See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/340870476

Evaluation of phytoremediation potential of Moringa oleifera and Moringa

stenopetala when grown in polluted soil with and without coal fly ash.

Article · April 2020

DOI: 10.12692/ijb/16.4.136-151

CITATIONS READS

2 810

4 authors:

Raviro Vurayai Baleseng Moseki

University of Botswana University of Botswana
6 PUBLICATIONS 129 CITATIONS 17 PUBLICATIONS 212 CITATIONS

SEE PROFILE SEE PROFILE

Bonang Nkoane Padmaja Chaturvedi

University of Botswana University of Botswana
12 PUBLICATIONS 120 CITATIONS 28 PUBLICATIONS 673 CITATIONS

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

ASSESSMENT OF PHYTOREMEDIATION POTENTIAL OF PLANTS: A CASE OF BCL COPPER/NICKEL MINE, SELIBE-PHIKWE, BOTSWANA View project

RPC-BCA View project

All content following this page was uploaded by Raviro Vurayai on 23 April 2020.

The user has requested enhancement of the downloaded file.

 Int. J. Biosci. 2020

International Journal of Biosciences | IJB |

ISSN: 2220-6655 (Print), 2222-5234 (Online)
http://www.innspub.net
Vol. 16, No. 4, p. 136-151, 2020

RESEARCH PAPER OPEN ACCESS

Evaluation of phytoremediation potential of Moringa oleifera

and Moringa stenopetala when grown in polluted soil with and
without coal fly ash

Raviro Vurayai1*, Baleseng Moseki1, Bonang Nkoane2, Padmaja Chaturvedi 1

1
Department of Biological Sciences, University of Botswana, Gaborone, Botswana
2
Department of Chemistry, University of Botswana, Gaborone, Botswana

Key words: Heavy metals, Phytoremediation, Coal fly ash, Moringa oleifera, Moringa stenopetala.

http://dx.doi.org/10.12692/ijb/16.4.136-151 Article published on April 14, 2020

Abstract
Phytoremediation potential of Moringa oleifera and Moringa stenopetala on polluted lands of east and west of
Bamangwato Concessions Limited Cu/Ni mine smelter, Selebi-Phikwe, Botswana was evaluated. Plants were
raised in greenhouse (pots) in soils collected 2.5 km east, 2.5 km west, 20 km west, and 55 km west (control) of
the mine smelter, which were supplemented with and without coal fly ash. Without ash, both species did not
survive in soils collected 2.5 and 20 km west. In soils from 2.5 km east, soil acidity and heavy metal stress
reduced vegetative growth and total dry weight. Both species accumulated more metals in roots than in shoots,
and failed to hyperaccumulate any metal. In soils collected 2.5 and 20 km west, coal fly ash enabled plants to
survive, reduced accumulation of majority of heavy metals and increased vegetative growth and total dry weight.
Heavy metal accumulation, translocation and bioaccumulation factors of plants grown in soils with ash generally
followed the order 55 km west> 2.5 km east> 20 km west> 2.5 km west. Moringa oleifera and Moringa
stenopetala are not good candidates for phytoextraction of heavy metals east and west of mine smelter, but have
a potential for phytostabilisation with the help of coal fly ash.
* Corresponding Author: Raviro Vurayai  [email protected]

136 Vurayai et al.

 Int. J. Biosci. 2020

Introduction phytoremediation technologies uses many

The occurrence of Cu-, Ni- and Zn- sulphidic ores in mechanisms (phytoextraction, phytostabilisation,
Selebi-Phikwe, Botswana (Barrie et al., 1993) and rhizofiltration, and phytovolatilization) and one of
their mining and smelting by the Bamangwato them is phytoextraction. The metals are taken up by
Concessions Limited (BCL) Cu/Ni mine has resulted the root system and transported to the stems and
in soil pollution. Soils around mine are enriched with leaves (phytoextraction) without showing a toxicity
heavy metals and have very low pH (Ekosse et al., syndrome (Cardwell et al., 2002; Chatterjee and
2003; 2004; Schippers et al., 2007, Vurayai et al., Chatterjee, 2000) thereby reducing the concentration
2015). The soils are contaminated through deposition of metals in contaminated soils to regulatory levels
of particulate air matter (PAM), wash-offs, and other within a reasonable time frame. Phytoextraction
wastes due to mining and smelting activities (Ekosse seems to be the most promising technique and has
et al., 2005; Ekosse and Ngole, 2012 and Likuku et received increasing attention from researchers since it
al., 2013). The mineral contents identified in the PAM was proposed by Chaney (1983). It depends on the
in Selebi-Phikwe are reflective of acidic nature and ability of plants to grow and accumulate metals in
these minerals combine with H2S, SO2 and harvestable tissues. The success of phytoextraction
atmospheric moisture to release H2SO4. These depends on the identification and selection of plants
reactions result in acid rain which lowers the pH of with exceptional, natural metal-accumulating
the soil and destroys vegetation. Vegetation around capacity: the so-called hyperaccumulators. Screening
the mine, mainly on the western side, was depleted and selection of plants species with superior
resulting on formation of dead zones (Ekosse et al., remediation properties is therefore very important
2005). (Prasad and Freitas, 2003).

