Skip to main content

Molecular and Spectroscopic Evaluation of the Effects of Coumarin on Lentil


This study examines the genotoxic effects of coumarin (2H-1-benzopyran-2-one) on Lens culinaris Medik cv Sultan in terms of DNA polymorphism and protein quantification. Effective concentration values were calculated according to a probit model, which is a type of regression where the dependent variable can take only two values, i.e., life or death, after 48 or 72 h of treatment. Based on this analysis, the effective concentration value of EC50 identified as approximately 278 μM, and then adjusted to 300 μM. The bulb roots were treated with 300 μM (EC50), 600 μM (2X EC50) concentrations and the Hoagland was used in the control group.10 RAPD primers were used and as a result of the analysis, it was observed that 2 monomorphic and 8 polymorphic band profile primers for treatment of 600 and 300 μM, coumarin concentration according to the control group. Total of 39 polymorphic bands were detected from total of 97 bands and the percentage of polymorphism was detected as 48.75%. SDS-PAGE analysis for total protein profile showed that there were differences between the treatment groups. In addition to PCR analysis, the root samples were examined by Fourier transform infrared (FT-IR) spectroscopy in order to determine the effects of coumarin on the quantity of biomolecules structure of L. culinaris One-way analysis of variance (ANOVA) was used in the calculation of statistical differences between the groups (P < 0.05).

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

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


  1. Ana, A.S., Penelope, G.A., Hugo, M., et al., The use of FTIR spectroscopy to monitor modifications in plant cell wall architecture caused by cellulose biosynthesis inhibitors, Plant Signal. Bah., 2011, vol. 6, pp. 1104–1110.

    CAS  Article  Google Scholar 

  2. Atienza, F.A., Conradie, M., Evenden, A.J., et al., 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, Env. Tox. Chem., 1999, vol. 18, pp. 2275–2282.

    Article  Google Scholar 

  3. Atienzar, F.A., Venier, P., Jha, A.N., et al., Evaluation of the random amplified polymorphic DNA (RAPD) assay for the detection of DNA damage and mutations, Mut. Res./Gen Tox. Env. Mut., 2002, vol. 521, pp. 151–163.

    CAS  Article  Google Scholar 

  4. Atienzar, F.A. and Jha, A.N., The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: a critical review, Mut. Res. Rev., 2006, vol. 613, pp. 76–102.

    CAS  Article  Google Scholar 

  5. Barun, S.G., Bjorn, P.J., Tao, G., et al., Application of ATR-FTIR spectroscopy to compare the cell materials of wood decay fungi with wood mould fungi, Int. J. Spectrosc., 1999, vol. 2015, pp. 1–8.

    CAS  Article  Google Scholar 

  6. Borgaud, F., Hehn, A., Larbat, R., et al., Biosynthesis of coumarins in plants: a major pathway still to be unraveled for cytochrome P450 enzymes, Phytochem. Rev., 2006, vol. 5, pp. 293–308.

    CAS  Article  Google Scholar 

  7. Damjanovich, S., Gáspár, R., and Panyi, G., An alternative to conventional immuno suppression: small-molecule inhibitors of Kv1. 3 channels, Mol. Int., 2004, vol. 4, pp. 250–254.

    CAS  Article  Google Scholar 

  8. De Wolf, H., Blust, R., and Backeljau, T., The use of RAPD in ecotoxicology, Mut. Res./Rev. Mut. Res., 2004, vol. 566, pp. 249–262.

    CAS  Article  Google Scholar 

  9. Gnonlonfin, G.B., Sanni, A., and Brimer, L., Review scopoletin—a coumarin phytoalexin with medicinal properties, Crit. Rev. Plant Sci., 2012, vol. 31, pp. 47–56.

    CAS  Article  Google Scholar 

  10. Kawase, M., Sakagam, H., Motohash, N., et al., Coumarin derivatives with tumor-specific cytotoxicity and multi drug resistance reversal activity in vivo, IV. IIAR J., 2005, vol. 19, pp. 705–711.

