Environmental Science and Pollution Research

, Volume 25, Issue 36, pp 36117–36123 | Cite as

Cytogenetic and genotoxic effects of 2-chlorophenol on Allium cepa L. root meristem cells

  • Derya Küçük
  • Recep LimanEmail author
Research Article


2-Chlorophenol (2-CP), a class of chlorinated organic pollutants like other chlorophenols, is used as intermediate in the synthesis of the higher chlorinated congeners, certain dyes, preservatives, herbicides, fungicides, and plastics. In this study, cytotoxic and genotoxic effects of 2-CP were investigated on the root meristem cells of Allium cepa for its effects on root growth, mitotic index (MI), mitotic phases, chromosomal abnormalities (CAs), and DNA damage by using Allium anaphase-telophase and Comet assays. EC50 of 2-CP value was determined as approximately 25 mg/L by Allium root growth inhibition test. Three concentrations of 2-CP (12.5, 25, and 50 mg/L), distilled water (negative control), and methyl methane sulfonate (MMS, 10 mg/L, positive control) were applied to onion stem cells under different exposure periods (24, 48, 72, and 96 h). All the applied doses of 2-CP slightly decreased MIs. 2-CP induced total CAs such as disturbed anaphase-telophase, chromosome laggards, stickiness, and bridges and also DNA damage at significant levels. These results demonstrate that 2-CP has genotoxic effects in A. cepa root meristematic cells.


2-Chlorophenol Allium cepa Genotoxicity Toxicity Comet assay Chromosome aberration 


Funding information.

This work was financially supported by Usak University BAP (Project no. 2017/TP027).


