Faculty Profiles - ICHINOSE Yuki

MISC - ICHINOSE Yuki

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Induction of defense responses in pea tissues by inorganic phosphate

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

Journal of General Plant Pathology 72 ( 3 ) 129 - 136 2006.6

Language：English

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

DOI： 10.1007/s10327-005-0261-9

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

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

PHYTOPATHOLOGY 96 ( 6 ) S74 - S74 2006.6

植物の非宿主抵抗性

一瀬勇規

化学と生物 44 ( 8 ) 556 - 562 2006

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

松井英譲, 豊田和弘, 稲垣善茂, 白石友紀, 一瀬勇規

日本植物病理学会報 72 ( 4 ) 2006

(18) Rer1 Gene Homologues Expressed in Medicago truncatula and Barley Challenged with Virulent Pathogens(Abstracts of the Papers Presented at the 2005 Annual Meeting in Shizuoka)

Yoshihiro M., Hirose M., Fujita K., Inagaki Y., Ichinose Y., Toyoda K., Shiraishi T.

Annals of the Phytopathological Society of Japan 71 ( 3 ) 190 - 190 2005.8

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

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

Journal of General Plant Pathology 71 ( 4 ) 289 - 295 2005.8

Language：English

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

DOI： 10.1007/s10327-005-0200-9

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

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

Journal of General Plant Pathology 71 ( 4 ) 302 - 307 2005.8

Language：English

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

DOI： 10.1007/s10327-005-0201-8

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

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

Journal of General Plant Pathology 71 ( 4 ) 302 - 307 2005.8

Language：English

DOI： 10.1007/s10327-005-0201-8

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

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

Journal of General Plant Pathology 71 ( 4 ) 273 - 279 2005.8

Language：English

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

DOI： 10.1007/s10327-005-0197-0

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Identification of genes expressed during spore germination of Mycosphaerella pinodes

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

Journal of General Plant Pathology 71 ( 3 ) 190 - 195 2005.6

Language：English

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

DOI： 10.1007/s10327-005-0185-4

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

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

植物微生物研究会研究交流会講演要旨集 14th (JA)120,(EN)121 2005.3

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

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

Journal of General Plant Pathology 71 ( 1 ) 8 - 22 2005.2

Language：English

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

DOI： 10.1007/s10327-004-0168-x

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

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

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

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

PLANT AND CELL PHYSIOLOGY 46 S94 - S94 2005

Mechanism of flagellin-induced hypersensitive cell death in Arabidopsis thaliana

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

PLANT AND CELL PHYSIOLOGY 46 S94 - S94 2005

Pseudomonas syringaeのべん毛を介した植物相互作用

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

日本植物病理学会植物感染生理談話会論文集 ( 40 ) 13 - 24 2004.8

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

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

Journal of General Plant Pathology 70 ( 2 ) 93 - 96 2004.4

Language：English

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

DOI： 10.1007/s10327-003-0099-y

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

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

MOLECULAR GENETICS AND GENOMICS 271 ( 1 ) 1 - 10 2004.2

Authorship：Last author,　Corresponding author Language：English Publisher：SPRINGER-VERLAG

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

DOI： 10.1007/s00438-003-0948-6

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

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

PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY 64 ( 1 ) 17 - 25 2004.1

Language：English Publisher：ACADEMIC PRESS LTD ELSEVIER SCIENCE LTD

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

DOI： 10.1016/j.pmpp.2004.05.003

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耐病性のシグナリングモデル，新

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

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