Analysis of pyrophosphate regulation in plant

• Project/Area Number: 16K18687
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Applied biochemistry
• Research Institution: Nagoya University
• Principal Investigator: Shoji Segami 名古屋大学, 生命農学研究科, 研究員 (00765935)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: ピロリン酸 / 糖代謝 / 細胞壁 / 窒素 / シロイヌナズナ / 液胞膜プロトンポンプ / ピロリン酸代謝 / 細胞壁形成 / 植物 / 生理学

Outline of Final Research Achievements

Pyrophosphate (PPi) is a byproduct of essential biosynthesis reactions including nucleic acid and cell wall, therefore PPi have to be removed from cytosol for thermodynamic reason. However, PPi regulating mechanism in plant, which possess PPi utilizing enzymes and requires adequate PPi concentration, had been unknown.
We revealed that PPi degradation is predominantly assumed by vacuolar H+-pyrophosphatase, and subsidiarily by soluble pyrophosphatases which found all organisms. We developed multidisciplinary analysis for PPi metabolism e.g. supplied nitrogen affect PPi production in leaves and root tip columella specific accumulation of starch is concerned with PPi accumulation.

Academic Significance and Societal Importance of the Research Achievements

ピロリン酸(PPi)はすべての生物に存在する基本代謝物質である。植物ではPPiが生長調節因子、またエネルギードナーとして働いていることが考えられるが、PPiの濃度調節は合成側ではなく分解酵素であるH+-PPaseとsPPaseが主に担うことを明らかにした。遺伝子破壊株を用いた解析によりPPi濃度は糖代謝を変動させ、細胞壁・デンプン・貯蔵油脂の含量が変化することを見出した。これらの知見から、様々な細胞が持つ固有の代謝要求を満たすためPPi濃度がそれぞれの細胞に応じて調節されている可能性を検証しており、道半ばではあるがその証拠を掴みつつある。以降の研究により実証を目指している。

