The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed)

Tuğba Şentürk; Mustafa Oskay

doi:10.15832/ankutbd.1784347

The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed)

Abstract

The increasing presence of microplastics (MPs) and heavy metals (HMs) in aquatic ecosystems poses a growing concern due to their potential ecotoxicological effects. While the individual toxicity of MPs and HMs has been widely investigated, limited attention has been given to their combined effects on aquatic macrophytes. In this study, we aimed to evaluate the single and combined impacts of two types of MPs [polypropylene (PP) and acrylonitrile butadiene styrene (ABS)] at concentrations of 25, 50, and 100 mg L⁻¹, along with three typical HMs (Zn²⁺, Cu²⁺, and Ni²⁺), on the growth, biochemical components, and antioxidant activity of the model macrophyte Lemna gibba under laboratory conditions over a 7-day exposure period. The results revealed that both contaminants alone negatively impacted growth and biochemical performance, but the combined application caused a more pronounced decrease, suggesting a synergistic inhibitory effect on plant metabolism. The simultaneous application of ABS-MP with nickel, copper, and zinc resulted in more pronounced adverse effects on L. gibba growth parameters, photosynthetic pigments, and carbohydrate content compared to single-pollutant exposures. Co-application of copper and nickel induced pronounced oxidative stress in plant tissues, as evidenced by increased malondialdehyde levels. Furthermore, significant reductions were observed in total protein, total phenolic, and total flavonoid content across all treatments. Conversely, total antioxidant activity showed variable results dependent on the specific contaminant and concentration applied. These findings provide preliminary evidence that co-occurring MPs and HMs may exert additive or synergistic stress effects on aquatic macrophytes. In particular, the comparative evaluation of ABS and PP microplastics, along with Cu, Zn, and Ni treatments, highlights polymer- and metal-specific toxicity patterns and integrated antioxidant response profiles that have not been previously reported for L. gibba.

Keywords

References

Adeleye A T, Bahar M M, Megharaj M, Fang C & Rahman M M (2024). The unseen threat of the synergistic effects of microplastics and heavy metals in aquatic environments: a critical review. Current Pollution Reports 10: 478–497. https://doi.org/10.1007/s40726-024 00298-7
An Q, Wen C & Yan C (2024). Meta-analysis reveals the combined effects of microplastics and heavy metal on plants. Journal of Hazardous Materials 476: 135028
Anonymous (2024a). Plastics Europe, Association of Plastics Manufacturers; https://plasticseurope.org/plastics-explained/a-large-family/ accessed date: 15 September 2024.
Anonymous (2024b). Plastics Europe, Association of Plastics manufacturers; https://plasticseurope.lca data.com/datasetdetail/process.xhtml?uuid=864a6be9-1a0a-44fb-860a-199bdb66dd60&version=09.00.000 accessed date: 15 September 2024
Banu Doğanlar Z (2013). Metal accumulation and physiological responses induced by copper and cadmium in Lemna gibba, L. minor and Spirodela polyrhiza. Chemical Speciation & Bioavailability 25(2): 79-88
Blokhina O, Virolainen E & Fagerstedt K V (2003). Antioxidants, oxidative damage and oxygen deprivation stress: a review. Annals of Botany 91(2): 179-194. https://doi.org/10.1093/aob/mcf118
Bradney L, Wijesekara H, Palansooriya K N, Obadamudalige N & Bolan N S (2019). Particulate plastics as a vector for toxic trace-element uptake by aquatic and terrestrial organisms and human health risk. Environment International 131: 104937. https://doi.org/10.1016/j.envint.2019.104937
Cao L, Wu D, Liu P, Hu W & Xu L (2021). Occurrence, distribution and affecting factors of microplastics in agricultural soils along the lower reaches of Yangtze River, China. Science of the Total Environment 794: 148694. https://doi.org/10.1016/j.scitotenv.2021.148694

