Skip to main content

Synechocystis PCC6803 and PCC6906 dnaK2 expression confers salt and oxidative stress tolerance in Arabidopsis via reduction of hydrogen peroxide accumulation

Molecular Biology Reports volume 41, pages 1091–1101 (2014)Cite this article

Abstract

Abiotic stress slows plant growth and development. Because salt stress, particularly from NaCl, acts as an important limiting factor in agricultural productivity, the identification and manipulation of genes related to salt tolerance could improve crop productivity. Prokaryotic, heat shock protein (Hsp), DnaK from the ubiquitous Hsp70 family is upregulated in cells that are under abiotic stress. Synechocystis spp. cyanobacteria encode at least three potential DnaK proteins in their genome. Here, expressions of dnaK1s and dnaK2s from two Synechocystis spp. PCC6803 (Sy6803) and PCC6906 (Sy6906), enhanced salt tolerance in a dnaK-defective Escherichia coli strain. In contrast, dnaK3s in both strains were ineffective, indicating that dnaK3 is functionally different from dnaK1 and dnaK2 in Synechocystis spp. under salt stress. Ectopic expression of dnaK2s from Sy6803 and Sy6906 conferred salt tolerance in transgenic Arabidopsis plants, which exhibited greater root length, chlorophyll content, fresh weight, and survival rate than wild type plants, all in the presence of NaCl. In transgenic plants, hydrogen peroxide (H2O2) accumulation was reduced under NaCl stress and loss of chlorophyll content was reduced under H2O2 stress. Overall results suggest that dnaK2s from Sy6803 and Sy6906 confer salt and oxidative tolerance in transgenic plants by reduction of H2O2 accumulation.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Abbreviations

Hsp:

Heat shock protein

ROS:

Reactive oxygen species

Sy6803:

Synechocystis spp. PCC6803

Sy6906:

Synechocystis spp. PCC6906

References

  1. Abogadallah GM (2010) Antioxidative defense under salt stress. Plant Signal Behav 5:369–374

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. Ahmad R, Kim MD, Back KH, Kim HS, Lee HS, Kwon SY, Murata N, Chung WI, Kwak SS (2008) Stress-induced expression of choline oxidase in potato plant chloroplasts confers enhanced tolerance to oxidative, salt, and drought stresses. Plant Cell Rep 27:687–698

    CAS  Article  PubMed  Google Scholar 

  3. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    CAS  Article  PubMed  Google Scholar 

  4. Ashrafa M, Harris PJC, Baak MA (2008) Potential biochemical indicators of salinity tolerance in plants. Plant Sci 166:3–16

    Article  Google Scholar 

  5. Begcy K, Mariano ED, Gentile A, Lembke CG, Zingaretti SM, Souza GM, Menossi M (2012) A novel stress-induced sugarcane gene confers tolerance to drought, salt and oxidative stress in transgenic tobacco plants. PLoS One 7:e44697

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Borsani O, Valpuesta V, Botella MA (2001) Evidence for a role of salicylic acid in the oxidative damage generated by NaCl and osmotic stress in Arabidopsis seedlings. Plant Physiol 126:1024–1030

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. Brehmer D, Rudiger S, Gassler CS, Klostermeier D, Packschies L, Reinstein J, Mayer MP, Bukau B (2001) Tuning of chaperone activity of Hsp70 proteins by modulation of nucleotide exchange. Nat Struct Biol 8:427–432

    CAS  Article  PubMed  Google Scholar 

  8. Buchberger A, Gassler CS, Buttner M, McMacken R, Bukau B (1999) Functional defects of the DnaK756 mutant chaperone of Escherichia coli indicate distinct roles for amino- and carboxyterminal residues in substrate and co-chaperone interaction and interdomain communication. J Biol Chem 274:38017–38026

    CAS  Article  PubMed  Google Scholar 

  9. Bukau B, Weissman J, Horwich A (2006) Molecular chaperones and protein quality control. Cell 125:443–451

    CAS  Article  PubMed  Google Scholar 

  10. Chitnis PR, Nelson N (1991) Molecular cloning of the genes encoding two chaperone proteins of the cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem 266:58–65