Heavy metals cannot be chemically degraded and Another phytoremediation mechanism which can be
hence persist in the soil for thousands of years and used in areas around the BCL Cu/Ni mine is
pose numerous health dangers to higher organisms phytostabilisation. Phytostabilisation is the use of
(Roy et al., 2005) and so the remediation and plants to immobilize the contaminants in the soil and
revegetation of areas around the mine appears to be groundwater through absorption and accumulation in
the most suitable method for long term land plant tissues, adsorption onto roots, or precipitation
reclamation. Low-cost and ecologically sustainable within the root zone preventing their migration in
remedial options are required to restore soil, as well as their movement by erosion and
contaminated lands and due to the expensive nature deflation (Erdei, 2005; Erakhrumen and Agbontalor,
of conventional remediation methods of heavy metals 2007). Phytostabilization is a less invasive, low-cost
(Danh, 2009), biological treatments which employs phytotechnology which in combination with
the use of living organisms to remove heavy metals amendments (i.e. aided phytostabilisation) has a
from soil are continuously being researched. potential to restore the physical, chemical, and
Phytoremediation has thus become an effective and biological properties of polluted soils (Bolan et al.,
affordable technological solution used to extract and 2003; Kumpiene et al., 2008). Amendments such as
remove heavy metals from the soil. coal fly ash (alkaline) can improve phytostabilisation
(Bolan et al., 2003; Mench et al., 2010) by increasing
Phytoremediation is a process that uses plants to soil pH of acidic soils around mines. A lower soil pH
remove, transfer, or stabilize contaminants in soil, and redox potential enhances the mobility of most
sediment, and ground water (Yu and Gu, 2007). It is metals, rendering them more bioavailable (Mench et
also called green remediation, botano-remediation, al., 2003) and coal fly ash increases soil pH
agroremediation, or vegetative remediation (Pivetz, immobilizing heavy metals thus reducing their
2001). Heavy metal uptake by plants through availability to plants (Polat et al., 2002; Su and Wong,

137 Vurayai et al.

 Int. J. Biosci. 2020

2004; Tsang et al., 2014). This enables plants to Plant growth

survive, vegetate the area and stabilize the M. oleifera and M. stenopetala seeds were
contaminants in the soil. germinated at an average temperature of 28°c in
trays. After radical and apical meristems emerged,
The tropical plants of the family of Moringaceae are seedlings were transplanted onto the nursery bed
used as a source of water purifying agents in with potting soils and allowed to grow. After two
developing countries. Seeds of Moringa oleifera are weeks, healthy seedlings were selected as
used to detoxify water by absorbing heavy metals like experimental plants, and transplanted into polyvinyl
Cu, Pb, Cd, Cr, and Zn (Sajid et al., 2005; Nand, chloride (PVC) plastic pots (210 × 230 mm). The pots
2012). Seed powder of Moringa is considered as were filled with 17 kg of four different soils described
capable of absorbing Cd and Pd from polluted water above. The 7.5% coal fly ash was added per kg of dry
(Mataka et al., 2006; Mataka et al., 2010). Other soil. The experiment was arranged in a 2 × 2 × 4
studies have shown that M. oleifera (Amadi and factorial design in a completely randomized block
Tanee, 2014; Offor et al., 2014) and Moringa design with 5 replications. There were 3 factors: Plant
stenopetala (Yimer and Khan, 2016) accumulated species (M. oleifera and M. stenopetala), fly ash (with
heavy metals. Both species are characterized by high and without fly ash) and distance from the mine
biomass yield and can tolerate unfavourable smelter (2.5 km east, 2.5 km west, 20 km west and 55
environmental conditions. They are drought tolerant km west-control). The planted pots were spaced 30
and are found in locations with as little as 500 mm cm apart to preclude competition effects among
annual rainfall and can be grown in a wide range of treatments, and the plants were watered to reach
soils. They also tolerate light frost and require little 100% plant available water (Rosenthal et al., 1987)
care after planting (Bosch, 2004; Jiru et al., 2006). every 2 days. Plants were also watered with half
This makes them most suitable for use as strength Hoagland solution (Hoagland and Arnon,
phytoremedial plants in the semi-arid region of 1950 ) once every week and the experiment ran for 90
Selebi-Phikwe. days.

The aims of the presented study was (i) to determine Growth measurements
the morphological responses of M. oleifera and M. The total number of leaves was recorded 90 days after
stenopetala when grown in acidic and heavy metal seedling transplanting per species per treatment per
contaminated soil, (ii) to determine the heavy metal replication. Stem diameter was measured 20 cm from
decontaminative capacity of M. oleifera and M. the soil surface with a Vernier calipers. Plant height
stenopetala and (iii) to investigate use of coal fly ash was determined by measuring the distance from soil
in the remediation of polluted soil collected east and level to the top of plants with a millimetre ruler. For
west of the BCL Cu/Ni mine smelter in Selebi-Phikwe, total dry weight measurement, plants were harvested
Botswana. (uprooted) 90 days after seedling transplantation,
and washed with tap water to remove soil. The
Materials and methods harvested tissues were allowed to dry in an oven to
Soil and coal fly ash collection constant weight at 60°C, and their total dry weights
Soil was collected from four different sites east and were measured with an electric balance.
west of the BCL Cu/Ni mine in Selebi-Phikwe,
Botswana: 2.5 km east, 2.5 km west, 20 km west and Plant and soil heavy metal analyses
55 km west of the BCL Cu/Ni mine smelter as Heavy metal content in soil after harvest was
described by Vurayai et al. (2015). Coal fly ash was measured as described by Vurayai et al. (2015), while
collected from ash piles at Morupule power station, heavy metal content in plants was measured as
Palapye, Botswana. described by Vurayai et al. (2017).

138 Vurayai et al.

 Int. J. Biosci. 2020

Bioaccumulation factor TF>1 signifies that the plant effectively translocate

Bioaccumulation factor (BAF) of heavy metal was heavy metals from roots to shoots.
calculated according to Baker (1981) and Ma et al.
(2001). Statistical analysis
All the above experiments were repeated twice and
BAF = Cb/Cs pooled data is presented. Statistical analyses were
Where performed by ANOVA using IBM SPSS Statistics 22.
Cb = heavy metal concentration in shoots (mg/kg) Treatment means were compared using LSD at
Cs = heavy metal concentration in soil (mg/kg) probability level of 0.05.
BAF is categorized as excluder (< 1), accumulator (1-
10) and hyperaccumulater (>10). Results and discussion
Plant growth
Translocation factor M. oleifera and M. stenopetala did not survive in soil
Heavy metal translocation from root to shoot in collected from 2.5 km west and 20 km west of the
plants was calculated using the formulae by Baker mine smelter (Fig. 1 and 2). This might be attributed
and Brooks (1989). to an inability of the plants to overcome heavy metal
toxicity and soil acidity. Vurayai et al. (2015)
Translocation factor (TF) = Cs/Cr indicated that the soil is polluted with heavy metals
Where and also has a low soil pH of 3.36 (2.5 km west) and
Cs = metal concentration in shoot (mg/kg) 4.3 (20 km west). Moringa is said to grow well in soil
Cr = metal concentration in roots (mg/kg) pH between 6 and 7 (Jiru et al., 2006).

