Document Type : Original Research Paper

Authors

1 Institute for the Application of Nuclear Energy-INEP

2 Faculty of Agriculture, University of Belgrade

3 Institute for the Application of Nuclear Energy-INEP, University of Belgrade

4 3Open Science, Association for Scientific Research, Educational and Cultural Activities, Jane Sandanski, Ohrid

10.22044/jme.2026.16918.3338

Abstract

Permanent mining generates substantial amounts of flotation tailings with highly unfavourable physical and chemical properties, often devoid of vegetation. Their stabilization relies on phytoremediation, particularly through the establishment of grass cover. Successful revegetation requires sufficient nutrient availability and the activity of soil microorganisms that transform nutrients into plant-accessible forms. However, the interactions between plants, nutrients, and microflora during this process remain poorly understood. This study aimed to investigate the temporal dynamics and interrelationships within the plant–nutrient–microorganism system during the revegetation of flotation waste using four grass species—tall fescue, red fescue, meadow fescue, and perennial ryegrass. Plants were grown under controlled conditions on flotation tailings with different fertilizer treatments: organic (NPK 4:4:4) and mineral (NPK 20:20:20) fertilizers at varying concentrations (1% and 2% O; 0.25% and 0.5% M) and irrigation levels (50% and 75% of field water capacity). Microbial diversity (culturable bacteria, ammonifiers, fungi, and actinomycetes) was used as an indicator of remediation efficiency. Organic fertilization had the most pronounced effect, improving plant height, biomass yield, and microbial activity, particularly in tall fescue. Bacteria and ammonifiers responded positively to mineral fertilization under higher irrigation in red fescue and to organo-mineral treatment under lower irrigation in perennial ryegrass. The highest abundance of actinomycetes occurred under reduced irrigation in red fescue and perennial ryegrass. Overall, perennial ryegrass demonstrated the strongest correlation between cultivation conditions, microbial activity, and phytoremediation potential, highlighting its suitability for the ecological rehabilitation of flotation tailings.

