Skip to main content
SpringerLink
Log in

Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays

  • Original Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC50 and 2×EC50 values were 8 and 16 mg/ml concentrations of T. turcica aqueous extracts, respectively. The negative control (distilled water), positive control (methyl methane sulfonate, 10 mg/l) and 8 and 16 mg/ml concentrations of T. turcica extracts were introduced to the roots of onion bulbs for 24 and 96 h. The root growth, DNA damage in root cells and randomly amplified polymorphic DNA (RAPD) profiles of root tissue were used as endpoints of the genotoxicity. The comet assay clearly indicated that dose-dependent single strand DNA breaks in the root nuclei of onions were determined for the treatment concentrations of T. turcica extracts. In comparison to RAPD profile of negative control group, RAPD polymorphisms became evident as disappearance and/or appearance of RAPD bands in treated roots. The diagnostic and phenetic numerical analyses of RAPD profiles obviously indicated dose-dependent genotoxicity induced by Thermopsis extracts. In conclusion, the results clearly indicated that water extract of T. turcica has genotoxic potential on the roots of onion bulbs as shown by comet assay and RAPD technique.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Akinboro A, Bakare AA (2007) Cytotoxic and genotoxic effects of aqueous extracts of five medicinal plants on Allium cepa Linn. J Ethnopharmacol 112:470–475. doi:10.1016/j.jep.2007.04.014

    Article  CAS  Google Scholar 

  • Alves dos Santos R, Cabral TR, Cabral IR, Antunes LM, Pontes Andrade C, Cerqueira dos Santos Cardoso P, de Oliveira Bahia M, Pessoa C, Martins do Nascimento JL, Rodríguez Burbano R, Takahashi CS (2008) Genotoxic effect of Physalis angulata L. (Solanaceae) extract on human lymphocytes treated in vitro. Biocell 32:195–200

  • Atienzar FA, Jha AN (2006) The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: acritical review. Mutat Res 613:76–102. doi:10.1016/j.mrrev.2006.06.001

    Article  CAS  Google Scholar 

  • 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. doi:10.1002/etc.5620181023

    Article  CAS  Google Scholar 

  • Atienzar FA, Cordi B, Donkin ME, Evenden AJ, Jha AN, Depledge MH (2000) Comparison of ultraviolet-induced genotoxicity detected by random amplified polymorphic DNA with chlorophyll fluorescence and growth in a marine macroalgae, Palmaria palmate. Aquat Toxicol 50:1–12. doi:10.1016/S0166-445X(99)00100-9

    Article  CAS  Google Scholar 

  • Bali EB, Açık L, Akca G, Sarper M, Elçi MP, Avcu F, Vural M (2014) Antimicrobial activity against periodontopathogenic bacteria, antioxidant and cytotoxic effects of various extracts from endemic Thermopsis turcica. Asian Pac J Trop Biomed 4:505–514. doi:10.12980/APJTB.4.2014APJTB-2013-0010

    Article  CAS  Google Scholar 

  • Basaran AA, Yu TW, Plewa MJ, Anderson D (1996) An investigation of some Turkish herbal medicines in Salmonella typhimurium and in the COMET assay in human lymphocytes. Teratog Carcinog Mutagen 16:125–138. doi:10.1002/(SICI)1520-6866

    Article  CAS  Google Scholar 

  • Çelik AT, Aslantürk ÖS (2010) Evaluation of cytotoxicity and genotoxicity of Inula viscosa leaf extracts with Allium test. J Biomed Res. doi:10.1155/2010/189252

  • Cenkci S, Kargıoğlu M, Dayan M, Konuk M (2007) Endangered status and propagation of an endemic plant species, Thermopsis turcica (Fabaceae). Asian J Plant Sci 6:288–293

    Article  Google Scholar 

  • Cenkci S, Kargıoğlu M, Dayan M, Konuk M (2008) In vitro propagation of an endangered plant species, Thermopsis turcica (Fabaceae). Biologia 63:652–657. doi:10.2478/s11756-008-0125-9

