研究者詳細 - 一瀬　勇規

論文 - 一瀬　勇規

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Positive chemotaxis to plant apoplastic fluids of Pseudomonas syringae pv. tabaci 6605 and metabolome analysis 査読

Yuta Watanabe, Stephany Angelia Tumewu, Hajime Yamada, Hidenori Matsui, Mikihiro Yamamoto, Yoshiteru Noutoshi, Kazuhiro Toyoda, Yuki Ichinose

Journal of General Plant Pathology 89 ( 4 ) 219 - 223 2023年6月

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

Abstract

Pseudomonas syringae pv. tabaci 6605 (Pta6605) is a causal agent of wildfire disease in host tobacco plants. Although chemotaxis has been shown to be necessary for Pta6605 in tobacco infection, the chemoattractants at the site of infection are unclear. Pta6605 was attracted to the apoplastic fluid from not only host tobacco leaves but also non-host plant leaves, indicating that Pta6605 is attracted to common plant metabolites. Metabolome analysis of apoplastic fluid from tobacco leaves revealed that amino acids including γ-aminobutyric acid and organic acids are abundant, suggesting that these compounds are potential chemoattractants.

DOI： 10.1007/s10327-023-01126-4

その他リンク： https://link.springer.com/article/10.1007/s10327-023-01126-4/fulltext.html

Requirement of chemotaxis and aerotaxis in host tobacco infection by Pseudomonas syringae pv. tabaci 6605 査読

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

Physiological and Molecular Plant Pathology 124 101970 - 101970 2023年3月

HopAZ1, a type-III effector of Pseudomonas amygdali pv. tabaci, induces a hypersensitive response in tobacco wildfire-resistant Nicotiana tabacum cv. N509. 査読

Kashihara, S., Nishimura, T., Noutoshi, Y., Yamamoto, M., Toyoda, K., Ichinose, Y. and Matsui, H.

Mol. Plant Pathol. 23 ( 6 ) 885 - 894 2022年6月

HopAZ1, a type III effector of Pseudomonas amygdali pv. tabaci , induces a hypersensitive response in tobacco wildfire‐resistant Nicotiana tabacum ‘N509’

Sachi Kashihara, Takafumi Nishimura, Yoshiteru Noutoshi, Mikihiro Yamamoto, Kazuhiro Toyoda, Yuki Ichinose, Hidenori Matsui

Molecular Plant Pathology 23 ( 6 ) 885 - 894 2022年6月

Identification of aerotaxis receptor proteins involved in host plant infection by Pseudomonas syringae pv. tabaci 6605. 査読

Tumewu, S. A., F., Matsui, H., Yamamoto, M., Noutoshi, Y., Toyoda, K., Ichinose, Y.

Microbes and Environments 37 ME21076 2022年2月

Identification of Aerotaxis Receptor Proteins Involved in Host Plant Infection by Pseudomonas syringae pv. tabaci 6605.

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

Microbes and environments 37 ( 1 ) 2022年

記述言語：英語掲載種別：研究論文（学術雑誌）

Pseudomonas syringae pv. tabaci 6605 (Pta6605) is a foliar plant pathogen that causes wildfire disease on tobacco plants. It requires chemotaxis to enter plants and establish infection. While chemotactic signals appear to be the main mechanism by which Pta6605 performs directional movement, the involvement of aerotaxis or energy taxis by this foliar pathogen is currently unknown. Based on domain structures and similarity with more than 50 previously identified putative methyl-accepting chemotaxis proteins (MCPs), the genome of Pta6605 encodes three potential aerotaxis transducers. We identified AerA as the main aerotaxis transducer and found that it possesses a taxis-to-serine-and-repellent (Tsr)-like domain structure that supports a periplasmic 4HB-type ligand-binding domain (LBD). The secondary aerotaxis transducer, AerB, possesses a cytosolic PAS-type LBD, similar to the Aer of Escherichia coli and Pseudomonas aeruginosa. Aerotaxis ability by single and double mutant strains of aerA and aerB was weaker than that by wild-type Pta6605. On the other hand, another cytosolic PAS-type LBD containing MCP did not make a major contribution to Pta6605 aerotaxis in our assay system. Furthermore, mutations in aerotaxis transducer genes did not affect surface motility or chemotactic attraction to yeast extract. Single and double mutant strains of aerA and aerB showed less colonization in the early stage of host plant infection and lower biofilm production than wild-type Pta6605. These results demonstrate the presence of aerotaxis transducers and their contribution to host plant infection by Pta6605.

