Subject Area 5.
3 Ethidium Bromide
Subject Area 5.3: Phytoremediation and ecosystem restoration
Research Article
Phytoremediation Potentials of Selected Tropical Plants for Ethidium Bromide*
Raynato B. Uera1, Annie Melinda Paz-Alberto1 and Gilbert C. Sigua2**
1 Environmental Management Department, Institute of Graduate Studies, Central Luzon State University, Science City of Muñoz,
Nueva Ecija, Philippines 3120
2 Subtropical Agricultural Research Station, Agricultural Research Service, United States Department of Agriculture, Brooksville, FL USA 34601
**Corresponding author (gcsigua@ifas.ufl.edu)
DOI: http://dx.doi.org/10.1065/espr2007.02.391
Please cite this paper as: Uera RB, Paz-Alberto AM, Sigua GC
(2007): Phytoremediation Potentials of Selected Tropical Plants
for Ethidium Bromide. Env Sci Pollut Res 14 (7) 505–509
Abstract
Background, Aims and Scope. Research and development has its
own benefits and inconveniences. One of the inconveniences is the
generation of enormous quantity of diverse toxic and hazardous
wastes and its eventual contamination to soil and groundwater re-
sources. Ethidium bromide (EtBr) is one of the commonly used sub-
stances in molecular biology experiments. It is highly mutagenic and
moderately toxic substance used in DNA-staining during electro-
phoresis. Interest in phytoremediation as a method to solve chemi-
cal contamination has been growing rapidly in recent years. The
technology has been utilized to clean up soil and groundwater from
heavy metals and other toxic organic compounds in many countries
like the United States, Russia, and most of European countries.
Phytoremediation requires somewhat limited resources and very
useful in treating wide variety of environmental contaminants. This
study aimed to assess the potential of selected tropical plants as
phytoremediators of EtBr.
Materials and Methods. This study used tomato (Solanum lycopersi-
cum), mustard (Brassica alba), vetivergrass (Vetiveria zizanioedes),
cogongrass (Imperata cylindrica), carabaograss (Paspalum conjuga-
tum), and talahib (Saccharum spontaneum) to remove EtBr from
laboratory wastes. The six tropical plants were planted in individual
plastic bags containing soil and 10% EtBr-stained agarose gel. The
plants were allowed to establish and grow in soil for 30 days. Ethi-
dium bromide content of the test plants and the soil were analyzed
before and after soil treatment. Ethidium bromide contents of the
plants and soils were analyzed using an UV VIS spectrophotometer.
Results. Results showed a highly significant (p≤0.001) difference in
the ability of the tropical plants to absorb EtBr from soils. Mustard
registered the highest absorption of EtBr (1.4±0.12 μg kg–1) followed
by tomato and vetivergrass with average uptake of 1.0±0.23 and
0.7±0.17 μg kg–1 EtBr, respectively. Cogongrass, talahib, and cara-
baograss had the least amount of EtBr absorbed (0.2±0.6 μg kg–1).
Ethidium bromide content of soil planted to mustard was reduced
by 10.7%. This was followed by tomato with an average reduction
of 8.1%. Only 5.6% reduction was obtained from soils planted to
vetivergrass. Soils planted to cogongrass, talahib, and carabaograss
had the least reduction of 1.52% from its initial EtBr content.
Discussion. In this study, mustard, tomato, and vetivergrass have shown
their ability to absorb EtBr from contaminated soil keeping them from
expanding their reach into the environment and preventing further
contamination. Its downside, however, is that living creatures includ-
ing humans, fish, and birds, must be prevented from eating the plants
that utilized these substances. Nonetheless, it is still easier to isolate,
cut down, and remove plants growing on the surface of the contami-
nated matrices, than to use strong acids and permanganates to chemi-
cally neutralize a dangerous process that can further contaminate the
environment and pose additional risks to humans. Though this alter-
Env Sci Pollut Res 14 (7) 505 – 509 (2007)
© 2007 ecomed publishers (Verlagsgruppe Hüthig Jehle Rehm GmbH), D-86899 Landsberg and Tokyo • Mumbai • Seoul • Melbourne • Paris
native method does not totally eliminate eventual environmental con-
tamination, it is by far produces extremely insignificant amount of
by-products compared with the existing processes and technologies.
