinositide dependent protein kinase orthologs reduces hypersensitive cell death in Nicotiana benthamiana. Reviewed

    Akinori Kiba, Kotoko Fukui, Maki Mitani, Ivan Galis, Yuko Hojo, Tomonori Shinya, Kouhei Ohnishi, Yasufumi Hikichi

    Plant Biotechnology (Tokyo, Japan)   37 ( 3 )   363 - 367   2020.9

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    Phosphatidic acid plays an important role in plant immune responses against phytopathogenic bacteria in Nicotiana benthamiana. Here we focused on phosphoinositide dependent protein kinases (PDKs) as a candidate required for phosphatidic acid signaling. Based on Arabidopsis PDK sequences, we identified four putative PDK orthologs in N. benthamiana genome. To address the role of PDKs in plant defense responses, we created all four NbPDKs-silenced plants by virus-induced gene silencing. the NbPDKs-silenced plants showed a moderately reduced growth phenotype. Induction of hypersensitive cell death was compromised in the NbPDKs-silenced plants challenged with Ralstonia solanacearum. The hypersensitive cell death induced by bacterial effectors was also reduced in the NbPDKs-silenced plants. the NbPDKs-silenced plants showed decreased production of salicylic acid, jasmonic acid and jasmonoyl-L-isoleucine, as well as hydrogen peroxide after inoculation with R. solanacearum. These results suggest that NbPDKs might have an important role in the regulation of the hypersensitive cell death via plant hormone signaling and oxidative burst.

    DOI: 10.5511/plantbiotechnology.20.0511b

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  • Nonglandular silicified trichomes are essential for rice defense against chewing herbivores Reviewed

    Joackin B. Andama, Kadis Mujiono, Yuko Hojo, Tomonori Shinya, Ivan Galis

    Plant, Cell & Environment   43 ( 9 )   2019 - 2032   2020.9

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    Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1111/pce.13775

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pce.13775

  • Stomatal immunity against fungal invasion comprises not only chitin-induced stomatal closure but also chitosan-induced guard cell death. Reviewed International journal

    Wenxiu Ye, Shintaro Munemasa, Tomonori Shinya, Wei Wu, Tao Ma, Jiang Lu, Toshinori Kinoshita, Hanae Kaku, Naoto Shibuya, Yoshiyuki Murata

    Proceedings of the National Academy of Sciences of the United States of America   117 ( 34 )   20932 - 20942   2020.8

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    Many pathogenic fungi exploit stomata as invasion routes, causing destructive diseases of major cereal crops. Intensive interaction is expected to occur between guard cells and fungi. In the present study, we took advantage of well-conserved molecules derived from the fungal cell wall, chitin oligosaccharide (CTOS), and chitosan oligosaccharide (CSOS) to study how guard cells respond to fungal invasion. In Arabidopsis, CTOS induced stomatal closure through a signaling mediated by its receptor CERK1, Ca2+, and a major S-type anion channel, SLAC1. CSOS, which is converted from CTOS by chitin deacetylases from invading fungi, did not induce stomatal closure, suggesting that this conversion is a fungal strategy to evade stomatal closure. At higher concentrations, CSOS but not CTOS induced guard cell death in a manner dependent on Ca2+ but not CERK1. These results suggest that stomatal immunity against fungal invasion comprises not only CTOS-induced stomatal closure but also CSOS-induced guard cell death.

    DOI: 10.1073/pnas.1922319117

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  • Phosphatidylinositol-phospholipase C2 regulates pattern-triggered immunity in Nicotiana benthamiana. Reviewed International journal

    Akinori Kiba, Masahito Nakano, Miki Hosokawa, Ivan Galis, Hiroko Nakatani, Tomonori Shinya, Kouhei Ohnishi, Yasufumi Hikichi

    Journal of Experimental Botany   71 ( 16 )   5027 - 5038   2020.8

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    Phospholipid signaling plays an important role in plant immune responses against phytopathogenic bacteria in Nicotiana benthamiana. Here, we isolated two phospholipase C2 (PLC2) orthologs in the N. benthamiana genome, designated as PLC2-1 and 2-2. Both NbPLC2-1 and NbPLC2-2 were expressed in most tissues and were induced by infiltration with bacteria and flg22. NbPLC2-1 and NbPLC2-2 (NbPLC2s) double-silenced plants showed a moderately reduced growth phenotype. The induction of the hypersensitive response was not affected, but bacterial growth and the appearance of bacterial wilt were accelerated in NbPLC2s-silenced plants when they were challenged with a virulent strain of Ralstonia solanacearum that was compatible with N. benthamiana. NbPLC2s-silenced plants showed reduced expression levels of NbPR-4, a marker gene for jasmonic acid signaling, and decreased jasmonic acid and jasmonoyl-L-isoleucine contents after inoculation with R. solanacearum. The induction of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) marker genes was reduced in NbPLC2s-silenced plants after infiltration with R. solanacearum or Pseudomonas fluorescens. Accordingly, the resistance induced by flg22 was compromised in NbPLC2s-silenced plants. In addition, the expression of flg22-induced PTI marker genes, the oxidative burst, stomatal closure, and callose deposition were all reduced in the silenced plants. Thus, NbPLC2s might have important roles in pre- and post-invasive defenses, namely in the induction of PTI.

    DOI: 10.1093/jxb/eraa233

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  • Affinity Labeling and Purification of Plant Chitin-Binding LysM Receptor with Chitin Octasaccharide Derivatives. International journal

    Tomonori Shinya, Naoto Shibuya, Hanae Kaku

    Methods in molecular biology (Clifton, N.J.)   2132   401 - 412   2020

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    Authorship:Lead author   Language:English  

    Lysin motif (LysM) is a carbohydrate-binding modules found in all kingdoms. LysM binds to N-acetylglucosamine-containing molecules such as peptidoglycan, chitin, Nod factor, and Myc factor and is found in peptidoglycan hydrolases, chitinases, and plant pathogen effectors and plant receptor/co-receptor for defense and symbiosis signaling. This chapter describes the synthesis of a nonradioactive chitin ligand, biotinylated chitin octasaccharide, (GlcNAc)8-Bio, and its application for the detection and characterization of chitin-binding LysM receptor CEBiP in the microsomal membrane fraction of rice suspension-cultured cells by affinity labeling. We also describe the purification of CEBiP from the plasma membrane of the rice cells by affinity chromatography with the synthesized (GlcNAc)8-APEA-CH-Sepharose as an affinity matrix.

    DOI: 10.1007/978-1-0716-0430-4_39

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  • Brown planthopper honeydew-associated symbiotic microbes elicit momilactones in rice Reviewed International journal

    David Wari, Kabir Md Alamgir, Kadis Mujiono, Yuko Hojo, Akio Tani, Tomonori Shinya, Hiroko Nakatani, Ivan Galis

    Plant Signaling & Behavior   14 ( 11 )   1655335 - 1655335   2019.11

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Informa UK Limited  

    Plants use many natural products to counter pests and diseases in nature. In rice, direct defense mechanisms include broad range of secondary metabolites, such as phenolamides (PA), diterpene phytoalexins, and flavonoid sakuranetin. Recently, accumulation of PAs in rice was shown to be under control of microbial symbionts in honeydew (HD), digestive waste from the rice brown planthopper (Nilaparvata lugens; BPH), but whether HD microbiota can also promote diterpene phytoalexins, momilactone A (MoA) and MoB, has not been reported. Here, we demonstrate that crude HD, but not a filtered one, induces MoA and MoB in rice, suggesting the involvement of BPH-HD endosymbionts. Consequently, microbial strains previously isolated from HD could promote MoA and MoB levels in wounded rice leaves, suggesting that rice indeed responds to BPH by cumulative chemical defense that involves both PA and diterpene phytoalexin pathways.

