Making virus tolerant plants by using Dicer and dsRNA-binding protein genes

• Project/Area Number: 21580411
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied molecular and cellular biology
• Research Institution: Tokyo University of Agriculture and Technology
• Principal Investigator: FUKUHARA Toshiyuki 東京農工大学, 大学院・農学研究院, 教授 (90228924)
• Co-Investigator(Renkei-kenkyūsha): NATSUAKI Tomohide 宇都宮大学, 農学部, 教授 (10134264) ; MORIYAMA Hiromitsu 東京農工大学, 大学院・農学研究院, 准教授 (20392673)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 細胞機能 / ウイルス / RNA干渉 / 2本鎖RNA / ダイサー / 2本鎖RNA結合タンパク質 / シロイヌナズナ / シロイヌナ / カブモザイクウイルス

Research Abstract

Inoculation experiments with Turnip mosaic virus on Arabidopsis plants with over-expressing Dicer-like protein(DCL1-4) or dsRNA-binding protein(DRB1-5) genes were carried out. Infected plants with over-expressing DRB2, DRB3 or DRB4 tended to exhibit mild symptom. Furthermore, a biochemical procedure for evaluating the dicer activity(dsRNA-cleaving activity) is established by using crude extracts from Arabidopsis seedlings.

Research Products

• [Journal Article] Specific requirement of DRB4, a dsRNA-binding protein, for the in vitro dsRNA-cleaving activity of Arabidopsis Dicer-like 4 (2011)
  • Author(s): Fukudome A., Kanaya A., Egami M., Nakazawa Y., Hiraguri A., Moriyama H. and Fukuhara T
  • Journal Title: RNA Volume: 17(4) Issue: 4 Pages: 750-760
  • DOI: 10.1261/rna.2455411
  • Peer Reviewed
• [Journal Article] An Arabidopsis RNase III-like protein, AtRTL2, cleaves double-stranded RNA in vitro (2011)
  • Author(s): Kiyota E., Okada R., Kondo N., Hiraguri A., Moriyama H. and Fukuhara T
  • Journal Title: Journal of Plant Research Volume: 124 Issue: 3 Pages: 405-414
  • DOI: 10.1007/s10265-010-0382-x
  • NAID: 10028258238
  • Peer Reviewed
• [Journal Article] Bell pepper endornavirus : molecular and biological properties and occurrence in the genus Capsicum (2011)
  • Author(s): Okada R., Kiyota E., Sabanadzovic S., Moriyama H., Fukuhara T., Saha P., Roossinck M. J., Severin A. and Valverde R. A.
  • Journal Title: Journal of General Virology Volume: 92(11) Issue: 11 Pages: 2664-2673
  • DOI: 10.1099/vir.0.034686-0
  • Peer Reviewed
• [Journal Article] The presence of double-stranded RNAs in Alternaria alternata Japanese pear pathotype is associated with its morphological changes (2011)
  • Author(s): Fuke K., Takeshita K., Aoki N., Fukuhara T., Egusa M., Kodama M. and Moriyama H
  • Journal Title: Journal of General Plant Pathology Volume: 77 Issue: 4 Pages: 248-252
  • DOI: 10.1007/s10327-011-0315-0
  • Peer Reviewed
• [Journal Article] Specific requirement of DRB4, a dsRNA-binding protein, for the in vitro dsRNA-cleaving activity of Arabidopsis Dicer-like 4 (2011)
  • Author(s): Fukudome T., Kanaya A., EgamiM., Nakazawa Y, Hiraguri A., Moriyama H., Fukuhara T.
  • Journal Title: RNA Volume: 17 Pages: 750-760
  • Peer Reviewed
• [Journal Article] An Arabidopsis RNase III-like protein, AtRTL2, cleaves double-stranded RNA in vitro (2011)
  • Author(s): Kiyota E., Okada R., Kondo N., Hiraguri A., Moriyama H., Fukuhara T.
  • Journal Title: Journal of Plant Research Volume: (印刷中)
  • NAID: 10028258238
  • Peer Reviewed
• [Journal Article] Knockdown of OsDCL2 in rice negatively affects maintenance of the endogenous dsRNA virus, Oryza sativa endornavirus (2010)
  • Author(s): Urayama S., Moriyama M., Aoki N., Nakazawa Y., Okada R., Kiyota E., Miki D., Shimamoto K. and Fukuhara T
  • Journal Title: Plant and Cell Physiology Volume: 51(1) Issue: 1 Pages: 58-67
  • DOI: 10.1093/pcp/pcp167