    CAS  PubMed  Google Scholar 

  11. Chung JS, Zhu JK, Bressan RA, Hasegawa PM, Shi H (2008) Reactive oxygen species mediate Na+ -induced SOS1 mRNA stability in Arabidopsis. Plant J 53:554–565

    CAS  Article  PubMed  Google Scholar 

  12. Fulda S, Mikkat S, Huang F, Huckauf J, Marin K, Norling B, Hagemann M (2006) Proteome analysis of salt stress response in the cyanobacterium Synechocystis sp. strain PCC 6803. Proteomics 6:2733–2745

    CAS  Article  PubMed  Google Scholar 

  13. Genevaux P, Georgopoulos C, Kelley WL (2007) The Hsp70 chaperone machines of Escherichia coli: a paradigm for the repartition of chaperone functions. Mol Microbiol 66:840–857

    CAS  Article  PubMed  Google Scholar 

  14. Georgopoulos CP, Lam B, Lundquist-Heil A, Rudolph CF, Yochem J, Feiss M (1979) Identification of the E. coli dnaK (groPC756) gene product. Mol Gen Genet 172:143–149

    CAS  Article  PubMed  Google Scholar 

  15. Gomez JM, Hernandez JA, Jimenez A, del Rio LA, Sevilla F (1999) Differential response of antioxidative enzymes of chloroplasts and mitochondria to long-term NaCl stress of pea plants. Free Radic Res 31(Suppl):S11–S18

    CAS  Article  PubMed  Google Scholar 

  16. Hernandez JA, Ferrer MA, Jimenez A, Barcelo AR, Sevilla F (2001) Antioxidant systems and O2 −/H2O2 production in the apoplast of pea leaves. Its relation with salt-induced necrotic lesions in minor veins. Plant Physiol 127:817–831

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  17. Huang L, McCluskey MP, Ni H, LaRossa RA (2002) Global gene expression profiles of the cyanobacterium Synechocystis sp. strain PCC 6803 in response to irradiation with UV-B and white light. J Bacteriol 184:6845–6858

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  18. Jungkunz I, Link K, Vogel F, Voll LM, Sonnewald S, Sonnewald U (2011) AtHsp70-15-deficient Arabidopsis plants are characterized by reduced growth, a constitutive cytosolic protein response and enhanced resistance to TuMV. Plant J 66:983–995

    CAS  Article  PubMed  Google Scholar 

  19. Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, Kimura T, Hosouchi T, Matsuno A, Muraki A, Nakazaki N, Naruo K, Okumura S, Shimpo S, Takeuchi C, Wada T, Watanabe A, Yamada M, Yasuda M, Tabata S (1996) Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res 3:109–136

    CAS  Article  PubMed  Google Scholar 

  20. Kanesaki Y, Suzuki I, Allakhverdiev SI, Mikami K, Murata N (2002) Salt stress and hyperosmotic stress regulate the expression of different sets of genes in Synechocystis sp. PCC 6803. Biochem Biophys Res Commun 290:339–348

    CAS  Article  PubMed  Google Scholar 

  21. Kim J, Yi H, Choi G, Shin B, Song PS, Choi G (2003) Functional characterization of phytochrome interacting factor 3 in phytochrome-mediated light signal transduction. Plant Cell 15:2399–2407

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. Latijnhouwers M, Xu XM, Moller SG (2010) Arabidopsis stromal 70-kDa heat shock proteins are essential for chloroplast development. Planta 232:567–578

    CAS  Article  PubMed  Google Scholar 

  23. Lee BH, Hibino T, Jo J, Viale AM, Takabe T (1997) Isolation and characterization of a dnaK genomic locus in a halotolerant cyanobacterium Aphanothece halophytica. Plant Mol Biol 35:763–775

    CAS  Article  PubMed  Google Scholar 

  24. Lee JH, Schöffl F (1996) AnHsp70 antisense gene affects the expression of HSP70/HSC70, the regulation of HSF, and the acquisition of thermotolerance in transgenic Arabidopsis thaliana. Mol Gen Genet 252:11–19

    CAS  PubMed  Google Scholar 

  25. Li H, Xie P, Zhang D, Chen J (2009) The first study on the effects of microcystin-RR on gene expression profiles of antioxidant enzymes and heat shock protein-70 in Synechocystis sp. PCC6803. Toxicon 53:595–601