    DOI: 10.1080/15592324.2019.1655335

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  • Honeydew-associated microbes elicit defense responses against brown planthopper in rice. Reviewed International journal

    David Wari, Md Alamgir Kabir, Kadis Mujiono, Yuko Hojo, Tomonori Shinya, Akio Tani, Hiroko Nakatani, Ivan Galis

    Journal of experimental botany   70 ( 5 )   1683 - 1696   2019.3

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    Feeding of sucking insects, such as the rice brown planthopper (Nilaparvata lugens; BPH), causes only limited mechanical damage on plants that is otherwise essential for injury-triggered defense responses against herbivores. In pursuit of complementary BPH elicitors perceived by plants, we examined the potential effects of BPH honeydew secretions on the BPH monocot host, rice (Oryza sativa). We found that BPH honeydew strongly elicits direct and putative indirect defenses in rice, namely accumulation of phytoalexins in the leaves, and release of volatile organic compounds from the leaves that serve to attract natural enemies of herbivores, respectively. We then examined the elicitor active components in the honeydew and found that bacteria in the secretions are responsible for the activation of plant defense. Corroborating the importance of honeydew-associated microbiota for induced plant resistance, BPHs partially devoid of their microbiota via prolonged antibiotics ingestion induced significantly less defense in rice relative to antibiotic-free insects applied to similar groups of plants. Our data suggest that rice plants may additionally perceive herbivores via their honeydew-associated microbes, allowing them to discriminate between incompatible herbivores-that do not produce honeydew-and those that are compatible and therefore dangerous.

    DOI: 10.1093/jxb/erz041

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  • Integration of danger peptide signals with herbivore-associated molecular pattern signaling amplifies anti-herbivore defense responses in rice. Reviewed International journal

    Tomonori Shinya, Shigetaka Yasuda, Kiwamu Hyodo, Rena Tani, Yuko Hojo, Yuka Fujiwara, Kei Hiruma, Takuma Ishizaki, Yasunari Fujita, Yusuke Saijo, Ivan Galis

    The Plant journal : for cell and molecular biology   94 ( 4 )   626 - 637   2018.5

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    Plant defense against herbivores is modulated by herbivore-associated molecular patterns (HAMPs) from oral secretions (OS) and/or saliva of insects. Furthermore, feeding wounds initiate plant self-damage responses modulated by danger-associated molecular patterns (DAMPs) such as immune defense-promoting plant elicitor peptides (Peps). While temporal and spatial co-existence of both patterns during herbivory implies a possibility of their close interaction, the molecular mechanisms remain undetermined. Here we report that exogenous application of rice (Oryza sativa) peptides (OsPeps) can elicit multiple defense responses in rice cell cultures. Specific activation of OsPROPEP3 gene transcripts in rice leaves by wounding and OS treatments further suggests a possible involvement of the OsPep3 peptide in rice-herbivore interactions. Correspondingly, we found that simultaneous application of OsPep3 and Mythimna loreyi OS significantly amplifies an array of defense responses in rice cells, including mitogen-activated protein kinase activation, and generation of defense-related hormones and metabolites. The induction of OsPROPEP3/4 by OsPep3 points to a positive auto-feedback loop in OsPep signaling which may contribute to additional enhancement of defense signal(s). Finally, the overexpression of the OsPep receptor OsPEPR1 increases the sensitivity of rice plants not only to the cognate OsPeps but also to OS signals. Our findings collectively suggest that HAMP-DAMP signal integration provides a critical step in the amplification of defense signaling in plants.

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  • Phosphoinositide 3-kinase participates in L-methionine sulfoximine-induced cell death via salicylic acid mediated signaling in Nicotiana benthamiana Reviewed

    Sayuri Sumida, Makoto Ito, Ivan Galis, Hiroko Nakatani, Tomonori Shinya, Kouhei Ohnishi, Yasufumi Hikichi, Akinori Kiba

    Journal of Plant Physiology   218   167 - 170   2017.11

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    Pseudomonas syringae pv. tabaci causes wildfire disease by the action of tabtoxinine-β-lactam (TβL), a non-specific bacterial toxin. To better understand the molecular mechanisms of wildfire disease and its development, we focused on the phosphoinositide 3-kinase in Nicotiana benthamiana (NbPI3K) and its potential role in the disease outbreak, using L-methionine sulfoximine (MSX) as an easily accessible mimic of the TβL action. The NbPI3K-silenced plants showed accelerated induction of cell death and necrotic lesion formation by MSX, and the expression of hin1, marker gene for the programmed cell death, was strongly induced in the plants. However, the accumulation of ammonium ions, caused by MSX inhibition of glutamine sythetase activity, was not affected by the NbPI3K-silencing. Interestingly, the expression of PR-1a, a marker gene for salicylic acid (SA) innate immunity signaling, and accumulation of SA were both enhanced in the NbPI3K-silenced plants. Accordingly, the acceleration of MSX-induced cell death by NbPI3K-silencing was reduced in NahG plants, and by double silencing of NbPI3K together with the NbICS1 encoding a SA-biosynthetic enzyme. As silencing of NbPI3K accelerated the TβL-induced necrotic lesions, and lesions of wildfire disease caused by P. syringae pv. tabaci, these results suggest that the NbPI3K-related pathway might act as a negative regulator of cell death during development of wildfire disease that involves SA-dependent signaling pathway downstream of TβL action in N. benthamiana.

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  • Oral Secretions Affect HIPVs Induced by Generalist (Mythimna loreyi) and Specialist (Parnara guttata) Herbivores in Rice. Reviewed International journal

    Islam S Sobhy, Atsushi Miyake, Tomonori Shinya, Ivan Galis

    Journal of chemical ecology   43 ( 9 )   929 - 943   2017.9

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    Plants synthesize variable mixtures of herbivore-induced plant volatiles (HIPVs) as part of their evolutionary conserved defense. To elucidate the impact of chewing herbivores with different level of adaptation on HIPV profiles in rice, we measured HIPVs released from rice seedlings challenged by either the generalist herbivore Mythimna loreyi (MYL) or the specialist Parnara guttata (PAG). Both herbivores markedly elicited the emission of HIPVs, mainly on the second and third days after attack compared to control plants. In addition, side-by-side HIPV comparisons using MYL and PAG caterpillars revealed that generalist feeding induced comparably more HIPVs relative to specialist, particularly on day two as highlighted by multivariate analysis (PLS-DA) of emitted HIPVs, and further confirmed in mimicked herbivory experiments. Here, mechanically wounded plants treated with water (WW) released more VOCs than untreated controls, and on top of this, oral secretions (OS) from both herbivores showed differential effects on volatile emissions from the wounded plants. Similar to actual herbivory, MYL OS promoted higher amounts of HIPVs relative to PAG OS, thus supporting disparate induction of rice indirect defenses in response to generalist and specialist herbivores, which could be due to the differential composition of their OS. (196 words).

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  • Molecular evidence for biochemical diversification of phenolamide biosynthesis in rice plants Reviewed

    Kimiaki Tanabe, Yuko Hojo, Tomonori Shinya, Ivan Galis

    JOURNAL OF INTEGRATIVE PLANT BIOLOGY   58 ( 11 )   903 - 913   2016.11

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    Two phenolamides (PAs), p-coumaroylputrescine and feruloylputrescine strongly accumulate in rice (Oryza sativa cv. Nipponbare) leaves subjected to attack of chewing and sucking herbivores. Here we identified and characterized in vitro three novel rice genes that mediated coumaroyl-CoA/feruloyl-CoA conjugation to polyamines, putrescine and agmatine. Interestingly, two genes were highly specific for their polyamine substrates, encoding putrescine N-hydroxycinnamoyltransferase and agmatine N-hydroxycinnamoyltransferase, while the third enzyme could use both polyamines and it was therefore annotated as putrescine/agmatine N-hydroxycinnamoyltransferase. All genes were preferentially expressed in rice roots and developing flowers, and in addition, the putrescine/agmatine N-hydroxycinnamoyltransferase transcripts were strongly induced by wounding in the young rice leaves. Because the wound response of this gene was only partially suppressed in the jasmonoyl-L-isoleucine deficient plants (Osjar1), it suggests that its upregulation (as well as inducible PAs in rice) may be largely independent of jasmonoyl-L-isoleucine signaling pathway. The finding of three closely related genes with a similar and/or overlapping activity in PA biosynthesis provides another striking example of rapid diversification of plant metabolism in response to environmental stresses in nature.