DOI： 10.1264/jsme2.ME21076

Ralstonia solanacearum type III effector RipAC targets SGT1 to suppress effector-triggered immunity. 招待

Nakano, M., Ichinose, Y., Mukaihara, T.

Plant Cell Physiol. 61 ( 12 ) 2067 - 2076 2021年12月

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

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

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

Microbiol. Res. 253 126869 2021年8月

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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. 査読国際誌

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

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

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

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

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

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

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

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

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

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.

DOI： 10.1111/j.1364-3703.2012.00789.x

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

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.

DOI： 10.3390/genes2040788

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

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

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

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

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

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

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

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

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 ( 1 ) 11 - 17 2009年

Pseudomonas syringae の鞭毛を介した植物相互作用の研究

一瀬勇規

日本植物病理学会報 75 ( 3 ) 147 - 149 2009年

記述言語：日本語出版者・発行元：日本植物病理學會

植物病原細菌Pseudomonas syringaeには宿主や病徴を異にする多くの病原型が存在し、宿主植物の葉や茎、果実の傷口、気孔などから侵入後、細胞間隙で増殖・移動し、主に斑点・褐変・枯死を引き起こす。これまでに50以上の病原型が報告され、それら病原型はそれぞれ固有の宿主植物に感染が可能である。例えば、タバコ野火病菌（以下Pta6605と略す）はタバコ葉に淡黄色水浸状小斑点を生じ、その周辺に黄化を引き起こすものの、ダイズやトマト、シロイヌナズナには非病原性である。植物病原細菌の非宿主植物に対する過敏感反応（HR）誘導能と宿主植物に対する病原性に必要な領域として、hrp遺伝子クラスターが知られている。hrp遺伝子クラスターには、Type III分泌経路を構成するタンパク質遺伝子、hrp遺伝子の制御因子遺伝子の他、非宿主植物に過敏感反応を誘導するタンパク質エリシターharpinをコードするhrpZなどが存在する。私たちはP. syringaeのいくつかの病原型細菌においてhrpZ遺伝子を調べたところ、Pta6605のhrpZは中央部に326塩基の内部欠損とそれに伴うフレームシフトを起こしており、HR誘導能を欠失していることを見出した。HarpinはP. syringaeの非宿主植物に対する主要なHR誘導因子であると考えられていたが、Pta6605を非宿主植物に高濃度で接種するとHRが誘導されることより、Pta6605にはharpin以外のエリシターが存在すると推察された。そこで、Pta6605の培養上清からトマトに対し、HRを誘導する分子の検索を行い、鞭毛構成タンパク質フラジェリンを同定した。

DOI： 10.3186/jjphytopath.75.147

その他リンク： http://agriknowledge.affrc.go.jp/RN/2010780609

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

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

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

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

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

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

Distinct Infection Mechanisms of Rhizoctonia solani AG-1 IA and AG-4 HG-I+II in Brachypodium distachyon and Barley

Niranjan Mahadevan, Rozi Fernanda, Yusuke Kouzai, Natsuka Kohno, Reiko Nagao, Khin Thida Nyein, Megumi Watanabe, Nanami Sakata, Hidenori Matsui, Kazuhiro Toyoda, Yuki Ichinose, Keiichi Mochida, Hiroshi Hisano, Yoshiteru Noutoshi