    CAS  Article  PubMed  Google Scholar 

  26. Lindquist S, Craig EA (1988) The heat shock proteins. Annu Rev Genet 22:631–677

    CAS  Article  PubMed  Google Scholar 

  27. Mary I, Tu CJ, Grossman A, Vaulot D (2004) Effects of high light on transcripts of stress-associated genes for the cyanobacteria Synechocystis sp. PCC 6803 and Prochlorococcus MED4 and MIT9313. Microbiology 150:1271–1281

    CAS  Article  PubMed  Google Scholar 

  28. Mayer MP, Bukau B (2005) Hsp70 chaperones: cellular functions and molecular mechanism. Cell Mol Life Sci 62:670–684

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    CAS  Article  Google Scholar 

  30. Nimura K, Yoshikawa H, Takahashi H (1994) Identification of dnaK multigene family in Synechococcus sp. PCC7942. Biochem Biophys Res Commun 201:466–471

    CAS  Article  PubMed  Google Scholar 

  31. Nimura K, Yoshikawa H, Takahashi H (1996) DnaK3, one of the three DnaK proteins of cyanobacterium Synechococcus sp. PCC7942, is quantitatively detected in the thylakoid membrane. Biochem Biophys Res Commun 229:334–340

    CAS  Article  PubMed  Google Scholar 

  32. Ono K, Hibino T, Kohinata T, Suzuki S, Tanaka Y, Nakamura T, Takabe T (2001) Overexpression of DnaK from a halotolerant cyanobacterium Aphanothece halophytica enhances the high-temperatue tolerance of tobacco during germination and early growth. Plant Sci 160:455–461

    CAS  Article  PubMed  Google Scholar 

  33. Packschies L, Theyssen H, Buchberger A, Bukau B, Goody RS, Reinstein J (1997) GrpE accelerates nucleotide exchange of the molecular chaperone DnaK with an associative displacement mechanism. Biochemistry 36:3417–3422

    CAS  Article  PubMed  Google Scholar 

  34. Paek KH, Walker GC (1987) Escherichia coli dnaK null mutants are inviable at high temperature. J Bacteriol 169:283–290

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Park HS, Jeong WJ, Kim E, Jung Y, Lim JM, Hwang MS, Park EJ, Ha DS, Choi DW (2012) Heat shock protein gene family of the Porphyra seriata and enhancement of heat stress tolerance by PsHSP70 in Chlamydomonas. Mar Biotechnol (NY) 14:332–342

    CAS  Article  Google Scholar 

  36. Rupprecht E, Duppre E, Schneider D (2010) Similarities and singularities of three DnaK proteins from the cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 51:1210–1218

    CAS  Article  PubMed  Google Scholar 

  37. Rupprecht E, Gathmann S, Fuhrmann E, Schneider D (2007) Three different DnaK proteins are functionally expressed in the cyanobacterium Synechocystis sp. PCC 6803. Microbiology 153:1828–1841

    CAS  Article  PubMed  Google Scholar 

  38. Sato M, Nimura-Matsune K, Watanabe S, Chibazakura T, Yoshikawa H (2007) Expression analysis of multiple dnaK genes in the cyanobacterium Synechococcus elongatus PCC 7942. J Bacteriol 189:3751–3758

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  39. Scarpeci TE, Zanor MI, Carrillo N, Mueller-Roeber B, Valle EM (2008) Generation of superoxide anion in chloroplasts of Arabidopsis thaliana during active photosynthesis: a focus on rapidly induced genes. Plant Mol Biol 66:361–378

    CAS  Article  PubMed  Google Scholar 

  40. Smirnoff N (1998) Plant resistance to environmental stress. Curr Opin Biotech 9:214–219

    CAS  Article  PubMed  Google Scholar 

  41. Sugino M, Hibino T, Tanaka Y, Nii N, Takabe T (1999) Overexpression of DnaK from a halotolerant cyanobacterium Aphanothece halophytica acquires resistance to salt stress in transgenic tobacco plants. Plant Sci 137:81–88

    Article  Google Scholar 

  42. Suzuki I, Simon WJ, Slabas AR (2006) The heat shock response of Synechocystis sp. PCC 6803 analysed by transcriptomics and proteomics. J Exp Bot 57:1573–1578