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  • Modulation of plant defense responses to herbivores by simultaneous recognition of different herbivore-associated elicitors in rice Reviewed

    Tomonori Shinya, Yuko Hojo, Yoshitake Desaki, John T. Christeller, Kazunori Okada, Naoto Shibuya, Ivan Galis

    SCIENTIFIC REPORTS   6   2016.9

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    Induced plant defense responses against insect herbivores are triggered by wounding and/or perception of herbivore elicitors from their oral secretions (OS) and/or saliva. In this study, we analyzed OS isolated from two rice chewing herbivores, Mythimna loreyi and Parnara guttata. Both types of crude OS had substantial elicitor activity in rice cell system that allowed rapid detection of early and late defense responses, i.e. accumulation of reactive oxygen species (ROS) and defense secondary metabolites, respectively. While the OS from M. loreyi contained large amounts of previously reported insect elicitors, fatty acid-amino acid conjugates (FACs), the elicitor-active P. guttata's OS contained no detectable FACs. Subsequently, elicitor activity associated with the high molecular mass fraction in OS of both herbivores was identified, and shown to promote ROS and metabolite accumulations in rice cells. Notably, the application of N-linolenoyl-Gln (FAC) alone had only negligible elicitor activity in rice cells; however, the activity of isolated elicitor fraction was substantially promoted by this FAC. Our results reveal that plants integrate various independent signals associated with their insect attackers to modulate their defense responses and reach maximal fitness in nature.

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  • Colletotrichum higginsianum extracellular LysM proteins play dual roles in appressorial function and suppression of chitin-triggered plant immunity Reviewed

    Hiroyuki Takahara, Stephane Hacquard, Anja Kombrink, H. Bleddyn Hughes, Vivek Halder, Guillaume P. Robin, Kei Hiruma, Ulla Neumann, Tomonori Shinya, Erich Kombrink, Naoto Shibuya, Bart P. H. J. Thomma, Richard J. O'Connell

    NEW PHYTOLOGIST   211 ( 4 )   1323 - 1337   2016.9

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    The genome of the hemibiotrophic anthracnose fungus, Colletotrichum higginsianum, encodes a large repertoire of candidate-secreted effectors containing LysM domains, but the role of such proteins in the pathogenicity of any Colletotrichum species is unknown. Here, we characterized the function of two effectors, ChELP1 and ChELP2, which are transcriptionally activated during the initial intracellular biotrophic phase of infection. Using immunocytochemistry, we found that ChELP2 is concentrated on the surface of bulbous biotrophic hyphae at the interface with living host cells but is absent from filamentous necrotrophic hyphae. We show that recombinant ChELP1 and ChELP2 bind chitin and chitin oligomers invitro with high affinity and specificity and that both proteins suppress the chitin-triggered activation of two immune-related plant mitogen-activated protein kinases in the host Arabidopsis. Using RNAi-mediated gene silencing, we found that ChELP1 and ChELP2 are essential for fungal virulence and appressorium-mediated penetration of both Arabidopsis epidermal cells and cellophane membranes invitro. The findings suggest a dual role for these LysM proteins as effectors for suppressing chitin-triggered immunity and as proteins required for appressorium function.

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  • Systematic analysis of rice (Oryza sativa) metabolic responses to herbivory Reviewed

    Kabir Md Alamgir, Yuko Hojo, John T. Christeller, Kaori Fukumoto, Ryutaro Isshiki, Tomonori Shinya, Ian T. Baldwin, Ivan Galis

    PLANT CELL AND ENVIRONMENT   39 ( 2 )   453 - 466   2016.2

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    Plants defend against attack from herbivores by direct and indirect defence mechanisms mediated by the accumulation of phytoalexins and release of volatile signals, respectively. While the defensive arsenals of some plants, such as tobacco and Arabidopsis are well known, most of rice's (Oryza sativa) defence metabolites and their effectiveness against herbivores remain uncharacterized. Here, we used a non-biassed metabolomics approach to identify many novel herbivory-regulated metabolic signatures in rice. Most were up-regulated by herbivore attack while only a few were suppressed. Two of the most prominent up-regulated signatures were characterized as phenolamides (PAs), p-coumaroylputrescine and feruloylputrescine. PAs accumulated in response to attack by both chewing insects, i. e. feeding of the lawn armyworm (Spodoptera mauritia) and the rice skipper (Parnara guttata) larvae, and the attack of the sucking insect, the brown planthopper (Nilaparvata lugens, BPH). In bioassays, BPH insects feeding on 15% sugar solution containing p-coumaroylputrescine or feruloylputrescine, at concentrations similar to those elicited by heavy BPH attack in rice, had a higher mortality compared to those feeding on sugar diet alone. Our results highlight PAs as a rapidly expanding new group of plant defence metabolites that are elicited by herbivore attack, and deter herbivores in rice and other plants.

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  • Silencing of Nicotiana benthamiana SEC14 phospholipid transfer protein reduced jasmonic acid dependent defense against Pseudomonas syringae Reviewed

    Akinori Kiba, Yu Imanaka, Masahito Nakano, Ivan Galis, Yuko Hojo, Tomonori Shinya, Kouhei Ohnishi, Yasufumi Hikichi

    Plant Biotechnology   33 ( 2 )   111 - 115   2016

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    © 2016 The Japanese Society for Plant Cell and Molecular Biology. We previously identified SEC14, phospholipid transfer protein superfamily gene, in Nicotiana benthamiana (NbSEC14) that was closely related to phospholipid signaling as well as jasmonic acid-dependent defense responses during plant immune responses against Ralstonia solanacearum. To examine effect of NbSEC14-silencing on basal plant defenses, we used two other bacterial pathogens with different virulent strategies, Pseudomonas syringae pv. tabaci and pv. mellea. NbSEC14-silenced plants showed accelerated growth of P. syringae pv. tabaci and pv. mellea, and formation of necrotic lesions. Induction of JA-related PR-4 gene was compromised in NbSEC14-silenced plants, which was supported by reduced jasmonic acid levels in NbSEC14-silenced plants. These results suggested that NbSEC14 might be regulating plant basal resistance against plant pathogenic Pseudomonads via jasmonic acid-dependent signaling pathway.

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  • Chitin-mediated plant-fungal interactions: catching, hiding and handshaking Reviewed

    Tomonori Shinya, Tomomi Nakagawa, Hanae Kaku, Naoto Shibuya

    CURRENT OPINION IN PLANT BIOLOGY   26   64 - 71   2015.8

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    Plants can detect infecting fungi through the perception of chitin oligosaccharides by lysin motif receptors such as CEBiP and CERK1. A major function of CERK1 seems to be as a signaling molecule in the receptor complex formed with ligand-binding molecules and to activate downstream defense signaling. Fungal pathogens, however, have developed counter strategies to escape from the chitin-mediated detection by using effectors and/or changing their cell walls. Common structural features between chitin and Nod-/Mycfactors and corresponding receptors have suggested the close relationships between the chitin-mediated immunity and rhizobial/arbuscular mycorrhizal symbiosis. The recent discovery of the dual function of OsCERK1 in both plant immunity and mycorrhizal symbiosis sheds new light on the evolutionary relationships between defense and symbiotic systems in plants.