Keywords

Main Subjects

[1]. Pouresmaieli, M., Ataei, M., & Qarahasanlou, A.N. (2023). A scientometrics view on sustainable development in surface mining: Everything from the beginning. Resources policy, 82, 103410.
[2]. Sonter, L.J., Moran, C.J., Barrett, D.J., & Soares-Filho, B.S. (2014). Processes of land use change in mining regions. Journal of Cleaner Production, 84, 494–501.
[3]. Cacciuttolo, C., Cano, D., & Custodio, M. (2023). Socio-Environmental risks linked with mine tailings chemical composition: Promoting Responsible and Safe Mine Tailings Management Considering Copper and Gold Mining Experiences from Chile and Peru. Toxics 11(5), 462.
[4]. Pouresmaieli, M., Ataei, M., Forouzandeh, P., Azizollahi, P., & Mahmoudifard, M. (2022). Recent progress on sustainable phytoremediation of heavy metals from soil. Journal of Environmental Chemical Engineering, 10(5), 108482.
[5]. Ghotbi-Ravandi, A.A., Pouresmaieli, M., Noorbakhsh, N., Khayati, N., Ekrami, E., & Zarezadeh, S. (2025). Innovative biosrption techniques for heavy metal removal using microalgae: A comprehensive review. Plant, Algae and Environment, 9(2), 71-112.
[6]. Razzak, S.A., Faruque, M.O., Alsheikh, Z., Alsheikhmohamad, L., Alkuroud, D., Alfayez, A., Hossain, S.M.Z., & Hossin, M.M. (2022). A Comprehensive review on conventional and biological-driven heavy metals removal from industrial wastewater. Environmental Advances, 7, 100168.
[7]. Sarathchandra, S.S., Rengel, Z., & Solaiman, Z.M. (2023). A review on remediation of iron ore mine tailings via organic amendments coupled with phytoremediation. Plants, 12, 1871.
[8]. Salehi, S., Pouresmaieli, M., & Qarahasanlou, A.N. (2025). A sustainable way to prevent oral diseases caused by heavy metals with phytoremediation. Case Studies in Chemical and Environmental Engineering, 11, 101106.
[9]. Martínez-Sánchez, M. J., García-Lorenzo, M. L., Pérez-Sirvent, C., & Bech, J. (2012). Trace element accumulation in plants from an aridic area affected by mining activities. Journal of Geochemical Exploration, 123, 8-12
[10]. Bakshe, P., & Jugade, R. (2023). Phytostabilization and rhizofiltration of toxic heavy metals by heavy metal accumulator plants for sustainable management of contaminated industrial sites: A comprehensive review. Journal of Hazardous Materials Advances, 10, 100293.
[11]. Pires-Lira, M. F., Castro, E. M., Lira, J. M., Oliveira, C., Pereira, F. J., & Pereira, M. P. (2020). Potential of Panicum aquanticum Poir. (Poaceae) for the phytoremediation of aquatic environments contaminated by lead. Ecotoxicology and Environment Safety 193, 110336.
[12]. Nefed’eva, E.E., Sevriukova, G. A., Zheltobryukhov, V. F., Gracheva, N. V., & Abdulabbas, A. Y. A. (2020). Assortment of herbaceous plants for remediation of soils contaminated with oil products and heavy metals. In IOP Conference Series: Earth and Environmental Science, 421(6) IOP Publishing.
[13]. Sladkovska, T., Wolski, K., Bujak, H., Radkowski, A., & Sobol, Ł. (2022). A review of research on the use of selected grass species in removal of heavy metals. Agronomy, 12, 2587.
[14]. Salmonova, H., & Bunešova, V. (2017). Methods of studying diversity of bacterial communities: a review. Scienia Agriculturae Bohemica, 48, 154-165.
[15]. De Souza, P.A., Ramos, J.N., Vasconcellos, L., Costa, L.V., Forsythe, S.J., & Brandão, M.L.L. (2025). Application and limitations of 16S rRNA gene sequencing for identifying WHO priority pathogenic Gram-negative bacilli. Infection and Drug Resistance, 18, 6353-6375.
[16]. Ramos, J.N., Costa, V.L., Vieira, V.V., & Brandão, M.L.L. (2025). Challenges in the identification of environmental bacterial isolates from a pharmaceutical industry facility by 16S rRNA gene sequences. DNA, 5(3), 33.
[17]. Balser, T.C., Liang, C., & Gutknecht, J.L.M. (2019). Linking microbial community analysis and ecosystem studies: A rapid lipid analysis protocol for high throghput. Soil Ecology Letters, 1, 22-32.
[18]. Verma, P., Pandey, V., Seleyi, S.C., Alagarsamy, A., & Dharani, G. (2024). Exploring the hidden trasures: Deep-sea bacterial community structure in the Bay of Bengal and their metabolic profile. Frontiers in Marine Science,10, 1308953.
