Faculty Profiles - YOSHIOKA, Hirofumi

Papers - YOSHIOKA, Hirofumi

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Molecular mechanisms of the radical burst in plant immunity Invited

Yoshioka H, Asai S, Miyagawa N, Ichikawa T, Yoshioka M, Kobayashi M

In Molecular Plant-Microbe Interactions. (Bouarab, K., Brisson, N. and Daayf, F. eds), CAB International, Oxfordshire, UK page： 59 - 74 2009

Nitric oxide as a partner of reactive oxygen species participates in disease resistance to necrotrophic pathogen Botrytis cinerea in Nicotiana benthamiana. Reviewed

Asai, S, Yoshioka, H

Molecular Plant-Microbe Interactions Vol. 22 page： 619-629 2009

Molecular Mechanisms of the Radical Burst in Plant Immunity

Yoshioka, H; Asai, S; Miyagawa, N; Ichikawa, T; Yoshioka, M; Kobayashi, M

MOLECULAR PLANT-MICROBE INTERACTIONS page： 59 - 74 2009

MAPK signaling regulates nitric oxide and NADPH oxidase-dependent oxidative bursts in Nicotiana benthamiana

Asai, S; Ohta, K; Yoshioka, H

PLANT CELL Vol. 20 ( 5 ) page： 1390 - 1406 2008.5

Involvement of NbNOA1 in NO production and defense responses in INF1-treated Nicotiana benthamiana

Kato, H; Asai, S; Yamamoto-Katou, A; Yoshioka, H; Doke, N; Kawakita, K

JOURNAL OF GENERAL PLANT PATHOLOGY Vol. 74 ( 1 ) page： 15 - 23 2008.2

*MAPK signaling regulates nitric oxide and NADPH oxidase-dependent oxidative bursts in Nicotiana benthamiana. Reviewed

Asai, S, Ohta, K, Yoshioka, H

The Plant Cell Vol. 20 page： 1390-1406 2008

Discovery of oxidative burst in the field of plant immunity: Looking back at the early pioneering works and towards the future development. Reviewed

Yoshioka, H, Bouteau, F, Kawano, T

Plant Signaling and Behavior Vol. 3 page： 153-155 2008

The role of radical burst via MAPK signaling in plant immunity. Invited Reviewed

Asai, S, Yoshioka, H

Plant Signaling and Behavior Vol. 3 page： 920-922 2008

Induction of a small heat shock protein and its functional roles in Nicotiana plants in the defense response against Ralstonia solanacearum(1[W]) Reviewed

Milimo Maimbo, Kouhei Ohnishi, Yasufumi Hikichi, Hirofumi Yoshioka, Akinori Kiba

PLANT PHYSIOLOGY Vol. 145 ( 4 ) page： 1588 - 1599 2007.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PLANT BIOLOGISTS

In tobacco (Nicotiana tabacum), Ralstonia solanacearum OE1-1 (RsOE1-1) is pathogenic, whereas R. solanacearum 8107 (Rs8107) is nonpathogenic and induces the hypersensitive response (HR). To elucidate the molecular mechanisms of plant-R. solanacearum interactions, we used differential display to isolate a cDNA fragment, A6, regulated in tobacco by inoculation with RsOE1-1. The deduced amino acid sequence predicted from full-length A6-cDNA showed similarity to small heat shock proteins from Arabidopsis (Arabidopsis thaliana; hypothetical protein), Medicago truncatula, and Cucumis melo; we therefore designated A6 to correspond to Ntshsp17 (for tobacco small heat shock protein 17). Recombinant Ntshsp17 overproduced in Escherichia coli exhibited molecular chaperone function. Expression of Ntshsp17 was increased in tobacco leaves inoculated with both RsOE1-1 and Rs8107. Expression was induced by heat treatment and by treatment with aminocyclopropane carboxylic acid, hydrogen peroxide, methyl jasmonate, and salicylic acid. Ntshsp17 expression was induced by inoculation with a HR and pathogenicity gene mutant of Rs8107 that does not induce the HR, but not by Agrobacterium-mediated transient expression of INF1, an HR elicitor. In Nbshsp17-silenced plants (an Ntshsp17 ortholog in Nicotiana benthamiana), expression of ETHYLENE-RESPONSE ELEMENT-BINDING PROTEIN, PATHOGENESIS-RELATED1a (PR1a), and PR4 genes was compromised, but expression of ELONGATION FACTOR1 alpha was scarcely affected. Appearance of the HR was not affected in the silenced plants. In the silenced plants, growth of Rs8107 was accelerated. Bacterial growth and wilt symptoms elicited by RsOE1-1 were also accelerated in the silenced plants. These results indicate that this small heat shock protein might have a role in HR-independent defenses in Nicotiana plants.

