This study investigated the use of Leucaena leucocephala as a potential plant species for phytoremediation of heavy metal contaminated soils. Tailings from the Sansu Tailings Dam was amended with top soil from Mampanhwe and three supplements to produce 10 treatments regimes.
Treatment soils of 5 kg were put in poly-pots. Each of the 10 treatments was replicated 6 times and harvesting was done twice at 45 and 75 days after transplanting. A total of 120 poly-pots were prepared. The concentrations of six heavy metals (As, Fe, Pb, Zn, Cd and Cu) were determined, in samples of shoots and roots from each harvest, using an Atomic Absorption Spectrometer.
The levels of heavy metals were highest in the roots than in the shoot. The fertilizer (NPK) and the organic manure (PKC) did not have any effect on the biomass.
In general the levels of heavy metal accumulation increased with the addition of the supplements (Chelator, PKC and NPK) and performed much better than the tailings/soil mixtures.
The results show that the plant is a phytoextractor and when aided with the addition of supplements, could be more effective in accumulation of heavy metals as a hyperaccumulator on long term cultivation.
INTRODUCTION
Background of Study
- Economic, agricultural and industrial developments are often linked to polluting the environment which include environmental soils.
- Sources of metal enrichment of soil include incinerators, fertilizers, urban compost, car exhausts, cement factories, residues from mining and smelting industries, sludge and sewage.
- Heavy metals are usually associated with pollution and toxicity although some of these elements (essential metals) are required by organisms at low concentrations (Adriano, 2001).
INTRODUCTION cont’d
- Elements associated with gold mining waste includes arsenic(As), cadmium (Cd), copper (Cu), lead (Pb), antimony (Sb) and zinc (Zn) (Ferreira da Silva et al., 2004).
- Phytoremediation involves using plants and their associated micro biota to solve pollution problems and in this case metal polluted soils.
INTRODUCTION cont’d
Justification
- Accumulation of heavy metals in the environment pose a threat to both human health and the natural environment.
- Conventional remediation technologies (solidification and stabilization, soil flushing, electrokinetics, chemical reduction/oxidation, soil washing, low temperature thermal desorption, incineration, vitrification, pneumatic fracturing, excavation/retrieval and landfill disposal) are expensive and destructive (Mellem, 2008).
- In places where the heavy metal contaminated waste are contained, according to Renault et. al (2005) wind and water can physically move tailings off-site causing contamination of adjacent areas.
INTRODUCTION cont’d
- Phytoremediation appears as a valid option since it is best suited for the remediation of these diffusely polluted areas and at much lower costs than other methods (Kumar et al., 1995).
- Lucaena leucocephala is a nitrogen fixing plant which will further add nitrogen to the soil .
- This study therefore seeks to contribute to the search for plants for phytoremediation.
INTRODUCTION cont’d
Aim and Objectives
The aim is to determine the capability of Leucaena leucocephala
in phytoremediation of heavy metal contaminated soils.
Specific Objectives
- To determine the levels of heavy metals accumulation in the Leucaena leucocephala.
- To determine the effect of inorganic fertilizer (NPK) on heavy metal accumulation by the Leucaena leucocephala.
- To determine the effect of organic manure (PKC) on enhancing phytoremediation of heavy metals by Leucaena leucocephala.
- To determine the potential of Leucaena leucocephala as a hyperaccumulator for specific heavy metals
MATERIALS AND METHODS
Study Site
Nursery and revegetation unit of AngloGold Ashanti in Obuasi
Collection of Tailings Soil Samples
Eastern part of the Sansu tailings dam. An area of 40m2 was divided into 8 equal zones and further divided into 5 subzones where 20kg of tailing soil was collected from each subzone at a depth of 30cm with soil auger. 100kg of tailing soil was collected from each zone making a total 800kg of tailing soil collected in sacks.
MATERIALS AND METHODS cont’d
Control soil was obtained from Mampanhwe. An area of 20m2 was selected and divided into 5 equal zones with each zone having an area of 4m2. 6 spots were then randomly selected from each zone and 10kg of soil was collected from each spot. The soil was taking at a depth of 40cm and a total of 300kg of top soil was taking as control.