[19]. Wang, X., Meng, D., Li, J., Lu, Z., Zhangm Z., Zhang, C., Song, S., Peng, Y., & Xia, L. (2023). Composition and dynamics of bacterial communities during flotation in coal preparation plant. Journal of Cleaner production, 385, 13569.
[20]. Zhang, Y., Wang, X., & Hongbing, J. (2020). Co-remediation of Pb Contaminated Soils by Heat Modifed Sawdust and Festuca arundinacea. Scientific Reports, 10, 4663.
[21]. Zhao, Y., Yao, J., Li, H., Sunahara, G., Li, M., Tang, C., Duran, R., Ma, B., Liu, H., Feng, L., Zhu, J., & Wu, J. (2024). Effects of three plant growth-promoting bacterial symbiosis with ryegrass for remediation of Cd, Pb and Zn soil in a mining area. Journal of Environmental Management, 353, 120167. 
[22]. Jiménez-Venegas, J., Zamora-Levia, L., Univaso, L., Soto, J., Tapia, Y., & Paneque, M. (2024). Profile of bacterial commumities in copper mine tailings revealed through high-throughput sequencing. Microorganisms, 12, 1820.
[23]. Qian, L., Lin, H., Li, B., & Dong, Y. (2023). Physicochemical characteristics and microbial communities of rhizosphere in complex amendment-assisted soilless revegetation of gold mine tailings. Chemosphere, 320, 138052.
[24]. Brajević, S., Simić, A., Andrejić, G., & Dželetović, Ž. (2023). Phytoremediation abilities of species from the Festuca genus. “The environment - research, charge, administration 15-16 june 2023” Book of proceedings; 49-53. 
[25]. Visconti, D., Caporale, A. G., Pontoni, L., Ventorino, V., Fagnano, M., Adamo, P., Pepe, O., Woo, S. L., & Fiorentino, N. (2020). Securing of an industrial soil using turfgrass assisted by biostimulants and compost amendment. Agronomy, 10, 1310.
[26]. Mohammad, S.J., Ling, Y.E., Halim, K.A., Sani, B.S., & Abdullahi, N.I. (2025). Heavy metal pollution and transformation in soil: A comprehensive review of natural bioremediation strategies. Journal of Umm-Al-Qura University for Applied Scienes, 11, 528-544.
[27]. Ventorino, V., Faraco V., Romano, I., & Pepe, O. (2018). Responses of bacterial community structure and diversity to soil eco-friendly bioremediation treatments of two multi-contaminated fields. Italian Journal of. Agronomy, 13(1), 53-58.
[28]. Liu, M., Li, Y., Che, Y., Deng, S., & Xiao, Y. (2017). Effects of different fertilizers on growth and nutrient uptake of Lolium multiflorum grown in Cd-contaminated soils. Environmental Science and Pollution Research, 24(29), 23363-23370.
[29]. Williams, D.J., & Currey, N.A. (2002). Engineering closure of an open pit gold operation in a semi-arid climate. International Journal of Mining, Reclamation and Environment, 16, 270–288.
[30]. Andrejić G., Brajević S., Simić A., Dželetović Ž., Aleksić U., & Sokolović D. (2022): Accumulation of heavy metals in root and shoot of red fescue grown at the flotation tailings dump. Book of proceedings XI International Symposium on Agricultural Sciences AgroReS 2022, Trebinje 26-28 May, 2022, 64-72.
[31]. Radulović, D.S., Stojanović, J., Bogdanović, G., Jovanović, V., Todorović, D., Ivošević, B., & Simić, V. (2025). Study of mineralogical and physicochemical properties of samples of flotation tailings "Rudnik“ mine. In: Štirbanović Z and Trumić M (Eds), XVI International Mineral Processing and Recycling Conference, IMPRC (28-30 May 2025, Belgrade, Serbia), University of Belgrade, Technical Faculty, Bor, 575-580 (ISBN 978-86-6305-158-4).
[32]. Pepper, I. L., Gerba, C. P., & Brendecke, J. W. (1995). Environmental microbiology: a laboratory manual. Academic Press.
[33]. Marjanović, M., Mihailović, I., & Spasić, K. (2016). Statistics in Economics and Business with a Collection of Solved Problems. Textbook; College of Applied Business Studies in Leskovac.
[34]. Cotrina-Teatino, M.A., Marquina-Araujo, J.J., Avalos-Murga, J.A., & Carrion-Villlacorta, F.L. (2025). Flotation of mine tailings: A bibliometric analysis and systematic literature review. Journal of Environmental Chemical Engineering, 13(2), 116136.
[35]. Vujović, N., Alivojvodić, V., Radovanović, D., Štulović, M., Sokić, M., & Kokalj, F. (2025). Towards circularity  in Serbian mining: unlocking the potential of flotation tailings and fly ash. Minerals, 15, 254.
