Analysis of mesophilic cyanobacterium mutant strain that has acquired a high temperature tolerance comparable to moderately thermophilic bacteria

• Project/Area Number: 15K07368
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied microbiology
• Research Institution: Shizuoka University (2017); Tokyo University of Agriculture (2015-2016)
• Principal Investigator: Kanesaki Yu 静岡大学, グリーン科学技術研究所, 特任助教 (70380293)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 適応進化 / シアノバクテリア / ゲノム解析 / 高温ストレス / トランスクリプトーム / 突然変異 / NGS / 長期培養 / 高温耐性 / リシーケンス解析 / リシークエンス解析

Outline of Final Research Achievements

The cyanobacterium Synechococcus elongatus PCC 7942 is the mesophilic photosynthetic organism. We tried long-term adaptive evolution to obtain mutants which are able to proliferate more than a growth limit of high temperature, 43℃. We successfully obtained several mutants which are able to growth more than 45℃ on the BG-11 agar plate. These mutants showed unique transcriptional profile comparing to that of wild-type cells. We identified all spontaneous mutated loci in the genome of the mutant cells by resequence analysis using NGS technology. By verification of the mutations, We found that two genes are associated to the phenotype of cellular thermo-tolerance.

Research Products

• [Int'l Joint Research] University of Freiburg(Germany)
• [Journal Article] Transcriptional activation of glycogen catabolism and oxidative pentose phosphate pathway by NrrA facilitates cell survival under nitrogen starvation in the cyanobacterium Synechococcus sp. strain PCC 7002. (2018)
  • Author(s): Shimmori Y, Kanesaki Y, Nozawa M, Yoshikawa H, and Ehira S.
  • Journal Title: Plant and Cell Physiology Volume: 印刷中 Issue: 6 Pages: 1225-1233
  • DOI: 10.1093/pcp/pcy059
  • Peer Reviewed
• [Journal Article] Complete Genome Sequence of the Nonheterocystous Cyanobacterium Pseudanabaena sp. ABRG5-3 (2018)
  • Author(s): Tajima Naoyuki、Kanesaki Yu、Sato Shusei、Yoshikawa Hirofumi、Maruyama Fumito、Kurokawa Ken、Ohta Hiroyuki、Nishizawa Tomoyasu、Asayama Munehiko、Sato Naoki
  • Journal Title: Genome Announcements Volume: 6 Issue: 6
  • DOI: 10.1128/genomea.01608-17
  • NAID: 120006457123
  • Peer Reviewed / Open Access
• [Journal Article] Draft Genome Sequence of the Nitrogen-Fixing and Hormogonia-Inducing Cyanobacterium Nostoc cycadae Strain WK-1, Isolated from the Coralloid Roots of Cycas revoluta. (2018)
  • Author(s): Kanesaki Y, Hirose M, Hirose Y, Fujisawa T, Nakamura Y, Watanabe S, Matsunaga S, Uchida H, and Murakami A.
  • Journal Title: Genome Announcements Volume: 6 Issue: 7
  • DOI: 10.1128/genomea.00021-18
  • NAID: 120006488529
  • Peer Reviewed / Open Access
• [Journal Article] Mutations responsible for alcohol tolerance in the mutant of Synechococcus elongatus PCC 7942 (SY1043) obtained by single-cell screening system. (2018)
  • Author(s): Hirokawa Y, Kanesaki Y, Arai S, Saruta F, Hayashihara K, Murakami A, Shimizu K, Honda H, Yoshikawa H, and Hanai T
  • Journal Title: Journal of Bioscience and Bioengineering Volume: 125 Issue: 5 Pages: 572-577
  • DOI: 10.1016/j.jbiosc.2017.11.012
  • NAID: 40021552647
  • Peer Reviewed
• [Journal Article] ParA-like protein influences the distribution of multi-copy chromosomes in cyanobacterium Synechococcus elongatus PCC 7942. (2018)
  • Author(s): Watanabe S, Noda A, Ohbayashi R, Uchioke K, Kurihara A, Nakatake S, Morioka S, Kanesaki Y, Chibazakura T, Yoshikawa H
  • Journal Title: Microbiology Volume: 164 Issue: 1 Pages: 45-56
  • DOI: 10.1099/mic.0.000577
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of mycosporine-like amino acids in the cyanobacterium <i>Nostoc verrucosum</i> (2018)
  • Author(s): Inoue-Sakamoto, K., Nazifi, E., Tsuji, C., Asano, T., Nishiuchi, T., Matsugo, S., Ishihara, K., Kanesaki, Y., Yoshikawa, H. and Sakamoto, T.
