Screening of plant signlas inducing hrp reguon in Ralstonia solanacearum

• Project/Area Number: 17K07667
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Plant protection science
• Research Institution: Kochi University
• Principal Investigator: Ohnishi Kouhei 高知大学, 教育研究部総合科学系生命環境医学部門, 教授 (50211800)
• Co-Investigator(Kenkyū-buntansha): 木場 章範 高知大学, 教育研究部総合科学系生命環境医学部門, 教授 (50343314) ; 曵地 康史 高知大学, 教育研究部総合科学系生命環境医学部門, 教授 (70291507)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 植物シグナル / レポーター株 / GFP / LacZ / 静止菌体 / 遺伝子発現解析 / 植物病原細菌 / hrp / レポーター遺伝子 / 3型分泌装置 / アンタゴニスト / 青枯病菌 / 二成分制御系

Outline of Final Research Achievements

Genes encoding the type 3 secretion system and the type 3 effector, which are the major virulence factors of bacterial wilt disease in Ralstonia solanacarum, are subjected to unified expression regulation as a hrp regulon. The master operon of the hrp regulon is hrpB, and hrpB gene expression occurs only in planta. The purpose of this study is to search for plant signals that induce hrpB gene expression. In the process, we revealed phosphorylation of the response regulator HrpG, which is a two-component regulatory system upstream of hrpB, for the first time, and identified the candidates of corresponding sensor kinases. Moreover, by constructing and using the hrpB-lacZ reporter strain, it was possible to establish a route to search for plant signals that induce hrpB expression.

Academic Significance and Societal Importance of the Research Achievements

青枯病菌は、主要な栽培作物を始め、２００種類以上の宿主植物に感染し、委ちょう症状を引き起こす難防除病害菌である。青枯病菌の感染時に特異的に働くhrpレギュロンを抑制することができれば、完全に防除が可能である。本研究は、にhrpレギュロンを誘導する植物シグナルのアンタゴニストの開発につながり社会的意義は大きい。