Collection of Planting Material
Seeds were collected from the tailings dam at a distance of 130m from the dam.
Nursing and Transplanting
Nursery beds were watered each morning. Seedlings were nursed for 3 weeks at the nursery. After 3 weeks they were transplanted.
MATERIALS AND METHODS cont’d
Experimental Design
- Layout of the experiment was Randomised Complete Block Design.
- 120 poly-pots of size 8 x 10 inches were filled with 5kg of treatment soil.
- 10 treatments with each treatment replicated 6 times for the two harvest periods.
Treatments used
Treatment 1- T1 (Tailings soil alone)
Treatment 2 - T2 (Tailing soil + chelator (EDTA))
MATERIALS AND METHODS cont’d
Treatment 3 - T3 (Tailing soil + Fertilizer (NPK))
Treatment 4 - T4 (Tailing soil + Fertilizer + Chelator (EDTA)) Treatment 5 - T5 (Tailing soil + Palm kernel Cake)
Treatment 6 - T6 (Tailing soil + Palm kernel Cake + Chelator (EDTA))
Treatment 7 - T7 (Tailing soil + Topsoil) (3:2)
Treatment 8 - T8 (Tailing soil + Topsoil) (2:3)
Treatment 9 - T9 (Tailing soil + Topsoil) (1:1)
Treatment 10 - T10 (Topsoil or Control)
MATERIALS AND METHODS cont’d
- Chelator (EDTA) was prepared by dissolving 60g of EDTA salt in 500ml of distilled water. The concentration used was 0.3M of which 25 ml was added a week before harvesting to prevent loss of shoots which might be concentrated with lead. According to Larson et al., (2007), chelates should be applied within one week of treatment to avoid loss of shoots.
- Treatment with inorganic fertilizer (NPK) was prepared by dissolving 370g (equivalent to 2 full milk tins of NPK) in 6 litres of water of which 150ml was mixed with tailings.
- Treatment with organic manure (PKC) was prepared by mixing 5kg of tailing soil with 120g of palm kernel cake (PKC).
MATERIALS AND METHODS cont’d
Harvesting
- First harvest was done 45 days after transplanting and 7 days after EDTA application.
- Samples were washed with distilled water and separated into above shoots and roots.
- The final harvest was done 30 days after the first harvest.
- Treatment soils were analysed for pH and heavy metals present after each harvest.
MATERIALS AND METHODS cont’d
Soil analysis
- NPK and particle size determination for tailing soil and control soil
- pH and heavy metal (As, Fe, Pb, Zn, Cd and Cu) analysis were done for treatments before transplanting, after first harvest and after second harvest.
Plant Analysis
- Heavy metal content in shoots and roots were determined before transplanting, after first harvest and after second harvest.
- Fresh and dry weights in whole plant were determined after first and second harvest.
MATERIALS AND METHODS conťd
Accumulation ratios
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Bioaccumulation ratios
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Translocation Ratio
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Percentage Reduction
RESULTS AND DISCUSSION
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Table 1. Physichochemical parameters of soil
Physichochemical parameters of soil
According to Suttie (2005) L. leucocephala does best on deep, well drained, neutral to calcareous soils. However, it grows on a wide variety of soil types including mildly acid soils.
Low nutrient in tailings accounts for the slow growth observed
Table 2. Levels of heavy metals in treatment soils before transplanting
Mean ± SD in the same column with different letters differ significantly (p < 0.05).
RESULTS AND DISCUSSION cont’d
- As in all the treatment soils exceeded the normal concentrations allowed due the underground arsenic bearing rock called Arsenopyrite.
- Iron (Fe), Lead (Pb) and Cd exceeded the normal concentrations values in soils in all treatments.
- Zinc concentration was generally below the normal values in
soils according to the European Union Regulatory Standards.
Copper (Cu) the normal values allowed in soils according to European Union Regulatory Standards was exceeded by treatments that contained the tailings soil (T1 to T9).
RESULTS AND DISCUSSION cont’d
Fig. 1 Levels of metals in plant shoot and roots before transplanting
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RESULTS AND DISCUSSION cont’d
Table 3. Metal concentration in shoots at 1st harvest
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Mean ± SD in the same column with different letters differ significantly (p < 0.05).