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  • Uptake of chitosan-derived D-glucosamine oligosaccharides in Streptomyces coelicolor A3(2) Reviewed

    Pascal Viens, Marie-Pierre Dubeau, Akane Kimura, Yoshitake Desaki, Tomonori Shinya, Naoto Shibuya, Akihiro Saito, Ryszard Brzezinski

    FEMS MICROBIOLOGY LETTERS   362 ( 9 )   2015.5

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    The csnR gene, localized at the beginning of an operon, csnR-K, which organization is conserved through many actinomycete genomes, was previously shown to repress the transcription of the chitosanase gene csnA in Streptomyces lividans. However, knowledge on the function of the whole csnR-K operon in the metabolism of chitosan (an N-deacetylated derivative of chitin) remained limited. Mutants of S. coelicolor A3(2) harboring partial or total deletions of the csnR-K operon were analyzed for their capacity to uptake glucosamine oligosaccharides (GlcN)(n). The csnR-K operon was autoregulated by CsnR repressor and its transcription was inducible by GlcN oligosaccharides. The operon controlled the uptake of GlcN oligosaccharides in S. coelicolor A3(2), with a minor contribution to the consumption of monomeric GlcN but not chitin-related N-acetylated derivatives. The deletion of the whole operon abolished the uptake of GlcN oligosaccharides. The CsnEFG transporter encoded by this operon is the front door for the assimilation of chitosan-derived hydrolysis products in S. coelicolor A3(2). The ATP-binding component MsiK was essential for CsnEFG transport function. Also, deletion of msiK abolished the induction of csnA transcription by GlcN oligosaccharides.

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  • Selective regulation of the chitin-induced defense response by the Arabidopsis receptor-like cytoplasmic kinase PBL27 Reviewed

    Tomonori Shinya, Koji Yamaguchi, Yoshitake Desaki, Kenta Yamada, Tomoko Narisawa, Yoshihiro Kobayashi, Kanako Maeda, Maruya Suzuki, Takumi Tanimoto, Jun Takeda, Masato Nakashima, Ryota Funama, Mari Narusaka, Yoshihiro Narusaka, Hanae Kaku, Tsutomu Kawasaki, Naoto Shibuya

    PLANT JOURNAL   79 ( 1 )   56 - 66   2014.7

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    Recognition of microbe-associated molecular patterns (MAMPs) initiates pattern-triggered immunity in host plants. Pattern recognition receptors (PRRs) and receptor-like cytoplasmic kinases (RLCKs) are the major components required for sensing and transduction of these molecular patterns. However, the regulation of RLCKs by PRRs and their specificity remain obscure. In this study we show that PBL27, an Arabidopsis ortholog of OsRLCK185, is an immediate downstream component of the chitin receptor CERK1 and contributes to the regulation of chitin-induced immunity in Arabidopsis. Knockout of PBL27 resulted in the suppression of several chitin-induced defense responses, including the activation of MPK3/6 and callose deposition as well as in disease resistance against fungal and bacterial infections. On the other hand, the contribution of PBL27 to flg22 signaling appears to be very limited, suggesting that PBL27 selectively regulates defense signaling downstream of specific PRR complexes. In vitro phosphorylation experiments showed that CERK1 preferentially phosphorylated PBL27 in comparison to BIK1, whereas phosphorylation of PBL27 by BAK1 was very low compared with that of BIK1. Thus, the substrate specificity of the signaling receptor-like kinases, CERK1 and BAK1, may determine the preference of downstream RLCKs.

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  • Presence of LYM2 dependent but CERK1 independent disease resistance in Arabidopsis Reviewed

    Yoshihiro Narusaka, Tomonori Shinya, Mari Narusaka, Noriko Motoyama, Hikaru Shimada, Kosuke Murakami, Naoto Shibuya

    Plant Signaling and Behavior   8 ( 9 )   e25345   2013.9

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    Plants have the ability to detect invading fungi through the perception of chitin fragments released from the fungal cell walls. Plant chitin receptor consists of two types of plasma membrane proteins, CEBiP and CERK1. However, the contribution of these proteins to chitin signaling is different between Arabidopsis and rice. In Arabidopsis, it seems CERK1 receptor kinase is enough for both ligand perception and signaling, whereas both CEBiP and OsCERK1 are required for chitin signaling in rice. Here we report that Arabidopsis CEBiP homolog, LYM2, is not involved in chitin signaling but contributes to resistance against a fungal pathogen, Alternaria brassicicola, indicating the presence of a novel disease resistance mechanism in Arabidopsis. © 2013 Landes Bioscience.

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  • Functional Characterization of CEBiP and CERK1 Homologs in Arabidopsis and Rice Reveals the Presence of Different Chitin Receptor Systems in Plants Reviewed

    Tomonori Shinya, Noriko Motoyama, Asahi Ikeda, Miyuki Wada, Kota Kamiya, Masahiro Hayafune, Hanae Kaku, Naoto Shibuya

    PLANT AND CELL PHYSIOLOGY   53 ( 10 )   1696 - 1706   2012.10

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    Chitin is a representative microbe-associated molecular pattern (MAMP) molecule for various fungi and induces immune responses in many plant species. It has been clarified that the chitin signaling in rice requires a receptor kinase OsCERK1 and a receptor-like protein (Os)CEBiP, which specifically binds chitin oligosaccharides. On the other hand, Arabidopsis requires a receptor kinase (At)CERK1 for chitin signaling but it is not clear whether the plant also requires a CEBiP-like molecule for chitin perception/signaling. To clarify the similarity/difference of the chitin receptor in these two model plants, we first characterized CEBiP homologs in Arabidopsis. Only one of three CEBiP homologs, AtCEBiP (LYM2), showed a high-affinity binding for chitin oligosaccharides similar to rice CEBiP. AtCEBiP also represented the major chitin-binding protein in the Arabidopsis membrane. However, the single/triple knockout (KO) mutants of Arabidopsis CEBiP homologs and the overexpressor of AtCEBiP showed chitin-induced defense responses similar to wild-type Arabidopsis, indicating that AtCEBiP is biochemically functional as a chitin-binding protein but does not contribute to signaling. Studies of the chitin binding properties of the ectodomains of At/OsCERK1 and the chimeric receptors consisting of ecto/cytosolic domains of these molecules indicated that AtCERK1 is sufficient for chitin perception by itself.

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  • Effector-Mediated Suppression of Chitin-Triggered Immunity by Magnaporthe oryzae Is Necessary for Rice Blast Disease Reviewed

    Thomas A. Mentlak, Anja Kombrink, Tomonori Shinya, Lauren S. Ryder, Ippei Otomo, Hiromasa Saitoh, Ryohei Terauchi, Yoko Nishizawa, Naoto Shibuya, Bart P. H. J. Thomma, Nicholas J. Talbot

    PLANT CELL   24 ( 1 )   322 - 335   2012.1

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    Plants use pattern recognition receptors to defend themselves from microbial pathogens. These receptors recognize pathogen-associated molecular patterns (PAMPs) and activate signaling pathways that lead to immunity. In rice (Oryza sativa), the chitin elicitor binding protein (CEBiP) recognizes chitin oligosaccharides released from the cell walls of fungal pathogens. Here, we show that the rice blast fungus Magnaporthe oryzae overcomes this first line of plant defense by secreting an effector protein, Secreted LysM Protein1 (Slp1), during invasion of new rice cells. We demonstrate that Slp1 accumulates at the interface between the fungal cell wall and the rice plasma membrane, can bind to chitin, and is able to suppress chitin-induced plant immune responses, including generation of reactive oxygen species and plant defense gene expression. Furthermore, we show that Slp1 competes with CEBiP for binding of chitin oligosaccharides. Slp1 is required by M. oryzae for full virulence and exerts a significant effect on tissue invasion and disease lesion expansion. By contrast, gene silencing of CEBiP in rice allows M. oryzae to cause rice blast disease in the absence of Slp1. We propose that Slp1 sequesters chitin oligosaccharides to prevent PAMP-triggered immunity in rice, thereby facilitating rapid spread of the fungus within host tissue.

