Effects of Copper and Cadmium on the Protein Profile and DNA Pattern of Marine Microalgae Chlorella salina and Nannochloropsis salina

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Toxicity of copper (Cu+2) and cadmium (Cd+2), as pollutants, in sea water was evaluated using two microalgae, Chlorella salina and Nannochloropsis salina. Experiments were carried out for 96 h with the two heavy metals causing changes in the growth, protein pattern and DNA profile of the studied algae. Lethal concentration (LC50) values for Cu+2 and Cd+2 indicated that both C. salina and N. salina are affected by Cu+2 than Cd+2. Polyacrylamide gel electrophoresis (SDS PAGE) was used for the separation and identification of proteins. The treatment of the two marine microalgae with Cu+2 and Cd+2 prompted the disappearance of a single band of C. salina and four bands of N. salina protein profiles, compared to control. From the phylogenetic tree that resulted, moderate genetic similarity was detected for untreated and heavy metals treated C. salina samples (52%), while low genetic similarity was detected for untreated and heavy metals treated N. salina samples (10%). Random amplified polymorphic DNA (RAPD) technique was used for investigating variability in genetic pattern of DNA in both C. salina and N. salina. In this regard, from the genetic pattern of DNA, it was concluded that both copper and cadmium induced some genotoxic influences in both algal species, with more genotoxic effect of copper than cadmium.

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  1. Al-Qurainy F, Alameri AA, Khan S (2010) RAPD profile for the assessment of genotoxicity on a medicinal plant: Eruca sativa. J Med Plant Res 4:579–586.

  2. Andrade LR, Farina M, Amado GM (2004) Effects of copper on Enteromorpha flexuosa (Chlorophyta) in vitro. Ecotoxicol Environ Saf 58:117–125.

  3. Atienzar FA, Jha AN (2006) The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: a critical review. Mutat Res 613(2–3):76–102

  4. Atienzar FA, Conradi M, Evenden AJ, Jha AN, Depledge MH (1999) Qualitative assessment of genotoxicity using random amplified polymorphic DNA: comparison of genomic template stability with key fitness parameters in Daphnia magna exposed to benzo[a]pyrene. Environ Toxicol Chem 18:2275–2282.

  5. Atienzar FA, Venier P, Jha AN, Depledge MH (2002) Evaluation of the random amplified polymorphic DNA(RAPD) assay for the detection of DNA damage and mutations. Mutat.Res. 521:151–163.

  6. Aydin SS, Basaran E, Cansaran-Duman D, Aras S (2013) Genotoxic effect of cadmium in okra seedlings: comparative investigation with population parameters and molecular markers. J Environ Biol 34(6):985–990

  7. Balaji S, Kalaivani T, Rajasekaran C, Shalini M, Siva R, Singh RK, Akthar M (2014) Arthrospira (Spirulina) species as bio adsorbents for lead, chromium and cadmium removal – a comparative study. Clean-Soil Air Water 42(12):1790–1797

  8. Balaji S, Kalaivani T, Rajasekaran C, Shalini M, Vinodhini S, Priyadharshini SS, Vidya AG (2015) Removal of heavy metals from tannery effluents of Ambur industrial area, Tamilnadu by Arthrospira (Spirulina). Environ Monit Assess 187(6):325.

  9. Balaji S, Kalaivani T, Shalini M, Sankari M, Priya RR, Siva R, Rajasekaran C (2016) Biomass characterisation and phylogenetic analysis of microalgae isolated from estuaries: role in phycoremediation of tannery effluent. Algal Res 14:92–99.

  10. Baumann HA, Morrison L, Stengel DB (2009) Metal accumulation and toxicity measured by PAM-chlorophyll fluorescence in seven species of marine macroalgae. Ecotoxicol Environ Saf 72:1063–1075.

  11. Cansaran-Duman D, Atakol O, Aras S (2011) Assessment of air pollution genotoxicity by RAPD in Evernia prunastri from around iron steel factory in Karabuk, Turkey. J Environ Sci 23:1171–1178.

  12. Cenkci S, Yildiz M, Cigerci IH, Konuk M, Bozdag A (2009) Toxic chemicals-induced genotoxicity detected by random amplified polymorphic DNA (RAPD) in bean (Phaseolus vulgaris L.) seedlings. Chemosphere 76(7):900–906.

  13. Collén J, Pinto E, Pedersen M, Colepicolo P (2003) Induction of oxidative stress in the red macroalga Gracilaria tenuistipitata by pollutants metals. Arch Environ Contam Toxicol 45:337–342.

  14. Donahue BA, Yin S, Taylor JS, Reines D, Hanawalt PC (1999) Transcript cleavage by RNA polymerase II arrested by a cyclobutane pyrimidine dimer in the DNA template. Proc Natl Acad Sci U S A 91:8502–8506.