Research Products

• [Journal Article] Expression of Ralstonia solanacearum type III secretion system is dependent on a novel type 4 pili (T4P) assembly protein (TapV) but is T4P independent (2020)
  • Author(s): Zhang Yong、Han Liangliang、Zhang Lichun、Xu Changzheng、Shi Xiaojun、Hikichi Yasufumi、Ohnishi Kouhei
  • Journal Title: Molecular Plant Pathology Volume: 21 Issue: 6 Pages: 777-793
  • DOI: 10.1111/mpp.12930
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional characterization of two putative DAHP synthases of AroG1 and AroG2 and their links with type III secretion system in Ralstonia solanacearum (2019)
  • Author(s): Zhang, W., Li, J., Shi, X., Hikichi, Y., Zhang, Y. and Ohnishi, K.
  • Journal Title: Frontiers in Microbiology Volume: 10 Pages: 183-183
  • DOI: 10.3389/fmicb.2019.00183
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Specific Reconstruction on pRC Plasmid to Facilitate Its Universal Chromosomal Integration in Different Ralstonia solanacearum Species Complex Strains (2019)
  • Author(s): Zhang Yong、Gao Shengsheng、Li Peng、Ohnishi Kouhei
  • Journal Title: Molecular Plant-Microbe Interactions Volume: 32 Issue: 9 Pages: 1063-1066
  • DOI: 10.1094/mpmi-01-19-0004-le
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Involvement of a PadR regulator PrhP on virulence of Ralstonia solanacearum by controlling detoxification of phenolic acids and type III secretion system (2019)
  • Author(s): Zhang, Y., Zhang, W., Han, L., Li, J., Shi, X., Hikichi, Y. and Ohnishi, K.
  • Journal Title: Molecular Plant Pathology Volume: 20 Issue: 11 Pages: 1477-149
  • DOI: 10.1111/mpp.12854
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Major exopolysaccharide, EPS I, is associated with the feedback loop in the quorum sensing of Ralstonia solanacearum strain OE1-1 (2019)
  • Author(s): Hayashi, K., Senuma, W., Kai, K., Kiba, A., Ohnishi, K. and Hikichi, Y
  • Journal Title: Molecular Plant Patholog Volume: 20 Issue: 12 Pages: 1740-1747
  • DOI: 10.1111/mpp.12870
  • Peer Reviewed / Open Access
• [Journal Article] Activation of ralfuranone/ralstonin production by plant sugars functions in the virulence of Ralstonia solanacearum (2019)
  • Author(s): Ishikawa, Y., Murai, Y., Sakata, M., Mori, S., Shoma Matsuo, S., Senuma, W., Ohnishi, K., Hikichi, Y. and Kai, K.
  • Journal Title: ACS Chemical Biology Volume: 14 Issue: 7 Pages: 1546-1555
  • DOI: 10.1021/acschembio.9b00301
  • Peer Reviewed / Open Access
• [Journal Article] Contribution of a lectin, LecM, to the quorum sensing signalling pathway of Ralstonia solanacearum strain OE1-1 (2018)
  • Author(s): Hayashi Kazusa、Kai Kenji、Mori Yuka、Ishikawa Shiho、Ujita Yumeto、Ohnishi Kouhei、Kiba Akinori、Hikichi Yasufumi
  • Journal Title: Molecular Plant Pathology Volume: 20 Issue: 3 Pages: 334-345
  • DOI: 10.1111/mpp.12757
  • Peer Reviewed / Open Access
• [Journal Article] Profiling of heavy metal(loid)-resistant bacterial community structure by metagenomic-DNA fingerprinting using PCR?DGGE for monitoring and bioremediation of contaminated environment (2018)
  • Author(s): Bhakta Jatindra N.、Lahiri Susmita、Bhuiyna Feroze A.、Rokunuzzaaman Md.、Ohonishi Kouhei、Iwasaki Kozo、Jana Bana B.
  • Journal Title: Energy, Ecology and Environment Volume: 3 Issue: 2 Pages: 102-109
  • DOI: 10.1007/s40974-017-0079-2
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Involvement of avirulence genes avrA and popP1 of Japanese Ralstonia solanacearum strains in the pathogenicity to tobacco. (2018)
  • Author(s): Chen, L., Dahal, A., Zhang, Y., Rokunuzzaman, Md., Kiba, A., Hikichi, Y. and Ohnishi, K.
  • Journal Title: Physiological and Molecular Plant Pathology Volume: 102 Pages: 154-162
  • DOI: 10.1016/j.pmpp.2017.12.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A putative LysR-type transcriptional regulator PrhO positively regulates the type III secretion system and contributes to the virulence of Ralstonia solanacearum (2018)
