植物の組織形成を規定するリン酸とピロリン酸の濃度バランス

• Project Area: Multidimensional Exploration of Logics of Plant Development
• Project/Area Number: 26113506
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagoya University
• Principal Investigator: 前島 正義 名古屋大学, 生命農学研究科, 教授 (80181577)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥11,440,000 (Direct Cost: ¥8,800,000、Indirect Cost: ¥2,640,000); Fiscal Year 2015: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2014: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
• Keywords: 組織形成 / ピロホスファターゼ / 代謝 / プロトンポンプ / ピロリン酸 / 形態形成

Outline of Annual Research Achievements

2015年度、下記の成果を得た。
(1) H+-PPase機能欠失株では、硝酸のみが窒素源の場合は本葉が顕著に萎縮するが、アンモニアを添加すると表現型は軽減する。壊死の起こる位置はPPiが多く発生する細胞分裂の盛んな組織であり、表現型は酵母のsPPaseにより相補されることから、アンモニアを含まない培地では、とくにH+-PPaseが葉のPPi代謝において重要な役割を担うことが明らかにした。
(2) H+-PPaseは、植物生育初期のPPi代謝において大きな役割をもち、機能欠損は子葉や胚軸に表現型を示す。加えて、破壊株及び過剰発現株を用いた解析により、後期成長においてもPPiの分解を介して植物の生育に寄与していることを明らかにした。
(3) 細胞質可溶性PPaseの多重破壊株には顕著な表現型がなく、細胞質PPiの分解においてはH+-PPaseの働きに優位性があることが明らかとなった。顕著な表現型を示すfugu5-1ppa1二重破壊株では、低浸透圧による根端の破裂がみられ、細胞壁の強度が低下し形態への影響が発生していると推測している。さらに、fugu5-1ppa1では異所的にデンプンが蓄積していることを発見し、特に外部よりショ糖を添加した場合はデンプンの量が著しく増加していた。WT、fugu5-1、ppa1、fugu5-1ppa1株の胚軸のメタボローム解析を行い、有意差のある代謝産物を多数同定した。その結果より、糖新生、細胞壁合成に用いられるべき糖ヌクレオチドがPPiの蓄積によって阻害され、表現型につながっている、という仮説を立てた。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] チューリッヒ大学(スイス)
• [Int'l Joint Research] ボーフム大学(ドイツ)
• [Int'l Joint Research] ポハン科学技術大学(韓国)
• [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
• [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] The histidine-rich loop of Arabidopsis vacuolar membrane zinc transporter AtMTP1 is involved in sensing cytosolic level of zinc. (2015)
  • Author(s): Tanaka, N., Fujiwara, T., Tomioka, R., Krämer, U., Kawachi, M., and Maeshima, M.
  • Journal Title: Plant and Cell Physiology Volume: 56 Issue: 3 Pages: 510-519
  • DOI: 10.1093/pcp/pcu194
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of a unique CDF member containing the long N-terminal region in Arabidopsis thaliana. (2015)
  • Author(s): Fujiwara, T., Mori, H., Kawachi, M., and Maeshima, M.
  • Journal Title: FEBS Journal Volume: 282 Pages: 1965-1979
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] GFPの二量体化を原因とする細胞内オルガネラの異常構造の形成とその予防策 (2015)
  • Author(s): 瀬上紹嗣、前島正義
  • Journal Title: 化学と生物 Volume: 10 Pages: 703-708
  • NAID: 130005416123
  • Peer Reviewed
• [Journal Article] Identification of a unique CDF member containing the long N-terminal region in Arabidopsis thaliana (2015)
  • Author(s): Fujiwara, T., Mori, H., Kawachi, M., and Maeshima, M.
  • Journal Title: FEBS Journal Volume: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Characteristics of a root hair-less line of Arabidopsis thaliana under physiological stresses. (2014)
  • Author(s): Tanaka, N., Kato, M., Tomioka, R., Kurata, R., Fukao, Y., Aoyama, T., and Maeshima, M.
  • Journal Title: Journal of Experimental Botany Volume: 65 Issue: 6 Pages: 1497-1512
  • DOI: 10.1093/jxb/eru014
  • Peer Reviewed / Open Access
• [Journal Article] Roles of the vacuolar H+-PPase in seed storage oil mobilization and plant development (2014)
  • Author(s): Ferjani, A., Segami, S., Horiguchi, G., Muto, Y., Maeshima, M., and Tsukaya, H.
  • Journal Title: Plant Morphology Volume: 26 Pages: 45-51
  • NAID: 130005066549
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Dynamics of vacuoles and H+-pyrophosphatase visualized by monomeric green fluorescent protein in Arabidopsis: Artifactual bulbs and native intravacuolar spherical structures. (2014)