  15. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

  16. Duque D, Montoya C, Botero LR (2019) Cadmium (Cd) tolerance evaluation of three strains of microalgae of the genus Ankistrodesmus, Chlorella and Scenedesmus. Revista Facultad de Ingeniería, Universidad de Antioquia 92:60–69.

  17. El-Naggar AH, Sheikh HM (2014) Response of the green microalga Chlorella vulgaris to the oxidative stress caused some heavy metals. Life Sci J 11:1349–1357

  18. Enan MR (2006) Application of random amplified polymorphic DNA (RAPD) to detect the genotoxic effect of heavy metals. Biotechnol Appl Biochem 43:147–154.

  19. Exss-Sonne P, Tolle J, Bader KP, MichelK P (2000) The Idil a protein of Synechocystis sp functions in protecting the acceptor side of photosystem II under oxidative stress. Photosynth Res 63:145–157.

  20. Finney DJ (1971) Probit Analysis, 3rd edn. Combrige Press, New York, 668p

  21. Franklin NM, Stauber JL, Lim RP, Petocz P (2002) Toxicity of metal mixtures to a tropical freshwater alga (chlorella sp.): the effect of interactions between copper, cadmium and zinc on metal cell binding and uptake. Environ Toxicol Chem 21:2412–2422

  22. Fulda S, Mikkat S, Schoder W, Hagemann M (1999) Isolation of salt induced periplasmic proteins from Synechocystis sp. Arch.Microbiol. 171:214–217.

  23. Gauch JR (1995) Classification. In: Gauch JR (ed) Community ecology. Cambridge University Press, Cambridge, pp 173–210

  24. Gerofke A, Kamp P, McLachlan MS (2005) Bioconcentration of persistent organic pollutants in four species of marine phytoplankton. Environ Toxcol Chem 24:2908–2917.

  25. Gianazza E, Wait R, Sozzi A, Regondi S, Saco D, Labra M, Agradi E (2007) Growth and protein profile changes in Lepidium sativum L. plantlets exposed to cadmium. Environ Exp Bot 59(2):179–187.

  26. Gjorgieva D, Panovska TK, Ruskovska T, BaIeva K, Stafilov T (2013) Influence of heavy metal stress on antioxidant status and DNA damage in Urtica dioica. Biomed Res Int 2013:1–6.

  27. Guillard RRI (1975) Culture of marine phytoplankton for feeding marine invertebrates. In: Smith WL, Chanley MH (eds) Culture of marine invertebrate animals. Plenum Press, New York, pp 26–60.

  28. Gupta M, Sarin NB (2009) Heavy metal induced DNA changes in aquatic macrophytes: random amplified polymorphic DNA analysis and identification of sequence characterized amplified region marker. J Environ Sci 21:686–690.

  29. Ide H, Petrullo LA, Hataet Z, Wallace SS (1991) Processing of DNA base damage by DNA polymerases. Dihydrothymine and beta-ureidoisobutyric acid as models for instructive and noninstructive lesions. J Biol Chem 266:1469–1477

  30. Ismail M, Phang SM, Tong SL, Brown MT (2002) A modified toxicity testing method using tropical marine microalgae. Environ Monit Assess 75:145–154.

  31. Jiang H, Gao B, Li W, Zhu M, Zheng C, Zheng Q, Wang C (2013) Physiological and biochemical responses of Ulva prolifera and U. linza to cadmium stress. Sci World J:1–11.

  32. John R, Ahmad P, Gadgil K, Sharma S (2008) Effect of cadmium and lead on growth, biochemical parameters and uptake in Lemna polyrrhiza L. Plant Soil Environ 54(6):262–270

  33. Jong LW, Thien VY, Yong YS, Rodrigues KF, Yong WTL (2015) Micropropagation and protein profile analysis by SDS-PAGE of Gracilaria changii (Rhodophyta, Solieriaceae). Aquac Rep 1:10–14.

  34. Körpe DA, Aras S (2011) Evaluation of copper-induced stress on eggplant (Solanum melongena L.) seedlings at the molecular and population levels by use of various biomarkers. Mutat Res 719(1–2):29–34.

  35. Lee MY, Shin HW (2003) Cadmium-induced changes in antioxidant enzymes from the marine alga Nannochloropsis oculata. J Appl Phycol 15:13–19.