Table 1. Ionic concentration (mg/kg Dw-1) of roots and shoots of M. oleifera. The % RSD for all metals was <
4.58 % (n=5).
Distance from Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
Mine smelter
2.5km East With ash Roots 2.3 156 1.44 1.1 0.94 0.09 0.8 2.99 0.15 0.95 2.1 0.6 0.11 0.24 0.28
Shoots 1.2 393 3.37 1.42 0.6 0.04 0.51 1.69 0.09 0.2 1.31 0 0.05 0.11 0.16
Without Roots 5.38 540 4 1.86 1.1 0.47 1.2 4.7 0.37 1.87 3.1 0.99 0.32 0.46 0.43
ash Shoots 3.8 341 2.87 0.87 0.78 0.33 0.91 3.24 0.31 0.41 2.3 0 0.21 0.3 0.3
55km West With ash Roots 3.53 202 2.2 1.29 1.1 0.14 1.02 3.3 0.27 1.12 2.78 0.9 0.17 0.32 0.43
Shoots 2.24 590 4.07 1.85 0.8 0.08 0.74 2.3 0.19 0.24 1.92 0 0.09 0.23 0.28
Without Roots 6.58 607 4.2 2.52 1.2 0.83 1.48 4.85 0.85 2.05 3.33 1.16 0.58 0.48 0.57
ash Shoots 5.39 492 3.99 1.36 1.1 0.76 1.33 4.02 0.68 0.72 3.13 0 0.45 0.39 0.48
20km West With ash Roots 1.94 122 1.1 0.98 0.83 0.04 0.24 2.2 0.14 0.82 1.95 0.47 0.08 0.2 0.3
Shoots 0.78 227 2.2 1.07 0.56 0.2 0.11 0.97 0.06 0.07 1.04 0 0.02 0.08 0.09
2.5km West With ash Roots 1.3 94 0.9 0.42 0.9 0.04 0.28 1.7 0.09 0.46 1.85 0.29 0.05 0.09 0.2

Shoots 0.42 120 1.2 0.58 0.44 0.01 0.03 0.41 0.02 0.03 0.79 0 0 0.02 0.04

In general, plant growth is limited in acid soils by a (Mench et al., 2003). If in excess, heavy metals affect
variety of factors including direct effect of pH (excess various physiological processes, ultimately causing
H+ concentration), which if in excess causes cell death cell and plant death (Shahid et al., 2015).
and consequent plant death. Moreover, the soil acidic
pH induces heavy metal toxicity and causes plant Both M. oleifera and M. stenopetala were able to
death, since low soil pH enhances the mobility of grow in soil with coal fly ash (Figs 1-4). Coal fly ash
heavy metals, rendering them more bioavailable used in this study was alkaline (pH 10.3) therefore

139 Vurayai et al.

 Int. J. Biosci. 2020

was able to increase soil pH of both soils (2.5 km and H+ by alkali salts and also due to basic solubilisation
20 km west of mine smelter) to above neutral 7 of basic metals oxides of fly ash in soil (Khan and
(Vurayai et al., 2017). Increase in soil pH after adding Khan, 1996).
alkaline ash can be attributed to the neutralisation of

Table 2. Ionic concentration (mg/kg Dw-1) of roots and shoots of M. stenopetala. The %RSD for all metals was
<4.8 % (n=5).
Distance from Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
Mine smelter
2.5km East With ash Roots 2.5 162 1.58 1.3 0.81 0.09 0.97 3.3 0.17 1.05 1.82 0.9 0.08 0.24 0.32
Shoots 1.68 428 3.2 1.66 0.56 0.06 0.63 1.98 0.09 0.16 1.15 0 0.04 0.17 0.08
Without ash Roots 5.3 584 4.72 1.22 1.29 0.53 0.96 5.01 0.37 1.77 3.9 1.06 0.37 0.48 0.19
Shoots 3.92 352 2.2 0.68 0.93 0.41 0.85 3.56 0.3 0.47 2.93 0 0.29 0.35 0.12
55km West With ash Roots 4.36 180 2.1 1.39 0.91 0.16 1.08 3.5 0.34 1.35 1.74 0.99 0.15 0.38 0.26
Shoots 3.06 610 4.66 1.95 0.68 0.12 0.88 2.18 0.24 0.29 1.29 0 0.07 0.31 0.04
Without ash Roots 7.19 680 5.1 2.78 1.43 0.9 1.21 4.18 0.89 2.17 3.42 1.85 0.59 0.49 0.18
Shoots 6.06 478 4.42 1.95 1.3 0.83 1.18 4.17 0.74 0.71 3.11 0 0.51 0.48 0.04
20km West With ash Roots 1.84 135 1.59 0.72 1.22 0.05 0.29 1.93 0.14 0.94 2.11 0.4 0.11 0.16 0.69
Shoots 0.97 340 2.28 1.3 0.63 0.03 0.17 0.97 0.05 0.08 1.26 0 0.05 0.1 0.09
2.5km West With ash Roots 1.4 76 0.92 0.39 1.1 0.05 0.38 2.1 0.09 0.51 1.62 0.16 0.38 0.07 0.72
Shoots 0.51 166 1.09 0.5 0.38 0.01 0.03 0.56 0.04 0.05 0.91 0 0.02 0.03 0.23

The current study seeks to identify plants which can the area (Baker et al., 1994; Ebbs and Kochian 1997;
be used for phytoremediation of land east and west of 1998). Biomass production by plants with
mine smelter in Selebi-Phikwe Cu/Ni mine and so phytoextraction (removal of metals by plants) abilities
plants which are capable of producing a high biomass is therefore a significant factor contributing to the
in the presence of heavy metal and soil acidity stress success of phytoremediation.
will be most suitable for use in phytoremediation of