  • NAID: 10027345912
  • Peer Reviewed
• [Journal Article] Mycoviruses related to chrysovirus affect vegetative growth in the rice blast fungus Magnaporthe oryzae (2010)
  • Author(s): Urayama S., Kato S., Suzuki Y., Aoki N., Le Minh T., Arie T., Teraoka T., Fukuhara T. and Moriyama H
  • Journal Title: Journal of General Virology Volume: 91(12) Issue: 12 Pages: 3085-3094
  • DOI: 10.1099/vir.0.025411-0
  • Peer Reviewed
• [Journal Article] Mycoviruses related to chrysovirus affect vegetative growth in the rice blast fungus Magnaporthe oryzae (2010)
  • Author(s): Urayama S, Kato S, Suzuki Y, Aoki N, Le MT, Arie T, Teraoka T, Fukuhara T, Moriyama H.
  • Journal Title: Journal of General Virology Volume: 91 Pages: 3085-3094
  • Peer Reviewed
• [Journal Article] Knockdown of OsDCL2 in rice negatively affects maintenance of the endogenous dsRNA virus, Oryza sativa endornavirus. (2010)
  • Author(s): Urayama U., Moriyama M., Aoki N., Nakazawa Y., Okada R., Kiyota E., Miki D., Shimamoto K., Fukuhara T.
  • Journal Title: Plant and Cell Physiology 51 Pages: 58-67
  • NAID: 10027345912
  • Peer Reviewed
• [Presentation] シロイヌナズナのダイサーDCL3およびDCL4の生化学的解析 (2012)
  • Author(s): 長野秀昭、福留章仁、中澤悠宏、平栗章弘、森山裕充、福原敏行
  • Organizer: 第53回日本植物生理学会年会
  • Place of Presentation: 京都産業大学(京都府)
  • Year and Date: 2012-03-16
• [Presentation] 福原敏行シロイヌナズナのダイサーDCL3およびDCL4の生化学的解析 (2012)
  • Author(s): 長野秀昭、福留章仁、中澤悠宏、平栗章弘、森山裕充
  • Organizer: 第53回日本植物生理学会年会
  • Place of Presentation: 京都産業大学
• [Presentation] A dsRNA-binding protein DRB4 is essential for the in vitro activity of Arabidopsis Dicer-like 4 (2011)
  • Author(s): Fukudome A, Nagano H., Hiraguri A, Haas G, Keller M, Moriyama H, Fukuhara T
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2011-12-13
• [Presentation] Involvement of DRB proteins in cell-to-cell spread of silencing signal (2011)
  • Author(s): Takahashi T, Usui T, Fukudome A, Moriyama H, Fukuhara T
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2011-12-13
• [Presentation] イネ内在性2本鎖RNAレプリコン(Oryza sativa endornavirus)の遺伝様式と宿主RNAサイレンシング機構との関係 (2011)
  • Author(s): 福原敏行、浦山俊一、岡田亮、森山裕充
  • Organizer: 日本遺伝学会第83回大会
  • Place of Presentation: 京都大学(京都府)(招待講演)
  • Year and Date: 2011-09-21
• [Presentation] in vitro dicing activity of Arabidopsis Dicer-like 4 requiring a dsRNA-binding protein DRB4 (2011)
  • Author(s): Fukudome A., Kanaya A., Egami M., Nakazawa Y., Hiraguri A., Moriyama H., Fukuhara T.
  • Organizer: International Plant RNA Workshop 2011
  • Place of Presentation: 理研横浜研究所(神奈川県)
  • Year and Date: 2011-06-20
• [Presentation] The role of dsRNA-binding protein DRB4 in the Dicer activity of DCL4 complex in Arabidopsis thaliana (2011)
  • Author(s): Fukudome A., Kanaya A., Egami M., Nakazawa Y., Hiraguri A., Moriyama H., Fukuhara T.
  • Organizer: The 16^<th> Annual Meeting of the RNA Sciety
  • Place of Presentation: 京都国際会議場(京都府)
  • Year and Date: 2011-06-17
• [Presentation] Endogenous dsRNA replicons and dsRNA-binding proteins in plants (2011)
  • Author(s): Toshiyuki Fukuhara
  • Organizer: The 1499th Biological Symposium in NIG
  • Place of Presentation: 遺伝学研究所(静岡県)(招待講演)
  • Year and Date: 2011-04-13
• [Presentation] イネ内在性2本鎖RNAレプリコン(Oryza sativa endornavirus)の遺伝様式と宿主RNAサイレンシング機構との関係 (2011)
  • Author(s): 福原敏行、浦山俊一、岡田亮、森山裕充
  • Organizer: 日本遺伝学会第83回大会シンポジウム
  • Place of Presentation: 京都大学
• [Presentation] The role of dsRNA-binding protein DRB4 in the Dicer activity of DCL4 complex in Arabidopsis thaliana (2011)
  • Author(s): Fukudome A., Kanaya A., Egami M., Nakazawa Y., Hiraguri A., Moriyama H. and Fukuhara T