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  • Biochemical and phylogenetic analysis of CEBiP-like LysM domain-containing extracellular proteins in higher plants. Reviewed International journal

    Judith Fliegmann, Sandra Uhlenbroich, Tomonori Shinya, Yves Martinez, Benoit Lefebvre, Naoto Shibuya, Jean-Jacques Bono

    Plant physiology and biochemistry : PPB   49 ( 7 )   709 - 20   2011.7

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    The chitin elicitor-binding protein (CEBiP) from rice was the first plant lysin motif (LysM) protein for which the biological and biochemical function had been established. It belongs to a plant-specific family of extracellular LysM proteins (LYMs) for which we analyzed the phylogeny. LYMs are present in vascular plants only, where an early gene duplication event might have resulted in two types which were retained in present day genomes. LYMs consist of a signal peptide, three consecutive LysMs, separated by cysteine pairs, and a C-terminal region without any known signature, whose length allows the distinction between the two types, and which may be followed by a glycosylphosphatidylinositol (GPI) anchor motif. We analyzed a representative of each type, MtLYM1 and MtLYM2, from Medicago truncatula at the biochemical level and with respect to their expression patterns and observed some similarities but also marked differences. MtLYM1 and MtLYM2 proved to be very different with regard to abundance and apparent molecular mass on SDS-PAGE. Both undergo several post-translational modifications, including N-glycosylation and the addition of a GPI anchor, which would position the proteins at the outer face of the plasma membrane. Only MtLYM2, but not MtLYM1, showed specific binding to biotinylated N-acetylchitooctaose in a manner similar to CEBiP, which belongs to the same type. We postulate that LYM2-type proteins likely function in the perception of chitin-related molecules, whereas possible functions of LYM1-type proteins remain to be elucidated.

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  • Conserved Fungal LysM Effector Ecp6 Prevents Chitin-Triggered Immunity in Plants Reviewed

    Ronnie de Jonge, H. Peter van Esse, Anja Kombrink, Tomonori Shinya, Yoshitake Desaki, Ralph Bours, Sander van der Krol, Naoto Shibuya, Matthieu H. A. J. Joosten, Bart P. H. J. Thomma

    SCIENCE   329 ( 5994 )   953 - 955   2010.8

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    Multicellular organisms activate immunity upon recognition of pathogen-associated molecular patterns (PAMPs). Chitin is the major component of fungal cell walls, and chitin oligosaccharides act as PAMPs in plant and mammalian cells. Microbial pathogens deliver effector proteins to suppress PAMP-triggered host immunity and to establish infection. Here, we show that the LysM domain-containing effector protein Ecp6 of the fungal plant pathogen Cladosporium fulvum mediates virulence through perturbation of chitin-triggered host immunity. During infection, Ecp6 sequesters chitin oligosaccharides that are released from the cell walls of invading hyphae to prevent elicitation of host immunity. This may represent a common strategy of host immune suppression by fungal pathogens, because LysM effectors are widely conserved in the fungal kingdom.

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  • Glyco-conjugates as elicitors or suppressors of plant innate immunity Reviewed

    Alba Silipo, Gitte Erbs, Tomonori Shinya, J. Maxwell Dow, Michelangelo Parrilli, Rosa Lanzetta, Naoto Shibuya, Mari-Anne Newman, Antonio Molinaro

    GLYCOBIOLOGY   20 ( 4 )   406 - 419   2010.4

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    Innate immunity is the first line of defense against invading microorganisms in vertebrates and the only line of defense in invertebrates and plants. Bacterial glyco-conjugates, such as lipopolysaccharides (LPS) from the outer membrane of Gram-negative bacteria and peptidoglycan (PGN) from the cell walls of both Gram-positive and Gram-negative bacteria, and fungal and oomycete glycoconjugates such as oligosaccharides derived from the cell wall components beta-glucan, chitin and chitosan, have been found to act as elicitors of plant innate immunity. These conserved indispensable microbe-specific molecules are also referred to as microbe-associated molecular patterns (MAMPs). Other glyco-conjugates such as bacterial extracellular polysaccharides (EPS) and cyclic glucan have been shown to suppress innate immune responses, thus conversely promoting pathogenesis. MAMPs are recognized by the plant innate immune system though the action of pattern recognition receptors (PRRs). A greater insight into the mechanisms of MAMP recognition and the description of PRRs for different microbial glyco-conjugates will have considerable impact on the improvement of plant health and disease resistance. Here we review the current knowledge about the bacterial MAMPs LPS and PGN, the fungal MAMPs beta-glucan, chitin and chitosan oligosaccharides and the bacterial suppressors EPS and cyclic glucan, with particular reference to the chemical structures of these molecules, the PRRs involved in their recognition (where these have been defined), and possible mechanisms underlying suppression.

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  • Characterization of receptor proteins using affinity cross-linking with biotinylated ligands. Reviewed

    Tomonori Shinya, Tomohiko Osada, Yoshitake Desaki, Masahiro Hatamoto, Yuko Yamanaka, Hisashi Hirano, Ryota Takai, Fang-Sik Che, Hanae Kaku, Naoto Shibuya

    Plant & cell physiology   51 ( 2 )   262 - 70   2010.2

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    The plant genome encodes a wide range of receptor-like proteins but the function of most of these proteins is unknown. We propose the use of affinity cross-linking of biotinylated ligands for a ligand-based survey of the corresponding receptor molecules. Biotinylated ligands not only enable the analysis of receptor-ligand interactions without the use of radioactive compounds but also the isolation and identification of receptor molecules by a simple affinity trapping method. We successfully applied this method for the characterization, isolation and identification of the chitin elicitor binding protein (CEBiP). A biocytin hydrazide conjugate of N-acetylchitooctaose (GN8-Bio) was synthesized and used for the detection of CEBiP in the plasma or microsomal membrane preparations from rice and carrot cells. Binding characteristics of CEBiP analyzed by inhibition studies were in good agreement with the previous results obtained with the use of a radiolabeled ligand. The biotin-tagged CEBiP could be purified by avidin affinity chromatography and identified by LC-MALDI-MS/MS after tryptic digestion. We also used this method to detect OsFLS2, a rice receptor-like kinase for the perception of the peptide elicitor flg22, in membrane preparations from rice cells overexpressing OsFLS2. This work demonstrates the applicability of this method to the purification and identification of plant receptor proteins.

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  • Suppression of a phospholipase D gene, OsPLDbeta1, activates defense responses and increases disease resistance in rice. Reviewed International journal

    Takeshi Yamaguchi, Masaharu Kuroda, Hiromoto Yamakawa, Taketo Ashizawa, Kazuyuki Hirayae, Leona Kurimoto, Tomonori Shinya, Naoto Shibuya

    Plant physiology   150 ( 1 )   308 - 19   2009.5

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    Phospholipase D (PLD) plays an important role in plants, including responses to abiotic as well as biotic stresses. A survey of the rice (Oryza sativa) genome database indicated the presence of 17 PLD genes in the genome, among which OsPLDalpha1, OsPLDalpha5, and OsPLDbeta1 were highly expressed in most tissues studied. To examine the physiological function of PLD in rice, we made knockdown plants for each PLD isoform by introducing gene-specific RNA interference constructs. One of them, OsPLDbeta1-knockdown plants, showed the accumulation of reactive oxygen species in the absence of pathogen infection. Reverse transcription-polymerase chain reaction and DNA microarray analyses revealed that the knockdown of OsPLDbeta1 resulted in the up-/down-regulation of more than 1,400 genes, including the induction of defense-related genes such as pathogenesis-related protein genes and WRKY/ERF family transcription factor genes. Hypersensitive response-like cell death and phytoalexin production were also observed at a later phase of growth in the OsPLDbeta1-knockdown plants. These results indicated that the OsPLDbeta1-knockdown plants spontaneously activated the defense responses in the absence of pathogen infection. Furthermore, the OsPLDbeta1-knockdown plants exhibited increased resistance to the infection of major pathogens of rice, Pyricularia grisea and Xanthomonas oryzae pv oryzae. These results suggested that OsPLDbeta1 functions as a negative regulator of defense responses and disease resistance in rice.