  • Author(s): Segami, S., Makino, S., Miyake, A., Asaoka, M., and Maeshima, M.
  • Journal Title: Plant Cell Volume: 26 Issue: 8 Pages: 3416-3434
  • DOI: 10.1105/tpc.114.127571
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification of the critical residues for the function of vacuolar H+-pyrophosphatase by amino acid screening based on the 3D structure. (2014)
  • Author(s): Asaoka, M., Segami, S., and Maeshima, M.
  • Journal Title: Journal of Biochemistry Volume: 156 Issue: 6 Pages: 333-344
  • DOI: 10.1093/jb/mvu046
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Cooperative works of H+-PPase and soluble PPases for PPi homeostasis in Arabidopsis. (2016)
  • Author(s): Segami, S., Tomoyama, T., Maeshima, M.
  • Organizer: Annual Meeting of Japan Society of Plant Physiologists
  • Place of Presentation: 岩手大学
  • Year and Date: 2016-03-18
• [Presentation] Contribution of PPi-hydrolysis function of H+-pyrophosphatase in vegetative growth of Arabidopsis thaliana. (2016)
  • Author(s): Asaoka, M., Segami, S., Ferjani, A., Maeshima, M.
  • Organizer: Annual Meeting of Japan Society of Plant Physiologists
  • Place of Presentation: 岩手大学
  • Year and Date: 2016-03-18
• [Presentation] Physiological effects of loss-of-function mutations of enzymes involved in pyrophosphate metabolism depending on nitrogen. (2016)
  • Author(s): Fukuda, M., Segami, S., Tomoyama, T., Gunji, S., Ferjanai, A., Maeshima, M.
  • Organizer: Annual Meeting of Japan Society of Plant Physiologists
  • Place of Presentation: 岩手大学
  • Year and Date: 2016-03-18
• [Presentation] Characterization of the histidine loop of Arabidopsis vacuolar membrane zinc transporter AtMTP1 as a sensor of cytosol zinc level. (2015)
  • Author(s): Tanaka, N., Fujiwara, T., Tomioka, R., Kraemer, U., Kawachi M. and Maeshima, M.:
  • Organizer: Vienna International Conference Series 2015
  • Place of Presentation: Wien, Austria
  • Year and Date: 2015-06-23
  • Int'l Joint Research
• [Presentation] ピロリン酸代謝関連酵素の機能欠失がピロリン酸濃度と生育に与える影響 (2015)
  • Author(s): 福田茉由、瀬上紹嗣、郡司玄、Ali Ferjani、前島正義
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 東京農業大学
  • Year and Date: 2015-03-16 – 2015-03-18
• [Presentation] The H+-pyrophosphatase is necessary for normal growth of Arabidopsis thaliana (2015)
  • Author(s): 浅岡真理子、瀬上紹嗣、Ali Ferjani, 前島正義
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 東京農業大学
  • Year and Date: 2015-03-16 – 2015-03-18
• [Presentation] PPi opposes gluconeogenesis progression during oilseed mobilization in Arabidopsis. (2015)
  • Author(s): Ferjani, A., Kawade, K.. Oikawa, A., Asaoka, Takahashi, K., Ishida, M., Maeshima, M., Yokota-Hirai, M., Saito, K. and Tsukaya, H.
  • Organizer: 日本植物生理学会年会
  • Place of Presentation: 東京農業大学
  • Year and Date: 2015-03-16 – 2015-03-18
• [Presentation] GFP二量体化による人為的な膜の接着と単量体化によって見えた真の液胞構造 (2014)
  • Author(s): 瀬上紹嗣、牧野沙知、三宅愛、浅岡真理子、前島正義
  • Organizer: 日本分子生物学会2014年度大会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] H+-pyrophosphatase is necessary for normal growth of Arabidopsis thaliana. (2014)
  • Author(s): saoka, M., Segami, S., Sato, C., Ferjani, A. and Maeshima, M.
  • Organizer: Plant Biology Europe FESPB/EPSO 2014 Congress.
  • Place of Presentation: Doublin, Irland
  • Year and Date: 2014-06-21 – 2014-06-27
• [Remarks] 細胞ダイナミクス研究室
  • URL: http://celld.agr.nagoya-u.ac.jp/index.html