Research Products

• [Int'l Joint Research] ハイデルベルグ大学(ドイツ)
• [Int'l Joint Research] ハイデルベルグ大学(ドイツ)
• [Journal Article] Polar vacuolar distribution is essential for accurate asymmetric division ofArabidopsiszygotes (2019)
  • Author(s): Kimata Yusuke、Kato Takehide、Higaki Takumi、Kurihara Daisuke、Yamada Tomomi、Segami Shoji、Morita Miyo Terao、Maeshima Masayoshi、Hasezawa Seiichiro、Higashiyama Tetsuya、Tasaka Masao、Ueda Minako
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 116 Issue: 6 Pages: 2338-2343
  • DOI: 10.1073/pnas.1814160116
  • Peer Reviewed / Open Access
• [Journal Article] Vacuolar H(+)-pyrophosphatase and cytosolic soluble pyrophosphatases cooperatively regulate pyrophosphate levels in Arabidopsis thaliana. (2018)
  • Author(s): Segami, S., Tomoyama, T., Sakamoto, S., Gunji, S., Fukuda, M., Kinoshita, S., Mitsuda, N., Ferjani, A., and Maeshima, M.
  • Journal Title: The Plant Cell Volume: 30 Issue: 5 Pages: 1040-1061
  • DOI: 10.1105/tpc.17.00911
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Biochemical, structural, and physiological characteristics of vacuolar H(+)-pyrophosphatase. (2018)
  • Author(s): Segami, S., Asaoka, M., Kinoshita, S., Fukuda, M., Nakanishi, Y., and Maeshima, M.
  • Journal Title: Plant & Cell Physiology Volume: 59 Pages: 1300-1308
  • DOI: 10.1093/pcp/pcy054
  • NAID: 120006529619
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functionally redundant LNG3 and LNG4 genes regulate turgor-driven polar cell elongation through activation of XTH17 and XTH24 (2018)
  • Author(s): Lee, Y.K., Rhee, J., Lee, S.H., Chung, G.C., Park, S.J., Segami, S., Maeshima, M., and Choi, G.
  • Journal Title: Plant Molecular Biology Volume: In press Issue: 1-2 Pages: 23-36
  • DOI: 10.1007/s11103-018-0722-0
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Vacuolar H+-pyrophosphatase and Cytosolic Soluble Pyrophosphatases Cooperatively Regulate Pyrophosphate Levels in Arabidopsis thaliana (2018)
  • Author(s): Segami, S., Tomoyama, T., Sakamoto, S., Gunji, S., Fukuda, M., Kinoshita, S., Mitsuda, N., Ferjani, A., and Maeshima, M.
  • Journal Title: The Plant Cell Volume: 30
  • Peer Reviewed
• [Journal Article] Biochemical, structural, and physiological characteristics of vacuolar H+-pyrophosphatase. (2018)
  • Author(s): Segami, S., Asaoka, M., Kinoshita, S., Fukuda, M., Nakanishi, Y., and Maeshima, M.
  • Journal Title: Plant & Cell Physiology Volume: 59
  • NAID: 120006529619
  • Peer Reviewed
• [Journal Article] Functionally redundant LNG3 and LNG4 genes regulate turgor-driven polar cell elongation mediated by activation of XTH gene family. (2018)
  • Author(s): Lee, Y.K., Rhee, J.Y., Chung, G.C., Park, S.J., Segami, S., Maeshima, M., Choi, G., Chung, W.I.
  • Journal Title: Plant Molecular Biology Volume: なし
• [Journal Article] Lack of H+-pyrophosphatase causes developmental damage in Arabidopsis leaves in ammonia-free culture medium. (2016)
  • Author(s): Fukuda, M., Segami, S., Tomoyama, T., Nakanishi, Y., Gunji, S., Ferjani, A., and Maeshima, M.
  • Journal Title: Frontiers of Plant Science Volume: 6 Pages: 819-819
  • DOI: 10.3389/fpls.2016.00819
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Contribution of PPi-hydrolyzing function of vacuolar H+-pyrophosphatase in vegetative growth of Arabidopsis: Evidenced by expression of uncoupling mutated enzymes. (2016)
  • Author(s): Asaoka, M., Segami, S., Ferjani, A., and Maeshima, M.
  • Journal Title: Frontiers in Plant Research Volume: 7 Pages: 415-415
  • DOI: 10.3389/fpls.2016.00415
  • NAID: 120005868253
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A novel-type phosphatidylinositol phosphate-interactive, Ca-binding protein PCaP1 in Arabidopsis thaliana: stable association with plasma membrane and partial involvement in stomata closure. (2016)