Cao Q, Tan A J, Lan Y, Zou W B & Yang G L (2025). Effects of Duckweed Species Diversity on Physiological Responses and Removal Efficiency under Cadmium Stress. Water, Air, & Soil Pollution 236(12): 1-15
Celekli A & Alkan E (2024). Effect of lead ions on biochemical behavior of Cladophora glomerata in sterilized and non-sterilized media. Protoplasma 261(1): 77-87
Celekli A & Bulut H (2020). Biochemical and morphological responses to cadmium-induced oxidative stress in Cladophora glomerata. Turkish Journal of Botany 44(3): 222-231
Ceschin S, Mariani F, Di Lernia D, Venditti I & Pelella E (2023). Effects of microplastic contamination on the aquatic plant Lemna minuta (least duckweed). Plants 12(1): 207. https://doi.org/10.3390/plants12010207
Chen Y, Leng Y, Liu X & Wang J (2020). Microplastic pollution in vegetable farmlands of suburb Wuhan, central China. Environmental Pollution 257: 113449. https://doi.org/10.1016/j.envpol.2019.113449
Dhameri S, Stallings Jr J, Fadhilah E, Ingram E, Leach M, Aronova A & Chwatko M (2025). Impact of Additives on Poly (acrylonitrilebutadiene-styrene) Membrane Formation Process Using Non-Solvent-Induced Phase Separation. Membranes 15(6): 181
Di Marzio, W D, Sáenz M E & Martinez R S (2024). Ecotoxicity, oxidative stress and phytoremediation of nickel on aquatic plants. In Lithium and nickel contamination in plants and the environment pp. 185-218
Dong C D, Chen C W, Chen Y C, Chen H H & Lee J S (2020). Polystyrene microplastic particles: In vitro pulmonary toxicity assessment. Journal of Hazardous Materials 385: 121575. https://doi.org/10.1016/j.jhazmat.2019.121575
Draper H H & Hadley M (1990). Malondialdehyde determination as index of lipid peroxidation. In Methods in Enzymology 186: 421-431. https://doi.org/10.1016/0076-6879(90)86135-I
DuBois M, Gilles K A, Hamilton J K, Rebers P T & Smith F (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28(3): 350-356. http://dx.doi.org/10.1021/ac60111a017
Fayshal M A (2024). Current practices of plastic waste management, environmental impacts, and potential alternatives for reducing pollution and improving management. Heliyon 10(23)
Frias J P & Nash R (2019). Microplastics: Finding a consensus on the definition. Marine Pollution Bulletin 138: 145-147. https://doi.org/10.1016/j.marpolbul.2018.11.022
Gao F, Li J, Sun C, Zhang L & Jiang F (2019). Study on the capability and characteristics of heavy metals enriched on microplastics in marine environment. Marine Pollution Bulletin 144: 61-67. https://doi.org/10.1016/j.marpolbul.2019.04.039
Gao X, Hassan I, Peng Y, Huo S & Ling L (2021). Behaviors and influencing factors of the heavy metals adsorption onto microplastics: A review. Journal of Cleaner Production 319: 128777. https://doi.org/10.1016/j.jclepro.2021.128777
Glenn E P & Doty M S (1992). Water motion affects the growth rates of Kappaphycus alvarezii and related red seaweeds. Aquaculture 108(3 4): 233-246. https://doi.org/10.1016/0044-8486(92)90109-X
Kalčíková G, Gotvajn AŽ, Kladnik A & Jemec A (2017). Impact of polyethylene microbeads on the floating freshwater plant duckweed Lemna minor. Environmental Pollution 230: 1108-1115. https://doi.org/10.1016/j.envpol.2017.07.050
Khellaf N & Zerdaoui M (2009). Growth response of the duckweed Lemna minor to heavy metal pollution. Journal of Environmental Health Science & Engineering 6(3): 161-166
Kruger N J (2009). The Bradford method for protein quantitation. The protein protocols handbook, 17-24. Third Edition. Edited by: J.M. Walker, Humana Press. https://doi.org/10.1385/0-89603-268-X:9
Lichtenthaler H (1987). Determination of total carotenoids and chlorophylls a and b of leaf in different solvents. Biochemical Society Transactions 11: 591-592. http://dx.doi.org/10.1042/bst0110591
Lilay G H, Thiébaut N, du Mee D, Assunção A G, Schjoerring J K, Husted S & Persson D P (2024). Linking the key physiological functions of essential micronutrients to their deficiency symptoms in plants. New Phytologist 242(3): 881-902
Liu S, Shi J, Wang J, Dai Y & Li H (2021). Interactions between microplastics and heavy metals in aquatic environments: a review. Frontiers in Microbiology 12: 652520. https://doi.org/10.3389/fmicb.2021.652520