Conclusions. Mustard had the highest potential as phytoremediator
of EtBr in soil. However, the absorption capabilities of the other test
plants may also be considered in terms of period of maturity and
productivity.
Recommendations and Perspectives. It is recommended that a more
detailed and complete investigation of the phytoremediation proper-
ties of the different plants tested should be conducted in actual field
experiments. Plants should be exposed until they reach maturity to
establish their maximum response to the toxicity and mutagenecity of
EtBr and their maximum absorbing capabilities. Different plant parts
should be analyzed individually to determine the movement and trans-
location of EtBr from soil to the tissues of plants. Since this study has
established that some plants can thrive and dwell in EtBr-treated soil,
an increased amount of EtBr application should be explored in future
studies. It is suggested therefore that a larger, more comprehensive
exploration of phytoremediation application in the management of
toxic and hazardous wastes emanating from biotechnology research
activities should be considered especially on the use of vetivergrass,
a very promising tropical perennial grass.
Keywords: Ethidium bromide (EtBr); hazardous; mutagenic; phyto-
remediation; tropical plants
1 Background, Aims and Scope
Around the world, land, surface waters, and groundwater
are increasingly affected by contamination from industrial,
research experiments, military and agricultural activities ei-
ther due to ignorance, lack of vision, carelessness, or high
cost of waste disposal and treatment. The rapid build-up of
toxic pollutants (metals, radionuclide, and organic contami-
nants in soil, surface water and groundwater) not only af-
fects natural resources but also causes a major strain on eco-
systems (Ona et al. 2006).
Research and development has its own benefits and inconve-
niences. One of the inconveniences is the generation of enor-
mous quantity of diverse toxic and hazardous wastes and its
eventual contamination to soil and groundwater resources.
Ethidium bromide (EtBr) is one of the commonly used sub-
stances in molecular biology experiments. It is highly mutagenic
and moderately toxic substance used in DNA-staining during
electrophoresis. It is a potent mutagen, capable of causing liv-
* ESS-Submission Editor: Professor Zhihong Xu
(zhihong.xu@griffith.edu.au; www.griffith.edu.au/centre/cfhr)
505
Ethidium Bromide
ing cell mutations and moderately toxic after an acute expo-
sure. It can be absorbed through the skin and also an irritant
to the skin, eyes, mouth, and upper respiratory tract. Although
there is no evidence at this time of human carcinogenicity or
teratogenicity, this material may still be considered a possible
carcinogen or teratogen (Indiana University Fact Sheet 1996).
Ethidium bromide is capable of disrupting efficient metabo-
lism and protein synthesis in plants and could have had af-
fected the growth and metabolism of the test plants during the
30 days exposure. The study of Zollinger and Morais (1979)
showed that EtBr decreased the rate of growth of infected
chick embryo cells and inhibited the synthesis of mitochon-
drial macromolecules. The DNA of a cell prevents the cell
from accessing its genome to carry out its necessary protein
synthesis to survive (Indiana University Chemical 1996).
Interest in phytoremediation as a method to solve chemical
contamination has been growing rapidly in recent years
(Clemens et al. 2002, Dhanker et al. 2002, Bettis 1997). The
technology has been utilized to clean up soil and groundwater
from heavy metals and other toxic organic compounds in many
countries like the United States, Russia, and most of Euro-
pean countries. Phytoremediation requires somewhat limited
resources and very useful in treating wide variety of environ-
mental contaminants. This study aimed to assess the potential
of selected tropical plants as phytoremediators of EtBr. The
technology has been utilized to clean up soil and groundwater
from heavy metals and other toxic organic compounds in many
countries like the United States and Russia. It requires limited
resources and very useful in treating wide variety of environ-
mental contaminants (Sigua 2005, Raskin and Salt 1997).
Phytoremediation involves growing plants in a contaminated
matrix, for a required growth period, to remove contami-
nants from the matrix, or facilitate immobilization (bind-
ing/containment) or degradation (detoxification) of the pol-
lutants. The plants can be subsequently harvested, processed
and disposed (Sigua et al. 2004b, Raskin and Salt 1997).
