Phormidium improves seed germination and growth parameters of berseem clover in hexadecane-contaminated soil

Petroleum is known as the main source of fuel and one of the most important environmental pollutants. Petroleum contaminated soil negatively influences plant growth and human health. Bioremediation of petroleum-contaminated soil could be achieved by adding cyanobacteria and photosynthetic microalgae in the soil. In the current study, the beneficial effects of cyanobacterium Phormidium sp. ISC108 treatment on seed germination and plant growth parameters were investigated on berseem clover plants exposed to 1% hexadecane- contaminated soil. For this purpose, after cultivation of berseem clover seeds in soil with 0 and 1% hexadecane, they were irrigated every two days by 3 ml Phormidium (OD600 = 0.7) and water for the cyanobacteria treatment and control, respectively. After 30 days, the biodegradability of hexadecane in the soil around the root was measured by gas chromatography mass-spectrometry (GC-MS). The results showed that Phormidium treatment accelerated germination of berseem clover seeds in both control and hexadecane- contaminated soil. Improvement of plant growth indices such as leaf area, plant fresh weight and leaf RWC due to cyanobacteria treatment was observed in hexadecane-contaminated soil. Hexadecane levels in the soil around the root of the plants irrigated by cyanobacteria were significantly decreased. In addition, the hexadecane degradation in the soil around the roots of berseem clover plant was increased in both control (water) treatment. In conclusion, due to positive effects of Phormidium treatment on seed germination, plant growth of berseem clover and hexadecane degradation, it can be effectively used to enhance plant capacity to cope with hexadecane toxicity in petroleum-contaminated soils.
Cyanobacteria, Hexadecane, Trifolium alexandrinum L.


Adam, G. and H. Duncan (2003). The Effect of Diesel Fuel on Common Vetch (Vicia Sativa L.) Plants. Environmental Geochemistry and Health, 25: 123–130.

Afzal, Peyman, et al. "Delineation of mineralization zones in porphyry Cu deposits by fractal concentration–volume modeling." Journal of Geochemical Exploration 108.3 (2011): 220-232.

Ahmad M, Usman AR, Al-Faraj AS, Ahmad M, Sallam A, Al-Wabel MI (2018) Phosphorus-loaded biochar changes soil heavy metals availability and uptake potential of maize (Zea mays L.). Chemosphere 194:327-339.

Ahmed Abdel – Megeed , Naif Al-Harbi , and salem Al-Deyab. Hexadecane degradation by bacterial strains isolated form contaminated soil. African Journal of Biotechnology 1 November, 2010;9 (44):7487-7494

Anjum, N. A., Arena, c. and Singhgill, S. (2014) Reactive Oxygen species (ROS) and response of antioxidants as ROSscavengers during enviromental stress in plant. Frontiers in Enviromental Science 2: 1-13

Atagana, Harrison Ifeanyichukwu. "Bioremediation of co-contamination of crude oil and heavy metals in soil by phytoremediation using Chromolaena odorata (L) King & HE Robinson." Water, Air, & Soil Pollution 215.1-4 (2011): 261-271

Baker, J. M. "The effects of oils on plants." Environmental Pollution (1970) 1.1 (1970): 27-44.

Besalatpour, A., Khoshgoftarmanesh, A. H., Hajabbasi, M. A., & Afyuni, M. (2008). Germination and growth of selected plants in a petroleum contaminated calcareous soil. Soil & Sediment Contamination, 17(6), 665-676.

Bossert, Ingeborg, and Richard Bartha. "Plant growth in soils with a history of oily sludge disposal." Soil Science 140.1 (1985): 75-77.

Creelman, Robert A., et al. "Water deficit and abscisic acid cause differential inhibition of shoot versus root growth in soybean seedlings: analysis of growth, sugar accumulation, and gene expression." Plant Physiology 92.1 (1990): 205-214.

Cvjetko P, Tolić S, Šikić S, Balen B, Tkalec M, Vidaković-Cifrek Ž, Pavlica M (2010) Effect of copper on the toxicity and genotoxicity of cadmium in duckweed (Lemna minor L.). Arh Hig Rada Toksikol 61:287–296

El-Sharkawi, H. M., K. A. Farghali, and S. A. Sayed (1989) Interactive Effects of Water Stress, Temperature and Nutrients in Seed Germination of Tree Desert Plants. Journal of Arid Environments 17: 307-317.

