Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells

Burcu Yüksel; Duygu Aydın; Özlem Aksoy; Dilek Sezer

Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells

Abstract

Background: Kathon CG is a widely used isothiazolinone-based biocide, and its release into agricultural soils and surface waters raises concerns regarding its potential cytogenetic effects on non-target plant species.

Aims: This study evaluated the cytogenotoxic effects of Kathon CG and the potential protective role of Liquid Vermicompost (LVC) in root meristem cells of Pisum sativum var. arvense L.

Methods: Seeds were exposed for 48 h to Kathon CG at the half-maximal effective concentration (EC50): EC50/2 (1.5 ml L−1), EC50 (3 ml L−1), and 2 × EC50 (6 ml L−1), with or without 10% (v/v) LVC. Cytogenetic endpoints, including mitotic index (MI) and chromosomal and nuclear abnormalities, were scored in at least 5,000 cells per group. Concurrently, DNA strand breaks were quantified in isolated nuclei using the alkaline Comet assay.

Results: Kathon CG produced a severe, dose-dependent suppression of mitotic activity, with MI decreasing from 12.89% in the negative control to 1.20% at 6 ml L−1. This was accompanied by increased frequencies of chromosomal aberrations and lethal nuclear abnormalities (pyknotic nuclei and ghost cells), together with elevated DNA damage parameters, including % Tail DNA and Tail Moment. Co-application of 10% LVC partially restored the MI (to 7.68% at 6 ml L−1), markedly reduced severe nuclear lesions (pyknotic nuclei and ghost cells became almost absent, and the frequency of abnormal mitoses decreased by approximately 30–40%), and significantly reduced Comet assay parameters, particularly at the 1.5 ml L−1 dose. A mild increase in Comet assay parameters was also detected in the LVC-only control. as reactive oxygen species levels and antioxidant markers were not measured, this finding is tentatively interpreted as a low-amplitude, repairable response that may be consistent with the redox-priming activity reported for bioactive humic constituents.

Conclusion: This study provides cytogenetic and single-cell electrophoretic evidence supporting a genoprotective role of LVC against Kathon CG-induced damage. These findings demonstrate associations rather than experimentally validated mechanisms, and extend the evidence for LVC–mediated protection beyond simple recovery of mitotic activity, supporting the further evaluation of vermicompost-based biostimulants under biocide-induced stress.

Keywords

Supporting Institution

Kocaeli University Scientific Research Projects Coordination Unit

Project Number

2019/019

Ethical Statement

Not required.

Thanks

The authors thank HK Tohumculuk (Kütahya, Türkiye) for supplying the certified Pisum sativum seeds and Riverm Ltd. Şti. (Tekirdağ, Türkiye) for providing the liquid vermicompost used in this study. The authors also thank Dr. Orhan Aksoy for developing the ‘Etiketle’ cell-counting software.