  • Journal Title: The Journal of General and Applied Microbiology Volume: 64 Issue: 5 Pages: 203-211
  • DOI: 10.2323/jgam.2017.12.003
  • NAID: 130007503362
  • ISSN: 0022-1260, 1349-8037
  • Peer Reviewed / Open Access
• [Journal Article] Acidophilic green algal genome provides insights into adaptation to an acidic environment (2017)
  • Author(s): Hirooka, S., Hirose, Y., Kanesaki, Y., Higuchi, S., Fujiwara, T., Onuma, R., Era, A., Ohbayashi, R., Uzuka, A., Nozaki, H., Yoshikawa, H., and Miyagishima, S.
  • Journal Title: Proc. Natl. Acad. Sci. USA. Volume: 114 Issue: 39
  • DOI: 10.1073/pnas.1707072114
  • Peer Reviewed / Open Access
• [Journal Article] Conserved two-component Hik34-Rre1 module directly activates heat-stress inducible transcription of major chaperone and other genes in Synechococcus elongatus PCC 7942 (2017)
  • Author(s): Ikki Kobayashi, Satoru Watanabe, Yu Kanesaki, Tomohiro Shimada, Hirofumi Yoshikawa and Kan Tanaka
  • Journal Title: Mol. Microbiol. Volume: 104 Issue: 2 Pages: 260-277
  • DOI: 10.1111/mmi.13624
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] CyanoBase: a large-scale update on its 20th anniversary. (2017)
  • Author(s): Fujisawa, T., Narikawa, R., Maeda, S., Watanabe, S., Kanesaki, Y., Kobayashi, K., Nomata, J., Hanaoka, M., Watanabe, M., Ehira, S., Suzuki, E., Awai, K. and Nakamura, Y.
  • Journal Title: Nucleic Acids Research Volume: 45 Issue: D1 Pages: 551-554
  • DOI: 10.1093/nar/gkw1131
  • Peer Reviewed / Open Access
• [Journal Article] Variety of DNA Replication Activity Among Cyanobacteria Correlates with Distinct Respiration Activity in the Dark (2017)
  • Author(s): Ryudo Ohbayashi, Jun-ya Yamamoto, Satoru Watanabe, Yu Kanesaki, Taku Chibazakura, Shin-ya Miyagishima, Hirofumi Yoshikawa
  • Journal Title: Plant and Cell Physiology Volume: 58 (2) Pages: 279-286
  • DOI: 10.1093/pcp/pcw186
  • Peer Reviewed
• [Journal Article] Chloroplast-mediated regulation of CO2-concentrating mechanism by Ca2+-binding protein CAS in the green alga Chlamydomonas reinhardtii (2016)
  • Author(s): L. Wang, T. Yamano, S. Takane, Y. Niikawa, C. Toyokawa, S. Ozawa, R. Tokutsu, Y. Takahashi, J. Minagawa, Y. Kanesaki, H. Yoshikawa, and H. Fukuzawa
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 113 Issue: 44 Pages: 12586-12591
  • DOI: 10.1073/pnas.1606519113
  • NAID: 120006425865
  • Peer Reviewed / Open Access
• [Journal Article] Complete genome sequencing of cyanobacterium Leptolyngbya sp. NIES-3755 Journal of Biotechnology. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Genome Announcements Volume: 4 Issue: 2
  • DOI: 10.1128/genomea.00090-16
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Complete genome sequence of cyanobacterium Fischerella sp. NIES-3754, providing thermoresistant optogenetic tools. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Journal of Biotechnology Volume: 220 Pages: 45-46
  • DOI: 10.1016/j.jbiotec.2016.01.011
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Complete genome sequence of cyanobacterium Nostoc sp. NIES-3756, a potentially useful strain for phytochrome-based bioengineering. (2016)
  • Author(s): Hirose Y., Fujisawa T., Ohtsubo Y., Katayama M., Misawa N., Wakazuki S., Shimura Y., Nakamura Y., Kawachi M., Yoshikawa H., Eki T., and Kanesaki Y.
  • Journal Title: Journal of Biotechnology Volume: 220 Pages: 51-52
  • DOI: 10.1016/j.jbiotec.2015.12.002
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Diversification of DnaA dependency for DNA replication in cyanobacterial evolution. (2016)
  • Author(s): Ohbayashi R., Watanabe S., Ehira S., Kanesaki Y., Chibazakura T., and Yoshikawa H.