    Article  Google Scholar 

  • Cenkci S, Temel M, Kargıoğlu M, Dayan S (2009a) Propagation of endangered Thermopsis turcica Kit Tan, Vural and Küçüködük using conventional and in vitro techniques. Turk J Biol 33:327–333. doi:10.3906/biy-0811-1

    CAS  Google Scholar 

  • Cenkci S, Yıldız M, Ciğerci İH, Konuk M, Bozdağ A (2009b) Toxic chemicals-induced genotoxicity detected by random amplified polymorphic DNA (RAPD) in bean (Phaseolus vulgaris L.) seedlings. Chemosphere 76:900–906. doi:10.1016/j.chemosphere.2009.05.001

    Article  CAS  Google Scholar 

  • Cenkci S, Ciğerci İH, Yıldız M, Özay C, Bozdağ A, Terzi H (2010a) Lead contamination reduces chlorophyll biosynthesis and genomic template stability in Brassica rapa L. Environ Exp Bot 67:467–473. doi:10.1016/j.envexpbot.2009.10.001

    Article  CAS  Google Scholar 

  • Cenkci S, Yıldız M, Ciğerci İH, Bozdağ A, Terzi H, Terzi ESA (2010b) Evaluation of 2,4-D and dicamba genotoxicity in bean seedlings using comet and RAPD assays. Ecotoxicol Environ Saf 73:1558–1564. doi:10.1016/j.envexpbot.2009.10.001

    Article  CAS  Google Scholar 

  • Ciğerci İH, Liman R, Özgül E, Konuk M (2013) Genotoxicity of indium tin oxide by Allium and comet tests. Cytotechnology. doi:10.1007/s10616-013-9673-0

    Google Scholar 

  • Collins AR (2004) The comet assay for DNA damage and repair. Mol Biotechnol 26:249–261. doi:10.1385/MB:26:3:249

    Article  CAS  Google Scholar 

  • Davis PH, Mill RR, Kit T (1988) Flora of Turkey and the East Aegean Islands, vol 10. Edinburgh University Press, Edinburgh, pp 111–112

    Google Scholar 

  • Dement WA, Mabry TJ (1975) Biological implication of flavonoid chemistry in Baptisia and Thermopsis. Biochem Syst Ecol 3:91–94. doi:10.1016/0305-1978(75)90047-2

    Article  CAS  Google Scholar 

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

    Google Scholar 

  • Gichner T, Patková Z, Száková J, Demnerová K (2006) Toxicity and DNA damage in tobacco and potato plants growing on soil polluted with heavy metals. Ecotoxicol Environ Saf 65:420–426. doi:10.1016/j.ecoenv.2005.08.006

    Article  CAS  Google Scholar 

  • Hong CE, Lyu SY (2011) Genotoxicity detection of five medicinal plants in Nigeria. J Toxicol Sci 36:87–93. doi:10.2131/jts.36.87

    Article  Google Scholar 

  • Kalantari H, Rezaei M, Mahdavinia M, Kalantar M, Amanpour Z (2012) Determination of the mutagenicity potential of supermint herbal medicine by single cell gel electrophoresis in rat hepatocytes. Adv Pharm Bull 2:245–248. doi:10.5681/apb.2012.037

    Google Scholar 

  • Keeler RF, Baker DC (1990) Myopathy in cattle induced by alkaloid extracts from Thermopsis montana, Laburnum anagyroides and a Lupinus sp. J Comp Pathol 103:169–182

    Article  CAS  Google Scholar 

  • Keeler RF, Johnson AE, Chase RL (1986) Toxicity of Thermopsis montana in cattle. Cornell Vet 76:115–127

    CAS  Google Scholar 

  • Khallef M, Liman L, Konuk M, Ciğerci İH, Benouareth D, Tabet M, Abda A (2013) Genotoxicity of drinking water disinfection by-products (bromoform and chloroform) by using both Allium anaphase–telophase and comet tests. Cytotechnology. doi:10.1007/s10616-013-9675-y

    Google Scholar 

  • Korcan SE, Cigerci IH, Dilek M, Kargioglu M, Cenkci S, Konuk M (2009) Antimicrobial activity of an endemic species, Thermopsis turcica, Turkey. Kuwait J Sci Eng 36:101–112