[36]. Błonska, A., Kompała-Bąbal, A., Sierka, E., Bierzal, W., Magurno, F., Besenyei, L., Ryśl, K., & Woźniak, G. (2019). Diversity of vegetation dominated by selected grass species on coal-mine spoil heaps in terms of reclamation of post-industrial areas. Journal of Ecological Engineering, 20(2), 209-217.
[37]. Van-Dongen, A., Samad, A., Heshka, N.E., Rathie, K., Martineau, C., Bruant, G., & Derenhardt, D.A. (2021). Deep look into the microbiology and chemistry of froth treatment tailings: a review. Microorganisms, 9(5), 1091.
[38]. Lin, H., Jiang, X., Li, B., Dong, Y., & Qian, L. (2021). An efficient means of improving physicochemical properties and reshamping microbial communities of high-salty gold mine tailings. Ecotoxicology an Environmental Safety, 207, 111246.
[39]. Sun, R.B., Zhang, X.X., Guo, X.S., Wang, D.Z., & Chu, H.Y. (2015). Bacterial diversity in soils subjected to long-term chemical fertilization can be more stably maintained with the addition of livestock manure than wheat straw. Soil Biology and Biochemistry, 88, 9–18.
[40]. Guo, Z.B., Wan, S.X., Hua, K.K., Yin, Y., Chu, H.Y., Wang, D.Z., & Guo, X.S. (2020). Fertilization regime has a greater effect on soil microbial community structure than crop rotation and growth stage in an agroecosystem. Applied Soil Ecology, 149, 103510.
[41]. Grzyb, A., Wolna-Maruwka, A., & Niewiadomska, A. (2020). Environmental factors affecting the mineralization of crop residues. Agronomy, 10, 1951.
[42]. Žurek, G., & Martyniak, D. (2025). Studies on grass germination and growth on post-flotation sediments. Sustainability, 17, 3438. 
[43]. Sarathchandra, S.S., Rengel, Z., & Solaiman, Z.M. (2024). Metal uptake from iron ore mine tailings by perennial ryegrass (Lolium perenne L.) is higher after wheat straw than wheat straw biochar amendment. Plant Soil, 502(1), 481-496.
[44]. Xue, J., Wang, W., He, M., You, J., & Han, H. (2022). Study on the effect of the copper tailing substrate with different treatments on the growth of tall fescue (Festuca arundinacea). Sustainability, 14, 15387.
[45]. Vidler, A.M. (2022). Water retention properties of engineered soils for mine rehabilitation. PhD thesis. Faculty of Engineering and Built Environment, University of Newwcastle, Australia.
[46]. Pandey, P., Verma, M. K., Mukhopadhyay, R., & De, N. (2016). Biological properties of selected overburdens of Singrauli coalfields. Nature Environment and Pollution Technology, 15, 853–858.
[47]. Chung, A.P., Coimbra, C., Farias, P., Francisco R., Branco, R., Simao, F.V., Gomes, E., Pereira, A., Fiuza, A., Mortensen, M.S., Sorensen, S.J., & Morais, P.V. (2019). Tailings microbial community profile and prediction of its functionality in basins of tungsten mine. Scientific Reports, 9, 19596.
[48]. Naylor, D., DeGraaf, S., Purdom, E, & Coleman-Derr, D. (2017). Drought and host selection influence bacterial community dynamics in the grass root microbiome. International Society for Microbial Ecology Journal, 11, 2691-2704.
[49]. De Silva, S., Kariyawasam Hetti Gamage, L., & Thapa, V.R. (2025). Impact of drought on soil microbial communities.  Microorganisms, 13(7), 1625.
[50]. Metze, D., Schnecker, J., Canarini, A., Fuchslueger, L., Koch, B.J., Stone, B,W., Hungate, B.A., Hausmann, B., Schmidt, H., Schaumberger, A., Bahn, M., Kaiser, C., & Richter, A. (2023). Microbial growth under drought is confined to district taxa and modified by potential future climate conditions. Nature Communications, 14, 5895.
[51]. Ait-El-Mokhtar, M., Meddich, A., & Baslam, M. (2023). Plant-microbiome interactions under drought-insights from the molecular machinist's toolbox. Frontiers in Sustainable Food Systems, 7, 1253735.
[52]. Santos-Medellín, C., Edwards, J., Liechty, Z., Nguyen, B., & Sundaresan, V. (2017). Drought stress results in a compartment-specific restructuring of the rice root associated microbiomes. MBio, 8, 1-15.
[53]. Xu, L., Dong, Z., Chiniquy, D., Pierroz, G., Deng, S., Gao, C., Diamond, S., Simmons, T., Wipf, H.M.L., Caddel, D., Varoquaux, N., Madera, M.A., Hutmacher, R., Deutcshbauer, A., Dahlberg, J., Guerinot, M.L., Purdom, E., Banfield, J.F., Taylor, J.W., Lemaux, P.G., & Coleman-Derr, D. (2021). Genome resolved metagenomics reveals role of iron metabolism in drought-induced rhizosphere microbiome dynamics. Nature Communications, 12, 553.