DOI： 10.1104/pp.107.105353

Web of Science

PubMed

Regulation of rice NADPH oxidase by binding of Rac GTPase to its N-terminal extension Reviewed

Hann Ling Wong, Reinhard Pinontoan, Kokoro Hayashi, Ryo Tabata, Takashi Yaeno, Kana Hasegawa, Chojiro Kojima, Hirofumi Yoshioka, Koh Iba, Tsutomu Kawasaki, Ko Shimamoto

PLANT CELL Vol. 19 ( 12 ) page： 4022 - 4034 2007.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PLANT BIOLOGISTS

Reactive oxygen species (ROS) produced by NADPH oxidase play critical roles in various cellular activities, including plant innate immunity response. In contrast with the large multiprotein NADPH oxidase complex of phagocytes, in plants, only the homologs of the catalytic subunit gp91(phox) and the cytosolic regulator small GTPase Rac are found. Plant homologs of the gp91phox subunit are known as Rboh (for respiratory burst oxidase homolog). Although numerous Rboh have been isolated in plants, the regulation of enzymatic activity remains unknown. All rboh genes identified to date possess a conserved N-terminal extension that contains two Ca2+ binding EF-hand motifs. Previously, we ascertained that a small GTPase Rac (Os Rac1) enhanced pathogen-associated molecular pattern-induced ROS production and resistance to pathogens in rice (Oryza sativa). In this study, using yeast two-hybrid assay, we found that interaction between Rac GTPases and the N-terminal extension is ubiquitous and that a substantial part of the N-terminal region of Rboh, including the two EF-hand motifs, is required for the interaction. The direct Rac-Rboh interaction was supported by further studies using in vitro pull-down assay, a nuclear magnetic resonance titration experiment, and in vivo fluorescence resonance energy transfer (FRET) microscopy. The FRET analysis also suggests that cytosolic Ca2+ concentration may regulate Rac-Rboh interaction in a dynamic manner. Furthermore, transient coexpression of Os Rac1 and rbohB enhanced ROS production in Nicotiana benthamiana, suggesting that direct Rac-Rboh interaction may activate NADPH oxidase activity in plants. Taken together, the results suggest that cytosolic Ca2+ concentration may modulate NADPH oxidase activity by regulating the interaction between Rac GTPase and Rboh.

DOI： 10.1105/tpc.107.055624

Web of Science

PubMed

Involvement of mitogen-activated protein kinase in the induction of StrbohC and StrbohD genes in response to pathogen signals in potato Reviewed