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  • The msiK gene, encoding the ATP-hydrolysing component of N,N '-diacetylchitobiose ABC transporters, is essential for induction of chitinase production in Streptomyces coelicolor A3(2) Reviewed

    Akihiro Saito, Takeshi Fujii, Tomonori Shinya, Naoto Shibuya, Akikazu Ando, Kiyotaka Miyashita

    MICROBIOLOGY-SGM   154 ( Pt 11 )   3358 - 3365   2008.11

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    The dasABC genes encode an ATP-binding cassette (ABC) transporter, which is one of the uptake systems for N,N'-diacetylchitobiose [(GlcNAC)(2)] in Streptomyces coelicolor A3(2), although the gene encoding the ABC subunit that provides ATP hydrolysis for DasABC has not been identified. In this study, we disrupted the sequence that is highly homologous to the msiK gene, the product of which is an ABC subunit assisting several ABC permeases in other Streptomyces species. Disruption of msiK severely affected the ability of S. coelicolor A3(2) to utilize maltose, cellobiose, starch, cellulose, chitin and chitosan, but not glucose. The msiK null mutant lacked (GlcNAC)(2)-uptake activity, but GlcNAc transport activity was unaffected. The data indicated that msiK is essential for (GlcNAC)(2) uptake, which in S. coelicolor A3(2) is governed by ABC transporters including the DasABC-MsiK system, in contrast to Escherichia coli and Serratia marcescens, in which (GlcNAC)(2) uptake is mediated by the phosphotransferase system. Interestingly, the induction of chitinase production by (GlcNAC)(2) or chitin was absent in the msiK null mutant, unlike in the parent strain M145. The defect in chitinase gene induction was rescued by expressing the His-tagged MsiK protein under the control of the putative native promoter on a multicopy plasmid. The data suggest that uptake of (GlcNAC)(2) is necessary for induction of chitinase production. The msiK gene was constitutively transcribed, whereas the transcription of dasA [(GlcNAC)(2)-binding protein gene], malE (putative maltose-binding protein gene), cebE1 (putative cellobiose-binding protein gene) and bxlE1 (putative xylobiose-binding protein gene) was induced by their corresponding sugar ligands. This is believed to be the first report to indicate that (GlcNAC)(2) uptake mediated by ABC transporters is essential for chitinase production in streptomycetes, which are known to be the main degraders of chitin in soil.

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  • CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis Reviewed

    Ayako Miya, Premkumar Albert, Tomonori Shinya, Yoshitake Desaki, Kazuya Ichimura, Ken Shirasu, Yoshihiro Narusaka, Naoto Kawakami, Hanae Kaku, Naoto Shibuya

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   104 ( 49 )   19613 - 19618   2007.12

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    Chitin is a major component of fungal cell walls and serves as a microbe-associated molecular pattern (MAMP) for the detection of various potential pathogens in innate immune systems of both plants and animals. We recently showed that chitin elicitor-binding protein (CEBiP), plasma membrane glycoprotein with LysM motifs, functions as a cell surface receptor for chitin elicitor in rice. The predicted structure of CEBiP does not contain any intracellular domains, suggesting that an additional component(s) is required for signaling through the plasma membrane into the cytoplasm. Here, we identified a receptor-like kinase, designated CERK1, which is essential for chitin elicitor signaling in Arabidopsis. The KO mutants for CERK1 completely lost the ability to respond to the chitin elicitor, including MAPK activation, reactive oxygen species generation, and gene expression. Disease resistance of the KO mutant against an incompatible fungus, Alternaria brassicicola, was partly impaired. Complementation with the WT CERK1 gene showed cerk1 mutations were responsible for the mutant phenotypes. CERK1 is a plasma membrane protein containing three LysM motifs in the extracellular domain and an intracellular Ser/Thr kinase domain with autophosphorylation/myelin basic protein kinase activity, suggesting that CERK1 plays a critical role in fungal MAMP perception in plants.

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  • Comprehensive analysis of glucan elicitor-regulated gene expression in tobacco BY-2 cells reveals a novel MYB transcription factor involved in the regulation of phenylpropanoid metabolism Reviewed

    Tomonori Shinya, Ivan Galis, Tomoko Narisawa, Mami Sasaki, Hiroo Fukuda, Hideaki Matsuoka, Mikako Saito, Ken Matsuoka

    PLANT AND CELL PHYSIOLOGY   48 ( 10 )   1404 - 1413   2007.10

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    We previously demonstrated that a -1,3-, 1,6-oligoglucan (AaGlucan) from the fungus Alternaria alternata 102 shows strong elicitor activity in tobacco BY-2 cells. We have used cDNA microarray analysis to monitor global changes in gene expression in tobacco cells treated with this A. alternata fraction or with laminarin. In total, we identified 265 genes that were induced 1 h after treatment with an AaGlucan-enriched fraction or laminarin. Among them, we characterized in detail a novel tobacco R2R3 MYB-type transcription factor homolog (NtMYBGR1) and two DC1 domain-containing genes (NtDC1A and NtDC1B). Microarray data, together with overexpression and metabolic analyses, indicated that NtMYBGR1, but not the NtDC1 proteins, primarily targets the phenylpropanoid synthesis-related genes PAL and 4CL. These results suggest that NtMYBGR1 specifically regulates defense responses in BY-2 cells by enhancing phenylpropanoid metabolism in response to AaGlucan and laminarin elicitors.

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  • The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N '-diacetylchitobiose in Streptomyces coelicolor A3(2)

    Akihiro Saito, Tomonori Shinya, Katsushiro Miyamoto, Tomofumi Yokoyama, Hanae Kaku, Eiichi Minami, Naoto Shibuya, Hiroshi Tsujibo, Yoshiho Nagata, Akikazu Ando, Takeshi Fujii, Kiyotaka Miyashita

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY   73 ( 9 )   3000 - 3008   2007.5

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    N,N'-Diacetylchitobiose [(GlcNAc)(2)] induces the transcription of chitinase (chi) genes in Streptomyces coelicolor A3(2). Physiological studies showed that (GlcNAc)(2) addition triggered chi expression and increased the rate of (GlcNAc)(2) concentration decline in culture supernatants of mycelia already cultivated with (GlcNAc)(2), suggesting that (GlcNAc)(2) induced the synthesis of its own uptake system. Four open reading frames (SCO0531, SCO0914, SC02946, and SCO5232) encoding putative sugar-binding proteins of ABC transporters were found in the genome by probing the 12-bp repeat sequence required for regulation of chi transcription. SCO5232, named dasA, showed transcriptional induction by (GlcNAc)(2) and N,N',N ''-triacetylchitotriose [(GlcNAc)(3)]. Surface plasmon resonance analysis showed that recombinant DasA protein exhibited the highest affinity for (GlcNAc)(2) (equilibrium dissociation constant [K(D)] = 3.22 x 10(-8)). In the dasA-null mutant, the rate of decline of the (GlcNAc)(2) concentration in the culture supernatant was about 25% of that in strain M145. The in vitro and in vivo data clearly demonstrated that dasA is involved in (GlcNAc)(2) uptake. Upstream and downstream of dasA, the transcriptional regulator gene (dasR) and two putative integral membrane protein genes (dasBC) are located in the opposite and same orientations, respectively. The expression of dasR and dasB, which seemed independent of dasA transcription, was also induced by (GlcNAc)(2) and (GlcNAc)(3).

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  • Characterization of NtChitIV, a class IV chitinase induced by beta-1,3-, 1,6-glucan elicitor from Alternaria alternata 102: Antagonistic effect of salicylic acid and methyl jasmonate on the induction of NtChitIV Reviewed

    Tomonori Shinya, Kazunari Hanai, Ivan Galis, Kaoru Suzuki, Ken Matsuoka, Hideaki Matsuoka, Mikako Saito

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   353 ( 2 )   311 - 317   2007.2

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    Tobacco BY-2 class IV chitinases (TBC-1, TBC-3) were rapidly and transiently induced by the beta-1,3-, 1,6-glucan elicitor from Alternaria alternata 102 (AaGlucan). The full-length cDNA and 5'-flanking region of a gene encoding class IV chitinases were isolated on the basis of the amino acid sequence of TBC-1. Sequence analysis indicated that NtChitIV encoded TBC-1, TBC-3, or both. Since purified TBC-1 and TBC-3 from BY-2 cells lack a chitin binding domain in the N-terminal region, these enzymes suggested to be derived from NtChitIV by post-translational proteolytic processing. The transcripts of NtChitIV accumulated rapidly within I h after treatment with AaGlucan. Accumulation was maximal 3 h after treatment. Reporter gene assays were used to analyze the promoter regions involved in the transcriptional control of NtChitIV, and these assays revealed that the 1.89-kb NtChitIV promoter was activated by AaGlucan but not by salicylic acid (SA) or methyl jasmonate (MeJA). The AaGlucan-induced transcriptional activation via 1.89-kb NtChitIV promoter was attenuated by pretreatment with SA or MeJA. These results suggest that NtChitIV expression is particularly induced by AaGlucan and that the AaGlucan-dependent signaling pathway is different from the SA- and MeJA-dependent signaling pathways. (c) 2006 Elsevier Inc. All rights reserved.