References

Afanasieva, K., & Sivolob, A. (2018). Physical principles and new applications of the comet assay. Biophysical Chemistry, 238, 1–7. https://doi.org/10.1016/j.bpc.2018.04.003
Aksoy, O., Aydın, D., & Yüksel, B. (2022). The healing effect of liquid vermicompost against Kathon CG application in Pisum sativum spp. arvense. Acta Physiologiae Plantarum, 44, 5. https://doi.org/10.1007/s11738-021-03340-z
Alias, C., Zerbini, I., Zani, C., & Feretti, D. (2024). The Allium cepa comet assay for environmental sample assessment: a scoping review. Mutagenesis, 39(4–5), 219–237. https://doi.org/10.1093/mutage/geae020
Aydın, D., & Yüksel, B. (2022). Lessening the toxic effect of methylisothiazolinone via vermicompost tea on Pisum sativum. Environmental Science and Pollution Research, 29, 50443–50453. https://doi.org/10.1007/s11356-022-19396-4
Azqueta, A., & Collins, A. R. (2013). The essential comet assay: A comprehensive guide to measuring DNA damage and repair. Archives of Toxicology, 87, 949–968. https://doi.org/10.1007/s00204-013-1070-0
Bhat, S. A., Singh, S., Singh, J., Kumar, S., & Vig, A. P. (2018). Bioremediation and detoxification of industrial wastes by earthworms: Vermicompost as a powerful crop nutrient in sustainable agriculture. Bioresource Technology, 252, 172–179. https://doi.org/10.1016/j.biortech.2018.01.003
Blouin, M., Barrere, J., Meyer, N., Lartigue, S., Barot, S., & Mathieu, J. (2019). Vermicompost significantly affects plant growth: A meta-analysis. Agronomy for Sustainable Development, 39, 34. https://doi.org/10.1007/s13593-019-0579-x
De Oliveira Sátiro, J. N., García, A. C., de Souza, A. F. F., de Oliveira, C. C. L., Nogueira, A. L. S. P., dos Santos, R. V., … Santos, L. A. (2026). Humic acid induces an adaptive stress response during early signaling in rice. Physiologia Plantarum, 178(2), e70877. https://doi.org/10.1111/ppl.70877