Table 3. Translocation factor (TF) from root to shoot of M. oleifera grown on heavy metal contaminated soil
amended with coal fly ash. The % RSD for all metals was < 6.07 % (n=5).
Distance from Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
Mine smelter
2.5km East With ash 0.52 2.52 2.34 1.29 0.64 0.44 0.64 0.57 0.6 0.21 0.62 0 0.46 0.46 0.57
Without Ash 0.71 0.63 0.72 0.47 0.71 0.7 0.76 0.69 0.84 0.22 0.74 0 0.66 0.65 0.7
55km West With ash 0.63 2.92 1.85 1.43 0.73 0.57 0.73 0.7 0.7 0.21 0.69 0 0.53 0.72 0.65
Without ash 0.82 0.81 0.95 0.54 0.92 0.92 0.9 0.83 0.8 0.35 0.94 0 0.78 0.81 0.84
20km West With ash 0.4 1.86 2 1.09 0.67 0.5 0.46 0.44 0.43 0.09 0.53 0 0.25 0.4 0.3
2.5km West With Ash 0.32 1.28 1.33 1.38 0.49 0.25 0.11 0.24 0.22 0.07 0.43 0 0 0.22 0.2

Heavy metal and acidity stress in the soils collected oleifera and M. stenopetala, respectively, when
2.5 km east, 2.5, and 20 km west of the mine smelter grown in soil collected from 2.5 km east of mine
significantly reduced total dry weight of all the plant smelter. Therefore a degree of reduction followed the
species as compared to the control soil collected from order M. stenopetala < M. oleifera in this case.
the 55 km west (p<0.05) (Fig. 5). As compared to the Reduction in total dry weight can be ascribed to
dry weights of plants grown in the control soil, 36 and reduction in vegetative growth (leaf number, stem
27% reduction in total dry weight was observed for M. diameter and plant height) (Figs 6, 7, and 8).

140 Vurayai et al.

 Int. J. Biosci. 2020

Table 4. Translocation factor (TF) from root to shoot of M. stenopetala grown on heavy metal contaminated soil
amended with coal fly. The % RSD for all metals was < 4.65 % (n=5).
Distance from Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
mine smelter
2.5km East With ash 0.67 2.64 2.03 1.28 0.69 0.67 0.65 0.6 0.53 0.15 0.63 0 0.5 0.71 0.25
Without Ash 0.74 0.6 0.47 0.56 0.72 0.77 0.89 0.71 0.81 0.27 0.75 0 0.78 0.73 0.63
With ash 0.7 3.39 2.22 1.4 0.75 0.75 0.81 0.62 0.71 0.21 0.74 0 0.47 0.82 0.15
55km West Without ash 0.84 0.7 0.87 0.7 0.91 0.92 0.98 0.87 0.83 0.33 0.91 0 0.87 0.98 0.22
20km West With ash 0.53 2.52 1.44 1.81 0.52 0.6 0.59 0.5 0.36 0.09 0.6 0 0.45 0.63 0.13
2.5km West With Ash 0.36 2.18 1.18 1.28 0.35 0.2 0.08 0.27 0.44 0.1 0.56 0 0.05 0.43 0.32

Leaf number, stem diameter and plant height of both soil acidity and high heavy metal content. Prolonged
M. oleifera and M. stenopetala were significantly exposure of roots to low pH leads to suppression of
reduced when grown in soil collected 2.5 km east as lateral root development and in extreme cases this
compared to the control soil from 55 km west leads to death of the root tips (Khan et al., 2013).
(p<0.05). Leaf number decreased by 28 % in M. Availability of most plant essential elements depends
oleifera and 33 % in M. stenopetala (Fig. 6). Stem on soil pH and so the contents of mineral nutrients in
diameter decreased by 35 % in M. stenopetala and 55 plants decrease with the decrease of pH (Pessarakli,
% in M. oleifera while plant height decreased by 22 % 1999). Heavy metal also affect biological and
in M. stenopetala and 25 % in M. oleifera (Figs 7 and physiological functions of plants which results in a
6).These observations suggest that vegetative growth decrease in vegetative growth or biomass production
of M. oleifera and M. stenopetala was reduced due to as shown in (Figs 6–8).

Table 5. Bioaccumulation factor (BAF) of M. oleifera. The %RSD for all metals was < 7.14 % (n=5).
Distance from Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
mine smelter
2.5km East With ash 0.09 0.39 0.22 0.44 0.22 0.13 0.18 0.13 0.05 0.2 0.12 0 0.07 0.06 0.18
Without ash 0.3 0.34 0.19 0.32 0.28 1.18 0.32 0.28 0.18 2.28 0.22 0 0.31 0.35 0.48
55km West With ash 2.4 0.85 0.24 0.74 0.42 0.23 0.36 0.19 0.12 0.28 0.72 0 0.38 0.82 0.46
Without ash 5.92 0.74 0.25 0.55 0.63 2.11 0.69 0.37 0.56 3.43 1.39 0 2.14 1.34 0.92
20km West With ash 0.05 0.12 0.17 0.24 0.14 0.07 0.05 0.08 0.03 0.64 0.08 0 0.1 0.09 0.06
2.5km West With ash 0.004 0.03 0.03 0.18 0.14 0.02 0.04 0.02 0.01 0.18 0.01 0 0 0.02 0.05

Addition of alkaline coal fly ash increased soil pH to of alkaline coal fly ash increases soil pH and
above neutral 7 (Vurayai et al., 2017), significantly immobilizes heavy metals thus reducing their
increased vegetative growth which culminated into availability to plants (Polat et al., 2002; Su and Wong,
increase in total dry weight in M. oleifera and M. 2004; Tsang et al., 2014), leading to improvement in
stenopetala grown in soil collected 2.5 km east and 55 plant growth. Incorporation of coal fly ash also
km west (Figs 5–8). In soil collected 2.5 km east, M. enabled M. oleifera and M. stenopetala to survive in
oleifera had the greater enhancement in total dry soil collected 2.5 km and 20 km west of the mine
weight as compared to M. stenopetala (Fig. 5). smelter (Figs 1-4). This shows that coal fly ash has
Enhanced growth after addition of fly ash has been potential for use as an aid in phytostabilisation.
reported in many plants (Jala and Goyal 2006;
Agrawal et al. 2006). Mittra et al (2005) Heavy metal uptake
demonstrated increased growth of rice plants on soil Subsequently, the ability of plants in accumulating
amended with fly ash, and Yunusa et al (2009) heavy metals under heavy metal and low pH stress
observed increases in dry weight in canola with was evaluated. M. oleifera and M. stenopetala
application of coal fly ash on acidic soil. Incorporation accumulated heavy metals with varying degree

