Bioremoval of hazardous cobalt, nickel, chromium, copper and cadmium compounds from contaminated soil by Nicotiana tabacum plants and associated microbiome

  • О. A. Havryliuk Zabolotny Institute of Microbiology and Virology of the NASU
  • V. M. Hovorukha Zabolotny Institute of Microbiology and Virology of the NASU
  • A. V. Sachko Yuriy Fedkovych Chernivtsi National University
  • G. V. Gladka Zabolotny Institute of Microbiology and Virology of the NASU
  • I. O. Bida Zabolotny Institute of Microbiology and Virology of the NASU
  • O. B. Tashyrev Zabolotny Institute of Microbiology and Virology of the NASU
Keywords: thermodynamic prognosis; heavy metals contamination; metal-resistant microorganisms; environmental biotechnology.


Contamination of soils with heavy metals leads to reduction of soil fertility, destruction of natural ecosystems and detrimental effects on the health of society by increasing content of metals in the food chains from microorganisms to plants, animals and humans. Bioremediation is one of the most promising and cost-effective methods of cleaning soils polluted with toxic metals. According to current researchers, microorganisms and plants have the genetic potential to remove toxic metals from contaminated sites. The method of thermodynamic prediction was used to theoretically substantiate the mechanisms of interaction of soil microorganisms and plants with heavy metals. According to the our prediction, exometabolite chelators of anaerobic microorganisms may increase the mobility of metals and thereby contribute to the active transport of metals and their accumulation in plants. Plants of Nicotiana tabacum L. of Djubek cultivar were used as plant material for the current investigation. The examined toxicants were heavy metals, namely cobalt (II), nickel (II), chromium (VI), copper (II) and cadmium (II). The aqueous solutions of metal salts were added to the boxes after two months of plants growing to the final super-high concentration – 500 mg/kg of absolutely dry weight of soil. Quantitative assessments of copper and chromium-resistant microorganisms were made by cultivation on agar nutrient medium NA with a gradient of Cu(II) and Cr(VI). The concentration of metals in soil and plant material (leaves, stems and roots) was determined by atomic absorption method. The study revealed that heavy metals inhibited the growth of the examined tobacco plants. This was expressed by the necrosis of plant tissues and, ultimately, their complete death. Despite this, all investigated heavy metals were accumulated in plant tissues during 3–7 days before death of plants. The uptake of metals was observed in all parts of plants – leaves, stems and roots. The highest concentrations of Co(II), Ni(II), Cd(II), Cr(VI) were found in the leaves, Cu(II) – in the roots. The results show that the bioremoval efficiency of the investigated metals ranged 0.60–3.65%. Given the super-high initial concentration of each of the metals (500 mg/kg), the determined removal efficiency was also high. Cadmium was the most toxic to plants. Thus, the basic points of the thermodynamic prognosis of the possibility of accumulation of heavy metals by phytomicrobial consortium were experimentally confirmed on the example of N. tabacum plants and metal-resistant microorganisms. The study demonstrated that despite the high initial metals concentration, rate of damage and death of plants, metals are accumulated inplant tissues in extremely hight concentrations. Soil microorganisms were observed to have high adaptation potencial to Cu(II) and Cr(VI). In anaerobic conditions, microorganisms presumably mobilize heavy metals, which later are absorbed by plants. The obtained results are the basis for the development of environmental biotechnologies for cleaning contaminated soils from heavy metal compounds.


Aleksandra, S.-N. (2011). Phytotechnologies: Importance in remediation of heavy metal-contaminated soils. In: Khan, M., Zaidi, A., Goel, R., & Musarrat, J. (Eds.). Biomanagement of metal-contaminated soils. Environmental Pollution, Springer, Dordrecht. Pp. 277–295.

Bhuiyan, M. A. H., Parvez, L., Islam, M. A., Dampare, S. B., & Suzuki, S. (2010). Heavy metal pollution of coal mine-affected agricultural soils in the northern part of Bangladesh. Journal of Hazardous Materials, 173, 384–392.

Brygadyrenko, V., & Ivanyshyn, V. (2015). Changes in the body mass of Megaphyllum kievense (Diplopoda, Julidae) and the granulometric composition of leaf litter subject to different concentrations of copper. Journal of Forest Science, 61(9), 369–376.

Dhiman, S. S., Zhao, X., Li, J., Kim, D., Kalia, V. C., Kim, I., Kim, J. Y., & Lee, J. (2017). Metal accumulation by sunflower (Helianthus annuus L.) and the efficacy of its biomass in enzymatic saccharification. PLoS One, 12(4), e0175845.

Dinu, C., Vasile, G.-G., Buleandra, M., Popa, D. E., Gheorghe, S., & Ungureanu, E.-M. (2020). Translocation and accumulation of heavy metals in Ocimum basilicum L. plants grown in a mining-contaminated soil. Journal of Soils and Sediments, 20(4), 2141–2154.

Dong, G., Wang, Y., Gong, L., Wang, M., Wang, H., He, N., Zheng, Y., & Li, Q. (2013). Formation of soluble Cr(III) end-products and nanoparticles during Cr(VI) reduction by Bacillus cereus strain XMCr-6. Biochemical Engineering Journal, 70, 166–172.