  • Author(s): Zhang Yong、Li Jiaman、Zhang Weiqi、Shi Hualei、Luo Feng、Hikichi Yasufumi、Shi Xiaojun、Ohnishi Kouhei
  • Journal Title: Molecular Plant Pathology Volume: 19 Issue: 8 Pages: 1808-1819
  • DOI: 10.1111/mpp.12660
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Involvement of ralfuranones in the quorum sensing signaling pathway and virulence of Ralstonia solanacearum strain OE1-1 (2017)
  • Author(s): Mori, M., Ishikawa, S., Ohnishi, H., Shimatani, M., Morikawa, Y., Hayashi, K., Ohnishi, K., Kiba, A., Kai K. and Hikichi, Y.
  • Journal Title: Molecular Plant Pathology Volume: 印刷中 Issue: 2 Pages: 454-463
  • DOI: 10.1111/mpp.12537
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Fixation of carbon dioxide by chemoautotrophic bacteria in grassland soil under dark conditions (2017)
  • Author(s): Jun Yang, Yumei Kang, Katsutoshi Sakurai & Kouhei Ohnishi
  • Journal Title: Acta Agriculturae Scandinavica, Section B ― Soil & Plant Science Volume: 1 Issue: 4 Pages: 362-371
  • DOI: 10.1080/09064710.2017.1281433
  • Peer Reviewed
• [Journal Article] RAPD marker assisted evaluation of chloroplast DNA variation in twelve hosta taxa (2017)
  • Author(s): Mehraj Hasan、Sharma Subarna、Ohnishi Kouhei、Shimasaki Kazuhiko、
  • Journal Title: Plant Omics Volume: 10 Issue: 03 Pages: 146-152
  • DOI: 10.21475/poj.10.03.17.pne506
  • Peer Reviewed
• [Journal Article] Phosphoinositide 3-kinase participates in l -methionine sulfoximine-induced cell death via salicylic acid mediated signaling in Nicotiana benthamiana (2017)
  • Author(s): Sumida Sayuri、Ito Makoto、Galis Ivan、Nakatani Hiroko、Shinya Tomonori、Ohnishi Kouhei、Hikichi Yasufumi、Kiba Akinori
  • Journal Title: Journal of Plant Physiology Volume: 218 Pages: 167-170
  • DOI: 10.1016/j.jplph.2017.07.016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Regulation Involved in Colonization of Intercellular Spaces of Host Plants in Ralstonia solanacearum (2017)
  • Author(s): Hikichi Yasufumi、Mori Yuka、Ishikawa Shiho、Hayashi Kazusa、Ohnishi Kouhei、Kiba Akinori、Kai Kenji
  • Journal Title: Frontiers in Plant Science Volume: 8 Pages: 967-967
  • DOI: 10.3389/fpls.2017.00967
  • Peer Reviewed / Open Access
• [Journal Article] Efficient renaturation of inclusion body proteins denatured by SDS (2017)
  • Author(s): He Chuan、Ohnishi Kouhei
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 490 Issue: 4 Pages: 1250-1253
  • DOI: 10.1016/j.bbrc.2017.07.003
  • Peer Reviewed
• [Journal Article] Ferulic Acid, But Not All Hydroxycinnamic Acids, Is a Novel T3SS Inducer of Ralstonia solanacearum and Promotes Its Infection Process in Host Plants under Hydroponic Condition (2017)
  • Author(s): Zhang Yong、Li Jing、Zhang Weiqi、Wang Rongsheng、Qiu Qiaoqing、Luo Feng、Hikichi Yasufumi、Ohnishi Kouhei、Ding Wei
  • Journal Title: Frontiers in Plant Science Volume: 8 Pages: 1595-1595
  • DOI: 10.3389/fpls.2017.01595
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of an Alteromonas long-type ulvan lyase involved in the degradation of ulvan extracted from Ulva ohnoi (2017)
  • Author(s): He Chuan、Muramatsu Hisashi、Kato Shin-ichiro、Ohnishi Kouhei
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 81 Issue: 11 Pages: 2145-2151
  • DOI: 10.1080/09168451.2017.1379352
  • Peer Reviewed
• [Journal Article] Involvement of NpdA, a Putative 2-Nitropropane Dioxygenase, in the T3SS Expression and Full Virulence in Ralstonia solanacearum OE1-1 (2017)
  • Author(s): Zhang Weiqi、Li Jing、Tang Yu、Chen Kai、Shi Xiaojun、Ohnishi Kouhei、Zhang Yong
  • Journal Title: Frontiers in Microbiology Volume: 8
  • DOI: 10.3389/fmicb.2017.01990
  • Peer Reviewed / Open Access
