Journal of Applied Phycology

, Volume 31, Issue 2, pp 1185–1196 | Cite as

Proteomic analysis of the salt-adapted and directly salt-(NaCl and NaCl+Na2SO4 mixture) stressed cyanobacterium Anabaena fertilissima

  • Ashwani K. RaiEmail author
  • Prashant Swapnil


Salinity is a serious threat to agriculture productivity. Beneficial microbes could be simple and low-cost biological methods to mitigate the salt toxicity. The present study describes proteome dynamics and salt stress tolerance of the salt-primed (pre-exposed to salt) and directly salt-stressed cyanobacterium Anabaena fertilissima. The difference in the proteome of differently salt-treated and the control cells was the abundance of proteins. Out of 130 proteins resolved in control, 51.8% remained constant, 25.5% were upregulated, and 22.7% protein spots were downregulated in salt-adapted cells (exposed to 500 mM NaCl). However, in cells exposed to 250 mM NaCl, percentage of constant, upregulated, and downregulated proteins was 56.8, 16, and 27.2, whereas in the cells exposed to equimolar NaCl+Na2SO4 mixture, these values were 41.8, 29.4, and 28.7, respectively. This indicated that an altered protein expression occurred in large number of protein species under different salt-stress regimes. Four proteins showing significant and reproducible changes under salt stress showed close homology with photosystem I reaction center subunit XII, epoxyqueuosine reductase, response regulator protein VraR, and molybdopterin biosynthesis protein. The kinetic analysis revealed that salt treatment increased the abundance of all the four identified proteins. Despite the exposure to higher NaCl concentration (500 mM), salt-adapted cells showed minimal hyper accumulation of these four proteins, followed by NaCl and NaCl+Na2SO4 mixture. Lesser accumulation of these identified proteins in salt mixture stressed cells than that of NaCl stressed cells revealed that the availability of sulfur during salt-stress relieved the salt-toxicity, while Cl increased it.


Protein abundance Salt acclimation Salt mixture Cyanobacterium Pre-exposure to salt 



Ashwani K Rai gratefully acknowledges the National Academy of Sciences, India, for awarding NASI-Senior Scientist Fellowship.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10811_2018_1607_MOESM1_ESM.docx (3 mb)
ESM 1 (DOCX 3039 kb)