Life 15 ( 2 ) 235 - 235 2025年2月

掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

Rhizoctonia solani is a basidiomycete phytopathogenic fungus that causes rapid necrosis in a wide range of crop species, leading to substantial agricultural losses worldwide. The species complex is divided into 13 anastomosis groups (AGs) based on hyphal fusion compatibility and further subdivided by culture morphology. While R. solani classifications were shown to be independent of host specificity, it remains unclear whether different R. solani isolates share similar virulence mechanisms. Here, we investigated the infectivity of Japanese R. solani isolates on Brachypodium distachyon and barley. Two isolates, AG-1 IA (from rice) and AG-4 HG-I+II (from cauliflower), infected leaves of both plants, but only AG-4 HG-I+II infected roots. B. distachyon accessions Bd3-1 and Gaz-4 and barley cultivar ‘Morex’ exhibited enhanced resistance to both isolates compared to B. distachyon Bd21 and barley cultivars ‘Haruna Nijo’ and ‘Golden Promise’. During AG-1 IA infection, but not AG-4 HG-I+II infection, resistant Bd3-1 and Morex induced genes for salicylic acid (SA) and N-hydroxypipecolic acid (NHP) biosynthesis. Pretreatment with SA or NHP conferred resistance to AG-1 IA, but not AG-4 HG-I+II, in susceptible B. distachyon Bd21 and barley Haruna Nijo. On the leaves of susceptible Bd21 and Haruna Nijo, AG-1 IA developed extensive mycelial networks with numerous infection cushions, which are specialized infection structures well-characterized in rice sheath blight. In contrast, AG-4 HG-I+II formed dispersed mycelial masses associated with underlying necrosis. We propose that the R. solani species complex encompasses at least two distinct infection strategies: AG-1 IA exhibits a hemibiotrophic lifestyle, while AG-4 HG-I+II follows a predominantly necrotrophic strategy.

DOI： 10.3390/life15020235

Nutrient requirements shape the preferential habitat of Allorhizobium vitis VAR03-1, a commensal bacterium, in the rhizosphere of Arabidopsis thaliana

Niarsi Merry Hemelda, Jiyuan Bao, Megumi Watanabe, Hidenori Matsui, Kazuhiro Toyoda, Yuki Ichinose, Yoshiteru Noutoshi

Plant And Cell Physiology 2024年8月

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

Abstract

A diverse range of commensal bacteria inhabit the rhizosphere, influencing host plant growth and responses to biotic and abiotic stresses. While root-released nutrients can define soil microbial habitats, the bacterial factors involved in plant-microbe interactions are not well characterized. In this study, we investigated the colonization patterns of two plant disease biocontrol agents, Allorhizobium vitis VAR03-1 and Pseudomonas protegens Cab57, in the rhizosphere of Arabidopsis thaliana using Murashige and Skoog (MS) agar medium. VAR03-1 formed colonies even at a distance from the roots, preferentially in the upper part, while Cab57 colonized only the root surface. The addition of sucrose to the agar medium resulted in excessive proliferation of VAR03-1, similar to its pattern without sucrose, whereas Cab57 formed colonies only near the root surface. Overgrowth of both bacterial strains upon nutrient supplementation inhibited host growth, independent of plant immune responses. This inhibition was reduced in the VAR03-1 ΔrecA mutant, which exhibited increased biofilm formation, suggesting that some activities associated with the free-living lifestyle rather than the sessile lifestyle may be detrimental to host growth. VAR03-1 grew in liquid MS medium with sucrose alone, while Cab57 required both sucrose and organic acids. Supplementation of sugars and organic acids allowed both bacterial strains to grow near and away from Arabidopsis roots in MS agar. These results suggest that nutrient requirements for bacterial growth may determine their growth habitats in the rhizosphere, with nutrients released in root exudates potentially acting as a limiting factor in harnessing microbiota.

DOI： 10.1093/pcp/pcae091

Rhizoviticin is an alphaproteobacterial tailocin that mediates biocontrol of grapevine crown gall disease

Tomoya Ishii, Natsuki Tsuchida, Niarsi Merry Hemelda, Kirara Saito, Jiyuan Bao, Megumi Watanabe, Atsushi Toyoda, Takehiro Matsubara, Mayuko Sato, Kiminori Toyooka, Nobuaki Ishihama, Ken Shirasu, Hidenori Matsui, Kazuhiro Toyoda, Yuki Ichinose, Tetsuya Hayashi, Akira Kawaguchi, Yoshiteru Noutoshi