Table 4. Metal concentration in shoots at 2nd harvest
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Mean ± SD in the same column with different letters differ significantly (p < 0.05).
RESULTS AND DISCUSSION cont’d
Table 5. Accumulať;ion ratios in shoots at 1st harvest
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Table 6. Accumulation ratios in shoots at 2nd harvest
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RESULTS AND DISCUSSION cont’d
- Generally the levels of metal concentrations and accumulation ratios in all the treatments increased from first to second harvest.
- The highest accumulation ratio recorded in T2 deviated from Dias et al. (2009) work which observed that As levels were relatively low in young leaves of L. leucocephala.
- The low accumulation ratio for Fe in T8 and Pb in T5 at the first harvest could be as a result of the fact that the treatment could not support their accumulations in the shoot earlier.
The low accumulation of Cu for T3, T5, T7, T8, T9 and T10 for the first harvest and T7 and T8 for the second harvest could be due to its slow Cu accumulation in shoots.
Table 7. Metal concentrati ratios in roots at lst harvest
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Table 8. Metal concentrationti atiou in roots at 2nd harvest
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RESULTS AND DISCUSSION cont’d
Tabe 9. Accumulation ratio in roots at 1st harvest
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Table 10. Accumulati°n ratio in roots at 2nd harvest
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RESULTS AND DISCUSSION cont’d
- The plant generally accumulated the metals in roots than the shoots.
- According to Dias et al. (2009), the highest As concentration in L. Leucocephala are mostly found in the roots and the study attested to that fact.
- The study confirms the proposition made by Saraswat and Rai (2011) that the plant accumulates Zn and Cd mostly in its roots.
RESULTS AND DISCUSSION cont’d
Table 11. Metals ac-c'uniulaUHl in whole plant at 1st harvest
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Table 12. Metals accumulated in whole plant at 2nd harvest
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RESULTS AND DISCUSSION cont’d
Table 13. Translocation factor at 1st harvest
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Table 13. Translocation factor at 2nd harvest
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RESULTS AND DISCUSSION cont’d
- Generally the plant was not translocating most metals to the shoots including those with chelator (T2 and T6).
- But it maintained high concentrations in the roots of most of the treatments. It can therefore be employed in regenerating heavy contaminated soils according to Baker (1981).
- Since the translocation ratios increased from first to second harvest, they could have increased if the duration for study was longer.
RESULTS AND DISCUSSION cont’d
Table 14. Bioaccuinulatiou factor at lst harvest
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Table 15. Bioaccuinulatiou factor at 2ml harvest
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RESULTS AND DISCUSSION cont’d
- Based on the duration and conditions provided for the study due to the bioaccumulation ratios, the plant can be said to be a poor hyperaccumulator of heavy metals.
- Gardezi et al. (2008) indicated that work done with L. leucocephala should be given a minimum of 1 year to allow the plant to mature so that it can reach its bioaccumulation capacity.
- Though the plant did not perform well based on its bioaccumulation factor, it was able to tolerate the tailing soil which is highly polluted with heavy metals and accumulated some biomass.
RESULTS AND DISCUSSION cont’d
Effect of Fertilizer (NPK) on Metal Concentration in Plants
- The effect of the fertilizer in increasing the biomass of the plant was not seen when compared to the control. The same incident of low biomass was also experienced by Aziz (2011).
- The accumulation ratios were all greater than 1 (>1).
- T4 performed better when it comes to the number of metals it was able to accumulate than T3 in the whole plant which could be due to the chelator.
- The biomass could have probably highly increased during the second harvest if fertilizer application was not one time and may be, the quantity varied.
RESULTS AND DISCUSSION cont’d
Effect of Chelator on Metal Concentration in Plants
- The accumulation ratios in all the treatments that had chelator added were greater than 1 (>1) in both harvests.
- Arsenic (As) was having the highest accumulation ratios in all the treatments for both harvests but the concentration was mostly in the roots except in T4.