  1. Allen MB, Arnon DI (1955) Studies on nitrogen-fixing blue-green algae. I. Growth and nitrogen fixation by Anabaena cylindrica Lemm. Plant Physiol 30:366–372CrossRefGoogle Scholar
  2. Amini S, Ghadiri H, Chen C, Marschner P (2016) Salt-affected soils, reclamation, carbon dynamics, and biochar: a review. J Soils Sediments 16:939–953CrossRefGoogle Scholar
  3. Beckers GJ, Conrath U (2007) Priming for stress resistance: from the lab to the field. Curr Opin Plant Biol 10:425–431CrossRefGoogle Scholar
  4. Belcheva A, Verma V, Golemi-Kotra D (2009) DNA-binding activity of the vancomycin resistance associated regulator protein VraR and the role of phosphorylation in transcriptional regulation of the vraSR operon. Biochemistry 48:5592–5601CrossRefGoogle Scholar
  5. Cameron JC, Pakrasi HB (2010) Essential role of glutathione in acclimation to environmental and redox perturbations in the cyanobacterium Synechocystis sp. PCC 6803. Plant Physiol 154:1672–1685CrossRefGoogle Scholar
  6. Chang C, Sommerfeldt TG, Carefoot JM, Schaalje GB (1983) Relationships of electrical conductivity with total dissolved salts and cation concentration of sulfate-dominant soil extracts. Can J Soil Sci 63:79–86CrossRefGoogle Scholar
  7. Chowdhury N, Marschner P, Burns RG (2011) Response of microbial activity and community structure to decreasing soil osmotic and matric potential. Plant Soil 344:241–254CrossRefGoogle Scholar
  8. Colla G, Rouphael Y, Rea E, Cardarelli M (2012) Grafting cucumber plants enhance tolerance to sodium chloride and sulfate salinization. Sci Hortic 135:177–185CrossRefGoogle Scholar
  9. Conrath U, Beckers GJ, Flors V, Garcia-Agustin P, Jakab G, Mauch F, Newman M-A, Pieterse CMJ, Poinssot B, Pozo MJ, Pugin A, Schaffrath U, Ton J, Wendehenne D, Zimmerli L, Mauch-Mani B (2006) Priming: getting ready for battle. Mol Plant-Microbe Interact 19:1062–1071CrossRefGoogle Scholar
  10. De Philippis R, Sili C, Paperi R, Vincenzini M (2001) Exopolysaccharide-producing cyanobacteria and their possible exploitation: a review. J Appl Phycol 13:293–299CrossRefGoogle Scholar
  11. Demetriou G, Neonaki C, Navakoudis E, Kotzabasis K (2007) Salt stress impact on the molecular structure and function of the photosynthetic apparatus—the protective role of polyamines. Biochim Biophys Acta 1767:272–280CrossRefGoogle Scholar
  12. Dubey AK, Rai AK (1995) Application of algal biofertilizers (Aulosira fertilissima tenuis and Anabaena doliolum Bhardawaja) for sustained paddy cultivation in Northern India. Isr J Plant Sci 43:41–51CrossRefGoogle Scholar
  13. El Yacoubi B, Bailly M, de Crécy-Lagard V (2012) Biosynthesis and function of posttranscriptional modifications of transfer RNAs. Annu Rev Genet 46:69–95CrossRefGoogle Scholar
  14. Fatma M, Asgher M, Masood A, Khan NA (2014) Excess sulfur supplementation improves photosynthesis and growth in mustard under salt stress through increased production of glutathione. Environ Exp Bot 107:55–63CrossRefGoogle Scholar
  15. Foyer C, Noctor G, Buchanan B, Dietz K, Pfannschmidt T (2009) Redox regulation in photosynthetic organisms: signaling, acclimation, and practical implications. Antioxid Redox Signal 11:861–905CrossRefGoogle Scholar
  16. Frazzon J, Dean DR (2002) Biosynthesis of the nitrogenase iron-molybdenum-cofactor from Azotobacter vinelandii. Met Ions Biol Syst 39:163–186Google Scholar
  17. Freedman B, Hutchinson TC (1980) Pollutant inputs from the atmosphere and accumulations in soils and vegetation near a nickel-copper smelter at Sudbury, Ontario, Canada. Can J Bot 58:108–132CrossRefGoogle Scholar
  18. 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 6:2733–2745CrossRefGoogle Scholar
  19. Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) A protein identification and analysis tools on the ExPASy Server. In: Walker JM (ed) The proteomics protocols handbook. Humana Press, New York, pp 571–607CrossRefGoogle Scholar