The ISME Journal 18 ( 1 ) 2024年1月

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

Abstract

Tailocins are headless phage tail structures that mediate interbacterial antagonism. Although the prototypical tailocins, R- and F-pyocins, in Pseudomonas aeruginosa, and other predominantly R-type tailocins have been studied, their presence in Alphaproteobacteria remains unexplored. Here, we report the first alphaproteobacterial F-type tailocin, named rhizoviticin, as a determinant of the biocontrol activity of Allorhizobium vitis VAR03-1 against crown gall. Rhizoviticin is encoded by a chimeric prophage genome, one providing transcriptional regulators and the other contributing to tail formation and cell lysis, but lacking head formation genes. The rhizoviticin genome retains a nearly intact early phage region containing an integrase remnant and replication-related genes critical for downstream gene transcription, suggesting an ongoing transition of this locus from a prophage to a tailocin-coding region. Rhizoviticin is responsible for the most antagonistic activity in VAR03-1 culture supernatant against pathogenic A. vitis strain, and rhizoviticin deficiency resulted in a significant reduction in the antitumorigenic activity in planta. We identified the rhizoviticin-coding locus in eight additional A. vitis strains from diverse geographical locations, highlighting a unique survival strategy of certain Rhizobiales bacteria in the rhizosphere. These findings advance our understanding of the evolutionary dynamics of tailocins and provide a scientific foundation for employing rhizoviticin-producing strains in plant disease control.

DOI： 10.1093/ismejo/wrad003

HexR Transcription Factor Contributes to Pseudomonas cannabina pv. alisalensis Virulence by Coordinating Type Three Secretion System Genes

Nanami Sakata, Takashi Fujikawa, Ayaka Uke, Takako Ishiga, Yuki Ichinose, Yasuhiro Ishiga

Microorganisms 2023年4月

CEP peptide, a family of conserved, secreted small peptides acts as an endogenous suppressor in Arabidopsis 招待査読

Toyoda, K, Fitrianti, A.N, Itoh, C, Hasegawa, H, Matsui, H, Noutoshi, Y, Yamamoto, M, Ichinose, Y, Shiraishi, T

125 102019 2023年4月

Time-series transcriptome of Brachypodium distachyon during bacterial flagellin-induced pattern-triggered immunity

Tsubasa Ogasahara, Yusuke Kouzai, Megumi Watanabe, Akihiro Takahashi, Kotaro Takahagi, June-Sik Kim, Hidenori Matsui, Mikihiro Yamamoto, Kazuhiro Toyoda, Yuki Ichinose, Keiichi Mochida, Yoshiteru Noutoshi

Frontiers in Plant Science 13 2022年9月

掲載種別：研究論文（学術雑誌）出版者・発行元：Frontiers Media {SA}

<jats:p>Plants protect themselves from microorganisms by inducing pattern-triggered immunity (PTI) <jats:italic>via</jats:italic> recognizing microbe-associated molecular patterns (MAMPs), conserved across many microbes. Although the MAMP perception mechanism and initial events during PTI have been well-characterized, knowledge of the transcriptomic changes in plants, especially monocots, is limited during the intermediate and terminal stages of PTI. Here, we report a time-series high-resolution RNA-sequencing (RNA-seq) analysis during PTI in the leaf disks of <jats:italic>Brachypodium distachyon</jats:italic>. We identified 6,039 differentially expressed genes (DEGs) in leaves sampled at 0, 0.5, 1, 3, 6, and 12 hours after treatment (hat) with the bacterial flagellin peptide flg22. The k-means clustering method classified these DEGs into 10 clusters (6 upregulated and 4 downregulated). Based on the results, we selected 10 PTI marker genes in <jats:italic>B. distachyon</jats:italic>. Gene ontology (GO) analysis suggested a tradeoff between defense responses and photosynthesis during PTI. The data indicated the recovery of photosynthesis started at least at 12 hat. Over-representation analysis of transcription factor genes and cis-regulatory elements in DEG promoters implied the contribution of 12 WRKY transcription factors in plant defense at the early stage of PTI induction.</jats:p>

DOI： 10.3389/fpls.2022.1004184

CEP peptide induces susceptibility of Arabidopsis thaliana to non-adapted pathogens. 査読

Fitrianti, A. N., Mai, T. L., Phuong, L. T., Monden, H., Shiiba, N., Matsui, H., Noutoshi, Y., Yamamoto, M., Ichinose, Y., Shiraishi, T. and Toyoda, K.