141 Vurayai et al.

 Int. J. Biosci. 2020

(Tables 1 and 2). Similar studies have shown that M. accumulated most metals in the roots as compared to
oleifera (Amadi and Tanee, 2014; Offor et al., 2014) shoots (Tables 1 and 2), showing that both plants are
and M. stenopetala (Yimer and Khan 2016) not favorable for phytoextraction, since successful
accumulated heavy metals. For plants to be phytoextraction depends on plants which can
successfully used for phytoextraction of heavy metals accumulate metals in harvestable shoots (Cardwell et
they have to accumulate metals in the shoots for al., 2002; Chatterjee and Chatterjee, 2000).
easiness of harvest. M. oleifera and M. stenopetala

Table 6. Bioaccumulation factor (BAF) of M. stenopetala. The %RSD for all metals was < 7. 45 % (n=5).
Distancefrom Cu Fe Mn Zn As Cd Co Cr Li Mo Ni Pb Pt Se Sn
Mine smelter
2.5km East With ash 0.13 0.42 0.21 0.52 0.2 0.2 0.22 0.15 0.05 0.16 0.11 0 0.06 0.1 0.09
Without ash 0.31 0.35 0.15 0.25 0.33 1.46 0.3 0.31 0.17 2.61 0.28 0 0.43 0.41 0.19
55km West With ash 3.29 0.88 0.27 0.78 0.36 0.34 0.43 0.18 0.15 0.34 0.48 0 0.29 1.1 0.07
Without ash 6.66 0.72 0.27 0.77 0.74 2.3 0.61 0.38 0.6 3.38 1.38 0 2.43 1.66 0.08
20km West With ash 0.07 0.19 0.17 0.3 0.15 0.11 0.08 0.08 0.03 0.73 0.1 0 0.24 0.11 0.06
2.5km West With ash 0.004 0.04 0.03 0.16 0.12 0.02 0.04 0.03 0.03 0.29 0.01 0 0.02 0.03 0.29

Translocation factors of heavy metals from root to stenopetala was however highest on soils without ash
shoot in both M. oleifera and M. stenopetala were except for Fe, Zn and Mn. Comparison of species has
less than 1 for most metals, except Fe, Mn and Zn shown that both M. oleifera and M. stenopetala were
where they were above 1 on soils where coal fly ash able to translocate Fe, Zn and Mn to a TF> 1 in soils
was added at all distances (Tables 3 and 4). with ash (Table 3 and 4).
Translocation factor for both M. oleifera and M.

Fig. 1. Moringa oleifera plants growing in soil collected from 2.5 km west of mine smelter with and without coal
fly ash.

142 Vurayai et al.

 Int. J. Biosci. 2020

The efficiency of phytoremediation is determined by M. stenopetala do not fulfil the requirements of a

the ability of the plant to translocate and concentrate good phytoremediator for land east and west of the
heavy metals into their shoots (Rai et al., 2004). BCL mine smelter.
These results therefore indicate that M. oleifera and

Fig. 2. Moringa stenopetala plants growing in soil collected from 2.5 km west of mine smelter with and without
coal fly ash.

M. oleifera was an excluder for most metals especially > Moringa stenopetala (7). Failure of M. oleifera and
on soils with coal fly ash except for Cu at 55 km west M. stenopetala to hyperaccumulate metals further
(Table 5). It is however an accumulator of Cu (55 km indicates that both species are not favourable for
west with and without ash), Cd (2.5 km east without phytoextraction of heavy metals of polluted soil east
ash; 55 km west without ash), Mo (2.5 km east and west of the BCL Cu/Ni mine since the success of
without ash; 55 km west without ash), Ni (55 km west phytoextraction depends on use of plant species
without ash), Pt (55 km west without ash), and Se (55 which are tolerant to pollution and have
km west without ash). M. stenopetala was also an hyperaccumulation capacities (Gonzalez, 2012). M.
excluder of most metals, except for Cu (55 km west oleifera and M. stenopetala failed to survive in soil
with and without ash), Cd (2.5 km without ash), Mo collected 2.5 km west and 20 km west of the mine
(2.5 km east without ash; 55 km west without ash), Ni smelter (Fig. 1-4) thus showing that they are both not
(55 km west without ash), Pt (55 km west without tolerant to pollution. M. oleifera and M. stenopetala
ash), and Se (55 km west with and without ash) accumulated more metals in soil without coal fly ash
(Table 6). Under ash condition, M. stenopetala was as compared to with fly ash and heavy metal levels in
an excluder for all metals except Cu and Se at 55 km control plants (55 km west) was generally higher than
west. M. oleifera and M. stenopetala however failed the other distances (Table 1; Table 2). Coal fly ash
to hyperaccumulate any metal and performance of immobilizes heavy metals in soil reducing their
species according to number of distances with BAF>1 availability to plants (Polat et al., 2002; Su and Wong,
(accumulators) follows the order Moringa oleifera (9) 2004; Tsang et al., 2014).

143 Vurayai et al.

 Int. J. Biosci. 2020

Fig. 3. Moringa oleifera plants growing in soil collected from 20 km west of mine smelter with and without coal
fly ash.

The reduction of heavy metal accumulation in plants increase in vegetative growth (leaf number, plant
growing in soil with ash therefore reduce the impact height and stem diameter) which increased biomass
of heavy metals on plant growth which resulted in the production (total dry weight) (Fig. 5; 6; 7; 8).

Fig. 4. Moringa stenopetala plants growing in soil collected from 20 km west of mine smelter with and without
coal fly ash.

144 Vurayai et al.

 Int. J. Biosci. 2020

The indicators of heavy metal accumulation, TF and east > 20 km west > 2.5 km east) in both soil with and
BAF generally followed the order 55 km west > 2.5 km without coal fly ash (Vurayai et al., 2015; Vurayai et
east for plants grown in soil without ash, and 55 km al., 2017). Heavy metal bioavailability increases at low
west > 2.5 km east > 20 km west for plants grown in soil pH (Mench et al., 2003) and so a combination of
soil with ash (Table 3-6). This is attributed to soil pH heavy metal and low pH stress may have impacted the
which followed the same trend (55 km west > 2.5 km efficiency of plants accumulate metals.