    CAS  Article  PubMed  Google Scholar 

  43. Takabe T, Uchida A, Shinagawa F, Terada Y, Kajita H, Tanaka Y, Takabe T, Hayashi T, Kawai T, Takabe T (2008) Overexpression of DnaK from a halotolerant cyanobacterium Aphanothece halophytica enhances growth rate as well as abiotic stress tolerance of poplar plants. Plant Growth Regul 56:265–273

    CAS  Article  Google Scholar 

  44. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  45. Uchida A, Hibino T, Shimada T, Saigusa M, Takabe T, Araki E, Kajita H, Takabe T (2008) Overexpression of DnaK chaperone from a halotolerant cyanobacterium Aphanothece halophytica increases seed yield in rice and tobacco. Plant Biotechnol 25:141–150

    CAS  Article  Google Scholar 

  46. Varvasovszki V, Glatz A, Shigapova N, Josvay K, Vigh L, Horvath I (2003) Only one dnaK homolog, dnaK2, is active transcriptionally and is essential in Synechocystis. Biochem Biophys Res Commun 305:641–648

    CAS  Article  PubMed  Google Scholar 

  47. Wang W, Vinocur B, Shoseyov O, Altman A (2004) Role of plant heat–shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci 9:244–252

    CAS  Article  PubMed  Google Scholar 

  48. Wu A, Allu AD, Garapati P, Siddiqui H, Dortay H, Zanor MI, Asensi-Fabado MA, Munne-Bosch S, Antonio C, Tohge T, Fernie AR, Kaufmann K, Xue GP, Mueller-Roeber B, Balazadeh S (2012) JUNGBRUNNEN1, a reactive oxygen species-responsive NAC transcription factor, regulates longevity in Arabidopsis. Plant Cell 24:482–506

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  49. Yoshimune K, Yoshimura T, Esaki N (1998) Hsc62, a new DnaK homologue of Escherichia coli. Biochem Biophys Res Comm 250:115–118

    CAS  Article  PubMed  Google Scholar 

  50. Zhu J, Fu X, Koo YD, Zhu JK, Jenney FE Jr, Adams MW, Zhu Y, Shi H, Yun DJ, Hasegawa PM, Bressan RA (2007) An enhancer mutant of Arabidopsis salt overly sensitive 3 mediates both ion homeostasis and the oxidative stress response. Mol Cell Biol 27:5214–5224

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  51. Zhu JK (2001) Plant salt tolerance. Trends Plant Sci 6:66–71

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant to J.R.L. from the Marine Extreme Genome Research Center, which is funded by the Korean Ministry of Marine Affairs and Fisheries; a grant to J.R.L. from the Technology Development Program for Agriculture and Forestry, which is funded by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries; and a grant to J.R.L. from the Korea Research Institute of Bioscience and Biotechnology for the Development of Cell Factory for Biomass and Bio-energy.

Author information

Affiliations

  1. Greenbio Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Taejon, 305-806, Korea

    Jonghyun Kim, Myung Suk Ahn, Young Min Park, Sung Ran Min, Weon Joong Jeong & Jang R. Liu

  2. Division of Biological Resource Management, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Taejon, 305-806, Korea

    Suk Weon Kim

  3. Division of Life Science, Korea Basic Science Institute (KBSI), 113 Gwahak-ro, Yuseong-gu, Taejon, 305-333, Korea

    Jonghyun Kim

Authors
  1. Jonghyun Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Myung Suk Ahn
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Young Min Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Suk Weon Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sung Ran Min
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Weon Joong Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jang R. Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jang R. Liu.

Additional information

J. Kim and M.S. Ahn have contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 25 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kim, J., Ahn, M.S., Park, Y.M. et al. Synechocystis PCC6803 and PCC6906 dnaK2 expression confers salt and oxidative stress tolerance in Arabidopsis via reduction of hydrogen peroxide accumulation. Mol Biol Rep 41, 1091–1101 (2014). https://doi.org/10.1007/s11033-013-2955-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-013-2955-y

Keywords

  • Arabidopsis thaliana
  • Hydrogen peroxide
  • Salt stress tolerance
  • Synechocystis
  • Transgenic plants