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Slr2019, lipid A transporter homolog, is essential for acidic tolerance in Synechocystis sp. PCC6803

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Abstract

Living organisms must defend themselves against various environmental stresses. Extracellular polysaccharide-producing cells exhibit enhanced tolerance toward adverse environmental stress. In Synechocystis sp. PCC6803 (Synechocystis), lipopolysaccharide (LPS) may play a role in this protection. To examine the relationship between stress tolerance of Synechocystis and LPS, we focused on Slr2019 because Slr2019 is homologous to MsbA in Escherichia coli, which is related to LPS synthesis. First, to obtain a defective mutant of LPS, we constructed the slr2019 insertion mutant (slr2019) strain. Sodium deoxycholate-polyacrylamide gel electrophoresis indicated that slr2019 strain did not synthesize normal LPS. Second, to clarify the participation of LPS in acid tolerance, wild type (WT) and slr2019 strain were grown under acid stress; slr2019 strain growth was significantly weaker than WT growth. Third, to examine influences on stress tolerance, slr2019 strain was grown under various stresses. Under salinity and temperature stress, slr2019 strain grew significantly slower than WT. To confirm cell morphology, cell shape and envelope of slr2019 strain were observed by transmission electron microscopy; slr2019 cells contained more electron-transparent bodies than WT cells. Finally, to confirm whether electron-transparent bodies are poly-3-hydroxybutyrate (PHB), slr2019 strain was stained with Nile Blue A, a PHB detector, and observed by fluorescence microscopy. The PHB granule content ratio of WT and slr2019 strain grown at BG-11 pH 8.0 was each 7.18 and 8.41 %. At pH 6.0, the PHB granule content ratio of WT and slr2019 strain was 2.99 and 2.60 %. However, the PHB granule content ratio of WT and slr2019 strain grown at BG-11N-reduced was 10.82 and 0.56 %. Because slr2019 strain significantly decreased PHB under BG-11N-reduced compared with WT, LPS synthesis may be related to PHB under particular conditions. These results indicated that Slr2019 is necessary for Synechocystis survival in various stresses.

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Abbreviations

EPS:

Extracellular polysaccharide

Kdo:

3-Deoxy-d-manno-oct-2-ulosonic acid

LPS:

Lipopolysaccharide

OM:

Outer membrane

PHB:

Poly-3-hydroxybutyrate

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

SDOC-PAGE:

Sodium deoxycholate-polyacrylamide gel electrophoresis

Synechocystis :

Synechocystis sp. PCC6803

WT:

Wild type

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Acknowledgments

This study was supported by the Program for Development of Strategic Research Center in Private Universities, which was supported by MEXT. The authors would like to thank Enago (www.enago.jp) for the English language review.

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Authors and Affiliations

  1. Graduate School of Mathematics and Science Education, Tokyo University of Science, Shinjuku-ku, Tokyo, 162-8601, Japan

    Ayumi Matsuhashi, Hiroko Tahara & Hisataka Ohta

  2. Department of Biological Science and Technology, Tokyo University of Science, Niijuku, Katsushika-ku, Tokyo, Japan

    Yutaro Ito

  3. Research Center for RNA Science, Research Institute for Science and Technology, Tokyo University of Science, Noda, Chiba, 278-8510, Japan

    Junji Uchiyama & Hisataka Ohta

  4. Laboratory of Electron Microscopy, School of Medicine, Mie University, Tsu, Mie, 514-8507, Japan

    Satoru Ogawa

  5. Department of Biology, Faculty of Science, Tokyo University of Science, Shinjuku-ku, Tokyo, 162-8601, Japan

    Hisataka Ohta

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  1. Ayumi Matsuhashi
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  2. Hiroko Tahara
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  3. Yutaro Ito
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  4. Junji Uchiyama
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Correspondence to Ayumi Matsuhashi.

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Matsuhashi, A., Tahara, H., Ito, Y. et al. Slr2019, lipid A transporter homolog, is essential for acidic tolerance in Synechocystis sp. PCC6803. Photosynth Res 125, 267–277 (2015). https://doi.org/10.1007/s11120-015-0129-6

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