El-Sheekh, M. M., et al. "Effect of cobalt on growth, pigments and the photosynthetic electron transport in Monoraphidium minutum and Nitzchia perminuta." Brazilian Journal of Plant Physiology 15.3 (2003): 159-166.

Endo, A., Sawada, Y., Takahashi, H. & Nambara, E. (2008) Drought induction of Arabidopsis 9-cis-epoxycarotenoid dioxygenase occurs in vascular parenchyma cells, Plant Physiology 147, 1984-1993.

Erdogan, E., et al. "Engineering carbon chains from mechanically stretched graphene-based materials." Physical Review B 83.4 (2011): 041401.

Fernet, J.L. (2008) Plant bacterial inoculants to remediate hydrocarbon polluted soil. Masters of Science Thesis, Department of Soil Science University of Saskatchewan.

Forooghi Nia F, Dezfulian M, Shokravi S, Harzandi N, Soltani N. Biodegradation effects of five species of isolated Cyanobacteria from oil contaminated areas of the southern part of Iran. J. Aqu. Eco.. 2013; 3 (1) :28-20.

Gamila, H. Ali, M. B. M. Ibrahim, and HH Abd El-Ghafar. "The role of cyanobacterial isolated strains in the biodegradation of crude oil." International journal of environmental studies 60.5 (2003): 435-444.

Gao, Y., Yu, X. Z., Wu, S. C., Cheung, K. C., Tam, N. F. Y., Qian, P. Y., & Wong, M. H. (2006). Interactions of rice (Oryza sativa L.) and PAH-degrading bacteria (Acinetobacter sp.) on enhanced dissipation of spiked phenanthrene and pyrene in waterlogged soil. Science of the Total Environment, 372(1), 1-11.

Johnsen, Anders R., Lukas Y. Wick, and Hauke Harms. "Principles of microbial PAH-degradation in soil." Environmental pollution 133.1 (2005): 71-84.

Kastner, M., Degradation of aromatic and polyaromatic compounds. Biotechnology, environmental processes. Germany: Wiley Vch; 2000.

Khoshgoftar B (1992) Berseem Clover. Publication of agriculture extention organization of Mazandaran province.

Kuiper, Irene, et al. "Rhizoremediation: a beneficial plant-microbe interaction." Molecular plant-microbe interactions 17.1 (2004): 6-15.

Kvesitadze, G., Khatisashvili, G., Sadunishvili, T., & Ramsden, J. J. (2006). Biochemical mechanisms of detoxification in higher plants: basis of phytoremediation. Springer Science & Business Media.

Li, J.H., Y. Gao, S.C. Wu, K.C. Cheung, X.R. Wang and M.H. Wong. 2008. Physiological and biochemical responses of rice (Oryza Sativa L.) to phenanthrene and pyrene. Int. J. Phytoremed. 10: 106-118.

Li, X., Feng, Y., & Sawatsky, N. (1997). Importance of soil-water relations in assessing the endpoint of bioremediated soils. Plant and Soil, 192(2), 219-226.

Ma, F., Shi, Sh., Sun, T.H., Li, A., Zhou, J.T., Qu, Y.Y. Biotransformation of benzene and toluene to catechols by phenol hydroxylase from Arthrobacter sp. W1. Applied Microbiology and Biotechnology, 97(11), pp, 5097-103, 2012.

Maguire JD (1962) Speed of Germination—Aid In Selection And Evaluation for Seedling Emergence And Vigor. Crop sci 2:176-177.

Marquardt, J., & Palinska, K. A. (2007). Genotypic and phenotypic diversity of cyanobacteria assigned to the genus Phormidium (Oscillatoriales) from different habitats and geographical sites. Archives of microbiology, 187(5), 397-413.

Marquardt, Jürgen, and Katarzyna A. Palinska. "Genotypic and phenotypic diversity of cyanobacteria assigned to the genus Phormidium (Oscillatoriales) from different habitats and geographical sites." Archives of microbiology 187.5 (2007): 397-413.

Martiniello, P., & Iannucci, A. (1998). Genetic variability in herbage and seed yield in selected half‐sib families of berseem clover, Trifolium alexandrinum L. Plant breeding, 117(6), 559-562.