  • Journal Title: ISME Journal Volume: 10 Issue: 5 Pages: 1113-1121
  • DOI: 10.1038/ismej.2015.194
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Intensive DNA replication and metabolism during the lag phase in cyanobacteria (2015)
  • Author(s): Watanabe S., Ohbayashi R., Kanesaki Y., Saito N., Chibazakura T., Soga T., and Yoshikawa H.
  • Journal Title: PLoS One Volume: 10 Issue: 9 Pages: e0136800-e0136800
  • DOI: 10.1371/journal.pone.0136800
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of spontaneous suppressor mutants from the photomixotrophically-grown pmgA-disrupted mutant in the cyanobacterium Synechocystis sp. PCC 6803 (2015)
  • Author(s): Nishijima, Y., Kanesaki, Y., Yoshikawa, H., Ogawa, T., Sonoike, K., Nishiyama, Y. and Hihara, Y.
  • Journal Title: Photosynth. Res. Volume: - Issue: 2-3 Pages: 465-475
  • DOI: 10.1007/s11120-015-0143-8
  • Peer Reviewed
• [Journal Article] Identification of centromere regions in chromosomes of a unicellular red alga, Cyanidioschyzon merolae (2015)
  • Author(s): Yu Kanesaki, Sousuke Imamura, Motomichi Matsuzaki, and Kan Tanaka
  • Journal Title: FEBS Lett. Volume: 589 Issue: 11 Pages: 1219-1224
  • DOI: 10.1016/j.febslet.2015.04.009
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] シアノバチルスにおけるシアノバクテリア遺伝子大規模発現の試み (2018)
  • Author(s): 美田 知也, 細村 匡太郎, 渡辺 智, 兼崎 友, 板谷 光泰, 朝井 計, 吉川 博文
  • Organizer: 日本農芸化学会2018年度大会
• [Presentation] クロロフィルdをもつシアノバクテリアAcaryochloris marina の橙色光への順化・適応機構の解析 (2018)
  • Author(s): 樫本友則, 兼崎友, 佐藤繭子, 渡邉麻衣, 渡辺智, 豊岡公徳, 池内昌彦, 成川礼
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] 網羅ゲノムシークエンスによるシアノバクテリアの補色応答の多様性の解析 (2018)
  • Author(s): 広瀬侑, 米川千夏, 藤澤貴智, 志村遥平, 兼崎友, 渡辺麻衣, 中村保一, 河地正伸, 池内昌彦, 浴俊彦
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] シアノバクテリアSynechocustis sp PCC 6803 における複製開始機構の解析 (2018)
  • Author(s): 山﨑 脩平, 大林龍胆, 松根 かおり, 兼崎 友, 渡辺 智
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] シアノバクテリアSynechocystis sp. PCC 6803 におけるRecA タンパク質の機能解析 (2018)
  • Author(s): 青柳 智大, 栗原 亜実, 大林龍胆, 兼崎 友, 渡辺 智
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] Synechocystis sp. PCC6803 のSll0914欠損株は酸性ストレスに感受性を示す (2018)
  • Author(s): 中原 凌波, 石川 晴菜, 板垣 文子, 甲賀 栄貴, 兼崎 友, 吉川 博文, 内山純爾, 太田尚孝
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] 陸棲シアノバクテリアであるイシクラゲ（Nostoc commune）のゲノム解析 (2018)
  • Author(s): 加藤 浩, 広瀬 侑, 兼崎 友, 藤澤 貴智, 中村 保一, 吉川 博文
  • Organizer: 第12回日本ゲノム微生物学会年会
• [Presentation] 有用微生物のゲノム解析実績と予想される今後の展開 (2018)
  • Author(s): 兼崎 友
  • Organizer: 東京農業大学生物資源ゲノム解析センター 2017年度シンポジウム 「生命科学と情報科学の融合による農学研究の拠点形成」
  • Invited
• [Presentation] シアノバクテリアにおける新規高温耐性株の取得とリシーケンス解析 (2017)
  • Author(s): 兼崎友、樋山智恵美、小澤啓吾、宮澤和己、渡辺智、吉川博文
  • Organizer: 日本農芸化学会 2017年度大会
  • Place of Presentation: 京都
  • Year and Date: 2017-03-17
