Pb phytostabilization by fast-growing trees inoculated with Pb-resistant plant growth-promoting endophytic bacterium

Document Type : Original Research Paper


1 Department of Agro-Industrial, Food, and Environmental Technology, Faculty of Applied Science, King Mongkut’s University of Technology North Bangkok, Bangkok, Thailand

2 School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University-Rangsit Campus, Pathum Thani 12120, Thailand

3 Department of Biology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand


Inoculation of endophytic bacteria has been accepted as a promising technique to assist phytostabilization of heavy metal-contaminated soils. This study investigated the effects of inoculating a bacterial strain closely related to Pseudomonas pyschrophila on the plant growth, and phytostabilization of fast-growing trees Acacia mangium and Eucalyptus camaldulensis, growing on artificial spiked soil with Pb up to 1500 mg/kg. After 60 days, the results showed that the strain closely related to P. pyschrophila slightly increased Pb bioavailability and Pb uptake by A. mangium, compared to non-inoculated controls. It slightly reduced Pb bioavailability in soil, but it did not affect the Pb uptake by E. camaldulensis, compared to non-inoculated controls. Interestingly, it was able to significantly increase Pb content in shoots by 3.07-fold in A. mangium and 2.95-fold in E. camaldulensis, compared to non-inoculated controls. Although the inoculation of the strain closely related to P. pyschrophila slightly increased the translocation factor (TF) of Pb in both tree species, their TF values were less than 1. This indicates that plants associated with the strain closely related to P. pyschrophila are suitable for phytostabilization of A. mangium, which may be used for cleaning up Pb contaminated sites. This strain displayed different influences on plant species and was found not suitable for phytostabilization of E. camaldulensis.