Dovgalyuk, А. (2013). Environmental pollution by toxic metals and its indication by plant test systems. Studia Biologica, 7(1), 197–204.

Fashola, M. O., Ngole-Jeme, V. M., & Babalola, O. O. (2016). Heavy metal pollution from gold mines: Environmental effects and bacterial strategies for resistance. International Journal of Environmental Research and Public Health, 13(11), 1047.

Gurajala, H. K., Cao, X., Tang, L., Ramesh, T. M., Lu, M., & Yang, X. (2019). Comparative assessment of Indian mustard (Brassica juncea L.) genotypes for phytoremediation of Cd and Pb contaminated soils. Environmental Pollution, 254, 113085.

Havryliuk, O., Hovorukha, V., Patrauchan, M., Youssef, N. H., & Tashyrev, O. (2020). Draft whole genome sequence for four highly copper resistant soil isolates Pseudomonas lactis strain UKR1, Pseudomonas panacis strain UKR2, and Pseudomonas veronii strains UKR3 and UKR4. Current Research in Microbial Sciences, 1, 44–52.

Husak, V. (2015). Copper and copper-containing pesticides: Metabolism, toxicity and oxidative stress. Journal of Vasyl Stefanyk Precarpathian National University, 2(1), 38–50.

Khan, F. I., Husain, T., & Hejazi, R. (2004). An overview and analysis of site remediation technologies. Journal of Environmental Management, 71(2), 95–122.

Khan, Z. I., Ugulu, I., Zafar, A., Mehmood, N., Bashir, H., Ahmad, K., & Sana, M. (2021). Biomonitoring of heavy metals accumulation in wild plants growing at Soon valley, Khushab, Pakistan. Pakistan Journal of Botany, 53(1), 247–252.

Kozak, V. M., & Brygadyrenko, V. V. (2018). Impact of cadmium and lead on Megaphyllum kievense (Diplopoda, Julidae) in a laboratory experiment. Biosystems Diversity, 26(2), 128–131.

Li, C., Zhou, K., Qin, W., Tian, C., Qi, M., & Yan, X. (2019). A review on heavy metals contamination in soil: Effects, sources, and remediation techniques, soil and sediment contamination. An International Journal, 28(4), 380–394.

Li, Z., Ma, Z., V. D. Kuijp, T. J., Yuan, Z., & Huang, L. (2014). A review of soil heavy metal pollution from mines in China: Pollution and health risk assessment. Science of the Total Environment, 468–469, 843–853.

Liu, L., Li, W., Song, W., & Guo, M. (2018). Remediation techniques for heavy metal-contaminated soils: Principles and applicability. Science of the Total Environment, 633, 206–219.

Mani, D., & Kumar, C. (2014). Biotechnological advances in bioremediation of heavy metals contaminated ecosystems: An overview with special reference to phytoremediation. International Journal of Environmental Science and Technology, 11(3), 843–872.

Mani, D., Sharma, B., Kumar, C., Pathak, N., & Balak, S. (2012). Phytoremediation potential of Helianthus annuus L in sewage-irrigated indo-gangetic alluvial soils. International Journal of Phytoremediation, 14(3), 235–246.

Marques, A. P. G. C., Rangel, A. O. S. S., & Castro, P. M. L. (2009). Remediation of heavy metal contaminated soils: Phytoremediation as a potentially promising clean-up technology. Critical Reviews in Environmental Science and Technology, 39(8), 622–654.

Masindi, V., & Muedi, K. L. (2018). Environmental contamination by heavy metals. In: El-Din, H., Saleh, M., & Aglan, R. F. (Eds). Heavy metals. IntechOpen. Pp. 115–133.

Mazurek, R., Kowalska, J., Gąsiorek, M., Zadrożny, P., Józefowska, A., Zaleski, T., Kepka, V., Tymczuk, M., & Orłowska, K. (2017). Assessment of heavy metals contamination in surface layers of Roztocze National Park forest soils (SE Poland) by indices of pollution. Chemosphere, 2016, 1–12.

Nahar, N., Rahman, A., Nawani, N. N., Ghosh, S., & Mandal, A. (2017). Phytoremediation of arsenic from the contaminated soil using transgenic tobacco plants expressing ACR2 gene of Arabidopsis thaliana. Journal of Plant Physiology, 218(8), 121–126.

Orell, A., Navarro, C. A., Arancibia, R., Mobarec, J. C., & Jerez, C. A. (2010). Life in blue: Copper resistance mechanisms of bacteria and Archaea used in industrial biomining of minerals. Biotechnology Advances, 28(6), 839–848.

Pogrzeba, M., Rusinowski, S., & Krzyżak, J. (2018). Macroelements and heavy metals content in energy crops cultivated on contaminated soil under different fertilization – case studies on autumn harvest. Environmental Science and Pollution Research, 25(12), 12096–12106.

Rajbanshi, A. (2009). Study on heavy metal resistant bacteria in Guheswori Sewage Treatment Plant. Our Nature, 6(1), 52–57.