- This could be attributed to the time of adding the chelator so if it had been added earlier it could have aided in translocating the metal to the shoots and also biomass was not enough.
- But because of the toxic effects, it is recommended that chelates should be applied only after a maximum amount of plant biomass has been produced and prompt harvesting (within one week of treatment) is required to minimize the loss of Pbladen shoots (Larson et al., 2007; Aziz, 2011).
RESULTS AND DISCUSSION cont’d
Effect of Palm Kernel Cake on Metal Concentration in Plants
- The idea behind the addition of the PKC was to help the plant to increase its biomass and also accumulation of the metals but the biomass obtained was less than the control.
- It could be due to the one time application and also the quantity which might not be enough. According to Kolade et al. (2005) PKC should be converted into compost and applied 4t/ha to obtain yields comparable to those of organo-minerals fertilizer and chemical fertilizers.
- The accumulation ratios for the treatments that contained PKC for both harvests in the whole plant were greater than 1 (>1). T5 was able to accumulate As, Fe and Pb higher than T6 which was able to accumulate Zn, Cd and Cu.
CONCLUSION AND RECOMMENDATION
Conclusion
- To survive high concentrations of heavy metals in soils, plants can either stabilize metal contaminants in the soil through avoidance or can take up the contaminants into their cellular structure by tolerating them as was described by other researchers. And so far as the plant has been able to grow and accumulate biomass and tolerate these metals, it has the capacity to take up the metals as well as tolerate the stress they gave it.
- The plant could also be seen to have slow growth due to the conditions that it found itself but when if the duration of study had increased it could have performed wonderfully. Gardezi et al. (2008) indicated that work done with L. Leucocephala should be given a minimum of 1 year to allow the plant to mature so that it can reach its bioaccumulation capacity
CONCLUSION AND RECOMMENDATION
Recommendations
- Duration of the study was not enough and future studies should consider at least one year as it has been talked about by other researchers.
- Chelator such as EDTA should be used for such a study by varying the quantity and also the time of addition to see its effect.
- Fertilizer (NPK) and PKC were added at once during the treatment preparation stage and this did not help in accumulating enough biomass which could be due to the quantities added or the one time application so future further studies should vary their quantities and also further subsequent additions with time.
REFERENCES
- Adriano, D.C. (2001). Cadmium. In Adriano D.C. (Ed.), Trace elements in terrestrial environments, biogeochemistry, bioavailability, and risks of metals. 2nd edition, Springer-Verlag, New York, pp. 264-314.
- Ferreira da Silva, E., Zhang, C., Serrano Pinto, L., Patinha, C. and Reis, P. (2004). Hazard assessment on arsenic and lead in soils of Castromil gold mining area, Portugal. Applied Geochemistry, 19(6): 887-898
- Mellem, J.J. (2010). Phytoremediation of heavy metals using Amaranthus dubius, Theses submitted to Department of Biotechnology and Food Technology, Durban University of Technology, Durban, South Africa.
- Renault, S. and Green, S. (2005). Phytoremediation and revegetation of mine tailings and bio-ore production: effects of paper mill sludge on plant growth in tailings from Central Manitoba (Au) minesite (NTS 52L13); In: Report of Activities 2005, Manitoba Industry, Economic Development and Mines, Manitoba Geological Survey, p. 167-169.
- Larson, S.L., Teeter, C. L., Medina, V.F. and Martin, W.A. (2007). Environmental Quality and Technology Program Treatment and Management of Closed or Inactive Small Arms Firing Ranges. US Army Kumar, N., Dushenkov, V, Motto, H. and Raskin, I. (1995). Phytoextraction: the use of plants to remove heavy metals from soils. Environmental Science and Technology 29.
Sutie, J.M. (2005). Food and Agriculture Organization (FAO), Leucaena leucocephala (Lam.) de Wit. http://www.fao.org/ag/AGP/AGPC/doc/Gbase/DATA/Pf000158.htm 23/06/2005 10:34:22
Dias, L.E., Melo, R.E., Vargas de Melo, J.W., Oliveira, J.A. and Daniels, W.L. (2010). Potential of three legume species for phytoremediation of Arsenic contaminated soils.Soil Dep. Universidade Federal de Viçosa - UFV, 36571-000, Viçosa-MG Brazil.