J. Gen. Plant Pathol. 88 ( 5 ) 287 - 292 2022年9月

CEP peptide induces susceptibility of Arabidopsis thaliana to non-adapted pathogens

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

Journal of General Plant Pathology 88 ( 5 ) 287 - 292 2022年9月

Surveillance of pathogenicity of Rhizoctonia solani Japanese isolates with varied anastomosis groups and subgroups on Arabidopsis thaliana. 査読

Abdelghany, M. M. A., Kurikawa, M., Watanabe, M., Yamamoto, M., Matsui, H., Ichinose, Y., Toyoda, K., Kouzai, Y. and Noutoshi, Y.

Life 12 76 2022年2月

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

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

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

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

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

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

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

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

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

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

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 217 ( 2 ) 771 - 783 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

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

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

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

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

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

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

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

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

植物細胞壁における病原体認識・情報伝達・応答のダイナミズム

豊田和弘, 田中佳織, 稲垣善茂, 一瀬勇規, 白石友紀

日本植物病理学会報 80 ( 3 ) 146 - 151 2014年

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

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

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

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

記述言語：日本語掲載種別：研究論文（大学，研究機関等紀要）出版者・発行元：岡山大学農学部

A plant growth-promoting fungus was isolated from an agricultural field in Okayama Prefecture, Japan. The strain FS2, which enhanced seed germination, root elongation and leaf growth of Brassica rapa var. perviridis, was identified as Talaromyces wortmannii based on ITS1 sequence and its morphology.
本研究では，実際の生産圃場から植物生育促進菌（PGPF）の探索を試み，コマツナの生育を促進するFS2株を分離した．FS2株の形態観察並びにのITS1領域の系統樹解析から本菌をTalaromyces wortmanniiと同定した．

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

記述言語：日本語掲載種別：研究論文（大学，研究機関等紀要）出版者・発行元：岡山大学農学部

A plant growth-promoting fungus, Talaromyces wortmannii strain FS2 was isolated from an agricultural field at Okayama Pref. FS2 enhanced seed germination, root elongation and leaf growth of Brassica rapa var perviridis (Komatsuna). Such plant growth-promoting effect was observed in the same sealed chamber where FS2 was cultured on PDA medium separated from seedlings, suggesting effective volatile compound(s). GC‒MS analysis showed that FS2 emitted at least seven terpenoids, of which a volatile was identified as β‒caryophyllene. β‒caryophyllene alone promoted the growth of cucumber, Nicotiana benthamiana and barley. Furthermore β‒caryophyllene increased the yield of cucumber fruits. Interestingly, we found that β‒caryophyllene conditioned these plants to be resistant to respective diseases caused by Colletotrichum orbiculare, Botrytis cinerea or Blumeria graminis f. sp hordei. The findings indicate that β‒caryophyllene has desirable dual features and therefore, it is available to cultivation of many crops.
岡山県総社市の圃場から分離した植物生育促進菌Talaromyces wortmannii FS2が生産するβ-caryophylleneは，コマツナ（アブラナ科）のみならず，キュウリ（ウリ科），タバコ（ナス科）およびオオムギ（イネ科）など広汎な植物に対して，生育促進作用および耐病性増進作用を示したことから，有用な農業資材として利用可能であるものと考察した．

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

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

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

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

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

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 ( 6 ) 473 - 484 2012年

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

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

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

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 -> 3)-alpha-L-Rhap-(1 -> 2)-alpha-L-Rhap-(1 -> 2)-alpha-L-Rha-(1 ->, as well a trisaccharide, 4,6-dideoxy-4-(3-hydroxybutanamido)-2-O-methyl-Glcp-(1 -> 3)-alpha-L-Rhap-(1 -> 2)-alpha-L-Rha-(1 ->,which was identified in a previous study. (C) 2009 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.carres.2009.07.004

我が国におけるブドウ根頭がんしゅ病菌の遺伝型とその分布

川口章, 澤田宏之, 一瀬勇規

土と微生物 62 ( 2 ) 143 - 143 2008年6月

[MAMP signal transduction].