Plants are unique organisms equipped with remarkable meta-
bolic and absorption capabilities, as well as transport sys-
tems that can take up nutrients or contaminants selectively
from the growth matrix, soil or water (Sigua et al. 2005,
Sigua et al. 2004a, Sigua et al. 2004b, USEPA 2001, Raskin
and Salt 1997). Phytoremediation of EtBr is relatively new
area of research and not very many useful and relevant in-
formation is available in the scientific community. Alterna-
tive and more practical methods of treating and disposing
hazardous wastes are highly needed. This study dealt with the
possibility of utilizing some plants with phytoremediation prop-
erties, which could be potential alternative to a costly and
dangerous hazardous waste treatment and disposal system of
EtBr-contaminated materials in the Philippines and elsewhere.
Finally, this study may pave the way for a larger and more
comprehensive exploration of phytoremediation application
utilizing plants in the management of toxic and hazardous
wastes emanating from biotechnology research activities.
2 Materials and Methods
2.1 Soil materials and ethidium bromide treatment
The soil used in this study was collected in Barangay Abar
1st, San jose City, Nueva Ecija and was analyzed at the
506
Subject Area 5.3
Agronomy and Soil Division, Philippine Rice Institute, Sci-
ence City of Muñoz, Nueva Ecija. The soil used had a me-
dium acidity of pH 5.94, organic matter content of 3.1%,
and a phosphorus content of 12.7 mg kg–1.
Ethidium bromide (EtBr)-stained agarose gels (2%) ready for
disposal were obtained from the Molecular Genetics Labora-
tory of the Philippine Rice Research Institute. Soil materials
(0–20 cm depth) were collected from actual agricultural fields
planted with tomato and mustard. The same types of soil were
collected from areas with vetivergrass, cogongrass, carabao-
grass, and talahib. Soil materials were air-dried, sieved, and
mixed together (composite). Black polyethylene bags with
3 kg capacity were used in the experiment. The untreated
soil (0% contamination) contained 2 kg of soil with no EtBr-
stained agarose gel. The treated soil (10% contamination),
had 200 grams of EtBr-stained agarose gel (approximately
28.5 μg kg–1 EtBr) and 1,800 grams of soil in each bag. All
treatments were replicated three times. All treatments were
given approximately three liters of distilled water every four
days to keep the moisture content of the soils at field capacity
(21%). Distilled water was used in the experiment to make
sure that water is free from impurities, chemicals and pollut-
ants, which are potential sources of contamination. Individual
bag was inspected almost daily for possible leakage that may
compromise the results of the study.
2.2 Plant materials
This study used tomato (Solanum lycopersicum), mustard
(Brassica alba), vetivergrass (Vetiveria zizanioedes), cogongrass
(Imperata cylindrica), carabaograss (Paspalum conjugatum),
and talahib (Saccharum spontaneum) to remove EtBr from
laboratory wastes. Seeds of tomato and mustard were ob-
tained from the Central Luzon State University Research,
Extension and Training Center (CLSU-RET) for this study.
Young seedlings of vetivergrass were secured from the Con-
servation Park of Environmental Management Institute,
Central Luzon State University while young seedlings of
cogongrass, carabaograss, and talahib were collected from
a nearby city. Tomato, mustard, and vetivergrass are plants
with phytoremediation properties. They can remove heavy
metals and other organic materials from contaminated soil
and water (Whitener 1993). Locally, cogongrass, carabaograss,
and talahib can grow anywhere and are easily available. They
are the most common and easily maintained grasses known
to human in the tropics (Watson and Dallwitz 1992).
Seeds of tomato and mustard were directly sown in straight
rows in an 18"x 12"x 6" plastic trays containing pure soil.
One-month old tomato and 21-day old mustard seedlings
were transplanted into previously prepared soil media (see
above). Young seedlings (approximately one-month old) of
vetivergrass, cogongrass, carabaograss and talahib were
transplanted in individual bags. All treatments in each plant
were given 50 ml tap water every other day. No fertilizer
was applied during the duration of the experiment.