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  • Novel beta-1,3-, 1,6-oligoglucan elicitor from Alternaria alternata 102 for defense responses in tobacco Reviewed

    T Shinya, R Menard, Kozone, I, H Matsuoka, N Shibuya, S Kauffmann, K Matsuoka, M Saito

    FEBS JOURNAL   273 ( 11 )   2421 - 2431   2006.6

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    A novel elicitor that induces chitinases in tobacco BY-2 cells was isolated from Alternaria alternata 102. Six other fungi, including A. alternata IFO 6587, could not induce, or weakly induce chitinase activity. The purified elicitor was soluble in 75% methanol and showed the chitinase-inducing activity when applied at concentrations of as low as 25 ng-mL(-1). Structural determination by methylation analysis, reducing-end analysis, MALDI-TOF/MS, and NMR spectroscopy indicated that the elicitor was a mixture of beta-1,3-, 1,6-oligoglucans mostly with a degree of polymerization of between 8 and 17. Periodate oxidation of the elicitor suggested that the 1,6-linked and nonreducing terminal residues are essential for the elicitor activity. Further analysis of the elicitor responses in BY-2 cells indicated that the activity of this beta-1,3-, 1,6-glucan elicitor was about 1000 times more potent than that of laminarin, which is a known elicitor of defense responses in tobacco. Analyzing the expression of defense-related genes indicated that a phenylalanine ammonia-lyase gene and a coumaroyl-CoA 0-methyltransferase gene were transiently expressed by this beta-1,3-, 1,6-glucan elicitor. The elicitor induced a weak oxidative burst but did not induce cell death in the BY-2 cells. In the tissue of tobacco plants, this beta-1,3-, 1,6-glucan elicitor induced the expression of basic PR-3 genes, the pherylpropanoid pathway genes, and the sesquiterpenoid pathway genes. In comparison with laminarin and laminarin sulfate, which are reported to be potent elicitors of defense responses in tobacco, the expression pattern of genes induced by the purified beta-1,3-, 1,6-glucan elicitor was more similar to that induced by laminarin than to that induced by laminarin sulfate.

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  • A novel chitinase isozyme in tobacco BY-2 cells induced by the autoclaved Alternaria alternata culture medium Reviewed

    Mikako Saito, Tomonori Shinya, Kazunari Hanai, Tetsuya Katagi, Hideaki Matsuoka

    Plant Science   167 ( 4 )   811 - 817   2004.10

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    BY-2 cells form a linear linkage of several cells. Such a unique configuration is thought to be suitable for the cell-to-cell communication analysis in vivo that is the basis of the elucidation of the systemic acquired resistance in plants. Since chitinase is one of the typical stress response proteins in plants, BY-2 cells were treated with various biotic and abiotic stresses to investigate if chitinase could be induced in BY-2 cells. Among 33 stresses, the autoclaved Alternaria alternata culture medium could induce at least three chitinase isozymes. The amount of each isozyme was very small but the most abundant one, TBC-1, could be successfully isolated at the yield of 2μg from 2.4kg wet matter of BY-2 cells. The N-terminal amino acid sequence of TBC-1 was analyzed, and 24 residues were determined. The sequence was homologous to those of class IV or II chitinases of other plants, but not homologous to those of already known tobacco chitinases. Therefore, TBC-1 was a novel isozyme of tobacco chitinase and the first chitinase found in a BY-2 cell line. © 2004 Elsevier Ireland Ltd. All rights reserved.

    DOI: 10.1016/j.plantsci.2004.05.018

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  • Cell-lytic activity of tobacco BY-2 induced by a fungal elicitor from alternaria alternata attributed to the expression of a class I beta-1,3-glucanase gene. Reviewed International journal

    Tomonori Shinya, Shinobu Gondo, Hiroshi Iijima, Kazunari Hanai, Hideaki Matsuoka, Mikako Saito

    Bioscience, biotechnology, and biochemistry   68 ( 6 )   1265 - 72   2004.6

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    Stress-induced cell-lytic activity was found in tobacco BY-2 cells treated with various stresses. Among 14 stresses, an elicitor fraction isolated from Alternaria alternata showed the highest inducing activity. Cell-lytic activity increased for 72 h even in the control sample, treated with distilled water, and several isozymes of beta-1,3-glucanases and chitinases were found to be involved in it. In contrast, cell-lytic activity in BY-2 cells treated with a fungal elicitor reached a higher level after 60 h. The principal enzymes specifically involved in this stress-induced portion are speculated to be basic beta-1,3-glucanases. A class I beta-1,3-glucanase gene (glu1) was found to be the specific gene for the stress-induced cell-lytic activity. Its expression became observable at 24 h, and the intensity reached a maximum at about 60-72 h. The glu1 was thus assigned as a late gene. Its role in the stress response is discussed in conjunction with earlier genes such as chitinases.

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  • Isolation of a novel isozyme of tobacco BY-2 chitinase induced by a fungal elicitor Reviewed

    Tomonori Shinya, Kazunari Hanai, Mikako Saito, Hideaki Matsuoka

    Plant Biotechnology   21 ( 2 )   155 - 158   2004.6

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    Three isozymes of chitinase were induced by a fungal elicitor in cultured tobacco BY-2 cells. The most acidic one, designated TBC-3, was described here. The molecular mass of TBC-3 was estimated to be 28.5 kDa. The N-terminal amino acid sequence of TBC-3 was analyzed and a homology search was performed. Fifteen amino acids at the N-terminus showed 60% homology to class II chitinases from other plants but not higher than 27% homology to already known chitinases from tobacco. Therefore, TBC-3 is thought to be a novel tobacco chitinase that may be classified into class II.

    DOI: 10.5511/plantbiotechnology.21.155

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  • Sensing of herbivores by damaged self in plants Invited Reviewed

    Tomonori Shinya, Toshihisa Kotake, Ivan Galis

    Japanese Journal of Pesticide Science   47 ( 2 )   74 - 77   2022.8

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    DOI: 10.1584/jpestics.w22-17

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  • 親和性標識実験

    新屋友規

    植物細胞壁実験法 (石井忠ら編)   374 - 376   2016.2

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  • Oligosaccharin receptors in plant immunity

    Tomonori Shinya, Yoshitake Desaki, Naoto Shibuya

    Research Progress in Oligosaccharins   29 - 39   2016.1

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    © Springer Science+Business Media New York 2016. It has been well known that oligosaccharides generated from fungal, bacterial, or plant cell walls can induce various defense responses in plant cells. Traditionally they have been called as “elicitors” or “general elicitors” that induce defense responses in a wide range of plant species. These molecules are now classifi ed as representative MAMPs/PAMPs (microbe/pathogen-associated molecular patterns) or DAMPs (damage/danger-associated molecular patterns) and believed to play important roles in plant immunity, as a trigger of the so-called pattern- triggered immunity (PTI). Biological activities of these oligosaccharide elicitors, including structure/function relationships, corresponding receptors, and downstream signaling, have been the subject of intense studies, by which these oligosaccharides became most well-characterized oligosaccharins. In this chapter, present knowledge on the plant receptors of carbohydrate elicitors generated from the cell walls of pathogenic microbes and also host plants is summarized. Mechanism of the activation of downstream signaling is discussed for chitin receptor, as this system has become the most well-characterized oligosaccharin receptor in plant immunity. It is also discussed how pathogenic microbes try to escape from the MAMP- mediated detection of their invasion by host plants.