Yuki Ichinose

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

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

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.

DOI： 10.1093/pcp/pcg042

エンドウのエリシター誘導性遺伝子発現におけるAAAGモチーフとPsDof1タンパク質の関与

關, 光, 丸谷, 瑞理, 稲垣, 善茂, 豊田, 和弘, 白石, 友紀, 一瀬, 勇規

岡山大學農學部學術報告 = Scientific report of the Faculty of Agriculture, Okayama University 92 21 - 26 2003年2月

記述言語：日本語掲載種別：研究論文（学術雑誌）出版者・発行元：岡山大學農學部

エリシターを処理したエンドウ上胚軸由来のRNAから作成されたcDNAライブラリーからエリシター処理により発現が増高する遺伝子候補のcDNAとしてPsDof1が単離された。大腸菌で生産されたGST-PsDof1融合タンパク質はAAAG配列をコアとするDNAに結合することが明らかにされている。本論文では、GST-PsDof1がエリシター応答性遺伝子の一つ、PsCHS1のプロモーター上のAAAGまたはCTTT配列を有する断片に特異的に結合することを明らかにした。更にAAAG配列のエリシター応答性シスエレメントとしての機能を解析するため、AAAG配列を4回繰り返したユニットをCaMV35Sの最小プロモーターとCATレポーター遺伝子に連結したキメラ遺伝子を構築し、エンドウプロトプラストにエレクトロポレーション法により導入した。CAT活性を指標にプロモーター活性を調べたところ、AAAG配列を有するプロモーターは、エリシター処理により活性化されることが明らかとなった。これらの結果はPsDof1がエリシター応答性防御遺伝子のプロモーター上のAAAG配列に結合し、転写を活性化させる可能性を示唆している。
identifier:670005
identifier:ZZ00015342

その他リンク： http://id.ndl.go.jp/bib/6470981

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

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

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

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.

DOI： 10.1016/S0014-5793(02)02697-2

初期・表層シグナル伝達系と防御システム

白石友紀, 豊田和弘, 木場章範, 一瀬勇規

化学と生物 39 ( 10 ) 686 - 692 2001年10月

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

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

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.

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

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

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.

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

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

H+-translocating Activity in Proteoliposomes Reconstituted with Pea Plasma Membrane ATPase and Its Inhibition by Fungal Suppressor from Mycosphaerella pinodes.

天野政史, 豊田和弘, 一瀬勇規, 山田哲治, 白石友紀

日本植物病理学会報 61 ( 4 ) 369 - 375 1995年

記述言語：英語出版者・発行元：The Phytopathological Society of Japan

エンドウ褐紋病菌のサプレッサーは宿主エンドウの原形質膜ATPase活性を抑制する。本報告では,可溶化・部分精製したエンドウ原形質膜ATPaseをリボソームに再構成し,ATPaseとプロトンポンプ活性に対するエリシターとサプレッサーの影響について調べた。再構成膜におけるプロトンポンプ活性はATPとMg²⁺に依存し,ATPaseの阻害剤で抑制されることからATPase依存性であることが推察できた。また,プロトンの勾配はプロトンイオノフォアやNH₄Clの添加で打ち消された。本活性に対するエリシターとサプレッサーの影響を調べたところ,エリシターは全く影響を及ぼさなかったが,サプレッサーは濃度依存的に活性を抑制した。さらに,ホスファチジルイノシトール2リン酸は本活性を上昇させたが,その作用点はサプレッサーの作用点とは異なることが示唆された。これらの結果から,エンドウ褐紋病菌はサプレッサーを分泌し,エンドウ細胞におけるホメオスタシスを攪乱することにより抵抗反応を抑制し,感染に成功するものと考察した。

DOI： 10.3186/jjphytopath.61.369

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

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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