Chihiro Yamamizo, Noriyuki Doke, Hirofumi Yoshioka, Kazuhito Kawakita

Journal of General Plant Pathology Vol. 73 ( 5 ) page： 304 - 313 2007.10

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

The respiratory burst oxidase homolog (rboh), a plant homolog of the catalytic subunit of phagocyte NADPH oxidase (gp91phox ), has been implicated in O2- generation. In the previous study, we reported that StrbohA is constitutively expressed at a low level, whereas StrbohB is upregulated after pathogen signals are perceived in potato tubers. To gain a better understanding about the role of rboh in the defense response in potato leaves, we investigated the regulatory mechanisms of the gene. StrbohB gene was activated in response to an avirulent race of Phytophthora infestans or to hyphal wall components (HWC) from the pathogen
however, levels of expression were very low. We isolated two novel rboh cDNAs, StrbohC and StrbohD, by virtue of the sequence information from the tomato whitefly-induced gene 1 (LeWfi1) and tomato TC121349, respectively. The expression of StrbohC and StrbohD was drastically upregulated in the potato leaves inoculated with P. infestans or treated with HWC elicitor. Mitogen-activated protein kinase (MAPK) is known to play a pivotal role in the oxidative burst. The transcript accumulation induced by HWC treatment was blocked by U0126, which is a specific inhibitor of MAPK kinase (MAPKK). Conversely, the StrbohC and StrbohD genes were upregulated, driven by a pathogen-inducible promoter, in transgenic potato leaves that carry a constitutively active mutant of the MAPKK, StMEK1 DD, that activates defense-related MAPKs. These results suggest that a MAPK cascade is involved in the process leading to the expression of the StrbohC and StrbohD genes. © 2007 The Phytopathological Society of Japan and Springer.

DOI： 10.1007/s10327-007-0020-1

Scopus

Calcium-dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase Reviewed

Michie Kobayashi, Ikuko Ohura, Kazuhito Kawakita, Naohiko Yokota, Masayuki Fujiwara, Ko Shimamoto, Noriyuki Doke, Hirofumi Yoshioka

PLANT CELL Vol. 19 ( 3 ) page： 1065 - 1080 2007.3

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PLANT BIOLOGISTS

Reactive oxygen species (ROS) are implicated in plant innate immunity. NADPH oxidase (RBOH; for Respiratory Burst Oxidase Homolog) plays a central role in the oxidative burst, and EF-hand motifs in the N terminus of this protein suggest possible regulation by Ca2+. However, regulatory mechanisms are largely unknown. We identified Ser-82 and Ser-97 in the N terminus of potato (Solanum tuberosum) St RBOHB as potential phosphorylation sites. An anti-phosphopeptide antibody (pSer82) indicated that Ser-82 was phosphorylated by pathogen signals in planta. We cloned two potato calcium-dependent protein kinases, St CDPK4 and St CDPK5, and mass spectrometry analyses showed that these CDPKs phosphorylated only Ser-82 and Ser-97 in the N terminus of St RBOHB in a calcium-dependent manner. Ectopic expression of the constitutively active mutant of St CDPK5, St CDPK5VK, provoked ROS production in Nicotiana benthamiana leaves. The CDPK-mediated ROS production was disrupted by knockdown of Nb RBOHB in N. benthamiana. The loss of function was complemented by heterologous expression of wild-type potato St RBOHB but not by a mutant (S82A/S97A). Furthermore, the heterologous expression of St CDPK5VK phosphorylated Ser-82 of St RBOHB in N. benthamiana. These results suggest that St CDPK5 induces the phosphorylation of St RBOHB and regulates the oxidative burst.

DOI： 10.1105/tpc.106.048884

Web of Science

PubMed

Non-destructive method to detect the cycle of lamination in sedimentary rocks: rhythmite sequence in Neoproterozoic Cap carbonates

Katsuta, N; Tojo, B; Takano, M; Yoshioka, H; Kawakami, S; Ohno, T; Kumazawa, M

RISE AND FALL OF THE EDIACARAN BIOTA Vol. 286 page： 27 - 34 2007

植物免疫におけるMAPキナーゼとラジカルバースト．植物における環境と生物ストレスに対する応答 Invited

吉岡博文, 浅井秀太

蛋白質核酸酵素増刊号 Vol. 52 page： 667-672 2007

植物病原細菌の感染を制御する宿主応答機構～ Ralstonia solanacearum- Nicotiana属植物相互作用の解析～ Invited

木場章範, Maimbo, M, 小森大輔, 張宛果, 富山裕美, 高橋彰子, 大西浩平, 吉岡博文, 曳地康史

植物－病原微生物の相互作用のダイナミズム Vol. 43 page： 55-65 2007

植物免疫におけるラジカルバーストの分子機構 Invited

吉岡博文, 浅井秀太, 間瀬圭介, 市川達士, 小林光智衣, 小八重善裕

植物－病原微生物の相互作用のダイナミズム Vol. 43 page： 37-47 2007

Nitrate reductase is responsible for elicitin-induced nitric oxide production in nicotiana benthamiana Reviewed