Ahemad, M. (2019) Remediation of metalliferous soils through the heavy metal resistant plant growth promoting bacteria: Paradigms and prospects. Arab. J. Chem., 12; 1365-1377.
Ahsan, M. T., Najam-ul-Haq, M., Idrees, M., Ullah, I. and Afzal, M. (2017). Bacterial endophytes enhance phytostabilization in soils contaminated with uranium and lead. Int. J. Phytorem., 19(10); 937-946.
Ali, H., Khan, E. and Sajad, M. A. (2013). Phytoremediation of heavy metals-Concepts and applications. Chemosphere, 91; 869-881.
Alkhatib, R., Bsoul, E., Blom, D. A., Ghoshroy, K., Creamer, R. and Ghoshroy, S. (2013). Microscopic analysis of lead accumulation in tobacco (Nicotiana tabacum var. Turkish) roots and leaves. J. Microsc. Ultrastruct., 1; 57-62.
Babu, A. G., Shea, P. J., Sudhakar, D., Jung, I. B. and Oh, B. T. (2015). Potential use of Pseudomonas koreensis AGB-1 in association with Miscanthus sinensis to remediate heavy metal(loid)-contaminated mining site soil. J .Environ .Manage., 151; 160-166.
Bressan, W. and Borges, M. T. (2004). Delivery methods for introducing endophytic bacteria into maize. BioControl, 49; 315-322.
Bray, R .H. and Kurtz, L. T .(1945) .Determination of total, organic and available forms of phosphorus in soils .Soil Sci., 59; 39-45.
Enamorado-Báez, S. M., Abril, J. M. and Gómez-Guzmán, J. M. (2013, June 27). Determination of 25 trace element concentrations in biological reference materials by ICP-MS following different microwave-assisted acid digestion methods based on scaling masses of digested samples. ISRN Analytical Chemistry, Article 851713. Retrieved November 3, 2014, from https://doi.org/10.1155/2013/851713
Fan, M., Liu, Z., Nan, L, Wang, E., Chen, W, Lin, Y. and Wei, G. (2018). Isolation, characterization, and selection of heavy metal-resistant and plant growth-promoting endophytic bacteria from root nodules of Robinia pseudoacacia in a Pb/Zn mining area. Microbiol. Res., 217; 51-59.
Fawcett, J. K. (1954). The semi-micro Kjeldahl method for the determination of nitrogen. J. Med. Lab. Technol., 12(1); 1-22.
He, H. D., Ye, Z. H., Yang, D. J., Yan, J. L., Xiao, L., Zhong, T., et al. (2013). Characterization of endophytic Rahnella sp. JN6 from Polygonum pubescens and its potential in promoting growth and Cd, Pb, Zn uptake by Brassica napus. Chemosphere 90; 1960-1965.
Henning, J. A., Weston, D. J., Pelletier, D. A., Timm, C. M., Jawdy, S. S. and Classen A.T. (2016,
Pollution, 6(4): 925-934, Autumn 2020
November 1). Root bacterial endophytes alter plant phenotype, but not physiology. PeerJ 4, Article e2606 Retrieved April 3, 2017, from https://doi.org/10.7717/peerj.2606.
Jan, R., Khan, M. A., Asaf, S., Lubna, Lee, I. J. and Kim, K. M. (2019, September 23). Metal resistant endophytic bacteria reduces cadmium, nickel toxicity, and enhances expression of metal stress related genes with improved growth of Oryza sativa, via regulating its antioxidant machinery and endogenous hormones. Plants 8, Article 363 Retrieved December 18, 2019, from https://doi.org/10.3390/plants8100363.
Jebara, S. H., Saadani, O., Fatnassi, I. C., Chiboub, M., Abdelkrim, S. and Jebara, M. (2015). Inoculation of Lens culinaris with Pb-resistant bacteria shows potential for phytostabilization. Environ. Sci. Pollut. Res., 22; 2537-2545.
Jiang, C. Y., Sheng, X. F., Qian, M. and Wang, Q.Y. (2008). Isolation and characterization of a heavy metal-resistant Burkholderia sp. from heavy metal-contaminated paddy field soil and its potential in promoting plant growth and heavy metal accumulation in metal-polluted soil. Chemosphere, 72(2); 157-64.
Li, H. Y., Wei, D. Q., Shen, M. and Zhou, Z. P. (2012). Endophytes and their role in phytoremediation. Fungal Divers., 54(1); 11-18.
Li, Z., Wang, P., Yue, X., Wang, J., Ren, B., Qu, L. and Han, H. (2019, September 30). Effects of Bacillus thuringiensis HC-2 combined with biochar on the growth and Cd and Pb accumulation of radish in a heavy metal-contaminated farmland under field conditions. Int J Environ Res Public Health, 16(19), Article 3676. Retrieved July 6, 2020, from https://doi.org/10.3390/ijerph16193676
Lindsay, W. L. and Norvell, W. A. (1978). Development of a DTPA test for zinc, iron, manganese, and copper. Soil Sci. Soc. Am. J., 42; 421-428.
Luo, S. L., Chen, L., Chen, J. L., Xiao, X., Xu, T. Y., Wan, Y., et al. (2011). Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation. Chemosphere, 85; 1130-1138.
Ma, Y., Rajkumar, M., Luo, Y. M. and Freitas, H. (2011). Inoculation of endophytic bacteria on host and non-host plants effects on plant growth and Ni uptake. J. Hazard. Mater., 196; 230-237.
Ma, Y., Rajkumar, M., Rocha, I., Oliveira, R. S. and Freitas, H. (2015, January 5). Serpentine bacteria influence metal translocation and bioconcentration of Brassica juncea and Ricinus communis grown in multi-metal polluted soils. Front. Plant Sci., 5, Article 775. Retrieved January 3, 2017, from https//:www.frontiersin.org/articles/10.3389/fpls.2014.00757/full.
Ma, Y., Rajkumar, M., Zhang, C. and Freitas, H. (2016). Beneficial role of bacterial endophytes in heavy metal phytoremediation. J. Environ. Manage., 174; 14-25.
Madhaiyan, M., Poonguzhali, S. and Sa, T. (2007). Metal tolerating methylotrophic bacteria reduces nickel and cadmium toxicity and promotes plant growth of tomato (Lycopersicon esculentum L.). Chemosphere, 69; 220-228.
Mahar, A., Wang, P., Ali, A., Awasthi, M. K., Lahori, A. H., Wang, Q., et al. (2016). Challenges and opportunities in the phytoremediation of heavy metals contaminated soils: A review. Ecotox. Environ. Safe., 126; 111-121.
