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

Year 2026, Volume: 32 Issue: 2, 408 - 422, 24.03.2026
https://doi.org/10.15832/ankutbd.1784347
https://izlik.org/JA34ZL69UJ

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. 

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
There are 67 citations in total.

Details

Primary Language English
Subjects Water Quality and Water Pollution
Journal Section Research Article
Authors

Tuğba Şentürk 0000-0002-9882-0079

Mustafa Oskay 0000-0001-8693-5621

Submission Date September 15, 2025
Acceptance Date December 24, 2025
Publication Date March 24, 2026
DOI https://doi.org/10.15832/ankutbd.1784347
IZ https://izlik.org/JA34ZL69UJ
Published in Issue Year 2026 Volume: 32 Issue: 2

Cite

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

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