  1. Agency for Toxic Substances and Disease Registry (ATSDR) (1999) Public Health Statement for Chlorophenols, U.S. Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry, Atlanta, GA. Assessed 20 April 2018
  2. Agency for Toxic Substances and Disease Registry (ATSDR) (2017) Priority List of Hazardous Substances, The ATSDR (2017) Substance Priority List, U.S. Department of Health and Human Services, Public Health Service, Atlanta, GA. Assessed 20 April 2018
  3. Aruoja V, Sihtmäe M, Dubourguier HC, Kahru A (2011) Toxicity of 58 substituted anilines and phenols to algae Pseudokirchneriella subcapitata and bacteria Vibrio fischeri: comparison with published data and QSARs. Chemosphere 84(10):1310–1320CrossRefGoogle Scholar
  4. Borzelleca JF, Condie LW, Hayes JR (1984). Toxicological evaluation of selected chlorinated phenols. In Proceedings of the fifth conference on water chlorination environ impact and health eff. Williamsburg, VA, pp 331–343Google Scholar
  5. Carita R, Marin-Morales M (2008) Induction of chromosome aberrations in the Allium cepa test system caused by the exposure of seeds to industrial effluents contaminated with azo dyes. Chemosphere 72:722–725CrossRefGoogle Scholar
  6. Čerňáková M (1994) Effect of chlorinated phenol derivatives on various cell models. Folia Microbiol 39(4):315–320CrossRefGoogle Scholar
  7. Chaparro TR, Botta CM, Pires EC (2010) Biodegradability and toxicity assessment of bleach plant effluents treated anaerobically. Water Sci Technol 62(6):1312–1319CrossRefGoogle Scholar
  8. Choi SH, Gu MB (2001) Phenolic toxicity—detection and classification through the use of a recombinant bioluminescent Escherichia coli. Environ Toxicol Chem 20(2):248–255Google Scholar
  9. Czaplicka M (2004) Sources and transformations of chlorophenols in the natural environment. Sci Total Environ 322(1–3):21–39CrossRefGoogle Scholar
  10. De Araujo BS, Dec J, Bollag JM, Pletsch M (2006) Uptake and transformation of phenol and chlorophenols by hairy root cultures of Daucus carota, Ipomoea batatas and Solanum aviculare. Chemosphere 63(4):642–651CrossRefGoogle Scholar
  11. Dhawan A, Anderson D (Eds.) (2016) The comet assay in toxicology. (Vol. 30) Roy Soc ChemGoogle Scholar
  12. El-Ghamery AA, Mousa MA (2017) Investigation on the effect of benzyladenine on the germination, radicle growth and meristematic cells of Nigella sativa L. and Allium cepa L. Ann Agric Sci 62(1):11–21Google Scholar
  13. El-Ghamery AA, El-Nahas AI, Mansour MM (2000) The action of atrazine herbicide as an indicator of cell division on chromosomes and nucleic acid content in root meristems of Allium cepa and Vicia faba. Cytologia 65:277–287CrossRefGoogle Scholar
  14. Ertürk MD, Saçan MT (2012) First toxicity data of chlorophenols on marine alga Dunaliella tertiolecta: correlation of marine algal toxicity with hydrophobicity and interspecies toxicity relationships. Environ Toxicol Chem 31(5):1113–1120CrossRefGoogle Scholar
  15. European Chemical Bureau (ECB) IUCLID Dataset, 2-Chlorophenol (2000) Available on line at: Accessed 16 March 2011
  16. Evseeva TI, Geras’kin SA, Shuktomova II, Taskaev AI (2005) Genotoxicity and cytotoxicity assay of water sampled from the underground nuclear explosion site in the north of the perm region (Russia). J Environ Radioact 80:59–74CrossRefGoogle Scholar
  17. Fernandes TCC, Mazzeo DEC, Marin-Morales MA (2007) Mechanism of micronuclei formation in polyploidizated cells of Allium cepa exposed to trifluralin herbicide. Pestic Biochem Physiol 88:252–259CrossRefGoogle Scholar
  18. Fiskesjö G (1985) The Allium test as a standard in environmental monitoring. Hereditas 102(1):99–112CrossRefGoogle Scholar
  19. Fiskesjö G, Levan A (1993) Evaluation of the first ten MEIC chemicals in the Allium test. ATLA 21:139–149Google Scholar
  20. Fusconi A, Repetto O, Bona E, Massa N, Gallo C, Dumas-Gaudot E, Berta G (2006) Effect of cadmium on meristem activity and nucleus ploidy in roots of Pisum sativum L. cv. Frisson seedlings. Environ Exp Bot 58:253–260CrossRefGoogle Scholar
  21. Gichner T, Znidar I, Wagner ED, Plewa MJ (2009) The use of higher plants in the Comet assay. Dhawan A, Anderson D (Eds.) The Comet Assay in Toxicology. Roy Soc Chem UK, pp 98–119Google Scholar
  22. Grant WF (1999) Higher plant assays for the detection of chromosomal aberrations and gene mutations—a brief historical background on their use for screening and monitoring environmental chemicals. Mutat Res 426(2):107–112CrossRefGoogle Scholar