[54]. Malik, A.A., & Bouskill, N.J. (2022). Drought impacts on microbial trait distribution and feedback to soil carbon cycling. Functional Ecology, 36, 1442-1456.
[55]. Liu, Y., Lan, X., Hou, H., Ji, J., Liu, X., & Lv, Z. (2024). Multifaceted Ability of Organic Fertilizers to Improve Crop Productivity and Abiotic Stress Tolerance: Review and Perspectives. Agronomy, 14, 1141.
[56]. Wang, W., Xue, J., Zhang, L., He, M., & You, J. (2024). Extraction of heavy metals from copper tailings by ryegrass (Lolium perenne L.) with the assistance of degradable chelating agents. Scientific Repports, 14, 7663.
[57]. Zhang, Q. C., Shamsi, I.H., Xu, D.-T., Wang, G.-H., Lin, X.-Y., Jilani, G., Hussain, N., & Chaudhry, A.N. (2012). Chemical fertilizer and organic manure inputs in soil exhibit a vice versa pattern of microbial community structure. Applied Soil Ecology, 57, 1-8.
[58]. Semenov, M. V., Krasnov, G. S., Semenov, V. M., & van Bruggen, A. (2022). Mineral and organic fertilizers distinctly affect fungal communities in the crop rhizosphere. Journal of Fungi, 8(3), 251.
[59]. Liu, H., Yao, J., Liu, B., Li, M., Liu, J., Jiang, S., Yu, W., Zhao, Y., & Duran, R. (2023). Active tailings disturb the surrounding vegetation soil fungal community: Diversity, assembly process and co-occurrence patterns. Science of the Total Environment, 865, 161133.
[60]. Xing, Y., Xie, Y., & Wang, X. (2025). Enhancing soil health through balanced fertilization: a patway to sustainable agriculture and food security. Frontiers in Microbiology, 16, 1536524.
[61]. Hartmann, M., & Six, J. (2023). Soil structure and microbiome functions in agroecosystems. Nature Reviews Earth & Environment, 4, 4-18.
[62]. Castellano-Hinojosa, A., Strauss, S.L., Gonzáles-López, J., & Bedmar, E.J. (2021). Changes in the diversity and predicted functional composition of the bulk and rhizosphere soil bacterial microbiomes of tomato and common bean after inorganic N-fertillization. Rhizosphere, 18, 100362.
[63]. Chen, X., Ma, X., Liu, Z., Gu, H., Fang, H., Shen, Z, Zhang, H., Wan, S., Li, W., Hao, X., Clarke, N.J., & Liu, J. (2025). Organic fertilizers increase microbial community diversity and stability down the transformation process of nutrient cycling. Environmental Microbiome, 20, 130.
[64]. Bo, H., Li, Z., Jin, D., Xu, M., & Zhang, Q. (2023). Fertilizer management methods affects bacterial community structure and diversity in the maize rhizosphere soil of a coal mine reclamation area. Annals of Microbiology, 73, 24.
[65]. Uddin, Md.K., Saha, B.K., Wong, V.N.L., & Patti, A.F. (2025). Organo-mineral fertilizer to sustain soil health and crop yield for reducing environmental impact: A comprehensive review. European Journal of Agronomy, 162, 127433.
[66]. Bo, H., Li, Z., Wang, W., Zhang, R., Wang, H., Jin, D., Xu, M., & Zhang, Q. (2024). Combining organic and inorganic fertilization enhances soil enzyme activity, the bacterial community, and molecular ecological network complexity in coal mine reclamation areas. Agronomy, 14, 1427.
[67]. Malal, H., Garcia, J.A., Marrs, A., Ait Hamza, M., Emerson, C., Nocco, M., Lakhtar, H., & Lazcano, C. (2025). Organic and inorganic fertilizers modulate the response of the soil microbiome to salinity stress. Frontiers in Microbiology, 16, 1551586.
[68]. Bai, H., He, S., Qin, T., Yan, D., Weng, B., Zhao, X., Li, X., Bai, Y., & Ma, J. (2019). Influences of irrigation amount on the rhizospheric microorganism composition and carbon dioxide flux of maize crops. Geoderma, 343, 1-9
[69]. Muhammad, I., Yang, L., Ahmad, S., Zeeshan, M., Farooq, S., Ali, I., Khan, A., & Zhou, X. B. (2022). Irrigation and nitrogen fertilization alter soil bacterial communities, soil enzyme activities and nutrient availability in maize crop. Frontiers in Microbiology, 13, 833758
[70]. Li, G., Wang, Z., Lv, Y., Jia, S., Chen, F., Liu, Y., & Huang, L. (2021). Effect of culturing ryegrass (Lolium perenne L.) on Cd and pyrene removal and bacteria variations in co-contaminated soil, Environmental Technology and Innovation, 24, 101963.
[71]. Zheng, X., Li, Q., Peng, Y., Wang, Z., & Chen, M. (2024). Phytoremediation of tungsten tailings under conditions of adding clean soil: microbiological research by metagenomic analysis. Sustainability, 16, 5715.