    CAS  Google Scholar 

  • Lamy E, Schröder J, Paulus S, Brenk P, Stahl T, Mersch-Sundermann V (2008) Antigenotoxic properties of Eruca sativa (rocket plant), erucin and erysolin in human hepatoma (HepG2) cells towards benzo (a) pyrene and their mode of action. Food Chem Toxicol 46:2415–2421. doi:10.1016/j.fct.2008.03.022

    Article  CAS  Google Scholar 

  • Liman R, Eren Y, Akyil D, Konuk M (2012) Determination of mutagenic potencies of aqueous extracts of Thermopsis turcica by Ames test. Turk J Biol 36:85–92. doi:10.3906/biy-1011-158

    Google Scholar 

  • Lynch M, Milligan B (1994) Analysis of population genetic structure with RAPD markers. Mol Ecol 3:91–99. doi:10.1111/j.1365-294X.1994.tb00109.x

    Article  CAS  Google Scholar 

  • Macêdo MFS, Sisenando HA, Queiroz JDF, Argolo AC, Saturnino ACRD, Coelho LC, Medeiros SR (2008) Determining the genotoxicity of an aqueous infusion of Bauhinia monandra leaves. Rev Bras Farmacogn 18:509–516. doi:10.1590/S0102-695X2008000400002

    Article  Google Scholar 

  • Maistro EL, Carvalho JCT, Mantovani MS (2004) Evaluation of the genotoxic potential of the Casearia sylvestris extract on HTC and V79 cells by the comet assay. Toxicol In Vitro 18:337–342. doi:10.1016/j.tiv.2003.10.002

    Article  CAS  Google Scholar 

  • McGrath-Hill CA, Vicas IM (1997) Case series of Thermopsis exposures. J Toxicol-Clin Toxicol 3:659–665

    Article  Google Scholar 

  • Medicinal Plants in Mongolia (2013) World Health Organization Regional Office for the Western Pacific. pp 210–211

  • Nitzsche D, Melzig MF, Arlt VM (2013) Evaluation of the cytotoxicity and genotoxicity of aristolochic acid I—a component of Aristolochiaceae plant extracts used in homeopathy. Environ Toxicol Pharmacol 35:325–334. doi:10.1016/j.etap.2013.01.007

    Article  CAS  Google Scholar 

  • Ohmiya S, Otomasu H, Murakoshi I, Haginiwa J (1974) N-Formylcytisine: a new alkoloid from Thermopsis chinensis. Phytochemistry 13:643–644. doi:10.1016/S0031-9422(00)91367-0

    Article  CAS  Google Scholar 

  • Page RDM (1996). TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358. doi:10.1093/bioinformatics/12.4.357

  • Pavlicek A, Hrda Š, Flegr J (1999) FreeTree—freeware program for construction of phylogenetic trees on the basis of distance data and for bootstrap/jackknife analysis of the trees robustness. Application in the RAPD analysis of genus Frenkelia. Folia Biol (Praha) 45:97–99

    CAS  Google Scholar 

  • Portmann E, Nigro MML, Reides CG, Llesuy S, Ricco RA, Wagner ML, Gurni AA, Carballo MA (2012) Aqueous extracts of Lippia turbinata and Aloysia citriodora (Verbenaceae): assessment of antioxidant capacity and DNA damage. Int J Toxicol 31:192–202. doi:10.1177/1091581812436726

  • Rank J (2003) The method of Allium anaphase–telophase chromosome aberration assay. Ekologija 1:38–42

    Google Scholar 

  • Rathnasamy S, Mohamed KB, Sulaiman SF, Akinboro A (2013) Evaluation of cytotoxic, mutagenic and antimutagenic potential of leaf extracts of three medicinal plants using Allium cepa chromosome assay. Int Curr Pharm J 2:131–140. doi:10.3329/icpj.v2i8.15588

    Article  Google Scholar 

  • Rucińska R, Waplak S, Gwóźdź EA (1999) Free radical formation and activity of antioxidant enzymes in lupin roots exposed to lead. Plant Physiol Biochem 37:187–194. doi:10.1016/S0981-9428(99)80033-3