Luo Y M, Shi H H, Tu C, Zhou Q & Ji R (2021). Research progresses and prospects of microplastics in the environment (in Chinese). Chinese Science Bulletin 66: 1547–1562. https://doi.org/10.1360/TB-2020-0979 Martinez R S. Sáenz M E, Alberdi J L & Di Marzio W D (2019). Comparative ecotoxicity of single and binary mixtures exposures of nickel and zinc on growth and biomarkers of Lemna gibba. Ecotoxicology 28(6): 686-697.
Mateos-Cárdenas A, Scott D T & Seitmaganbetova G (2019). Polyethylene microplastics adhere to Lemna minor (L.) yet have no effects on plant growth or feeding by Gammarus duebeni (Lillj.). Science of the Total Environment 689: 413-421. https://doi.org/10.1016/j.scitotenv.2019.06.359
Matešković A (2024). Utjecaj mikroplastike na rast i proces fotosinteze u vodenoj leći (Lemna minor) (Doctoral dissertation, University of Zagreb. Faculty of Science. Department of Biology)
Narwal N. Kakakhel M A, Katyal D, Yadav S, Rose P K & Rene E R (2024). Interactions between microplastic and heavy metals in the aquatic environment: Implications for toxicity and mitigation strategies. Water, Air, & Soil Pollution 235(9): 567
Nava V & Leoni B (2021). A critical review of interactions between microplastics, microalgae and aquatic ecosystem function. Water research 188, 116476.
OECD (2006). OECD guidelines for the testing of chemicals, revised proposal for a new guideline 221, Lemna sp. growth inhibition test.
Ogo H A, Tang N, Li X, Gao X & Xing W (2022). Combined toxicity of microplastic and lead on submerged macrophytes. Chemosphere 295: 133956. https://doi.org/10.1016/j.chemosphere.2022.133956.
Panda S K & Choudhury S (2005). Chromium stress in plants. Brazilian journal of Plant Physiology 17: 95-102. https://doi.org/10.1590/S1677 04202005000100008
Parlak K U (2016). Effects of copper on accumulation, antioxidant activity and mda content in Lemna minor, Lemna gibba and Spirodela polyrrhiza (L.). Erzincan University Journal of Science and Technology 9(2): 95-106. https://doi.org/10.18185/eufbed.73581
Parlak K U & Yilmaz D D (2012). Response of antioxidant defences to Zn stress in three duckweed species. Ecotoxicology and Environmental Safety 85: 52-58. https://doi.org/10.1016/j.ecoenv.2012.08.023
Prado F E, Boero C, Gallardo M R A & González J A (2000). Effect of NaCl on growth germination and soluble sugars content in Chenopodium quinoa Willd. seeds. Botanical Bulletin-Academia Sinica Taipei 41(1): 27-34
Prakash D, Singh B N & Upadhyay G (2007). Antioxidant and free radical scavenging activities of phenols from onion (Allium cepa). Food Chemistry 102(4): 1389-1393. https://doi.org/10.1016/j.foodchem.2006.06.063
Rahman S, Ahmad I & Nafees M (2023). Mitigation of heavy metal stress in maize (Zea mays L.) through application of silicon nanoparticles. Biocatalysis and Agricultural Biotechnology 50: 102757
Rao K M & Sresty T V S (2000). Antioxidative parameters in the seedlings of pigeonpea (Cajanus cajan (L.) Millspaugh) in response to Zn and Ni stresses. Plant Science 157(1): 113-128. https://doi.org/10.1016/s0168-9452(00)00273-9.
Riyazuddin R, Nisha N, Ejaz B, Khan M I R, Kumar M, Ramteke P W & Gupta R (2021). A comprehensive review on the heavy metal toxicity and sequestration in plants. Biomolecules 12(1): 43
Rozman U, Kokalj A J, Dolar A, Drobne D & Kalčíková G (2022). Long-term interactions between microplastics and floating macrophyte Lemna minor: The potential for phytoremediation of microplastics in the aquatic environment. Science of the Total Environment 831: 154866. http://dx.doi.org/10.1016/j.scitotenv.2022.154866
Saeed N, Khan M R & Shabbir M (2012). Antioxidant activity, total phenolic and total flavonoid contents of whole plant extracts Torilis leptophylla L. BMC Complementary and Alternative Medicine 12: 1-12. https://doi.org/10.1186/1472-6882-12-221
Santos-Echeandía J, Rivera-Hernández J R, Rodrigues J P & Molto V (2020). Interaction of mercury with beached plastics with special attention to zonation, degradation https://doi.org/10.1016/j.marchem.2020.103788 status and polymer type. Marine Chemistry 222: 103788.
Senturk T & Yildiz Ş (2016). Adsorbent effect of Chlorella vulgaris and Scenedesmus sp. (Chlorophyta) for the removal of some heavy metals and nutrients. Turkish Journal of Biochemistry 41(2): 87-95. https://doi.org/10.1515/tjb-2016-0015