  23. Hidalgo A, Gonzales-Reyes JA, Navas P, Garcia-Herdugo G (1989) Abnormal mitosis and growth inhibition in Allium cepa roots induced by propham and chlorpropham. Cytobios 57:7–14Google Scholar
  24. Jana A, Ghosh M, Sinha S, Jothiramajayam M, Nag A, Mukherjee A (2017) Hazard identification of coal fly ash leachate using a battery of cyto-genotoxic and biochemical tests in Allium cepa. Arch Agron Soil Sci 63(10):1443–1453CrossRefGoogle Scholar
  25. Jennings VL, Rayner-Brandes MH, Bird DJ (2001) Assessing chemical toxicity with the bioluminescent photobacterium (Vibrio fischeri): a comparison of three commercial systems. Water Res 35(14):3448–3456CrossRefGoogle Scholar
  26. Kaya N, Çakmak I, Akarsu E, Kaya B (2015) DNA damage induced by silica nanopartıcle. Fresenius Envıron Bull 24(12A):4478–4484Google Scholar
  27. Kaygisiz ŞY, Ciğerci İH (2017) Genotoxic evaluation of different sizes of iron oxide nanoparticles and ionic form by SMART, Allium and comet assay. Toxicol Ind Health 33(10):802–809CrossRefGoogle Scholar
  28. Koçyiğit A, Keles H, Selek S, Guzel S, Celik H, Erel O (2005) Increased DNA damage and oxidative stress in patients with cutaneous leishmaniasis. Mutat Res 585(1):71–78Google Scholar
  29. Kumari M, Mukherjee A, Chandrasekaran N (2009) Genotoxicity of silver nanoparticles in Allium cepa. Sci Total Environ 407(19):5243–5246CrossRefGoogle Scholar
  30. Liman R (2013) Genotoxic effects of bismuth (III) oxide nanoparticles by Allium and comet assay. Chemosphere 93(2):269–273CrossRefGoogle Scholar
  31. Liman R, Ciğerci İH, Öztürk NS (2015) Determination of genotoxic effects of Imazethapyr herbicide in Allium cepa root cells by mitotic activity, chromosome aberration, and comet assay. Pestic Biochem Physiol 118:38–42CrossRefGoogle Scholar
  32. Luo LZ, Werner KM, Gollin SM, Saunders WS (2004) Cigarette smoke induces anaphase bridges and genomic imbalances in normal cells. Mutat Res 554(1):375–385CrossRefGoogle Scholar
  33. Luo Y, Su Y, Lin RZ, Shi HH, Wang XR (2006) 2-Chlorophenol induced ROS generation in fish Carassius auratus based on the EPR method. Chemosphere 65(6):1064–1073CrossRefGoogle Scholar
  34. Luo Y, Sui YX, Wang XR, Tian Y (2008) 2-Chlorophenol induced hydroxyl radical production in mitochondria in Carassius auratus and oxidative stress–an electron paramagnetic resonance study. Chemosphere 71(7):1260–1268CrossRefGoogle Scholar
  35. Mangalampalli B, Dumala N, Grover P (2018) Allium cepa root tip assay in assessment of toxicity of magnesium oxide nanoparticles and microparticles. J Environ Sci 66:125–137CrossRefGoogle Scholar
  36. Mauro MO, Pesarini JR, Marin-Morales MA, Monreal MTFD, Monreal ACD, Mantovani MS, Oliveira RJ (2014) Evaluation of the antimutagenic activity and mode of action of the fructooligosaccharide inulin in the meristematic cells of Allium cepa culture. Genet Mol Res 13(3):4808–4819CrossRefGoogle Scholar
  37. Michałowicz J, Duda W (2007) Phenols--sources and toxicity. Pol J Environ Stud 16(3):347–362Google Scholar
  38. Ministry of Health Labor and Welfare of Japan (MHLW) (2001) 2,4-Dinitrophenol. Toxicity testing reports of environmental chemicals 8(1):7–36Google Scholar
  39. Moridani MY, Siraki A, O'Brien PJ (2003) Quantitative structure toxicity relationships for phenols in isolated rat hepatocytes. Chem Biol Interact 145(2):213–223CrossRefGoogle Scholar
  40. Morita T, Honma M, Morikawa K (2012) Effect of reducing the top concentration used in the in vitro chromosomal aberration test in CHL cells on the evaluation of industrial chemical genotoxicity. Mutat Res 741(1):32–56CrossRefGoogle Scholar
  41. Önfelt A (1987) Spindle disturbances in mammalian cells III. Toxicity, c-mitosis and aneuploidy with 22 different compounds. Specific and unspecific mechanisms. Mutat Res 182(3):135–154CrossRefGoogle Scholar
  42. Palmieri MJ, Andrade-Vieira LF, Trento MVC, de Faria Eleutério MW, Luber J, Davide LC, Marcussi S (2016) Cytogenotoxic effects of spent pot liner (SPL) and its main components on human leukocytes and meristematic cells of Allium cepa. Water Air Soil Pollut 227(5):156–166CrossRefGoogle Scholar
  43. Parida KM, Parija S (2006) Photocatalytic degradation of phenol under solar radiation using microwave irradiated zinc oxide. Sol Energy 80(8):1048–1054CrossRefGoogle Scholar