  20. Grattan SR, Grieve CM (1999) Mineral nutrient acquisition and response by plants grown in saline environments. In: Pessarakli M (ed) Handbook of plant and crop stress, 3rd edn. Marcel Dekker, New York, pp 203–229Google Scholar
  21. Hagemann M (2011) Molecular biology of cyanobacterial salt acclimation. FEMS Microbiol Rev 35:87–123CrossRefGoogle Scholar
  22. Hagemann M, Techel D, Rensing L (1991) Comparison of salt- and heat-induced alterations of protein synthesis in the cyanobacterium Synechocystis sp. PCC 6803. Arch Microbiol 155:587–592CrossRefGoogle Scholar
  23. Hagemann M, Fulda S, Schubert H (1994) DNA, RNA, and protein synthesis in the cyanobacterium Synechocystis sp. PCC 6803 adapted to different salt concentrations. Curr Microbiol 28:201–207CrossRefGoogle Scholar
  24. Hibino T, Lee BH, Rai AK, Ishikawa H, Kojima H, Tawada M, Shimoyama H, Takabe T (1996) Salt enhances photosystem I content and cyclic electron flow via NAD(P)H dehydrogenase in the halotolerant cyanobacterium Aphanothece halophytica. Aust J Plant Physiol 23:321–330Google Scholar
  25. Hille R (2002) Molybdenum and tungsten in biology. Trends Biochem Sci 27:360–367CrossRefGoogle Scholar
  26. Jeanjean R, Matthijs HCP, Onana B, Havaux M, Joset F (1993) Exposure of the cyanobacterium Synechocystis PCC6803 to salt stress induces concerted changes in respiration and photosynthesis. Plant Cell Physiol 34:1073–1079Google Scholar
  27. Johnson ZI, Zinser ER, Coe A, McNulty NP, Woodward EM, Chisholm SW (2006) Niche partitioning among Prochlorococcus ecotypes along ocean-scale environmental gradients. Science 311:1737–1740CrossRefGoogle Scholar
  28. Jones CG, Lawton JH, Shachak M (1994) Organisms as ecosystem engineers. Oikos 69:373–386CrossRefGoogle Scholar
  29. Jordan P, Fromme P, Witt HT, Klukas O, Saenger W, Krauß N (2001) Three-dimensional structure of cyanobacterial photosystem I at 2.5 Å resolution. Nature 411:909–917CrossRefGoogle Scholar
  30. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  31. Lawson DM, Smith BE (2002) Molybdenum nitrogenases: a crystallographic and mechanistic view. Met Ions Biol Syst 39:75–119Google Scholar
  32. Lessel U, Schomburg D (1994) Similarities between protein 3-D structures. Protein Eng 7:1175–1187CrossRefGoogle Scholar
  33. Mackay MA, Norton RS, Borowitzka LJ (1984) Organic osmoregulatory solutes in cyanobacteria. J Gen Microbiol 130:2177–2191Google Scholar
  34. Maiti R, Van Domselaar GH, Zhang H, Wishart DS (2004) SuperPose: a simple server for sophisticated structural superposition. Nucleic Acids Res 32:W590–W594CrossRefGoogle Scholar
  35. Masip L, Veeravalli K, Georgiou G (2006) The many faces of glutathione in bacteria. Antioxid Redox Signal 8:753–762CrossRefGoogle Scholar
  36. Messaoudi A, Belguith H, Ben HJ (2011) Three-dimensional structure of Arabidopsis thaliana lipase predicted by homology modeling method. Evol Bioinformatics Online 7:99–105Google Scholar
  37. Miyao M, Murata N (1983) Partial disintegration and reconstitution of the photosynthetic oxygen evolution system. Binding of 24 kilodalton and 18 kilodalton polypeptides. Biochim Biophys Acta 725:87–93CrossRefGoogle Scholar
  38. Murakami A, Kim SJ, Fujita Y (1997) Changes in photosystem stoichiometery in response to environmental conditions for cell growth observed with the cyanophyte Synechocystis PCC 6714. Plant Cell Physiol 38:392–397CrossRefGoogle Scholar
  39. Nagahara N, Wrobel M (2017) Atomic sulfur: an element for adaptation to an oxidative environment. Molecules 22:1821CrossRefGoogle Scholar
  40. Nishimura S (1983) Structure, biosynthesis, and function of queuosine in transfer RNA. Prog Nucleic Acid Res Mol Biol 28:49–73CrossRefGoogle Scholar
  41. Nriagu JO (1978) Production and uses of sulfur. In: Nriagu JO (ed) Sulfur in the environment, part 1: the atmospheric cycle. Wiley, New York, pp 1–21Google Scholar