Fig. 5. Effect of coal fly ash treatment (7.5%) of polluted soil on total dry weight of Moringa oleifera (Mo O) and
Moringa stenopetala (Mo S). Error bars indicate ± standard error (n=5) and p.d indicates that plants died.

Fig. 6. Effect of coal fly ash treatment (7.5%) of polluted soil on leaf number Moringa oleifera (Mo O) and
Moringa stenopetala (Mo S). Error bars indicate ± standard error (n=5) and p.d indicates that plants died.

145 Vurayai et al.

 Int. J. Biosci. 2020

The current study has indicated that M. oleifera and cannot be used for phytoremediation of the area east
M. stenopetala are not good candidates for and west of the BCL mine smelter.
phytoextraction but this does not mean that they

Fig. 7. Effect of coal fly ash treatment (7.5%) of polluted soil on stem diameter of Moringa oleifera (Mo O) and
Moringa stenopetala (Mo S). Error bars indicate ± standard error (n=5) and p.d indicates that plants died.

Fig. 8. Effect of coal fly ash treatment (7.5%) of polluted soil on plant height of Moringa oleifera (Mo O) and
Moringa stenopetala (Mo S). Error bars indicate ± standard error (n=5) and p.d indicates that plant.

Coal fly ash has been shown to impede accumulation phytostabilisation since it enabled M. oleifera and M.
of heavy metals in plant biomass and so can be stenopetala to survive polluted soils. Addition of coal
effective in situ remediation or aiding fly ash enables plants to survive and grow, and

146 Vurayai et al.

 Int. J. Biosci. 2020

immobilize the contaminants in the soil and Reports 5(12), 1263-1274.

groundwater through absorption and accumulation in http://dx.doi.org/10.1007/s00299-006-0204-8
plant tissues, adsorption onto roots, or precipitation
within the root zone preventing their migration in Amadi N, Tanee FBG. 2014. Efficacy of Moringa
soil, as well as their movement by erosion and oleifera as a phytoextraction plant in the remediation
deflation (Erdei, 2005; Erakhrumen and Agbontalor, of heavy metals polluted soil. African Journal Plant
2007). Science 8(12), 546-553.
http://dx.doi.org/10.5897/AJPS
Conclusion
M. oleifera and M. stenopetala failed to survive in Baker AJM. 1981. Accumulators and
polluted soils collected 2.5 and 20 km west of the excluders‐strategies in the response of plants to heavy
mine smelter, showing that both species are not good metals. Journal of Plant Nutrition 3(1-4), 643-654.
candidates for phytoremediation of the area. Soil http://dx.doi.org/10.1080/01904168109362867
acidity and heavy metal stress reduced vegetative
growth (leaf number, stem diameter, plant height) of Baker AJM, McGrath SP, Sidoli CMD, Reeves
both species, which resulted in decrease in total dry RD. 1994. The possibility of in situ heavy metal
weight. Both species also failed to effectively decontamination of polluted soils using crops of
translocate metals to shoots as they accumulated metal-accumulating plants. Resource Conservation
more metals in roots than in shoots. and Recycling 11(1-4), 41-49.
http://dx.doi.org/10.1016/0921-3449(94)90077-9.
They also both failed to hyperaccumulate any metal in
all polluted soils examined, further confirming that Baker AJM, Brooks RR. 1989. Terrestrial higher
both species are not good candidates for plants which hyperaccumulate metallic elements -A
phytoremediation of the area. Application of coal fly review of their distribution, ecology and
ash enabled M. oleifera and M. stenopetala to survive phytochemistry. Biorecovery 1, 81-126.
in soil collected 2.5 and 20 km west of mine smelter. http://dx.doi.org/10.1080/01904168109362867.
Coal fly ash reduced heavy metal accumulation in
both species at all distances, which led to reduction in Barrie CT, Ludden JN, Green TH. 1993.
translocation and bioaccumulation factors. Geochemistry of volcanic rocks associated with Cu-Zn
and Ni-Cu deposits in the Abitibi Subprovince.
Heavy metal accumulation generally followed the Economic Geology 88(6), 1341-1358.
order 55 km west> 2.5 km east> 20 km west> 2.5 km http://dx.doi.org/10.2113/gsecongeo.88.6.1341
east which is the same order for soil pH from highest
to lowest. Coal fly ash can therefore be used for aiding Bolan NS, Duraisamy VP. 2003. Role of inorganic
phytostabilisation as it enables plants to survive in and organic soil amendments on immobilisation and
polluted soil. M. oleifera and M. stenopetala are not phytoavailability of heavy metals: a review involving
good candidates for phytoextraction of heavy metals specific case studies. Soil Research 41(3), 533-555.
east and west of the BCL Cu/Ni mine smelter, but can http://dx.doi.org/10.1071/SR02122.
be used for phytostabilisation with the help of coal fly
ash. Bosch CH. 2004. Moringa oleifera Lam. PROTA
Foundation (pp392-395). Backhuys, Wageningen, the
References Netherlands.
Agarwal PK, Agarwal P, Reddy MK, Sopory
SK. 2006. Role of DREB transcription factors in Cardwell AJ, Hawker DW, Greenway M. 2002.
abiotic and biotic stress tolerance in plants. Plant Cell Metal accumulation in aquatic macrophytes from