  36. Liang Y, Wang S, Feng LX, Tian CY (2009) Effects of heavy metals stress on growth and chlorophyll fluorescence of Phaeodactylum tricornutum. Mar Environ Sci 28:374–382

  37. Manoj K, Padhy PK (2013) Oxidative stress and heavy metals: an appraisal with reference to environmental biology. Int Res J Biol Sci 2:91–101

  38. Mohy El-Din SM, Noaman NH, Zaky SH (2016) Effects of chloramphenicol, clofibric acid, acetyl salicylic acid, nonylphenol and bisphenol on the protein profile and ultrastructure of marine macroalgae Pterocladia capillacea and Ulva lactuca. Egypt J Bot 56(1):335–351.

  39. Nelson JR, Lawrence CW, Hinkle DC (1996) Thymine-thymine dimer bypass by yeast DNA polymerase zeta. Science. 272:1646–1649

  40. Padmesh P, Sabu KK, Seeni S, Pushpangadan P (1999) The use of RAPD in assessing genetic variability in Andrographis paniculate Nees, a hepatoprotective drug. Curr Sci 76:833–835

  41. Qian H, Li J, Sun L, Chen W, Sheng D, Liu W, Fu Z (2009) Combined effect of copper and cadmium on Chlorella vulgaris growth and photosynthesis-related gene transcription. Aqua.Toxico. 94:56–61.

  42. Raj A, Kumar S, Haq I, Kumar M (2014) Detection of tannery effluents induced DNA damage in mung bean by use of random amplified polymorphic DNA markers. ISRN Iotechnol 2014:1–8.

  43. Saleh B (2015) Detection of genetic variations in marine algae Ulva lactuca induced by heavy metal pollutants. J Stress Physiol Biochem 11:26–37

  44. Saleh B (2016) Genomic DNA changes in Ulva lactuca (Chlorophyta) under heavy metal stress Internat. J Environ Sci 7(3):245–255.

  45. Santana-Vieira DDS, Milori DMBP, Boas PRV, Silva MF, Santos MG, Gaiotto FA, Filho WSS, Gesteira AS (2014) Rapid differentiation of closely related Citrus genotypes by fluorescence spectroscopy. Adv Biosci Biotechnol 5:903–914.

  46. Sbihi K, Cherifi O, El Gharmali A, Oudra B, Aziz F (2012) Accumulation and toxicological effects of cadmium, copper and zinc on the growth and photosynthesis of the freshwater diatom Planothidium lanceolatum (Brébisson) Lange-Bertalot: a laboratory study. J Mater Environ Sci 3(3):497–506

  47. Sinha RP, Singh N, Kumar A, Kumar HD, Häder M, Häder D (1996) Effects of UV irradiation on certain physiological and biochemical processes in cyanobacteria. J Photoch Photobiol Biol 32:107–113.

  48. Torab-Mostaedi M, Asadollahzadeh M, Hemmati A, Khosravi A (2013) Equilibrium, kinetic, and thermodynamic studies for biosorption of cadmium and nickel on grapefruit peel. J Taiwan Inst Chem Eng 44:295–302.

  49. Torres MA, Barros MP, Campos SCG, Pinto E, Rajamani S, Sayre RT, Colepicolo P (2008) Biochemical biomarkers in algae and marine pollution: a review. Ecotoxicol Environ Safe 71:1–15.

  50. Unal D, Isik NO, Sukatar A (2010) Effects of chromium VI stress on green alga Ulva lactuca (L.). Turk J Biol 34:119–124.

  51. Wang JS, Chou H, Fan JJ, Chen CM (1998) Uptake and transfer of high PCB concentrations from phytoplankton to aquatic biota. Chemosphere. 36:1201–1210

  52. Williams J, Kubelik AR, Livak KJ (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535.

  53. Yoshida N, IKeda R, Okuno T (2006) Identification and characterization of heavy metal resistant unicellular alga isolated from soil and its potential for phytoremediation. Bioresour Technol 97:1843–1849.

  54. Yruela I (2005) Copper in plants. Braz J Plant Physiol 17:145–156.

  55. Yu Z, Zhang T, Hao R, Zhu Y (2019) Sensitivity of Chlamydomonas reinhardtii to cadmium stress is associated with phototaxis. Environ Sci Process Impacts 21:1011–1020.

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Soad M. Mohy El.Din put was involved in the design and implementation of the research, the analysis of the results, and the writing of the manuscript. Mohamed S. Abdel-Kareem critically reviewed the manuscript.

Correspondence to Soad M. Mohy El-Din.

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Mohy El-Din, S.M., Abdel-Kareem, M.S. Effects of Copper and Cadmium on the Protein Profile and DNA Pattern of Marine Microalgae Chlorella salina and Nannochloropsis salina. Environ. Process. (2020).

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  • Marine microalgae
  • Heavy metal stress
  • Lethal concentration
  • Protein profile
  • RAPD marker