Document Type : Research Paper


1 M.Sc. Graduate of Horticultural Science, Department of Horticultural Science, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran

2 Assistant Professor, Department of Horticultural Science, Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran


Background and Objectives
Among the major environmental pollutants, lead is the most substantial contaminant due to causing toxicity in plants and organisms. Recently, the remediation of these metals has been considered in plants. Plant in vitro culture is a key tool in phytoremediation research. In general, Plant tissue culture and in vitro selection techniques are used to increase the tolerance and the accumulation of heavy metals which have been reported in numerous plant species and populations. The application of ornamental plants for phytoremediation in metal-contaminated soil is a new alternative. Ornamental kale is an important bedding plant in many landscapes of cold temperate regions. The present study was aimed to evaluate the Pb resistance and remediation of Brassica OleraceaVar. Acephala affected by in vitro culture as a bedding plant. This experiment was carried out in 2016 at Arak University.
Materials and Methods
In this study, the stalk explants of Brassica OleraceaVar. Acephala was used for callus induction. Callus and plantlet were exposed to different concentrations of lead in the media. The Callus induction medium was MS (Murashige and Skoog) supplemented with 0.1 mgL-1 2,4-D and BA. The regeneration medium was MS medium supplemented with 0.5 mgL-1 NAA in combination with 1 mgL-1 BA. The rooting medium was half-strength MS medium without plant growth regulators and sucrose. All of the media containing 30 gL-1 sucrose, 7 gL-1 agar, and pH were adjusted to 5.8. All of the regeneration stages from callus induction to rooting and the acclimatization stages were done by different concentration of Pb(NO3)2 (0, 10, 25, and 50 mgL-1). The traits, such as callus browning, fresh and dry weight, callus formation, survival rate, root and shoot length, leaf number, and the lead accumulation, were evaluated during various stages of callus regeneration and rooting in plantlets. A Completely Randomized Design (CRD) arrangement with three replications was used in this experiment. Data were analyzed using the ANOVA procedure of SAS statistical software (version 9.2).
Based on the results, not only did different concentrations of lead not have the toxic effects but also they had a stimulation effect in some concentrations on callus growth and regeneration. Lead concentrations increased leaf number and length of shoots in the regenerated ornamental kale. Lead accumulation in cultures was increased by increasing the concentration of lead in the medium at all stages of the regeneration. The highest accumulation of Pb was obtained 2395.7 ppb in callus treated by 25 mg L-1 Pb in media.
The results of the present study showed the occurrence probability of the somaclonal variation due to 2,4-D supplemented in the media. In the regenerated plant, results revealed that the growth traits and Pb uptake were affected by Pb concentrations. Some in vitro derived plantlets showed an increase in uptake potential of lead in their organs (6.1 to 2.7 times compared to the control) and that it is remarkable and suitable for phytoremediation studies.