  • Author(s): Nagata, C., Miwa, C., Tanaka, N., Kato, M., Suito, M., Tsuchihira, A., Sato, Y., Segami, S. and Maeshima, M.
  • Journal Title: Journal of Plant Research Volume: In press Issue: 3 Pages: 539-550
  • DOI: 10.1007/s10265-016-0787-2
  • NAID: 120005868252
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 代謝の変動によるピロリン酸の濃度変化はピロホスファターゼの協働により抑制される (2020)
  • Author(s): Shoji Segami, Mayu Fukuda, Marika Mieda, Takaaki Tomoyama, Satoru Kinoshita, Ali Ferjani, Masayoshi Maeshima
  • Organizer: 第61回植物生理学会年会
• [Presentation] GFP やTagRFP によるオイルボディ、液胞の形態と植物生長への人為的影響 (2019)
  • Author(s): 瀬上 紹嗣，木下 悟，島田 貴士，嶋田 知生，西村 いくこ，前島 正義
  • Organizer: 日本植物学会第83回大会
• [Presentation] GFP やTagRFP によるオイルボディ、液胞の形態と植物生長への人為的影響 (2019)
  • Author(s): 瀬上 紹嗣，木下 悟，島田 貴士，嶋田 知生，西村 いくこ，前島 正義
  • Organizer: 第60回日本植物生理学会年会
• [Presentation] シロイヌナズナにおける初期胚のライブイメージング (2019)
  • Author(s): 植田 美那子，木全 祐資，田中 小百合，加藤 壮英，桧垣 匠，栗原 大輔，山田 朋美，安藤 奈央惠，森田（寺尾）美代，瀬上 紹嗣，前島 正義，馳澤 盛一郎，桑田 啓子，佐藤 綾人，鈴木 孝征，東山 哲也，田坂 昌生
  • Organizer: 第60回日本植物生理学会年会
• [Presentation] ピロリン酸が根端コルメラ細胞のデンプン蓄積に与える影響の解析 (2019)
  • Author(s): 木下悟、瀬上紹嗣、前島正義
  • Organizer: 第60回日本植物生理学会年会
• [Presentation] シロイヌナズナのコルメラ細胞におけるピロリン酸分解酵素の生理的重要性 (2018)
  • Author(s): 木下悟、瀬上紹嗣、前島正義
  • Organizer: 日本植物学会第82回大会
• [Presentation] 液胞膜H+-ピロホスファターゼと可溶性ピロホスファターゼが協働してピロリン酸レベルを調節する (2018)
  • Author(s): 瀬上紹嗣、巴山貴晶、坂本真吾、郡司玄、福田茉由、木下悟、Ali Ferjani、前島正義
  • Organizer: 日本植物学会第82回大会
• [Presentation] ピロリン酸が植物の形態形成と成長に与える影響の分子生物学的解析. (2018)
  • Author(s): 福田茉由、瀬上紹嗣、Ali Ferjani、前島正義
  • Organizer: 日本農芸化学会大会
• [Presentation] GFP等の融合タグによるオルガネラ形態と生長への人為的影響 (2018)
  • Author(s): 瀬上紹嗣、木下悟、島田貴士、嶋田知生、西村いくこ、前島正義
  • Organizer: 日本植物生理学会年会
• [Presentation] Physiological Importance of Pyrophosphatases in Lateral Nectary of Arabidopsis thaliana. (2018)
  • Author(s): 木下悟、瀬上紹嗣、前島正義
  • Organizer: 日本植物生理学会年会
• [Presentation] Physiological importance of gene suppression of H+-pyrophosphatase in columella cells of Arabidopsis thaliana. (2017)
  • Author(s): 木下悟、瀬上紹嗣、前島正義
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 鹿児島大学
  • Year and Date: 2017-03-16
• [Presentation] Contribution of soluble and H+ translocating pyrophosphatases to pyrophosphate homeostasis. (2017)
  • Author(s): 巴山貴晶、瀬上紹嗣、前島正義
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 鹿児島大学
  • Year and Date: 2017-03-16
• [Presentation] Lack of H+-pyrophosphatase prompts developmental damage in Arabidopsis leaves on ammonia-free (2016)
  • Author(s): Fukuda, M., Segami, S., Asaoka, M., Gunji, S., Ferjani, A., and Maeshima, M.
  • Organizer: 17th International Workshop on Plant Membrane Biology
  • Place of Presentation: Annapolis, Maryland, USA
  • Year and Date: 2016-06-05
  • Int'l Joint Research
• [Presentation] Dual functions of vacuolar H+-pyrophosphatase: vacuolar acidification and PPi homeostasis. (2016)
  • Author(s): Maeshima M, Segami S, Fukuda M, Asaoka M, Ferjani A
  • Organizer: 17th International Workshop on Plant Membrane Biology
  • Place of Presentation: Annapolis, Maryland, USA
  • Year and Date: 2016-06-05
  • Int'l Joint Research / Invited
• [Presentation] Dynamics of vacuoles and H+-pyrophosphatase visualized by monomeric green fluorescent protein in Arabidopsis. (2016)
  • Author(s): Segami, S. and Maesima, M.
  • Organizer: 17th International Workshop on Plant Membrane Biology
  • Place of Presentation: Annapolis, Maryland, USA
  • Year and Date: 2016-06-05
  • Int'l Joint Research
• [Remarks] 小さなエネルギー分子の代謝の仕組みを発見！～植物のピロリン酸代謝は2種類の酵素が協働～
  • URL: http://www.nagoya-u.ac.jp/about-nu/public-relations/researchinfo/upload_images/20180625_agr_1.pdf