2.3 Sample preparations for EtBr analysis
2.3.1 Plant sample
The different test plants were harvested 30 days after expo-
sure. The whole plants (roots, stems, and leaves) were used
Env Sci Pollut Res 14 (7) 2007
Subject Area 5.3
for the tissue analysis of EtBr. Plant samples were washed
with running tap water until soils and other dirt particles
were removed completely. Wet plant tissue samples were
wiped with clean tissue paper and allowed to air dry. The
samples were placed in properly labeled foil envelopes and
placed inside plastic bags, then kept in freezer before labo-
ratory analysis. Plant samples were extracted for EtBr fol-
lowing the extraction procedures developed by Washington
State University, Chemical Wastes Section. The required
amount of plant samples were weighed and treated with
Tris-Borate-EDTA (TBE: Tris base + Boric acid + Na4EDTA).
The plant tissue was heated at 65°C for 10 minutes and
then mixed thoroughly. Chloroform was added and the mix-
ture was heated again for 10 minutes 65°C. The solutions
were filtered using a Whatman 342 filter paper. This was
followed by the measurement of EtBr concentration using
an UV VIS spectrophotometer (see section 2.3.3).
2.3.2 Soil samples
Soil samples from each treatment were thoroughly mixed
until homogeneity was reached. A 20-gram composite soil
sample was placed in a 50-liter corning tubes and were kept
in a freezer before analysis. The concentration of EtBr in
soils planted to the different test plants after trials was de-
termined using an UV VIS spectrophotometer (see section
2.3.3). The reduction of EtBr in soil was computed by sub-
tracting the final concentration of EtBr in soil from the ini-
tial concentration of EtBr in soil divide by the initial con-
centration of EtBr in soil multiply by 100.
2.3.3 Spectrophotometric analysis of EtBr
Analysis of the EtBr content was done using an UV VIS spec-
trophotometer at the Food Protection Laboratory of the Bu-
reau of Post-Harvest Research and Extension. The wavelength
was set at 317 nm. The calibration curve was set using a series
of known concentration of EtBr solution set at 0.0, 0.2, 0.4,
0.6, 0.8 and 1.0 mg kg–1. Calibration curve was done by plot-
ting the absorbance against the concentration of EtBr stan-
dards. The concentration of the solution was computed based
on Beer's Law of Concentration Determination.
2.4 Statistical design and data analysis
This study was laid-out in a two-factor experiment in a com-
pletely randomized design. All treatments were replicated
three times (SAS, 2000). Where the F-test indicated a sig-
nificant (p≤0.05) effect, means were separated following the
procedures of Duncan Multiple Range Test (DMRT).
3 Results and Discussion
3.1 Plant uptake of etbr and EtBr reduction in soils
Three out of the six plants screened as possible phytoreme-
diator of EtBr-stained agarose gel in soil showed positive
and remarkable results. These three were mustard, tomato,
and vetivergrass (Fig. 1). Statistical analysis showed that the
differences among the plant's accumulation of EtBr were
highly significant (p≤0.001). A significant difference (p≤0.05)
was noted between the means of mustard and vetivergrass.
However, treatment means of mustard and tomato did not
differ significantly at 5% level significance. The same was
Env Sci Pollut Res 14 (7) 2007
Ethidium Bromide
1.80
1.60 a
LEVELS OF EtBr IN PLANTS (u g kg-1)
1.40
b
1.20
1.00
b
0.80
0.60
0.40
c c
c
0.20
0.00
TOMATO MUSTARD VETIVER TALAHIB CARABAO COGON
TROPICAL PLANTS
Fig. 1: Average levels of EtBr in plants at harvest. Concentrations of EtBr
among the different tropical plants are significantly different (p£0.05) when
superscripts located at top of bars are different
observed in tomato and vetivergrass (see Fig. 1). Different
plants may have different responses to a variety of environ-
mental stresses (e.g., EtBr treatment). Some plants can develop
tolerance or resistance along the way, but others may be com-
pletely affected. A stress may change metabolism and may
alter the morphology, therefore disrupting efficient growth and
development (Hale and Orcutt 1978). Deficiency or abundance
of an element can result from a number of conditions like the
amount and concentration present in the soil, the form in which
they exist, and the processes by which they become available
(Clarkson and Hanson 1980). Hale and Orcutt (1987) claimed
that slow rate of growth in plants is largely caused by envi-
ronmental stress particularly nutrient deficiency and chemi-
cal toxicity. Clark (1982) declared that tolerance to a cer-
tain stress, be it chemical or physical is brought about by
several factors. Some of these factors are: 1) the amount or
concentration present in the soil is not enough to cause stress;
2) the length of exposure is very limited and not adequate to
cause strain; 3) the plant has developed resistance or toler-
ance to the stress; and 4) inclusion of EtBr-stained agarose
gel introduced not only the toxic and mutagenic component
of EtBr, but also abundant supply of carbohydrates neces-
sary in plant's energy production.