Romantschuk, M., Sarand, I., Petänen, T., Peltola, R., Jonsson-Vihanne, M., Koivula, T., Yrjälä, K., & Haahtela, K. (2000). Means to improve the effect of in situ bioremediation ofcontaminated soil. Environmental Pollution, 107(2), 179–185.

Sathishkumar, K., Murugan, K., Benelli, G., Higuchi, A., & Rajasekar, A. (2017). Bioreduction of hexavalent chromium by Pseudomonas stutzeri L1 and Acinetobacter baumannii L2. Annals of Microbiology, 67(1), 91–98.

Say, R., Yilmaz, N., & Denizli, A. (2003). Removal of heavy metal ions using the fungus Penicillium canescens. Adsorption Science and Technology, 21(7), 643–650.

Shulman, M. V., Pakhomov, O. Y., & Brygadyrenko, V. V. (2017). Effect of lead and cadmium ions upon the pupariation and morphological changes in Calliphora vicina (Diptera, Calliphoridae). Folia Oecologica, 44(1), 28–37.

Sims, J. L., Sims, R. C., & Matthews, J. E. (1990). Approach to bioremediation of contaminated soil. Hazardous Waste and Hazardous Materials, 7(2), 117–149.

Sulaiman, M. B., Salawu, K., & Barambu, A. U. (2019). Assessment of concentrations and ecological risk of heavy metals at resident and remediated soils of uncontrolled mining site at Dareta Village, Zamfara, Nigeria. Journal of Applied Sciences and Environmental Management, 23(1), 189–193.

Sun, H., Sun, X., Wang, H., & Ma, X. (2020). Advances in salt tolerance molecular mechanism in tobacco plants. Hereditas, 157(1), 1–6.

Sun, Z., Xie, X., Wang, P., Hu, Y., & Cheng, H. (2018). Heavy metal pollution caused by small-scale metal ore mining activities: A case study from a polymetallic mine in South China. Science of the Total Environment, 639, 217–227.

Tang, Z., Chai, M., Cheng, J., Jin, J., Yang, Y., Nie, Z., & Huang, Q. (2017). Ecotoxicology and environmental safety contamination and health risks of heavy metals in street dust from a coal-mining city in Eastern China. Ecotoxicology and Environmental Safety, 138(2016), 83–91.

Tashyrev, O., Govorukha, V., Suslova, O., & Tashyreva, H. (2018). Thermodynamic prediction for development of novel environmental biotechnologies and valuable products from waste obtaining. Ecological Engineering and Environment Protection, 2018(1), 24–35.

Taştan, B. E., Ertuǧrul, S., & Dönmez, G. (2010). Effective bioremoval of reactive dye and heavy metals by Aspergillus versicolor. Bioresource Technology, 101(3), 870–876.

Taylor, J. (1962). The estimation of numbers of bacteria by tenfold dilution series. Journal of Applied Bacteriology, 25(1), 54–61.

Tripathi, D. K., Singh, V., Chauhan, D., Prasad, S., & Dubey, N. K. (2014). Recent advances and future prospective. In: Ahmad, P., Wani, M., Azooz, M., & Phan Tran, L. S. (Eds.). Improvement of crops in the era of climatic changes. Springer, New York. Pp. 197–216.

Ukah, B. U., Egbueri, J. C., Unigwe, C. O., & Ubido, O. E. (2019). Extent of heavy metals pollution and health risk assessment of groundwater in a densely populated industrial area, Lagos, Nigeria. International Journal of Energy and Water Resources, 3, 291–303.

Verma, T., Garg, S. K., & Ramteke, P. W. (2009). Genetic correlation between chromium resistance and reduction in Bacillus brevis isolated from tannery effluent. Journal of Applied Microbiology, 107(5), 1425–1432.

Volentini, S. I., Farías, R. N., Rodríguez-Montelongo, L., & Rapisarda, V. A. (2011). Cu(II)-reduction by Escherichia coli cells is dependent on respiratory chain components. BioMetals, 24(5), 827–835.

Wang, G., Zhang, S., Xu, X., Zhong, Q., Zhang, C., Jia, Y., Li, T., Deng, O., & Li, Y. (2016). Heavy metal removal by GLDA washing: Optimization, redistribution, recycling, and changes in soil fertility. Science of the Total Environment, 569–570, 557–568.

Wu, Z., Yang, J., Zhang, Y., Wang, C., & Guo, S. (2021). Growth responses, accumulation, translocation and distribution of vanadium in tobacco and its potential in phytoremediation. Ecotoxicology and Environmental Safety, 207, 111297.

Xiao, R., Wang, S., Li, R., Wang, J. J., & Zhang, Z. (2017). Soil heavy metal contamination and health risks associated with artisanal gold mining in Tongguan, Shaanxi, China. Ecotoxicology and Environmental Safety, 141, 17–24.

Yang, Y., Hu, M., Zhou, D., Fan, W., Wang, X., & Huo, M. (2017). Bioremoval of Cu2+ from CMP wastewater by a novel copper-resistant bacterium Cupriavidus gilardii CR3: Characteristics and mechanisms. RSC Advances, 7(30), 18793–18802.