• [Presentation] Flagellar gene expression in Ralstonia solanacearum is differently regulated in vitro and in planta (2020)
  • Author(s): 高毓蔚, 曵地康史, 木場章範, 大西浩平
  • Organizer: 日本農芸化学会中四国支部第56回講演会
• [Presentation] Regulation mechanism of two response regulators that regulate the expression of hrpB, a central transcriptional regulator for hrp regulon in Ralstonia solanacerum (2020)
  • Author(s): 朱超, 曵地康史, 木場章範, 大西浩平
  • Organizer: 日本農芸化学会中四国支部第56回講演会
• [Presentation] Regulation mechanism of two homologous response regulator genes for hrp regulon in Ralstonia solanacearum (2020)
  • Author(s): Zhu, C., Hikichi, Y., Kiba, A., Ohnishi, K.
  • Organizer: 令和2年度日本植物病理学会大会
• [Presentation] Regulation of flagellar regulon expression in Ralstonia solanacerum (2020)
  • Author(s): Gao, Y., Hikichi, Y., Kiba, A., Ohnishi, K.
  • Organizer: 令和2年度日本植物病理学会大会
• [Presentation] 青枯病菌のクオラムセンシングに対してクエンチング活性を示す化合物の作用機序 (2019)
  • Author(s): 竹村知夏, 吉原彩華, 林 一沙, 瀬沼和香奈, 木場章範, 大西浩平, 甲斐建次, 曵地康史
  • Organizer: 平成31年度日本植物病理学会大会
• [Presentation] cbhA遺伝子は青枯病菌OE1-1株の運動能に関わる (2019)
  • Author(s): 川本大輝, 林 一沙, 瀬沼和香奈, 木場章範, 大西浩平, 甲斐建次, 曵地康史
  • Organizer: 平成31年度日本植物病理学会大会
• [Presentation] Ralstonia solanacearumのクオラムセンシングは複数のシグナル伝達系から構成される (2019)
  • Author(s): 瀬沼和香奈, 林 一沙, 登 達也, 木場章範, 大西浩平, 甲斐建次, 津田賢一, 曵地康史
  • Organizer: 平成31年度日本植物病理学会大会
• [Presentation] Core type III effectors are necessary for full virulence in Ralstonia solanacearum (2019)
  • Author(s): Lei, N., Chen, L., Kiba, A., Hikichi, Y., and Ohnishi, K.
  • Organizer: 平成31年度日本植物病理学会大会
• [Presentation] Interaction of tobacco chloroplastic proteins with Ralstonia solanacearum type III effector RipAJ (2019)
  • Author(s): Kharel, L., Kiba, A., Hikichi, Y., and Ohnishi, K.
  • Organizer: 平成31年度日本植物病理学会大会
• [Presentation] Screening of tobacco proteins targeted by Ralstonia solanacearum type III effectors with yeast two hybrid system (2019)
  • Author(s): Laxmi Kharel, Akinori Kiba, Yasufumi Hikichi, Kouhei Ohnishi
  • Organizer: 第92回日本細菌学会総会
• [Presentation] Systematic multiple deletion analysis of T3E effectors in Ralstonia solanacerum (2019)
  • Author(s): Ni Lei, Li Chen, Akinori Kiba, Yasufumi Hikichi, Kouhei Ohnishi
  • Organizer: 第92回日本細菌学会総会
• [Presentation] Multiple deletion mutation analysis of T3E effectors in Ralstonia solanacearum (2019)
  • Author(s): Ni Lei, Li Chen, Akinori Kiba, Yasufumi Hikichi and Kouhei Ohnishi
  • Organizer: XVIII International Congress on Molecular Plant-Microbe Interactions
  • Int'l Joint Research
• [Presentation] The type III effectors of Ralstonia solanacearum interact with tobacco chloroplastic proteins (2019)
  • Author(s): Kouhei Ohnishi, Amol Dahal, Laxmi Kharel, Akinori Kiba and Yasufumi Hikichi
  • Organizer: XVIII International Congress on Molecular Plant-Microbe Interactions
  • Int'l Joint Research
• [Presentation] 海洋性細菌KUL106の有する多糖類のウルバンリアーゼに関する研究 (2018)
  • Author(s): 小林慎太郎，大西浩平
  • Organizer: 日本農芸化学会中四国支部第50回記念講演会
• [Presentation] Chloroplastic proteins are the targets for the Ralstonia solanacearum type III effectors (2018)
  • Author(s): Dahal A, Kharel L, Kiba A, Hikichi Y, Ohnishi K
  • Organizer: 日本植物病理学会大会
• [Presentation] 青枯病菌OE1-1株の主要な菌体外多糖ESPIは，クオラムセンシングにより制御される遺伝子発現に寄与する (2018)
  • Author(s): 林一沙，氏田夢斗，大西 浩平，木場 章範，甲斐 建次，曵地 康史
  • Organizer: 日本植物病理学会大会
• [Presentation] 青枯病菌OE1-1株の主要な菌体外多糖ESPIは，クオラムセンシングにより制御される遺伝子発現に寄与する (2018)
  • Author(s): 林一沙，甲斐 建次，木場 章範，大西 浩平，曵地 康史
  • Organizer: 日本植物病理学会大会
• [Presentation] Chloroplastic proteins are targets of the RipG effectors of Ralstonia solanacearum (2018)
  • Author(s): Amol Dahal，木場章範，曵地康史，大西浩平
  • Organizer: 第91回日本細菌学会総会
• [Presentation] ラルフラノン化合物は青枯病菌によるマッシュルーム型バイオフィルム形成に関与する (2018)
  • Author(s): 曵地康史，林一沙，井上加奈子，大西浩平，木場章範，甲斐建次
  • Organizer: 第91回日本細菌学会総会
• [Presentation] 二次代謝物質による青枯病菌のクオラムセンシングのフィードバック制御 (2018)
  • Author(s): 林一沙，甲斐 建次，大西 浩平，木場 章範，曵地 康史
  • Organizer: 第91回日本細菌学会総会