De Souza, A. F. F., García, A. C., de Oliveira Sátiro, J. N., de Lima, B. R., Fernandes, M. S., Berbara, R. L. L., & Santos, L. A. (2025). Humic acid regulates root growth through ROS-dependent pathway and hormone signalling in rice. Journal of Agricultural and Food Chemistry, 73(31), 18243–18256. https://doi.org/10.1021/acs.jafc.5c06288
Do, V. Q., Seo, Y. S., Park, J. M., Yu, J., Men, T. H. D., Nakai, J., Kim, S. K., Ahn, H. C., & Lee, M. Y. (2021). A mixture of chloromethylisothiazolinone and methylisothiazolinone impairs rat vascular smooth muscle by depleting thiols and elevating cytosolic Zn²⁺ and reactive oxygen species. Archives of Toxicology, 95, 541–556. https://doi.org/10.1007/s00204-020-02927-8
Domínguez, J., Aira, M., Crandall, K. A., & Pérez-Losada, M. (2021). Earthworms drastically change fungal and bacterial communities during vermicomposting of sewage sludge. Scientific Reports, 11, 15457. https://doi.org/10.1038/s41598-021-95099-z
Ferguson, L. R., Chen, H., Collins, A. R., Connell, M., Damia, G., Dasgupta, S., et al. (2015). Genomic instability in human cancer: Molecular insights and opportunities for therapeutic attack and prevention through diet and nutrition. Seminars in Cancer Biology, 35, S5–S24. https://doi.org/10.1016/j.semcancer.2015.03.005
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. Environmental and Experimental Botany, 58, 253–260. https://doi.org/10.1016/j.envexpbot.2005.09.008
García, A. C., Santos, L. A., Izquierdo, F. G., Sperandio, M. V. L., Castro, R. N., & Berbara, R. L. L. (2012). Vermicompost humic acids as an ecological pathway to protect rice plants against oxidative stress. Ecological Engineering, 47, 203–208. https://doi.org/10.1016/j.ecoleng.2012.06.011
Ghaffari, H., Tadayon, M. R., Bahador, M., & Razmjoo, J. (2022). Biochemical and yield response of sugar beet to drought stress and foliar application of vermicompost tea. Plant Stress, 5, 100087. https://doi.org/10.1016/j.stress.2022.100087
Gholami, H., Fard, F. R., Saharkhiz, M. J., & Ghani, A. (2018). Yield and physicochemical properties of inulin obtained from Iranian chicory roots under vermicompost and humic acid treatments. Industrial Crops and Products, 123, 610–616. https://doi.org/10.1016/j.indcrop.2018.07.041
Gichner, T., Patkova, Z., & Kim, J. K. (2003). DNA damage measured by the Comet assay in eight agronomic plants. Biologia Plantarum, 47, 185–188. https://doi.org/10.1023/B:BIOP.0000022248.36488.94
Jamil, A., Mehmood, R., Mehmood, F., Naveed, M., Mustafa, A., Sarfraz, S., & Faizan, M. (2025). Comparative cytotoxic and genotoxic assessment of methylparaben and propylparaben on Allium cepa root tips by Comet and Allium cepa assays. Environmental Toxicology, 40(12), 1417–1426. https://doi.org/10.1002/tox.24553
Karakas Zeybek, D., Ari, F., & Ulukaya, E. (2017). The MTT viability assay yields strikingly false-positive viabilities, although the cells are killed by some plant extracts. Turkish Journal of Biology, 41, 919–925. https://doi.org/10.3906/biy-1704-77
Kim, J., & Choi, J. (2023). Trans- and multigenerational effects of isothiazolinone biocide CMIT/MIT on genotoxicity and epigenotoxicity in Daphnia magna. Toxics, 11(4), 388. https://doi.org/10.3390/toxics11040388
Kim, M. K., Kim, K. B., Lee, J. Y., Kwack, S. J., Kwon, Y. C., Kang, J. S., Kim, H. S., & Lee, B.-M. (2019). Risk assessment of 5-chloro-2-methylisothiazol-3(2H)-one/2-methylisothiazol-3(2H)-one (CMIT/MIT) used as a preservative in cosmetics. Toxicological Research, 35, 103–117. https://doi.org/10.5487/TR.2019.35.2.103
Kresmann, S., Arokia, A. H. R., Koch, C., & Sures, B. (2018). Ecotoxicological potential of the biocides terbutryn, octhilinone and methylisothiazolinone: Underestimated risk from biocidal pathways? Science of the Total Environment, 625, 900–908. https://doi.org/10.1016/j.scitotenv.2017.12.297
Lee, J., Lee, H., Jang, S., Hong, S. H., Kim, W. J., Ryu, S. M., et al. (2019). CMIT/MIT induce apoptosis and inflammation in alveolar epithelial cells through the p38/JNK/ERK1/2 signalling pathway. Molecular & Cellular Toxicology, 15, 41–48. https://doi.org/10.1007/s13273-019-0005-0
Leme, D. M., & Marin-Morales, M. A. (2009). Allium cepa test in environmental monitoring: A review on its application. Mutation Research/Reviews in Mutation Research, 682, 71–81. https://doi.org/10.1016/j.mrrev.2009.06.002
Makkar, C., Singh, J., Parkash, C., Singh, S., Vig, A. P., & Dhaliwal, S. S. (2022). Vermicompost acts as a bio-modulator for plants under stress and non-stress conditions. Environment, Development and Sustainability, 25, 2006–2026. https://doi.org/10.1007/s10668-022-02132-w