Saraswat S. and Rai J.P. (2011). Prospective application of Leucaena leucocephala for phytoextraction of Cd and Zn and nitrogen fixation in metal polluted soils. Ecotechnology Laboratory, Department of Environmental Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, India
Baker, A.J.M. (1981). Accumulators and excluders - Strategies in the response of plants to heavy metals, J. Plant Nutr. 3(1-4): 643-654.
Aziz, F. (2011). Phytoremediation of Heavy Metal Contaminated Soil Using Chromolaena odorata and Lantana camara. Master’s Thesis for the Award of MSc. Environmental Science. Department of Theoretical and Applied Biology, KNUST Kumasi.
Gardezi, A.K., Barceló, I.D., García, A.E., Saenz, E.M., Saavedra, U.L., Sergio R., Márquez, B., Verduzco, C.E., Gardezi, H. and Talevera-Magaña, D. T. (2008). Cu2 + Bioaccumulation by Leucaena leucocephala in symbiosis with Glomus spp. and Rhizobium in Copper-containing soil.Colegio de Postgraduados. Instituto de Recursos Naturales, Programa Hidrociencias. Montecillo, Texcoco, Edo. de México
Frequently Asked Questions About Leucaena leucocephala and Phytoremediation of Heavy Metals
What is the background of this study?
This study investigates phytoremediation, using plants to solve pollution problems, specifically focusing on heavy metal polluted soils. Economic, agricultural, and industrial developments often lead to environmental pollution, including metal enrichment of soil from sources like incinerators, fertilizers, mining residues, and sewage. Heavy metals, while sometimes essential in low concentrations, are generally associated with pollution and toxicity. Elements associated with gold mining waste include arsenic (As), cadmium (Cd), copper (Cu), lead (Pb), antimony (Sb) and zinc (Zn). Phytoremediation offers a potentially cost-effective solution for remediating these polluted soils.
What is the justification for this study?
Accumulation of heavy metals poses a threat to both human health and the environment. Conventional remediation technologies are expensive and often destructive. Phytoremediation, particularly using Leucaena leucocephala, presents a valid alternative because it's suitable for diffusely polluted areas and is cost-effective. Leucaena leucocephala also fixes nitrogen, improving soil quality. This study aims to contribute to the search for effective phytoremediation plants.
What are the aims and objectives of the study?
The aim is to determine the capability of Leucaena leucocephala in phytoremediation of heavy metal contaminated soils. Specific objectives include:
- Determining heavy metal accumulation levels in Leucaena leucocephala.
- Determining the effect of inorganic fertilizer (NPK) on heavy metal accumulation.
- Determining the effect of organic manure (PKC) on enhancing phytoremediation.
- Determining the potential of Leucaena leucocephala as a hyperaccumulator for specific heavy metals.
Where was the study conducted, and how were samples collected?
The study was conducted at the nursery and revegetation unit of AngloGold Ashanti in Obuasi. Tailings soil samples were collected from the eastern part of the Sansu tailings dam. The area was divided into zones and subzones, with soil collected at a depth of 30cm. Control soil was obtained from Mampanhwe, with samples taken at a depth of 40cm. Seeds of Leucaena leucocephala were collected from the tailings dam area.
What was the experimental design and what were the treatments used?
The experimental design was a Randomized Complete Block Design using 120 poly-pots filled with treatment soil. Ten treatments were used, each replicated six times for two harvest periods. The treatments were:
- T1: Tailings soil alone
- T2: Tailing soil + chelator (EDTA)
- T3: Tailing soil + Fertilizer (NPK)
- T4: Tailing soil + Fertilizer + Chelator (EDTA)
- T5: Tailing soil + Palm kernel Cake
- T6: Tailing soil + Palm kernel Cake + Chelator (EDTA)
- T7: Tailing soil + Topsoil (3:2)
- T8: Tailing soil + Topsoil (2:3)
- T9: Tailing soil + Topsoil (1:1)
- T10: Topsoil or Control
How were the chelator, fertilizer, and organic manure treatments prepared?