Ayako Yamamoto-Katou, Shinpei Katou, Hirofumi Yoshioka, Noriyuki Doke, Kazuhito Kawakita

PLANT AND CELL PHYSIOLOGY Vol. 47 ( 6 ) page： 726 - 735 2006.6

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

Recent works have established a key role for nitric oxide (NO) in activating disease resistance in plants. Nitrate reductase (NR) is one of the enzymes that are capable of producing NO in plants. In a previous study, we reported that pathogen signals induce expression of NR genes in potato, suggesting the involvement of NR in NO production induced by pathogen signals. In this study, we cloned NR genes from Nicotiana benthamiana and investigated their involvement in NO production induced by INF1, a major elicitin secreted by Phytophthora infestans. Treatment of protoplasts prepared from N. benthamiana leaves with INF1 elevated NO production to a maximum level 1-3 h after treatment. INF1-induced NO generation was suppressed completely by an NO-specific scavenger, but partially by a nitric oxide synthase inhibitor. To investigate the involvement of NR in INF1-induced NO production, NR genes were silenced by virus-induced gene silencing. The NR-silenced plants showed yellowish leaves which resemble the characteristic of Arabidopsis NR double mutants. Silencing of NR genes significantly decreased both NO2--producing activity and INF1-induced NO production, indicating that NR is involved in INF1-induced NO production. In contrast, overexpression of NbNR1 encoding N. benthamiana NR by Agrobacterium-mediated transient expression elevated NO2--producing activity nine times over the control; however, INF1-induced NO production in protoplasts overexpressing NbNR1 was comparable with that in control protoplasts. These results suggest that NR is involved in INF1-induced NO production, and post-translational modification of NR or availability of substrate NO2- may be a rate-limiting step of NO production by NR.

DOI： 10.1093/pcp/pcj044

Web of Science

PubMed

Peroxynitrite generation and tyrosine nitration in defense responses in tobacco BY-2 cells

Syuhei Saito, Ayako Yamamoto-Katou, Hirofumi Yoshioka, Noriyuki Doke, Kazuhito Kawakita

PLANT AND CELL PHYSIOLOGY Vol. 47 ( 6 ) page： 689 - 697 2006.6

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

Peroxynitrite (ONOO-) is a compound formed by reaction of superoxide (O-2 (-)) with nitric oxide (NO) and is expected to possess characteristics of both O-2 (-) reactivity and NO mobility in order to function as a signal molecule. Although there are several reports that describe the role of ONOO- in defense responses in plants, it has been very difficult to detect ONOO- in bioimaging due to its short half-life or paucity of methods for ONOO--specific detection among reactive oxygen species or free radicals. Aminophenyl fluorescein (APF), a recently developed novel fluorophore for direct detection of ONOO- in bioimaging, was used for intracellular ONOO- detection. ONOO- generation in tobacco BY-2 cells treated with INF1, the major elicitin secreted by the late blight pathogen Phytophthora infestans, occurred within 1 h and reached a maximum level at 6-12 h after INF1 treatment. Urate, a ONOO- scavenger, abolished INF1-induced ONOO- generation. It is well known that ONOO- reacts with tyrosine residues in proteins to form nitrotyrosine in a nitration reaction as an ONOO--specific reaction. Western blot analysis using anti-nitrotyrosine antibodies recognized nitrotyrosine-containing proteins in 20 and 50 kDa bands in BY-2 protein extract containing SIN-1 [3-(4-morpholinyl) sydnonimine hydrochloride; an ONOO- donor]. These bands were also recognized in INF1-treated BY-2 cells and were found to be slightly suppressed by urate. Our study is the first to report ONOO- detection and tyrosine nitration in defense responses in plants.