  44. Patil BC, Bhat GI (1992) A comparative study of MH and EMS in the induction of chromosomal aberrations on lateral root meristem in Clitoria termata L. Cytologia 57:259–264CrossRefGoogle Scholar
  45. Perez-Moya M, Graells M, del Valle LJ, Centelles E, Mansilla HD (2007) Fenton and photo-Fenton degradation of 2-chlorophenol: multivariate analysis and toxicity monitoring. Catal Today 124(3–4):163–171CrossRefGoogle Scholar
  46. Rahman MM, Rahman MF, Nasirujjaman K (2017) A study on genotoxicity of textile dyeing industry effluents from Rajshahi, Bangladesh, by the Allium cepa test. Chem Ecol 33(5):434–446CrossRefGoogle Scholar
  47. Rajeshwari A, Roy B, Chandrasekaran N, Mukherjee A (2016) Cytogenetic evaluation of gold nanorods using Allium cepa test. Plant Physiol Biochem 109:209–219CrossRefGoogle Scholar
  48. Santos CL, Pourrut B, Oliveira JMP (2015) The use of comet assay in plant toxicology: recent advances. Front Genet 6:216CrossRefGoogle Scholar
  49. Saxena PN, Chauhan LKS, Gupta SK (2005) Cytogenetic effects of commercial formulation of cypermethrin in root meristem cells of Allium sativum: spectroscopic basis of chromosome damage. Toxicology 216(2–3):244–252CrossRefGoogle Scholar
  50. Sharma S, Vig AP (2012) Genotoxicity of atrazine, avenoxan, diuron and quizalofop-P-ethyl herbicides using the Allium cepa root chromosomal aberration assay. Terr Aquat Environ Toxicol 6(2):90–95Google Scholar
  51. Shehata M, Durner J, Thiessen D, Shirin M, Lottner S, Van Landuyt K, Reichl FX (2012) Induction of DNA double-strand breaks by monochlorophenol isomers and ChKM in human gingival fibroblasts. Arch Toxicol 86(9):1423–1429CrossRefGoogle Scholar
  52. Silveira GL, Lima MGF, dos Reis GB, Palmieri MJ, Andrade-Vieria LF (2017) Toxic effects of environmental pollutants: comparative investigation using Allium cepa L. and Lactuca sativa L. Chemosphere 178:359–367CrossRefGoogle Scholar
  53. Singh D, Roy BK (2017) Evaluation of malathion-induced cytogenetical effects and oxidative stress in plants using Allium test. Acta Physiol Plant 39(4):92–102CrossRefGoogle Scholar
  54. Soliman MI, Ghoneam GT (2004) The mutagenic potentialities of some herbicides using Vicia faba as a biological system. Biotechnology 3(2):140–154CrossRefGoogle Scholar
  55. Sudhakar R, Ninge Gowda KN, Venu G (2001) Mitotic abnormalities induced by silk dyeing industry effluents in the cells of Allium cepa. Cytologia 66(3):235–239CrossRefGoogle Scholar
  56. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35(3):206–221CrossRefGoogle Scholar
  57. Türkoğlu Ş (2015) Evaluation of genotoxic effects of five flavour enhancers (glutamates) on the root meristem cells of Allium cepa. Toxicol Ind Health 31(9):792–801CrossRefGoogle Scholar
  58. USEPA National Recommended Water Quality Criteria, United States Environment Protection Agency (2002) EPA Report EPA-822-R-02-047Google Scholar
  59. Vallejo M, Fernández-Castro P, San Román MF, Ortiz I (2015) Assessment of PCDD/fs formation in the Fenton oxidation of 2-chlorophenol: influence of the iron dose applied. Chemosphere 137:135–141CrossRefGoogle Scholar
  60. Ventura L, Giovannini A, Savio M, Donà M, Macovei A, Buttafava A, Carbonera D, Balestrazzi A (2013) Single cell gel electrophoresis (comet) assay with plants: research on DNA repair and ecogenotoxicity testing. Chemosphere 92:1–9CrossRefGoogle Scholar
  61. Vlastos D, Antonopoulou M, Konstantinou I (2016) Evaluation of toxicity and genotoxicity of 2-chlorophenol on bacteria, fish and human cells. Sci Total Environ 551:649–655CrossRefGoogle Scholar
  62. Wang W (1987) Root elongation method for toxicity testing of organic and inorganic pollutants. Environ Toxicol Chem 6(5):409–414CrossRefGoogle Scholar
  63. Webster PL, Macleod RD (1996) The root apical meristem and its magrin. In: Waishel Y, Eshel A, Kafkafi U (eds) Plant roots. The hidden half, 2nd edn. Marcel Dekker, New York, pp 51–76Google Scholar
  64. WHO (2003) Chlorophenols in drinking-water. Background document for preparation of WHO guidelines for drinking-water quality World Health Organization, Geneva (WHO/SDE/WSH/03.04/47)Google Scholar
  65. Yang Y, Liu Z, Zheng M, Fang Z, Wang L, Sun W, Ren J (2009) The joint toxicity in juvenile Carassius auratus exposed to 2-chlorophenol and 2,4-dichlorophenol. Fresenius Environ Bull 18(1):21–25Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Faculty of Arts and Sciences, Molecular Biology and Genetics DepartmentUşak UniversityUşakTurkey

Personalised recommendations