Singh H, Kumar D & Soni V (2022). Performance of chlorophyll a fluorescence parameters in Lemna minor under heavy metal stress induced by various concentration of copper. Scientific reports 12(1): 10620
Singh P, Mishra V K, Kashyap R & Rawat R (2022). Understanding the Proteomics of Medicinal Plants under Environmental Pollution: Challenges and Opportunities. Environmental Pollution and Medicinal Plants pp. 213-225
So W K, Chan K & Not C (2018). Abundance of plastic microbeads in Hong Kong coastal water. Marine Pollution Bulletin 133: 500-505. https://doi.org/10.1016/j.marpolbul.2018.05.066
Song G, Hou W, Wang Q, Wang J & Jin X (2006). Effect of low temperature on eutrophicated waterbody restoration by Spirodela polyrhiza. Bioresource Technology 97(15): 1865-1869. https://doi.org/10.1016/j.biortech.2005.08.012
Tanyolaç D, Ekmekci Y & Unalan Ş (2007). Changes in photochemical and antioxidant enzyme activities in maize (Zea mays L.) leaves exposed to excess copper. Chemosphere 67(1): 89-98. https://doi.org/10.1016/j.chemosphere.2006.09.052
Taş İ (2022). Nanoplastik stresi altında yetiştirilen buğday (Triticum aestivum) köklerinde biochar uygulamasının su durumu, antioksidan enzim/izozim sistemi ve lipid peroksidasyon üzerine etkileri (Yüksek Lisans Tezi); Fen Bilimleri Enstitüsü, Necmettin Erbakan Üniversitesi
Tkalec M, Vidaković-Cifrek Ž & Regula I (1998). The effect of oil industry “high density brines” on duckweed Lemna minor L. Chemosphere 37(13): 2703-2715. https://doi.org/10.1016/S0045-6535(98)00156-8
Tu C, Chen T, Zhou Q, Liu Y & Wei J (2020). Biofilm formation and its influences on the properties of microplastics as affected by exposure time and depth in the seawater. Science of the Total Environment 734: 139237
Vinodhini R & Narayanan M (2009). Heavy metal induced histopathological alterations in selected organs of the Cyprinus carpio L. (Common Carp). International Journal of Environmental Research 3(1):95-100
Vladimirova I N & Georgiyants V A (2014). Biologically active compounds from Lemna minor SF Gray. Pharmaceutical Chemistry Journal 47: 599-601. https://doi.org/10.1007/s11094-014-1016-8
Xiong X, Wang J, Liu J & Xiao T (2024). Microplastics and potentially toxic elements: A review of interactions, fate and bioavailability in the environment. Environmental pollution 340, 122754
Xue Z, Liao X, Hou J, Xu J & Lin D (2025). Tissue-specific responses of duckweed to cadmium stress under nanoplastic co-exposure: differential accumulation and toxicity in roots and fronds. Environmental Science: Nano 12, 4235-4246 https://doi.org/10.1039/D5EN00432B
Yang Y, Liu W, Zhang Z, Grossart H P & Gadd G M (2020). Microplastics provide new microbial niches in aquatic environments. Applied Microbiology and Biotechnology 104: 6501-6511. https://doi.org/10.1007/s00253-020-10704-x
Yang X, Liao H M, Tan A J, Gan S X & Yang G L (2023). Effects of microplastics and cadmium on growth rate, photosynthetic pigment content and antioxidant enzymes of duckweed (Lemma minor). Environmental Science and Pollution Research 30(42): 96181-96190
Yu T, Wang J, Ding N, Guo X, Wang M & Chen Y (2024). Tourmaline for heavy metals removal in wastewater treatment: A review. Journal of Industrial and Engineering Chemistry 131: 44-53
Zeng J, Tang J, Zhang F, Wang Y, Kang H & Chen G (2021). Ammonium regulates redox homeostasis and photosynthetic ability to mitigate copper toxicity in wheat seedlings. Ecotoxicology and Environmental Safety 226, 112825
Zhang D, Cui Y, Zhou H, Jin C, Yu X (2020). Microplastic pollution in water, sediment, and fish from artificial reefs around the Ma’an Archipelago, Shengsi, China. Science of the Total Environment 703: 134768. https://doi.org/10.1016/j.scitotenv.2019.134768
Zhao Z, Zheng X, Han Z, Yang S, Zhang H & Lin T (2023). Response mechanisms of Chlorella sorokiniana to microplastics and PFOA stress: Photosynthesis, oxidative stress, extracellular polymeric substances and antioxidant system. Chemosphere 323, 138256
Zhishen J, Mengcheng T & Jianming W (1999). The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chemistry 64(4): 555-559. https://doi.org/10.1016/S0308-8146(98)00102-2