• [Presentation] シアノバクテリアにおける複数コピーゲノムのDNA複製制御機構 (2017)
  • Author(s): 大林龍胆、中町愛、兼崎友、渡辺智、吉川博文、宮城島進也
  • Organizer: 第11回 日本ゲノム微生物学会
  • Place of Presentation: 藤沢
  • Year and Date: 2017-03-02
• [Presentation] アオコ原因シアノバクテリア Planktothrix agardhii NIES-204株の完全ゲノム解読 (2017)
  • Author(s): 志村遥平、藤澤貴智、広瀬侑、兼崎友、河地正伸
  • Organizer: 第11回 日本ゲノム微生物学会
  • Place of Presentation: 藤沢
  • Year and Date: 2017-03-02
• [Presentation] 繰り返し配列を介したゲノムシャッフリングによる好熱性シアノバクテリアのゲノム構造の進化 (2017)
  • Author(s): 前田海成、広瀬侑、藤澤貴智、兼崎友、吉川博文、池内昌彦
  • Organizer: 第11回 日本ゲノム微生物学会
  • Place of Presentation: 藤沢
  • Year and Date: 2017-03-02
• [Presentation] マルチコピーゲノムの複製制御機構からゲノムの存在意義を探る (2017)
  • Author(s): 大林龍胆, 兼崎友, 渡辺智, 吉川博文, 宮城島進也
  • Organizer: 藍藻の分子生物学会2017
• [Presentation] 非窒素固定型シアノバクテリアの窒素飢餓適応における糖異化の役割 (2017)
  • Author(s): 新森友香, 兼崎 友, 吉川博文, 得平茂樹
  • Organizer: 藍藻の分子生物学会2017
• [Presentation] 遠赤色光吸収クロロフィルdを持つシアノバクテリアAcaryochloris marinaの光質感知・順化・適応機構の解析 (2017)
  • Author(s): 樫本 友則, 兼崎 友, 佐藤 繭子, 伏見 圭司, 池内 昌彦, 豊岡 公徳, 渡辺 智, 成川 礼
  • Organizer: 日本植物学会第81回大会
• [Presentation] 新奇糸状性シアノバクテリアのSigB 相同性因子の特徴付けと発現解析 (2017)
  • Author(s): 菅原 卓也, 鎭西 真理子, 川又 透, 田島 直幸, 兼崎 友, 中平 洋一, 吉川 博文, 佐藤 直樹, 朝山 宗彦
  • Organizer: 第69回日本生物工学会大会
• [Presentation] シアノバクテリアにおける高温適応進化実験の試み (2017)
  • Author(s): 兼崎 友, 樋山 智恵美, 宮澤 和己, 渡辺 智, 吉川 博文
  • Organizer: 環境微生物系学会合同大会2017
• [Presentation] 長期高温培養によるシアノバクテリア新規高温適応株の取得 (2017)
  • Author(s): 兼崎 友, 樋山 智恵美, 宮澤 和己, 渡辺 智
  • Organizer: NGS現場の会 第五回研究会
• [Presentation] 重金属耐性を示す単細胞紅藻シアニジウムのオミクス解析 (2017)
  • Author(s): 兼崎友
  • Organizer: 東京工業大学シンポジウム「光合成とバイオマス」
  • Place of Presentation: 横浜
  • Invited
• [Presentation] Identification of the associated genes for substrain-specific phenotypes of a cyanobacterium, Synechococcus elongatus PCC 7942. (2015)
  • Author(s): Kanesaki Y., Ohbayashi R., Watanabe S., and Yoshikawa H.
  • Organizer: International Symposium on Phototrophic Prokaryotes
  • Place of Presentation: Tubingen, Germany
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Variety of dependency on DnaA protein for DNA replication among cyanobacteria (2015)
  • Author(s): Ohbayashi R., Watanabe S., Ehira S., Kanesaki Y., Chibazakura T., and Yoshikawa H.
  • Organizer: International Symposium on Phototrophic Prokaryotes
  • Place of Presentation: Tubingen, Germany
  • Year and Date: 2015-08-02
  • Int'l Joint Research
• [Presentation] Cell division-uncoupled DNA replication and metabolism in cyanobacteria Synechococcus elongatus PCC 7942. (2015)
  • Author(s): Watanabe S., Ohbayashi R., Kanesaki Y., Chibazakura T., and Yoshikawa H.
  • Organizer: International Symposium on Phototrophic Prokaryotes
  • Place of Presentation: Tubingen, Germany
  • Year and Date: 2015-08-02
  • Int'l Joint Research