DOI： 10.1093/pcp/pcj038

Web of Science

PubMed

Loss of AtPDR8, a plasma membrane ABC transporter of Arabidopsis thaliana, causes hypersensitive cell death upon pathogen infection Reviewed

Y Kobae, T Sekino, H Yoshioka, T Nakagawa, E Martinoia, M Maeshima

PLANT AND CELL PHYSIOLOGY Vol. 47 ( 3 ) page： 309 - 318 2006.3

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

Plants contain a large number of ATP-binding cassette (ABC) transporters belonging to different subclasses. AtPDR8 is the only member of the pleiotropic drug resistance (PDR) ABC transporter subclass in Arabidopsis that is constitutively highly expressed. In transgenic Arabidopsis plants harboring the AtPDR8 promoter fused to beta-glucuronidase (GUS), reporter expression was shown to he strong in the stomata and hydathode. In the stomata, transcripts of AtPDR8 were particularly frequent in the cells surrounding air spaces. Subcellular fractionation and immunochemical analysis showed that AtPDR8 was localized in the plasma membrane. When a knockout mutant of AtPDR8 (atpdr8) was infected with bacterial and oomycete pathogens, the plants exhibited chlorotic lesions and a hypersensitive response (HR)-like cell death. Cell death was detected in the atpdr8 mutants within 10 h of infection with the virulent bacterial pathogen, Pseudomonas syringae. As a result, the growth of P syringae in the leaves of the atpdr8 mutant was reduced to 1% of that in the wild type. The defense response genes, PR-1, PR-2, PR-5, VPE gamma, AtrbohD and AtrbohF were highly expressed when the mutant plants were grown under non-sterile conditions. The expression of the AtPDR8 gene was enhanced by infection of virulent and avirulent bacterial pathogens. Our results indicate that AtPDR8 is a key factor controlling the extent of cell death in the defense response and suggest that AtPDR8 transports some substance(s) which is closely related to the response of plants to pathogens.

DOI： 10.1093/pcp/pcj001

Web of Science

PubMed

Rewiring mitogen-activated protein kinase cascade by positive feedback confers potato blight resistance Reviewed

C Yamamizo, K Kuchimura, A Kobayashi, S Katou, K Kawakita, JDG Jones, N Doke, H Yoshioka

PLANT PHYSIOLOGY Vol. 140 ( 2 ) page： 681 - 692 2006.2

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC PLANT BIOLOGISTS

Late blight, caused by the notorious pathogen Phytophthora infestans, is a devastating disease of potato (Solanum tuberosum) and tomato (Solanum lycopersicum), and during the 1840s caused the Irish potato famine and over one million fatalities. Currently, grown potato cultivars lack adequate blight tolerance. Earlier cultivars bred for resistance used disease resistance genes that confer immunity only to some strains of the pathogen harboring corresponding avirulence gene. Specific resistance gene-mediated immunity and chemical controls are rapidly overcome in the field when new pathogen races arise through mutation, recombination, or migration from elsewhere. A mitogen-activated protein kinase (MAPK) cascade plays a pivotal role in plant innate immunity. Here we show that the transgenic potato plants that carry a constitutively active form of MAPK kinase driven by a pathogen-inducible promoter of potato showed high resistance to early blight pathogen Alternaria solani as well as P. infestans. The pathogen attack provoked defense-related MAPK activation followed by induction of NADPH oxidase gene expression, which is implicated in reactive oxygen species production, and resulted in hypersensitive response-like phenotype. We propose that enhancing disease resistance through altered regulation of plant defense mechanisms should be more durable and publicly acceptable than engineering overexpression of antimicrobial proteins.

DOI： 10.1104/pp.105.074906

Web of Science

PubMed

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Papers - YOSHIOKA, Hirofumi