  42. Oren A (2008) Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Systems 4:2CrossRefGoogle Scholar
  43. Pade N, Hagemann M (2015) Salt acclimation of cyanobacteria and their application in biotechnology. Life 5:25–49CrossRefGoogle Scholar
  44. Pandhal J, Snijders AP, Wright PC, Biggs CA (2008) A cross-species quantitative proteomic study of salt adaptation in a halotolerant environmental isolate using 15N metabolic labelling. Proteomics 8:2266–2284CrossRefGoogle Scholar
  45. Pitman MG, Lauchli A (2002) Global impact of salinity and agricultural ecosystems. In: Lauchli A, Lüttge U (eds) Salinity: environment-plants-molecules. Kluwer, Dordrecht, pp 3–20Google Scholar
  46. Qiao J, Huang S, Te R, Wang J, Chen L, Zhang W (2013) Integrated proteomic and transcriptomic analysis reveals novel genes and regulatory mechanisms involved in salt stress responses in Synechocystis sp. PCC 6803. Appl Microbiol Biotechnol 97:8253–8264CrossRefGoogle Scholar
  47. Rai AK (1990) Biochemical characteristics of photosynthetic response to various salinities in halotolerant and freshwater cyanobacteria. FEMS Microbiol Lett 69:177–180CrossRefGoogle Scholar
  48. Rai AK, Abraham G (1993) Salinity tolerance and growth analysis of the cyanobacterium Anabaena doliolum. Bull Environ Contam Toxicol 51:724–731Google Scholar
  49. Rai V, Rai AK (1999) Growth behaviour of Azolla pinnata at various salinity levels and induction of high salt tolerance. Plant Soil 206:79–84CrossRefGoogle Scholar
  50. Rajagopalan KV (1996) Biosynthesis of the molybdenum cofactor. In: Neidhardt FW, Curtiss IIIR, Ingraham JL, Lin ECC, Low KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella: cellular and molecular biology, 2nd edn. ASM Press, Washington DC, pp 674–679Google Scholar
  51. Rietz DN, Haynes RJ (2003) Effects of irrigation-induced salinity and sodicity on soil microbial activity. Soil Biol Biochem 35:845–854CrossRefGoogle Scholar
  52. Rodríguez AA, Stella AM, Storni MM, Zulpa G, Zaccaro MC (2006) Effects of cyanobacterial extracellular products and gibberellic acid on salinity tolerance in Oryza sativa L. Saline Systems 2:7CrossRefGoogle Scholar
  53. Rossi F, De Philippis R (2016) Exocellular polysaccharides in microalgae and cyanobacteria: chemical features, role and enzymes and genes involved in their biosynthesis. In: Borowitzka MA, Beardall J, Raven JA (eds) The physiology of microalgae. Springer, Dordrecht, pp 565–590CrossRefGoogle Scholar
  54. Rossi F, Li H, Liu Y, De Philippis R (2017) Cyanobacterial inoculation (cyanobacterisation): perspectives for the development of a standardized multifunctional technology for soil fertilization and desertification reversal. Earth Sci Rev 171:28–43CrossRefGoogle Scholar
  55. Rubio LM, Flores E, Herrero A (1999) Molybdopterin guanine dinucleotide cofactor in Synechococcus sp. nitrate reductase: identification of mobA and isolation of a putative moeB gene. FEBS Lett 462:358–362CrossRefGoogle Scholar
  56. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor, New YorkGoogle Scholar
  57. Schafer FQ, Buettner GR (2001) Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radic Biol Med 30:1191–1212CrossRefGoogle Scholar
  58. Schubert H, Hagemann M (1990) Salt effect on 77 K fluorescence and photosynthesis in the cyanobacterium Synechocystis spec. PCC 6803. FEMS Microbiol Lett 71:169–172CrossRefGoogle Scholar
  59. Schwede T, Kopp J, Guex N, Peitsch MC (2003) SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res 31:3381–3385CrossRefGoogle Scholar
  60. Şener MK, Jolley C, Ben-Shem A, Fromme P, Nelson N, Croce R, Schulten K (2005) Comparison of the light-harvesting networks of plant and cyanobacterial photosystem I. Biophys J 89:1630–1642CrossRefGoogle Scholar
  61. Sergeeva E, Liaimer A, Bergman B (2002) Evidence for production of the phytohormone indole-3-acetic acid by cyanobacteria. Planta 215:229–238CrossRefGoogle Scholar