Martiniello, P., and A. Iannucci. "Genetic variability in herbage and seed yield in selected half‐sib families of berseem clover, Trifolium alexandrinum L." Plant breeding 117.6 (1998): 559-562

Megharaj, Mallavarapu, et al. "Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil." Archives of Environmental contamination and Toxicology 38.4 (2000): 439-445

Merkl, Nicole, Rainer Schultze-Kraft, and Marianela Arias. "Effect of the tropical grass Brachiaria brizantha (Hochst. ex A. Rich.) Stapf on microbial population and activity in petroleum-contaminated soil." Microbiological Research 161.1 (2006): 80-91.

Morley, Tracey A., Daniel T. Wallace, and Christopher W. Maurer. "Bipolar cauterizing instrument." U.S. Patent No. 6,840,938. 11 Jan. 2005.

Muratova, A. Yu, et al. "Phytoremediation of oil-sludge–contaminated soil." International Journal of Phytoremediation10.6 (2008): 486-502.

Paul, Alison, Peter C. Griffiths, and Philippe G. Rogueda. "Towards an understanding of adsorption behaviour in non‐aqueous systems: adsorption of poly (vinyl pyrrolidone) and poly (ethylene glycol) onto silica from 2H, 3H‐perfluoropentane." Journal of pharmacy and pharmacology 57.11 (2005): 1383-1387.

Pena-Castro, J.M., Barrera-Figueroa, B.E., Fernandez, L.L., Ruiz, M.R., & Xoconostle, C.B. (2006) Isolation & identification of upregulated genes in bermudagrass roots (Cynodon dactylon L.) grown under petroleum hydrocarbon stress, Plant Science 170, 724-731.

Reynoso-Cuevas, L., et al. "In vitro evaluation of germination and growth of five plant species on medium supplemented with hydrocarbons associated with contaminated soils." Bioresource technology 99.14 (2008): 6379-6385.

Safari, M., 2013, Study of Antimicrobial activity and Biodegradation of crude oil by Cyanobacteria , Shahid Beheshti University

Schleucher, J., Schwendinger, M., Sattler, M., Schmidt, P., Schedletzky, O., Glaser, S. J., ... & Griesinger, C. (1994). A general enhancement scheme in heteronuclear multidimensional NMR employing pulsed field gradients. Journal of biomolecular NMR, 4(2), 301-306.

Setayesh Mehr, Z., and A. Ganjali. 2013. Study the effect of drought stress on growth and physiological characteristics of Anethum graveolens. J Hort. Sci. 27: 27-35. (In Persian).

Setti L, Lanzarini G , Pifferi PG , Spuqna G. n-alkanes in a heavy oil by a pure culture of a pseudomonas sp. Chemosphere 1993 Mar; 26(6):1101-70.

Udeani, T. K. C., et al. "Isolation of bacteria from mechanic workshops’ soil environment contaminated with used engine oil." African journal of Biotechnology 8.22 (2009).

Volke- Sepulveda TL. Gutierrez – Rojas M, Favela – Torres E. Biodegradation of hexadecane in liquid and soil – state fermentations by Aspergillus niger . bioresource Technology 2003 Mar ;87(1):81-60.

Wang, Yanxu, and Hiroshi Oyaizu. "Evaluation of the phytoremediation potential of four plant species for dibenzofuran-contaminated soil." Journal of hazardous materials 168.2-3 (2009): 760-764

Wang, Z.Y., Xu,Y., Zhao, J., Li, F.M., Gao, D.M. and Xing, B.S. Remediation of petroleum contaminated soils through composting and rhizosphere degradation, Journal of Hazardous Materials, 190, pp, 677–685, 2011.

Wardlaw, Sharon L. "Hypothalamic proopiomelanocortin processing and the regulation of energy balance." European journal of pharmacology 660.1 (2011): 213-219.

Waterbury, J.B., Stanier, R.Y., 1981. Isolation and growth of cyanobacteria from marine and hypersaline environments. In: Starr, M.P., Stolp, H., Trüper, H.G., Balows, A., Schlegel, H.G. (Eds.), The Prokaryotes, vol. 1. Springer-Verlag, Berlin, pp. 221–223.

Waterbury, John B., and Roger Y. Stanier. "Isolation and growth of cyanobacteria from marine and hypersaline environments." The prokaryotes. Springer, Berlin, Heidelberg, 1981. 221-223.

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