147 Vurayai et al.

 Int. J. Biosci. 2020

southeast Queensland, Australia. Chemosphere mineralogy of particulate air matter around Selebi
48(7), 653-663. PMID: 12201195 Phikwe nickel–copper plant, Botswana. Mineral
http://dx.doi.org/10.1016/s0045-6535(02)00164-9. Engineering 17(2), 349-353.
http://dx.doi.org/10.1016/j.mineng.2003.08.016.
Chaney RL. 1983. Plant uptake of inorganic waste
constituents. In: JF Parr, Marsh PB, Kla JM, Ed. Land Ekosse G, Van Den Heever D, De Jager L,
Treatment of Hazardous Wastes. New Jersey, USA: Totolo O. 2003. Environmental mineralogy of soils
Noyes Data Corp., p. 50-76. around Selebi Phikwe nickel-copper plant, Botswana.
International Journal of Environmental Studies 60
Chatterjee J, Chatterjee C. 2000. Phytotoxicity of (3), 251-262.
cobalt, chromium and copper in http://dx.doi.org/10.1080/00207230290034332
cauliflower. Environmental Pollution 109(1), 69-74.
PMID: 15092914. Ekosse GIE, Ngole VM. 2012. Copper, nickel and
http://dx.doi.org/10.1016/s0269-7491(99)00238-9. zinc contamination in soils within the precincts of
mining and landfilling environments. International
Danh LT, Truong P, Mammucari R, Tran T, Journal of Environmental Science and
Foster N. 2009. Vetiver grass, Vetiveria zizanioides: Technology 9(3), 485-494.
a choice plant for phytoremediation of heavy metals http://dx.doi.org/10.1007/s13762-012-0055-5.
and organic wastes. International Journal of
Phytoremediation 11(8), 664-691. Erakhrumen A, Agbontalor A. 2007. Review
http://dx.doi.org/10.1080/15226510902787302. Phytoremediation: an environmentally sound
technology for pollution prevention, control and
Ebbs SD, Kochian LV. 1997. Toxicity of zinc and remediation in developing countries. Educational
copper to Brassica species: implications for Research and Review 2(7), 151-156.
phytoremediation. Journal Environmental Quality 26
(3), 776-781. Erdei L, Mezˆ osi G, M´ecs I, Vass I, Fˆoglein
http://dx.doi.org/10.2134/jeq1997.00472425002600 F, Bulik L. 2005. Phytoremediation as a program for
030026x. decontamination of heavy-metal polluted
environment. Acta Biologica Szegediensis 49(1-2),
Ebbs SD, Kochian LV. 1998. Phytoextraction of 75-76.
zinc by oat (Avena sativa), barley (Hordeum
vulgare), and Indian mustard (Brassica González V, García I, Del Moral F, Simón M.
juncea). Environmental Science and 2012. Effectiveness of amendments on the spread and
Technology 32(6), 802-806. phytotoxicity of contaminants in metal–arsenic
http://dx.doi.org/10.1021/es970698p. polluted soil. Journal of hazardous materials 205, 72-
80.
Ekosse GIE, Ngila CJ, Forcheh N. 2005. http://dx.doi.org/10.1016/j.jhazmat.2011.12.011.
Multivariate analyses of heavy metals in soils and
Colophospermum mopane leaves around the Selebi Hoagland DR, Arnon DI. 1950. The Water-Culture
Phikwe nickel-copper mine and smelter/concentrator Method for Growing Plants without Soil. California
plant area, Botswana. Journal of Applied Science and Agricultural Experiment Station, Circular-347.
Environmental Management 9(1), 177–185.
Jala S, Goyal D. 2006. Fly ash as a soil ameliorant
Ekosse G, Van den Heever DJ, De Jager L, for improving crop production-a review. Bioresource
Totolo O. 2004. Environmental chemistry and Technology 97(9), 1136-1147.

148 Vurayai et al.

 Int. J. Biosci. 2020

http://dx.doi.org/10.1016/j.biortech.2004.09.004 contaminated water using Moringa Stenopetala and

Moringa oleifera seed powder. International Journal
of Environmental Science and Technology 3(2), 131-
Jiru D, Sonder K, Alemayehu L, Mekonen Y, 139.
Anjulo A. 2006. Leaf yield and nutritive value of http://dx.doi.org/10.1007/bf03325916.
Moringa stenopetala and Moringa oleifera
accessions: Its potential role in food security in Mataka LM, Sajidu SM, Masamba WRL,
constrained dry farming agroforestry Mwatseteza JF. 2010. Cadmium sorption by
system. Proceedings of the Moringa and Other Highly Moringa stenopetala and Moringa oleiferaseed
Nutritious Plant Resources: Strategies, Standards and powders: Batch, time, temperature, PH and
Markets for a Better Impact on Nutrition in Africa, adsorption isotherm studies. International Journal of
Accra, Ghana, 16-18. Water Resources and Environmental, Engineering
2(3), 50-59.
Khan MR, Wajid M. 1996. The effect of fly ash on
plant growth and yield of tomato. Environmental Mench M, Bussiere S, Boisson J, Castaing E,
Pollution 92(2), 105-111. Vangronsveld J, Ruttens A, Manceau A. 2003.
http://dx.doi.org/10.1016/0269-7491(95)00098-4. Progress in remediation and revegetation of the
barren Jales gold mine spoil after in situ
Khan S, Wang N, Reid BJ, Freddo A, Cai C. treatments. Plant and soil 249(1), 187-202.
2013. Reduced bioaccumulation of PAHs by Lactuca http://dx.doi.org/10.1023/a:1022566431272.
satuva L. grown in contaminated soil amended with
sewage sludge and sewage sludge derived Mench M, Manceau A, Vangronsveld J,
biochar. Environmental pollution 175, 64-68. Clijsters, H, Mocquot B. 2000. Capacity of soil
http://dx.doi.org/10.1016/j.envpol.2012.12.014. amendments in lowering the phytoavailability of
sludge-borne zinc. Agronomie 20(4), 383-397.
Kumpiene J, Lagerkvist A, Maurice C. 2008. http://dx.doi.org/10.1051/agro:2000135.
Stabilization of As, Cr, Cu, Pb and Zn in soil using
amendments–a review. Waste management 28(1), Mitra BN, Karmakar S, Swain DK, Ghosh BC.
215-225. 2005. Fly ash-a potential sourceof soil amendment
http://dx.doi.org/10.1016/j.wasman.2006.12.012. and a component of integrated plant nutrient
supplysystem. Fuel 84, 1447–1451.
Likuku AS, Mmolawa KB, Gaboutloeloe GK. http://dx.doi.org/10.1016/j.fuel.2004.10.019.
2013. Assessment of heavy metal enrichment and
degree of contamination around the copper-nickel Nand V, Maata M, Koshy K, Sotheeswaran S.
mine in the Selebi Phikwe Region, Eastern 2012. Water purification using Moringa oleifera and
Botswana. Environment and Ecology Research 1(2), other locally available seeds in Fiji for heavy metal
32-40. removal. International Journal of Applied Science
and Technology 2(5).
Ma LQ, Komar KM, Tu C, Zhang W, Cai Y,
Kennelley ED. 2001. A fern that hyperaccumulates Offor I, Ehiri R, Njoku C. 2014. Proximate
arsenic. Nature 409(6820), 579-579. nutritional analysis and heavy metal composition of
http://dx.doi.org/10.1038/35054664. dried Moringa oleifera leaves from Oshiri Onicha
LGA, Ebonyi State, Nigeria. Journal of Environmental
Mataka LM, Henry EMT, Masamba WRL, Science, Toxicology and Food Technology 8(1), 57-
Sajidu SM. 2006. Lead remediation of 62.