  • Organizer: The 16th Annual Meeting of the RNA Society
  • Place of Presentation: 京都国際会議場
• [Presentation] in vitro dicing activity of Arabidopsis Dicer-like 4 requiring a dsRNA-binding protein DRB4 (2011)
  • Author(s): Fukudome A., Kanaya A., Egami M., Nakazawa Y., Hiraguri A., Moriyama H. and Fukuhara T
  • Organizer: International Plant RNA Workshop 2011
  • Place of Presentation: 理研横浜研究所
• [Presentation] Involvement of DRB proteins in cell-to-cell spread of silencing signal (2011)
  • Author(s): Takahashi T, Usui T, Fukudome A, Moriyama H and Fukuhara T
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
• [Presentation] シロイヌナズナDicer-like 4のdsRNA切断活性におけるdsRNA結合タンパク質DRB4の機能解析 (2010)
  • Author(s): 福留章仁、金屋明宏、江上舞、中澤悠宏、平栗章弘、森山裕充、福原敏行
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2010-12-10
• [Presentation] 2本鎖RNA切断酵素AtRTL2の異所発現とウイルスに対する酵素活性 (2010)
  • Author(s): 清田依里、岡田亮、森山裕充、福原敏行
  • Organizer: 日本植物学会第74回大会
  • Place of Presentation: 中部大学
  • Year and Date: 2010-09-09
• [Presentation] Class 1 RNase III in Arabidopsis thaliana (AtRTL2) cleaves double-stranded RNAs specifically (2010)
  • Author(s): Kiyota E., Okada R., Hiraguri A., Moriyama H., Fukuhara T.
  • Organizer: 21th International Conference on Arabidopsis Research
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2010-06-07
• [Presentation] Arabidopsis Dicer-like 4 requires the dsRNA-binding protein DRB4 to cleave long-dsRNAs into 21 nt small RNA in vitro (2010)
  • Author(s): Fukudome A, Kanaya A., Egami M., Nakazawa Y, Hiraguri A., Moriyama H., Fukuhara T.
  • Organizer: 21th International Conference on Arabidopsis Research
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2010-06-07
• [Presentation] シロイヌナズナDCL4/DRB4複合体のダイサー活性におけるDRB4の機能解析 (2010)
  • Author(s): 福留章仁、金屋明宏、江上舞、中澤悠宏、平栗章弘、森山裕充、福原敏行
  • Organizer: 日本植物生理学会
  • Place of Presentation: 熊本大学
  • Year and Date: 2010-03-18
• [Presentation] シロイヌナズナの二本鎖RNA結合タンパク質のキメラ遺伝子過剰発現体を用いた解析 (2009)
  • Author(s): 江上舞、中澤悠宏、金屋明宏、平栗章弘、森山裕充、福原敏行
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-11
• [Presentation] シロイヌナズナにおけるDCL/DRBタンパク質複合体の二本鎖RNA切断活性の生化学的解析 (2009)
  • Author(s): 福留章仁、金屋明宏、江上舞、中澤悠宏、平栗章弘、森山裕充、福原敏行
  • Organizer: 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2009-12-11
• [Presentation] Biochemical characterization of class 1 RNase III in Alabidopsis thaliana(AtRTL2) (2009)
  • Author(s): 清田依里, 岡田亮, 平栗章弘, 森山裕充, 福原敏行
  • Organizer: 9th International Plant Molecular Biology Congress
  • Place of Presentation: St.Louis, MO, USA
  • Year and Date: 2009-10-18
• [Presentation] 2本鎖RNAを特異的に切断するシロイヌナズナのclass 1 RNase III(AtRTL2) (2009)
  • Author(s): 清田依里、近藤直子、岡田亮、熊志傑、平栗章弘、森山裕充、福原敏行
  • Organizer: 日本植物学会
  • Place of Presentation: 山形大学
  • Year and Date: 2009-09-17
• [Book] Endornavirus, Unassigned Genus (2012)
  • Author(s): Fukuhara T.
  • Publisher: Springer
• [Book] Family Endornaviridae (2012)
  • Author(s): Fukuhara T. and Gibbs M. J
  • Publisher: Ninth Report of the International Committee on Taxonomy of Viruses
• [Book] Virus Taxonomy : Ninth Report of the International Committee on Taxonomy of Viruses (2012)
  • Author(s): Fukuhara T., Gibbs M.J.
  • Total Pages: 1338
  • Publisher: Family Endornaviridae
• [Book] Detection of long and short double-stranded RNAs (2011)
  • Author(s): Fukuhara T., Urayama S., Okada R., Kiyota E. and Moriyama H
  • Publisher: Humana Press
• [Book] Methods in Molecular Biology 744 (2011)
  • Author(s): Fukuhara T., Urayama S., Okada R., Kiyota E., Moriyama H
  • Total Pages: 233
  • Publisher: Detection of long and short double-stranded RNAs