Mustard, registered as the most active phytoremediator of EtBr
in soil with 1.4±0.12 μg kg–1 after 30 days of exposure. This
plant is known to remove large quantities of chromium, lead,
copper, and nickel in soils (Wall Street 1996). This could be
attributed to its well-developed root system as observed visu-
ally during sampling. Another factor could be its physiology for
being an early maturing vegetable. As the length of exposure
can affect the amount of uptake, it can also be influenced by the
plant's maturity (Whitener 1993). In addition, mustard as a
leafy vegetable has a biomass composed of nearly 80% leaves
and 20% roots and stems. As such, mustard plant has more
room for storing foods and other non-essential elements. Mus-
tard can absorb lead, cadmium, chromium, nickel, zinc, and
copper up to 3.5% of its dry body weight (Wall Street 1996).
Tomato was second to mustard with 1.0±0.23 μg kg–1 of EtBr
on their tissues (see Fig. 1). This could probably be due to its
far-reaching root system observed visually during sampling and
its excellent tolerance under the stress. Its stem structure is com-
posed of 95% water and water-soluble materials enabling it to
507
Ethidium Bromide
hold and store larger amount of micro and macronutrients.
Tomato is also a known phytoremediator of chromium, lead,
copper, and nickel in soil (Wall Street 1996). The inhibitory
activity of toxic compounds is related to the amount absorbed
and translocated to the different parts of the plants (Hale and
Orcutt 1987, Fritz 1983). This may be the reason why mus-
tard has accumulated the largest amount of EtBr in soil and
exhibited cessation of growth among the other test plants.
Decrease in growth rate of the root system or complete cessa-
tion of its growth in developed and fruit-bearing tomato plants
is known phenomena. It was suggested that a limited supplyof
carbohydrates to this organ, due to its relative weakness in com-
petition with the flowers and developing fruitlets is the main
cause for these disorders (Pressman et al. 1997). Conversely,
growth responses of talahib, cogongrass and carabaograss were
the least affected or not affected by EtBr treatment.
Vetivergrass was able to absorb 0.7±0.17 μg kg–1 of EtBr
from the soil (see Fig. 1). Roots and shoots of vetivergrass
could accumulate five times more EtBr than the chromium
and zinc levels in the soil (Troung 1999). This small amount
of EtBr in vetivergrass might be a function of the length of
exposure that affected the absorption rate. It is possible that
vetivergrass being a perennial plant can still increase its up-
take if its exposure to the soil had been prolonged much
longer or until it reaches maturity. In general, mature (120
days of exposure) vetivergrass can absorb more heavy met-
als (Roongtanakiat 2001). Vetivergrass as compared with
talahib and cogongrass is characterized by its massive finely
structured, deep, and easily adoptive root system. In addi-
tion, vetivergrass is tolerant to extreme climatic variation
such as prolonged drought, flood, submergence, and extreme
temperature. It is also tolerant to wide range of soil pH,
from 3.0 to 10.5, highly tolerant to soil salinity, sodicity,
acidity, aluminum, and manganese toxicities, and heavy
metals such as arsenic, cadmium, chromium, nickel, lead,
zinc, mercury, selenium, and copper in the soil (Truong 1999).
Carabaograss was observed to have a very strong and ex-
tensive root system, yet it was able to absorb very little
amount of EtBr (0.2±0.06 μg kg–1) from the soil (see Fig. 1).