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  • Plant immunity and cell wall(<Feature Articles>The plant cell wall as an information processing system)

    Shinya Tomonori, Shibuya Naoto

    Regulation of Plant Growth & Development   50 ( 1 )   76 - 82   2015

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    <p>Plants evolved a multilayered immune system against pathogenic microbes and insect herbivores. Plant cell wall and apoplast consist an essential component of this immune system as the first line of defense, where plant cells contact with these invaders. Plant immune responses can be triggered by the recognition of conserved microbial signature called MAMP (Microbe-associated molecular pattern) at the apoplastic region. In the plant immune system, plant cell wall can function as a barrier against invasion and also a sensor for pathogen attack. Alteration of cell wall integrity and the generation of cell wall-derived fragments are both considered to induce plant immune responses. These MAMPs and damage signals (including DAMPs, Damage-associated molecular patterns) trigger the generation of anti-microbial compounds in the apoplastic region and the reinforcement of the cell wall. On the other hand, pathogenic microbes evolved effector systems to inhibit/escape from the host defense machinery and surveillance system. These battles at the molecular level between plants and microbes often take place in the apoplast. It has also been becoming clear that the plant cell walls are involved in the detection of and defense responses against insect herbivores. Thus plant cell wall and apoplast are not only important as the battle field where host plants and invading organisms compete for the detection and defense responses but also important as a sensor and barrier for these invaders.</p>

    DOI: 10.18978/jscrp.50.1_76

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  • キチン・キトサンの生物活性と利用

    新屋友規, 渋谷直人

    菌類の事典 人間社会編   532 - 533   2013

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  • 免疫装置としてのはたらき「植物の病原菌認識とシグナル伝達における細胞壁の役割」

    新屋友規, 渋谷直人

    生物の科学 遺伝   2012

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  • LysM受容体を介した植物免疫応答

    賀来華江, 新屋友規, 渋谷直人

    化学と生物   50 ( 1 )   52 - 58   2012

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    Language:Japanese   Publisher:Japan Society for Bioscience, Biotechnology, and Agrochemistry  

    DOI: 10.1271/kagakutoseibutsu.50.52

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  • キチン受容体をめぐる植物と微生物のせめぎあい

    新屋友規, 渋谷直人

    化学と生物   2010

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Research Projects

  • 植食性昆虫とその腸内細菌による危険信号分子の産生と植物の受容機構

    Grant number:21K05506  2021.04 - 2025.03

    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)

    新屋 友規

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    Grant amount:\4160000 ( Direct expense: \3200000 、 Indirect expense:\960000 )

    外敵との攻防において植物が自身を守るためには、外敵を適切に認識することが防御機構を効果的に駆動するうえで重要である。食害時の損傷にともない生じる植物由来の分子の一部がエリシターとして植物に認識されることが知られており、我々も植物細胞壁由来の新規糖鎖エリシターを、イネとクサシロキヨトウの相互作用系から見出していた。本研究は、細胞壁損傷における昆虫腸内細菌の関与に注目して、植物由来エリシターの食害時産生メカニズムを明らかにするとともに、植物の昆虫エリシター認識機構に迫る研究を行うことを目的としている。
    令和3年度は、腸内細菌の植物由来エリシター産生への関与の検証を行った。抗生物質カクテルを含む餌で昆虫腸内細菌数を減少させ、エリシターとなる細胞壁断片の産生に必要な細胞壁酵素が腸内細菌に由来することを確認した。さらに、腸内細菌数を減少させたのち回収した吐き戻し液の、防御応答誘導能への影響を解析した。現在、吐き戻し液中の当該糖鎖の直接解析をLC-MSを用いて試みており、検出系が構築できた場合は、腸内細菌数を減少させたのち回収した吐き戻し液において当該糖鎖が検出されるかどうかを調べる。一方で、当該糖鎖エリシターによる一連の応答解析はイネ培養細胞の実験系を用いて行われてきたが、令和3年度はイネリーフディスクアッセイ系を用いた応答解析を行っており、次年度以降も継続して応答解析を進める。また、当該糖鎖のエリシター活性に重要な構造を明らかにするために、類似構造を有する糖鎖の応答解析を行った。

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  • Sensing of herbivores by damaged-cell wall recognition in plants

    Grant number:18K05558  2018.04 - 2022.03

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

    Shinya Tomonori

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    Grant amount:\4420000 ( Direct expense: \3400000 、 Indirect expense:\1020000 )

    Molecules in insect oral secretions, which are deposited at the feeding area on plant leaves, play an important role in the recognition of herbivores by plants. In this study, we identified a plant cell wall derived-saccharide in the oral secretion of rice armyworm (Mythimna loreyi) larvae. The saccharide triggered an array of defense responses in rice-cultured cells. Further analysis implied that the cell wall degrading enzymes involved in the saccharide release is derived from the gut microbes of M. loreyi larvae.

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  • Elucidation of molecular mechanisms for transduction of jasmonate signal to fertility in rice

    Grant number:16K08143  2016.10 - 2019.03

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

    Galis Ivan, SHINYA tomonori

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    We characterized in detail hormonal contents in rice flowers, finding that high levels of JA-Ile coincide with anthesis stage in the rice monocot model. Next, we identified multiple novel targets of JA signaling in flowers by microarray analysis, some of which belong to regulatory class genes. Out of them, novel JA-controlled NAC, MADS-box and AP2 genes of yet unknown functions were depicted as potential master regulators of fertility in rice. Furthermore, we make progress in understanding of metabolic defense in rice that is controlled by JA pathway in vegetative tissues but apparently becomes uncoupled from JA signal during later reproductive stages. Overall, our research provides advances in understanding of cross-talk between growth and defense that is important for achieving optimal fitness and growth of plants in nature.

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  • Functional role of molecular pattern recognition in plant-herbivore interactions

    Grant number:15K18820  2015.04 - 2018.03

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

    Shinya Tomonori

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    Plants detect herbivory through the recognition of molecular patterns derived from herbivores and themselves, however little is known about the molecular interactions of such co-existing signals during insect attack. In this study, we show that rice can simultaneously recognize several molecular patterns from rice armyworm (Mythimna loreyi) larvae. These findings suggest that a concurrent perception of diverse molecular patterns in rice may be required for a more efficient activation of its defense against herbivores.

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  • Molecular interactions at plant-insect interface

    Grant number:24570026  2012.04 - 2015.03

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

    GALIS Ivan, SHINYA Tomonori, TANI Akio

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    Grant amount:\5460000 ( Direct expense: \4200000 、 Indirect expense:\1260000 )

    We established a novel system to study rice-insect interactions. We showed that the accumulation of active defense hormone jasmonoyl-L-isoleucine (JA-Ile) in rice depends on the function of OsJAR1 enzyme. JA and JA-Ile accumulations in rice were controlled by the availability of the main pathway substrate, α-linolenic acid. In comparison to wounding, rice plants specifically responded to elicitors present in chewing herbivore oral secretions. We identified two novel herbivory-related phenolamide phytoalexins. Using ROS and rice phenolamides as molecular markers, we identified and partially purified a new putative insect elicitor from Mythimna loreyi oral secretions.

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  • Similarities and differences in chitin-mediated recognition of microbes between plant species

    Grant number:24780334  2012.04 - 2015.03

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

    SHINYA TOMONORI

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    Grant amount:\4550000 ( Direct expense: \3500000 、 Indirect expense:\1050000 )

    Chitin is a major structural component of fungal cell walls and its perception triggers plant immune responses. Comparative analysis of chitin receptors in two model plants, Arabidopsis and rice, indicated species-specific differences in components of the receptor complex and their molecular characteristics. We also demonstrated that receptor-like cytoplasmic kinase PBL27 is an immediate downstream component of AtCERK1 signaling pathway in Arabidopsis.

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  • Special Seminar in Plant-Insect Interactions (2023academic year) Year-round  - その他

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  • Topics in Plant immunity (2020academic year) special  - その他

  • Plant physiology 2 (2020academic year) Fourth semester  - 金1,金2

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  • Seminar in Plant-insect interactions (2020academic year) Prophase  - その他

  • Seminar in Plant-Insect Interactions (2020academic year) Late  - その他

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