Olive, P. L., & Durand, R. E. (2005). Heterogeneity in DNA damage using the Comet assay. Cytometry Part A, 66A, 1–8. https://doi.org/10.1002/cyto.a.20154
Pan, B., Ning, P., & Xing, B. (2008). Part V—sorption of pharmaceuticals and personal care products. Environmental Science and Pollution Research, 16, 106–116. https://doi.org/10.1007/s11356-008-0052-x
Park, E. J., Kim, S., & Chang, J. (2018). Methylisothiazolinone may induce cell death and inflammatory responses through DNA damage in human liver epithelium cells. Environmental Toxicology, 33, 156–166. https://doi.org/10.1002/tox.22504
Park, E. J., Han, J. S., Seong, E., Park, E. J., Lee, B. S., Lee, S. J., & Lee, K. (2020). Inhaled Kathon may induce eosinophilia-mediated disease in the lung. Environmental Toxicology, 35, 27–36. https://doi.org/10.1002/tox.22839
Salter, C. E., & Edwards, C. A. (2010). The production of vermicompost aqueous solutions or teas. In C. A. Edwards, N. Q. Arancon, & R. Sherman (Eds.), Vermiculture technology: Earthworms, organic wastes, and environmental management (pp. 153–163). CRC Press.
Silva, V., Silva, C., Soares, P., Garrido, E. M., Borges, F., & Garrido, J. (2020). Isothiazolinone biocides: Chemistry, biological and toxicity profiles. Molecules, 25, 991. https://doi.org/10.3390/molecules25040991
Singh, A., Karmegam, N., Singh, G. S., Bhadauria, T., Chang, S. W., Awasthi, M. K., et al. (2020). Earthworms and vermicompost: An eco-friendly approach for repaying nature’s debt. Environmental Geochemistry and Health, 42, 1617–1642. https://doi.org/10.1007/s10653-019-00510-4
Sulaiman, I. S. C., & Mohamad, A. (2020). The use of vermiwash and vermicompost extract in plant disease and pest control. In M. Egbuna, B. Sawicka (Eds.), Natural remedies for pest, disease and weed control (pp. 187–201). Academic Press. https://doi.org/10.1016/B978-0-12-819304-4.00016-6
Türkoğlu, Ş. (2012). Determination of genotoxic effects of chlorfenvinphos and fenbuconazole in Allium cepa root cells by mitotic activity, chromosome aberration and Comet assays. Pesticide Biochemistry and Physiology, 103, 224–230. https://doi.org/10.1016/j.pestbp.2012.06.001
Tufail, M., Luqman, M., Mehmood, Z., Abbas, W., Iqbal, M., Shah, H., Awan, I.F., Sana, N., John , A., Rafiq, S., Riaz, S., Ahmad, A., Tahir, A., & Awan, M. U. F. (2026). Impact of food processing industry wastewater on root growth and DNA damage in Allium cepa L. as assessed by the comet assay. Water, 18(3), 435. https://doi.org/10.3390/w18030435
Xiao, Z., Liu, M., Jiang, L., Chen, X., Griffiths, B. S., Li, H., & Hu, F. (2016). Vermicompost increases defence against root-knot nematode (Meloidogyne incognita) in tomato plants. Applied Soil Ecology, 105, 177–186. https://doi.org/10.1016/j.apsoil.2016.04.003
Yatoo, A. M., Ali, M. N., Baba, Z. A., & Hassan, B. (2021a). Sustainable management of diseases and pests in crops by vermicompost and vermicompost tea. A review. Agronomy for Sustainable Development, 41, 39. https://doi.org/10.1007/s13593-021-00692-1
Yatoo, A. M., Rasool, S., Ali, S., Majid, S., Rehman, M. U., Ali, M. N., Eachkoti,R., Rasool, S., Rashid, S. M., & Farooq, S. (2021b). Vermicomposting: An eco-friendly approach for recycling/management of organic wastes. In K. Hakeem, R. Bhat, & H. Qadri (Eds.), Bioremediation and biotechnology (pp. 167–187). Springer. https://doi.org/10.1007/978-3-030-35691-0_8
Yıldız, M., Ciğerci, İ. H., Konuk, M., Fidan, A. F., & 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. https://doi.org/10.1016/j.chemosphere.2009.01.023
Younis, M. E., Abdel-Aziz, H. M. M., & Heikal, Y. M. (2019). Nanopriming technology enhances vigour and mitotic index of aged Vicia faba seeds using chemically synthesised silver nanoparticles. South African Journal of Botany, 125, 393–401. https://doi.org/10.1016/j.sajb.2019.08.013
Yüksel, B., & Aksoy, O. (2017). Cytological effects of coumarin on the mitosis of Lens culinaris Medik. Fresenius Environmental Bulletin, 26, 6400–6407.
Yüksel, B., Aydın, D., & Aksoy, O. (2022). The impact of vermicompost on Pisum sativum spp. arvense L. exposed to methylisothiazolinone. Biologia, 77, 1109–1119. https://doi.org/10.1007/s11756-021-00977-6
Zhang, H., Tan, S. N., Teo, C. H., Yew, Y. R., Ge, L., Chen, X., & Yong, J. W. H. (2015). Analysis of phytohormones in vermicompost using a novel combinative sample preparation strategy of ultrasound-assisted extraction and solid-phase extraction coupled with liquid chromatography-tandem mass spectrometry. Talanta, 139, 189–197. https://doi.org/10.1016/j.talanta.2015.02.052