Main Subjects

Akhtar, S., Niaz, M., Sajid-ur-Rahman, M. Y. and Zaffar, M. (2012). Somaclonal variation for development of salt tolerance in selected wheat (Triticum aestivum) cultivars. International Journal of Agriculture and Biology, 14(4), 600-604.
Alkorta, I. and Garbisu, C. (2001). Phytoremediation of organic contaminants. Bioresource Technology, 79(3), 273-276.
Al-Zahim, M., Ford, B. and Newbury, H. (1999). Detection of somaclonal variation in garlic (Allium sativum L.) using RAPD and cytological analysis. Plant cell Reports, 18(6), 473-477.
Ashwini, A., Waoo, S. and Sujata, G. (2014). Toxic effect of different lead concentrations on in-vitro culture of Datura inoxia. Journal of Scientific and Innovative Research, 3(5), 532-535.
Bojarczuk, K. (2004). Effect of toxic metals on the development of poplar (Populus tremula L. × P. alba L.) cultured in vitro. Polish Journal of Environmental Studies, 13(2), 115 120.
Doran, P. M. (2009). Application of Plant Tissue Cultures in Phytoremediation Research: Incentives and Limitations. Biotechnology and Bioengineering, 103(1), 60-76
Evans, D. A., Sharp, W. R. and Medina-Filho, A. P. (1984). Somaclonal variation and gametoclonal variation. American Journal of Botany, 71(6), 759-774.
Fassler, E., Evangelou, M. W., Robinson, B. H. and Schulin R. (2010). Effects of indole-3-acetic acid (IAA) on sunflower growth and heavy metal uptake in combination with ethylene diamine disuccinic acid (EDDS). Chemosphere, 80(8), 901-907.
Ghahreman, A. (1993). Plant systematic (2 Vol.). Tehran: Tehran University Press. [In Farsi]
Karimi, M., Taghizadeh, M., Sanati, M. H. and Solgi, M. (2015). Effect of salicylic acid and variety on seed priming of ornamental cabbage (Brassica oleracea var. acephala) under heavy metal stress. Second National Conference on conservation planning, environmental protection and sustainable development, Shahid Beheshti University, Tehran. [In Farsi]
Keaton, C. M. (1937). The influence of lead compounds on the growth of barley. Soil Science, 43(6), 401-411.
Larkin, P. J. and Scowcroft, W. R. (1988). Somaclonal variation- a novel source of variability from cell cultures for plant improvement. Theoretical and Applied Genetic, 60, 197-214.
Lone, M. I., Li, H., Zhen, P. J., Stoffella, E. and Yang, X. (2008). Phytoremediation of heavy metal polluted soils and water: Progresses and perspectives. Journal of Zhejiang University Science, 9(3), 210-220.
Morcillo, F., Gagneur, C., Adam, H., Richard, F. Singh, R., Cheah, S. C., Rival, A. Duval, Y. and Tregear, J. W. (2006). Somaclonal variation in micropropagated oil palm: Characterisation two novel genes with enhanced expression in epigenetically abnormal cell lines and in response to auxin. Tree Physiology, 26(5), 585-594.
Najeeb, U., Ahmad, W., Zia, M. H., Malik, Z., Zhou, W. (2017). Enhancing the lead phytostabilization in wetland plant Juncus effusus L. through somaclonal manipulation and EDTA enrichment. Arabian Journal of Chemistry, 10, 3310-3317.
Nehnevajova, E., Herzig, R., Erismann, K. H. and Schwitzguebel, J. P. (2007). In vitro breeding of Brassica juncea L. to enhance metal accumulation and extraction properties. Plant Cell Report, 26(4), 429-437.
Rahmani, H. R., Kalbasi, M. S. and Haj, R. (2000). Plant pollution by lead from automobile around the some of Iran highways. Journal of Environmental Studies, 26(26), 76-83.
Sharafi, Y. (2017). Effects of some heavy metals (Copper and Lead) on pollen germination and tube growth of some cherry (Pruons avium).Plant Productions, 39(4), 79-86.
Sharma, S. (2009). Study on impact of heavy metal accumulation in Brachythecium populeum (Hedw.) Ecological Indicators, 9(4), 807-811.
Sharma, S., Chatterjee, S., Datta, S., Mitra A., Vairale, M., Veer, V., Chourasia, A. and Gupta, D. K. (2014). In vitro selection of plants for the removal of toxic metals from contaminated soil: Role of genetic variation in phytoremediation. In D. K. Gupta, S. Chatterjee (Ed.), Heavy metal remediation: Transport and accumulation in plants (pp. 155-178). USA: Nova Science Publishers, Inc.
Strubinska, J. and Sniezko, R. (2002). Lead tolerance and the interactive effects of lead and IAA on direct root regeneration in the sunflower. Cellular and Molecular Biology Letters, 7:2002.
Taghizadeh, M. (2010). Assessment of turfgrass potential for Lead phytoremediation, In Vitrocally Inducing and Molecular tracing. Ph.D. Thesis of Horticultural Science, Tehran University, Tehran. [In Farsi]
Taghizadeh, M., Kafi, M. and Fattahi Moghadam, M. R. (2015). Breeding by In vitro Culture to Improve Tolerance and Accumulation of Lead in Cynodon Dactylon L. Journal Agriculture Science and Technology, 17, 1851-1860.
Taghizadeh, M., Mohtadi, A. and Asemaneh, T. (2017). Investigating of copper effect on growth and physiological characteristics of nasturtium officinale. Plant Productions, 39(4), 101-114.
Tu, C., Ma, L. Q. and Bondada, B. (2002). Arsenic accumulation in the hyperaccumulator chinese brake and its utilization potential for phytoremediation. Journal Environmental Quality, 31(5), 1671-1675.