Chelator (EDTA) was prepared by dissolving 60g of EDTA salt in 500ml of distilled water (0.3M), with 25 ml added a week before harvesting. Inorganic fertilizer (NPK) was prepared by dissolving 370g in 6 liters of water, with 150ml mixed with tailings. Organic manure (PKC) was prepared by mixing 5kg of tailing soil with 120g of palm kernel cake.
How was harvesting done and what analyses were performed?
The first harvest was done 45 days after transplanting, 7 days after EDTA application. The final harvest was done 30 days after the first harvest. Samples were separated into shoots and roots. Treatment soils were analyzed for pH and heavy metals after each harvest. Soil analysis included NPK and particle size determination. Plant analysis included heavy metal content in shoots and roots, and fresh and dry weights.
What were the key results regarding soil characteristics?
The tailings soil had low nutrient content, likely contributing to the slow growth observed. Arsenic (As), Iron (Fe), Lead (Pb) and Cadmium (Cd) levels exceeded normal concentrations in all treatment soils. Zinc (Zn) concentration was generally below normal values, while Copper (Cu) exceeded normal values in treatments containing tailings soil.
How did the plant accumulate metals in shoots and roots?
Generally, the levels of metal concentrations and accumulation ratios in all the treatments increased from first to second harvest. The plant generally accumulated more metals in the roots than in the shoots. Arsenic concentration was highest in the roots, confirming findings from other studies. The plant also accumulated Zinc and Cadmium mostly in its roots.
What was the effect of Fertilizer (NPK) on Metal Concentration in Plants?
The fertilizer did not significantly increase the biomass of the plant compared to the control. The accumulation ratios were generally greater than 1 (>1). Treatment 4 (Tailing soil + Fertilizer + Chelator (EDTA)) performed better than T3 (Tailing soil + Fertilizer (NPK)) in terms of the number of metals accumulated in the whole plant, potentially due to the chelator.
What was the effect of Chelator on Metal Concentration in Plants?
The accumulation ratios in all treatments that had chelator added were greater than 1 (>1) in both harvests. Arsenic (As) had the highest accumulation ratios in all treatments for both harvests, but the concentration was mostly in the roots except in T4. It is recommended that chelates should be applied only after a maximum amount of plant biomass has been produced and prompt harvesting (within one week of treatment) is required to minimize the loss of Pbladen shoots.
What was the effect of Palm Kernel Cake on Metal Concentration in Plants?
The biomass obtained with PKC was less than the control. The accumulation ratios for the treatments that contained PKC for both harvests in the whole plant were greater than 1 (>1). T5 (Tailing soil + Palm kernel Cake) accumulated As, Fe, and Pb higher than T6 (Tailing soil + Palm kernel Cake + Chelator (EDTA)), which accumulated Zn, Cd, and Cu higher.
What are the conclusions of the study?
To survive high concentrations of heavy metals in soils, plants can either stabilize metal contaminants in the soil through avoidance or can take up the contaminants into their cellular structure by tolerating them. Leucaena leucocephala was able to grow, accumulate biomass, and tolerate the metals, indicating its capacity for metal uptake and stress tolerance. The plant showed slow growth due to the conditions, suggesting that a longer duration of study could yield better results. Based on the study due to the bioaccumulation ratios, the plant can be said to be a poor hyperaccumulator of heavy metals. The plant can be said to be a poor hyperaccumulator of heavy metals, according to Gardezi et al. (2008) that works done with L. leucocephala should be given a minimum of 1 year to allow the plant to mature so that it can reach its bioaccumulation capacity. The study can also conclude that this plant was also able to accumulate the metals in roots than the shoots.
What are the recommendations for future research?
Recommendations include:
- Extending the duration of the study to at least one year.
- Varying the quantity and timing of chelator (EDTA) application to assess its effect.
- Modifying the quantity and timing of fertilizer (NPK) and PKC additions to optimize biomass accumulation.
- Quote paper
- Yaw Boateng (Author), 2014, Phytoremediation of Heavy Metal Contaminated Soil Using Leucaena Leucocephala, Munich, GRIN Verlag, https://www.grin.com/document/283856