    CAS  Google Scholar 

  11. Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 1970, vol. 227, pp. 680–685.

    CAS  Article  PubMed  Google Scholar 

  12. Liu, W., Yang, Y.S., Zhou, Q., et al., Impact assessment of cadmium contamination on rice (Oryza sativa L.) seedlings at molecular and population levels using multiple biomarkers, Chemosphere, 2007, vol. 67, pp. 1155–1163.

    CAS  Article  PubMed  Google Scholar 

  13. Malaiyandi, V., Sellers, E.M., and Tyndale, R.F., Implications of CYP2A6 genetic variation for smoking behaviors and nicotine dependence, Clin. Pharm. Ther., 2005, vol. 77, pp. 145–158.

    CAS  Article  Google Scholar 

  14. Nei, M., Genetic distance between populations, Am. Nat., 1972, vol. 106, pp. 283–292.

    Article  Google Scholar 

  15. Ozek, N., Bal, I., Sara, Y., et al., Structural and functional characterization of simvastatin-induced myotoxicity in different skeletal muscles, Biochim. Biophys. Acta, 2014, vol. 1840, pp. 406–415.

    CAS  Article  Google Scholar 

  16. Plumas, J., Drillat, P., Jacob, M., et al., Extra corporeal photochemotherapy for treatment of clonal T cell proliferations, Bul. Ducan., 2003, vol. 90, pp. 763–770.

    Google Scholar 

  17. Shinde, R.G., Khan, A.A., and Barik, A., Coumarin derivatives with antioxidant and anticancer potential: a review, Int. J. Med. Appl. Sci., 2014, vol. 3, pp. 165–184.

  18. Turker, S., Dogan, F.T., and Severcan, F., The characterization and differentiation of higher plants by Fourier transform infrared spectroscopy, Appl. Spectrosc., 2007, vol. 61, pp. 300–308.

    Article  Google Scholar 

  19. Venugopala, K.N., Rashmi, V., and Odhav, B., Review on natural coumarin lead compounds for their pharmacological activity, BioMed. Res. Int., 2013.

  20. Williams, J.G., Kubelik, A.R., Livak, K.J., et al., DNA polymorphisms amplified by arbitrary primers are useful as genetic markers, Nucleic Acids Res., 1990, vol. 18, pp. 6531–6535.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  21. Wulff, H., Rauer, H., Düring, T., et al., Alkoxypsoralens, novelnon peptide blockers of Shaker-type K+ channels: synthesis and photo reactivity, J. Med. Chem., 1998, vol. 41, pp. 4542–4549.

    CAS  Article  PubMed  Google Scholar 

  22. Xu, R.X., Gao, S., Zhao, Y., Lou, H.X., and Cheng, A.X., Functional characterization of a Mg2+-dependent O‑methyltransferase with coumarin as preferred substrate from the liverwort Plagiochasma appendiculatum, Plant Physiol. Biochem., 2016, vol. 106, pp. 269–277.

  23. Yuksel, B. and Aksoy, O., Cytological effects of coumann on the mitosis of Lens culinaris Medik, Fresemus Env. Bull., 2017, vol. 26, pp. 6400–6407.

    CAS  Google Scholar 

  24. Yuksel, B. and Aksoy, O., Analysis of the effects of coumarin on Lens culinaris Medik by some biochemical parameters using real-time polymerase chain reaction, Legume Res., 2019, vol. 42, pp. 640–645.

    Article  Google Scholar 

Download references


We would like to express our gratitude to Kocaeli University [BAP-2016-02] for their financial support during this study.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Burcu Yuksel.

Ethics declarations

The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

About this article

Verify currency and authenticity via CrossMark

Cite this article

Yuksel, B., Aksoy, O. & Kutluk, T. Molecular and Spectroscopic Evaluation of the Effects of Coumarin on Lentil. Cytol. Genet. 55, 274–282 (2021).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • Lentil
  • Coumarin
  • FT-IR