    Article  Google Scholar 

  • Saito K, Yamazaki M, Takamatsu S, Kawaguchi A, Murakoshi I (1989) Greening induced production of (+)-lupanine in tissue culture of Thermopsis lupinoides. Phytochemistry 28:2341–2344. doi:10.1016/S0031-9422(00)97980-9

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  Google Scholar 

  • Santos FV, Colus IMS, Silva MA, Vilegas W, Varanda EA (2006) Assessment of DNA damage by extracts and fractions of Strychnos pseudoquina, a Brazilian medicinal plant with antiulcerogenic activity. Food Chem Toxicol 44:1585–1589. doi:10.1016/j.fct.2006.03.012

    Article  CAS  Google Scholar 

  • Şekeroğlu ZA, Şekeroğlu V (2012) Effects of Viscum album L. extract and quercetin on methotrexate-induced cyto-genotoxicity in mouse bone-marrow cells. Mutat Res Genet Toxicol Environ Mutagen 746:56–59. doi:10.1016/j.mrgentox.2012.02.012

    Article  Google Scholar 

  • Şener B, Koyuncu M, Ergun F (1992) Determining anagyrine in Thermpsis turcica plants by HPLC. In: Başer KHC (ed) Proceedings of the 9th symposium on plant drugs Eskişehir (Turkey), 16–19 May, 1992, p 411

  • Singh KN (2012) Traditional knowledge on ethnobotanical uses of plant biodiversity: a detailed study from the Indian western Himalaya. Biodivers Res Conserv 28:63–77. doi:10.2478/v10119-012-0028-z

    Article  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191. doi:10.1016/0014-4827(88)90265-0

    Article  CAS  Google Scholar 

  • Skandrani I, Boubaker J, Bouhlel I, Limem I, Ghedira K, Chekir-Ghedira L (2010) Leaf and root extracts of Moricandia arvensis protect against DNA damage in human lymphoblast cell K562 and enhance antioxidant activity. Environ Toxicol Pharmacol 30:61–67. doi:10.1016/j.etap.2010.03.014

    Article  CAS  Google Scholar 

  • Tan K, Vural M, Küçüködük M (1983) An unusual new Thermopsis from Turkey. Notes RBG Edinb 40:515–518

    Google Scholar 

  • Tilley D (2012) Plant guide for mountain goldenbanner (Thermopsis montana). USDA-Natural Resources Conservation Service, Aberdeen Plant Materials Center, Aberdeen, Idaho 83210

  • Tucker SC (2003) Floral development in legumes. Plant Physiol 131:911–926. doi:10.1104/pp.102.017459

    Article  CAS  Google Scholar 

  • Yıldız M, Ciğerci İH, Konuk M, Fatih Fidan A, Terzi H (2009) Determination of genotoxic effects of copper sulphate and cobalt chloride in Allium cepa root cells by chromosome aberration and comet assays. Chemosphere 75:934–938. doi:10.1016/j.chemosphere.2009.01.023

    Article  Google Scholar 

Download references

Acknowledgments

The authors wish to thank Afyon Kocatepe University Research Fund (Project No: 11.FENED.05) for financial support.

Author information

Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Faculty of Science and Arts, Afyon Kocatepe University, 03200, Afyonkarahisar, Turkey

    İbrahim Hakkı Ciğerci, Süleyman Cenkci & Mustafa Kargıoğlu

  2. Molecular Biology and Genetics Department, Faculty of Engineering and Natural Sciences, Üsküdar University, Altunizade, 34662, Istanbul, Turkey

    Muhsin Konuk

Authors
  1. İbrahim Hakkı Ciğerci
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Süleyman Cenkci
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mustafa Kargıoğlu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Muhsin Konuk
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to İbrahim Hakkı Ciğerci.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ciğerci, İ.H., Cenkci, S., Kargıoğlu, M. et al. Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays. Cytotechnology 68, 829–838 (2016). https://doi.org/10.1007/s10616-014-9835-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-014-9835-8

Keywords

Search

Navigation