149 Vurayai et al.

 Int. J. Biosci. 2020

http://dx.doi.org/10.9790/2402-08115762. means of polyelectrolytes of the Moringa oleifera

whole seed kernel. WIT Transactions on Ecology and
Pessarakli M. 1999. Handbook of plant and crop the Environment, 80.
stress 2nded. New York, USA: Marcel Dekker Inc., p
720. Schippers A, Kock D, Schwartz M, Bo¨ttcher
ME, Vogel H, Hagger M. 2007.
Pivetz BE. 2001. Ground water issue: Geomicrobiological and geochemical investigation of
phytoremediation of contaminated soil and ground a pyrrhotite-containing mine waste tailings dam near
water at hazardous waste sites. National risk Selebi Phikwe in Botswana. Journal of Geochem
management research lab ADA OK, p 37. Exploration 92(2–3), 151–158.
http://dx.doi.org/10.1016/j.gexplo.2006.08.003.
Polat M, Lederman E, Pelly I, Cohen H. 2002.
Chemical neutralization of acidic wastes using fly ash Shahid M, Khalid S, Abbas G, Shahid N,
in Israel. Journal of Chemical Technology and Nadeem M, Sabir M. 2015. Heavy metal stress and
Biotechnology 77(3), 377-381. crop productivity. In: Hakeem KR, Ed. Crop
http://dx.doi.org/10.9790/2402-08115762. Production and Global Environmental Issues.Cham:
Springer International Publishing, p 1–25.
Prasad MNV, Freitas HMO. 2003. Metal
hyperaccumulation in plants: biodiversity prospecting Su DC, Wong JWC. 2004. Chemical speciation and
for phytoremediation technology. Electronic Journal phytoavailability of Zn, Cu, Ni and Cd in soil amended
of Biotechnology 6(3), 285-321. with fly ash-stabilized sewage sludge. Environment
http://dx.doi.org/10.2225/vol6-issue3-fulltext-6. International 29(7), 895-900.
http://dx.doi.org/10.1016/s0160-4120(03)00052-7.
Rai V, Vajpayee P, Singh SN, Mehrotra S. 2004.
Effect of chromium accumulation on photosynthetic Tsang DC, Yip AC, Olds WE, Weber PA. 2014.
pigments, oxidative stress defense system, nitrate Arsenic and copper stabilisation in a contaminated
reduction, proline level and eugenol content of soil by coal fly ash and green waste
Ocimum tenuiflorum L. Plant science 167(5), 1159- compost. Environmental Science and Pollution
1169. Research 21(17), 10194-10204.
http://dx.doi.org/10.1016/j.plantsci.2004.06.016. http://dx.doi.org/10.1007/s11356-014-3032-3.

Rosenthal WD, Arkin GF, Shouse PE, Jordan Vurayai R, Nkoane B, Moseki B, Chartuvedi P.
WR. 1987. Water deficit effects on transpiration and 2015. Assessment of heavy metal
leaf growth. Agronomy Journal 79(6), 1019-1026. pollution/contamination in soils east and west of the
http://dx.doi.org/10.2134/agronj1987.00021962007 Bamangwato Concessions Ltd (BCL) Cu/Ni mine
900060014x. smelter in Selebi-Phikwe, Botswana. Journal of
Biodiversity and Environmental Science 7(6), 111-
Roy S, Labelle S, Mehta P, Mihoc A, Fortin N, 120.
Masson C, Olsen C. 2005. Phytoremediation of
heavy metal and PAH-contaminated brownfield Vurayai R, Nkoane B, Moseki B, Chartuvedi P.
sites. Plant and Soil 272(1), 277-290. 2017. Phytoremediation potential of Jatropha curcas
http://dx.doi.org/10.1007/s11104-004-5295-9. and Pennisetum clandestinum grown in polluted soil
with and without coal fly ash: a case of BCL Cu/Ni
Sajidu SM, Henry EMT, Kwamdera G, Mataka mine, Selibe-Phikwe, Botswana. Journal of
L. 2005. Removal of lead, iron and cadmium ions by Biodiversity and Environmental Science 10(5),

150 Vurayai et al.

 Int. J. Biosci. 2020

193-206. http://dx.doi.org/10.1007/s10646-006-0129-6.

Yimer AM, Khan MA. 2016. Atomic Absorption Yunusa IA, Burchett MD, Manoharan V,
Spectrometric Determination of Some Heavy Metals DeSilva DL, Eamus D, Skilbeck CG. 2009.
from the Leaves of Moringa stenopetala Grown in Photosynthetic pigment concentrations, gas exchange
Gamo Gofa Zone, Ethiopia. British Journal of Applied and vegetative growth for selected monocots and
Science and Technology 13, 5. dicots treated with two contrasting coal fly
http://dx.doi.org/10.9734/bjast/2016/17726. ashes. Journal of environmental quality 38(4), 1466-
1472.
Yu XZ, Gu JD, Huang SZ. 2007. Hexavalent http://dx.doi.org/10.2134/jeq2008.0285.
chromium induced stress and metabolic responses in
hybrid willows. Ecotoxicology 16(3), 299-309.

151 Vurayai et al.

View publication stats