Details

Primary Language

English

Subjects

Plant Biotechnology

Journal Section

Research Article

Authors

Burcu Yüksel ^*
0000-0001-8691-6447
Türkiye

Duygu Aydın
0000-0003-0969-5171
Türkiye

Özlem Aksoy
0000-0003-4909-3562
Türkiye

Dilek Sezer This is me
0009-0008-3448-381X
Türkiye

Early Pub Date

July 26, 2026

Publication Date

-

Submission Date

May 1, 2026

Acceptance Date

July 7, 2026

Published in Issue

Year 2026 Number: Advanced Online Publication

IZ

https://izlik.org/JA89EL99GP

Cite

RIS / Bibtex

APA

Yüksel, B., Aydın, D., Aksoy, Ö., & Sezer, D. (2026). Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells. Trakya University Journal of Natural Sciences, Advanced Online Publication. https://izlik.org/JA89EL99GP

AMA

1.Yüksel B, Aydın D, Aksoy Ö, Sezer D. Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells. Trakya Univ J Nat Sci. 2026;(Advanced Online Publication). https://izlik.org/JA89EL99GP

Chicago

Yüksel, Burcu, Duygu Aydın, Özlem Aksoy, and Dilek Sezer. 2026. “Potential Protective Role of Liquid Vermicompost Against Kathon CG-Induced Cytogenotoxicity and DNA Damage in Pisum Sativum Var. Arvense L. Root Meristem Cells”. Trakya University Journal of Natural Sciences, no. Advanced Online Publication. https://izlik.org/JA89EL99GP.

EndNote

Yüksel B, Aydın D, Aksoy Ö, Sezer D (July 1, 2026) Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells. Trakya University Journal of Natural Sciences Advanced Online Publication

IEEE

[1]B. Yüksel, D. Aydın, Ö. Aksoy, and D. Sezer, “Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells”, Trakya Univ J Nat Sci, no. Advanced Online Publication, July 2026, [Online]. Available: https://izlik.org/JA89EL99GP

ISNAD

Yüksel, Burcu - Aydın, Duygu - Aksoy, Özlem - Sezer, Dilek. “Potential Protective Role of Liquid Vermicompost Against Kathon CG-Induced Cytogenotoxicity and DNA Damage in Pisum Sativum Var. Arvense L. Root Meristem Cells”. Trakya University Journal of Natural Sciences. Advanced Online Publication (July 1, 2026). https://izlik.org/JA89EL99GP.

JAMA

1.Yüksel B, Aydın D, Aksoy Ö, Sezer D. Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells. Trakya Univ J Nat Sci. 2026. Available at https://izlik.org/JA89EL99GP.

MLA

Yüksel, Burcu, et al. “Potential Protective Role of Liquid Vermicompost Against Kathon CG-Induced Cytogenotoxicity and DNA Damage in Pisum Sativum Var. Arvense L. Root Meristem Cells”. Trakya University Journal of Natural Sciences, no. Advanced Online Publication, July 2026, https://izlik.org/JA89EL99GP.

Vancouver

1.Burcu Yüksel, Duygu Aydın, Özlem Aksoy, Dilek Sezer. Potential protective role of liquid vermicompost against Kathon CG-induced cytogenotoxicity and DNA damage in Pisum sativum var. arvense L. root meristem cells. Trakya Univ J Nat Sci [Internet]. 2026 Jul. 1;(Advanced Online Publication). Available from: https://izlik.org/JA89EL99GP