Details

Primary Language

English

Subjects

Water Quality and Water Pollution

Journal Section

Research Article

Authors

Tuğba Şentürk ^*
0000-0002-9882-0079
Türkiye

Mustafa Oskay
0000-0001-8693-5621
Türkiye

Publication Date

March 24, 2026

Submission Date

September 15, 2025

Acceptance Date

December 24, 2025

Published in Issue

Year 2026 Volume: 32 Number: 2

DOI

https://doi.org/10.15832/ankutbd.1784347

IZ

https://izlik.org/JA34ZL69UJ

Cite

RIS / Bibtex

APA

Şentürk, T., & Oskay, M. (2026). The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed). Journal of Agricultural Sciences, 32(2), 408-422. https://doi.org/10.15832/ankutbd.1784347

AMA

1.Şentürk T, Oskay M. The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed). J Agr Sci-Tarim Bili. 2026;32(2):408-422. doi:10.15832/ankutbd.1784347

Chicago

Şentürk, Tuğba, and Mustafa Oskay. 2026. “The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna Gibba (Duckweed)”. Journal of Agricultural Sciences 32 (2): 408-22. https://doi.org/10.15832/ankutbd.1784347.

EndNote

Şentürk T, Oskay M (March 1, 2026) The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed). Journal of Agricultural Sciences 32 2 408–422.

IEEE

[1]T. Şentürk and M. Oskay, “The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed)”, J Agr Sci-Tarim Bili, vol. 32, no. 2, pp. 408–422, Mar. 2026, doi: 10.15832/ankutbd.1784347.

ISNAD

Şentürk, Tuğba - Oskay, Mustafa. “The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna Gibba (Duckweed)”. Journal of Agricultural Sciences 32/2 (March 1, 2026): 408-422. https://doi.org/10.15832/ankutbd.1784347.

JAMA

1.Şentürk T, Oskay M. The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed). J Agr Sci-Tarim Bili. 2026;32:408–422.

MLA

Şentürk, Tuğba, and Mustafa Oskay. “The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna Gibba (Duckweed)”. Journal of Agricultural Sciences, vol. 32, no. 2, Mar. 2026, pp. 408-22, doi:10.15832/ankutbd.1784347.

Vancouver

1.Tuğba Şentürk, Mustafa Oskay. The Effect of Single and Combined Stress of Microplastics and Heavy Metals on Growth, Biochemical Components and Antioxidant Activity of Lemna gibba (Duckweed). J Agr Sci-Tarim Bili. 2026 Mar. 1;32(2):408-22. doi:10.15832/ankutbd.1784347