Marcos, F. C. C., Iório, R. P. F., Silveira, A. P. D., Ribeiro, R. V., Machado, E. C. and Lagôa, A. M. M. A. (2016). Endophytic bacteria affect sugarcane physiology without changing plant growth. Bragantia, 7(1); 1-9. Mesa, J., Mateos-Naranjo, E., Caviedes, M. A., Redondo-Gómez, S., Pajuelo, E. and Rodríguez-Llorente, I. D. (2015, December 22). Endophytic cultivable bacteria of the metal bioaccumulator Spartina maritima improve plant growth but not metal uptake in polluted marshes soils. Front. Microbiol., 6, Article 1450. Retrieved April 20, 2018, from https://doi.org/10.3389/fmicb.2015.01450.
Meeinkuirt, W., Pokethitiyook, P., Kruatrachue, M., Tanhan, P. and Chaiyarat, R. (2012). Phytostabilization of a Pb-contaminated mine tailing by various tree species in pot and field trial experiments. Int. J. Phytorem., 14; 925-938.
Navarro-Torre, S., Mateos-Naranjo, E., Caviedes, M. A., Pajuelo, E. and Rodriguez-Llorente, I. D. (2016). Isolation of plant-growth promoting and metal resistant cultivable bacteria from Arthrocnemum macrostachyum in the Odiel marshes with potential use in phytoremediation. Mar. Pollut. Bull., 110; 133-142.
Pandey, V. C., Bajpai, O. and Singh, N. (2016). Energy crops in sustainable phytoremediation. Renew. Sust. Energ. Rev., 54; 58-73.
Pierzynski, G. M., Schnoor, J. L., Youngman, A., Licht, L. and Erickson, L. E. (2002). Poplar trees for phytostabilization of abandoned zinc-lead smelter. Pract. Period. Hazard. Toxic Radioact. Waste Manage., 6(3); 177-183.
Yongpisanphop, J., et al.
Pollution is licensed under a "Creative Commons Attribution 4.0 International (CC-BY 4.0)"
Rajkumar, M., Ma, Y. and Freitas, H. (2013). Improvement of Ni phytostabilization by inoculation of Ni resistant Bacillus megaterium SR28C. J. Environ. Manage., 128; 973-980.
Saadani, O., Fatnassi, I. C., Chiboub, M., Abdelkrim, S., Barhoumi, F., Jebara, M. and Jebara, S.H. (2016). In situ phytostabilisation capacity of three legumes and their associated Plant Growth Promoting Bacteria (PGPBs) in mine tailings of northern Tunisia. Ecotox. Environ. Safe., 130; 263-269.
Saifullah, R., Meers, E., Qadir, M., de Caritat, P., Tack, F. M. G., Du Laing, G. and Zia, M. H. (2009). EDTA-assisted Pb phytoextraction. Chemosphere, 74; 1279-1291.
Sessitsch, A., Kuffner, M., Kidd, P., Vangronsveld, J., Wenzel, W.W., Fallmann, K. and Puschenreiter, M. (2013). The role of plant-associated bacteria in the mobilization and phytoextraction of trace elements in contaminated soils. Soil Biol. Biochem., 60; 182-194.
Sheng, X. F., Xia, J. J., Jiang, C. Y., He, L. Y. and Qian, M. (2008). Characterization of heavy metal-resistant endophytic bacteria from rape (Brassica napus) roots and their potential in promoting the growth and lead accumulation of rape. Environ. Pollut., 156; 1164-1170.
Shin, M. N., Shim, J., You, Y., Myung, H., Bang, K. S., Cho, M., et al. (2012). Characterization of lead resistant endophytic Bacillus sp. MN3-4 and its potential for promoting lead accumulation in metal hyperaccumulator Alnus firma. J. Hazard. Mater., 199-200; 314-320.
Siripan, O., Arinthip, T. and Wunrada, S. (2018). Enhancement of the efficiency of Cd phytoextraction using bacterial endophytes isolated from Chromolaena odorata, a Cd hyperaccumulator. Int. J. Phytorem., 20(11); 1096-1105.
Sun, L. N., Zhang, Y. F., He, L.Y., Chen, Z. J., Wang, Q. Y., Qian, M. and Sheng, X. F. (2010). Genetic diversity and characterization of heavy metal-resistant-endophytic bacteria from two copper-tolerant plant species on copper mine wasteland. Bioresour. Technol., 101; 501-509.
Sylvain, B., Mikael, M. H., Florie, M., Emmanuel, J., Marilyne, S., Sylvain, B. and Domenico M. (2016). Phytostabilization of As, Sb and Pb by two willow species (S. viminalis and S. purpurea) on former mine technosols. Catena, 136; 44-52.
United States Environmental Protection Agency (U.S. EPA). 2007. Method 3051A (SW-846): Microwave assisted acid digestion of sediments, sludges, and oils [Electronic version]. Retrieved April 15, 2014, from: https://www.epa.gov/sites/production/files/2015-12/documents/3051a.pdf
Walkley, A. and Black, I. A. (1947). Chronic acid titration method for determination of soil organic matter. Soil Sci. Amer. Proc., 63; 257.
Yongpisanphop, J., Babel, S., Kurisu, F., Kruatrachue, M. and Pokethitiyook, P. (2019, May 20). Isolation and characterization of Pb-resistant plant growth promoting endophytic bacteria and their role in Pb accumulation by fast-growing trees. Environ. Technol., Retrieved June 6, 2019, fromhttps://doi.org/10.1080/09593330.2019.1615993
Yongpisanphop, J. and Babel, S. (2017, June). Assessment of Pb accumulation in roots of fast-growing trees inoculated with endophytic bacteria: Hydroponic culture (Paper presented at the 5th International Conference on Sustainable Solid Waste Management, Athens)
Zhang, Y. F., He, L.Y., Chen, Z. J., Zhang, W. H., Wang, Q. Y., Qian, M. and Sheng, X. F. (2011). Characterization of lead-resistant and ACC deaminase-producing endophytic bacteria and their potential in promoting lead accumulation of rape. J. Hazard. Mater., 186(2-3); 1720-1725.
Zulfiqara, U., Farooq, M., Hussain, S., Maqsood, M., Hussain, M., Ishfaq, M., et al. (2019, November 15). Lead toxicity in plants: Impacts and remediation. J. Environ. Manage., 250, Article 109557. Retrieved June 30, 2020, from https://doi.org/10.1016/j.jenvman.2019.109557