  62. Shevchenko A, Tomas H, Havli J, Olsen JV, Mann M (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860CrossRefGoogle Scholar
  63. Simon WJ, Hall JJ, Suzuki I, Murata N, Slabas AR (2002) Proteomic study of the soluble proteins from the unicellular cyanobacterium Synechocystis sp. PCC6803 using automated matrix-assisted laser desorption/ionization-time of flight peptide mass fingerprinting. Proteomics 2:1735–1742CrossRefGoogle Scholar
  64. Singh NK, Dhar DW (2010) Cyanobacterial reclamation of salt-affected soil. In: Lichtfouse E (ed) Genetic engineering, biofertilisation, soil quality and organic farming sustainable agriculture reviews, vol 4. Springer, Dordrecht, pp 243–275CrossRefGoogle Scholar
  65. Singh VP, Trehan K (1973) Effect of extracellular products of Aulosira fertilissima on the growth of rice seedlings. Plant Soil 38:457–464CrossRefGoogle Scholar
  66. Singh P, Yekondi S, Chen PW, Tsai CH, Yu CW, Wu K, Zimmerli L (2014) Environmental history modulates Arabidopsis pattern-triggered immunity in a histone acetyltransferase1-dependent manner. Plant Cell 26:2676–2688CrossRefGoogle Scholar
  67. Sonoike K, Terashima (1994) Mechanism of the photosystem I photoinhibition in leaves of Cucumis sativus L. Planta 194:287–293CrossRefGoogle Scholar
  68. Stirk WA, Ördög V, Van Staden J (1999) Identification of the cytokinin isopentenyladenine in a strain of Arthronema africanum (Cyanobacteria). J Phycol 35:89–92CrossRefGoogle Scholar
  69. Sudhir P, Pogoryelov D, Kovacs L, Garab G, Murthy SDS (2005) The effects of salt stress on photosynthetic electron transport and thylakoid membrane proteins in the cyanobacterium Spirulina platensis. J Biochem Mol Biol 38:481–485Google Scholar
  70. Swapnil P, Rai AK (2018) Physiological responses to salt stress of salt-adapted and directly salt (NaCl and NaCl+Na2SO4 mixture)-stressed cyanobacterium Anabaena fertilissima. Protoplasma 255:963–976CrossRefGoogle Scholar
  71. Swapnil P, Singh M, Singh S, Sharma NK, Rai AK (2015) Recombinant glycinebetaine improves metabolic activities, ionic balance and salt tolerance in diazotrophic freshwater cyanobacteria. Algal Res 11:194–203CrossRefGoogle Scholar
  72. Swapnil P, Yadav AK, Srivastav S, Sharma NK, Srikrishna S, Rai AK (2017) Biphasic ROS accumulation and programmed cell death in a cyanobacterium exposed to salinity (NaCl and Na2SO4). Algal Res 23:88–95CrossRefGoogle Scholar
  73. Takabe T, Rai AK, Akazawa T (1984) Interaction of constituent subunits in ribulose 1,5-bisphosphate carboxylase from Aphanothece halophytica. Arch Biochem Biophys 229:202–211CrossRefGoogle Scholar
  74. Terashima I, Funayama S, Sonoike K (1994) The site of photoinhibition in leaves of Cucumis sativus L. at low temperature is photosystem I, not photosystem II. Planta 193:300–306CrossRefGoogle Scholar
  75. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680CrossRefGoogle Scholar
  76. Türkan I, Demiral T (2009) Recent developments in understanding salinity tolerance. Environ Exp Bot 67:2–9CrossRefGoogle Scholar
  77. Wiederstein M, Sippl MJ (2007) ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins. Nucleic Acids Res 35:W407–W410CrossRefGoogle Scholar
  78. Wong VNL, Greene RSB, Dalal RC, Murphy BW (2010) Soil carbon dynamics in saline and sodic soils: a review. Soil Use Manag 26:2–11CrossRefGoogle Scholar
  79. Yang C, Zhang ZS, Gao HY, Fan XL, Liu MJ, Li XD (2014) The mechanism by which NaCl treatment alleviates PSI photoinhibition under chilling-light treatment. J Photochem Photobiol B 140:286–291CrossRefGoogle Scholar
  80. Yang J, Ma LA, Jiang H, Wu G, Dong H (2016) Salinity shapes microbial diversity and community structure in surface sediments of the Qinghai-Tibetan Lakes. Sci Rep 6:25078CrossRefGoogle Scholar

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© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of BotanyBanaras Hindu UniversityVaranasiIndia

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