Similar observation was found in talahib and cogongrass,
which also registered 0.2±0.55 μg kg–1 of EtBr after 30 days
of exposure (see Fig. 1). While a strong rooting system is
one of the required characters of a good phytoremediator, it
does not always mean it has inherent ability to accumulate
some chemicals and nutrients available in the soil. Also, plant
maturity period and type are to be considered. These grasses
are perennial plants that may take longer time to mature
and establish their rooting systems. Moreover, some plants
can accumulate high concentrations of some elements while
others may extrude them (Hale and Orcutt 1987). Absorp-
tion of materials by plants from the soil involves movement
from soil to roots, from the exterior of roots to interior, and
translocation within the plant. Furthermore, absorption and
uptake in phytoremediation are governed by physicochemi-
cal properties of the compounds, and the compound's solu-
bility to water (Raskin and Salt 1997). When inorganic nu-
trients and photosynthetic products are in water, these can
easily be conveyed to various parts of the plant. This is also
possible with EtBr, which is highly soluble in water and is
most likely bioavailable to vascular plants.
508
Subject Area 5.3
10.0
8.0
a
PLANT UPTAKE OF EtBr (g kg )
-1
6.0
b
b
4.0
2.0
c c c
0.0
TOMATO MUSTARD VETIVER TALAHIB CARABAO COGON
TROPICAL PLANTS
Fig. 2: Average uptake of EtBr by the different tropical plants. Uptake of
EtBr by different tropical plants are significantly different (p≤0.05) when
superscripts located at top of bars are different
Mustard had the highest plant uptake of EtBr (7.1 g kg–1)
compared with 5.1 g kg–1 by tomato and 3.1 g kg–1 taken
by vetivergrass. Talahib, cogongrass, and carabaograss re-
corded comparable uptake of 1.0 g kg–1 EtBr from the soil
(Fig. 2). Statistical analysis showed that the differences among
the test plants regarding their ability to accumulate EtBr
were highly significant at 1% level of significance. Com-
parison among treatment means also revealed that the re-
duction caused by mustard and tomato did not differ sig-
nificantly (p≤0.05). The same was observed in tomato and
vetivergrass. However, significant difference was noted be-
tween the means of mustard and vetiver grass (see Fig. 2).
The determination of plants that can work most efficiently
in a given application is the most important part in phyto-
remediation. Not all plant species can metabolize, volatil-
ize, or accumulate pollutants in the same manner. A good
candidate must grow quickly and consume large quantities
of water in a short time. It should also be able to remediate
more than one pollutant because pollution rarely occurs as
a single compound. Moreover, it must also find a way to
develop tolerance and must be able to absorb considerable
amount of pollutants in the least possible time of exposure.
Some studies were made on the subsequent result of plant
absorption of organic pollutants from soil. Organic pollut-
ants once absorbed by plants, can have multiple fates (Clemens
et al. 2002, Dhanker et al. 2002, Clarkson and Hanson 1980).
Many compounds are substantially bound into plant tissues
in a form that is less biologically available and may be un-
available to normal chemical metabolism. This may mean that
organic pollutants can be degraded or even mineralized by
plants or their associated microorganisms. Also, some are lost
with the aid of many physical factors such as temperature,
moisture, pH, organic matter content, and sunlight. The re-
duction of EtBr content of soil was highest where mustard
(10.7%) was planted (Table 1). This was followed by the soils
planted with tomato (8.1%) and vetivergrass (5.6%). Soils
planted with talahib, carabaograss, and cogongrass had the
lowest reduction of EtBr (see Table 1). Statistical analysis
showed highly significant differences (p≤0.001) among the
different test plants to reduce EtBr in soil. The reduction of
EtBr in soil by mustard, tomato, and vetivergrass were sta-
tistically different from one another (p≤0.05).
Env Sci Pollut Res 14 (7) 2007
Subject Area 5.3
Table 1: Levels of EtBr in soils and relative reduction of EtBr in soils after
30 days
Plants Initial Final Level
(treatments) Level in Soil in Soil Reduction
(μg kg–1) (μg kg–1)
Tomato 19.7 18.1±0.17
Mustard 19.7 17.6±0.23
Vetivergrass 19.7 18.6±0.23
Talahib 19.7 19.4±0.15
Carabaograss 19.7 19.4±0.20
Cogongrass 19.7 19.4±0.21
a
Means in column followed by a common letter(s) are not significantly
different from each other at p≤0.05
4 Conclusions
In this study, mustard, tomato, and vetivergrass have shown
their ability to absorb EtBr in contaminated soil keeping
them from expanding their reach into the environment and
preventing further contamination. Its downside, however, is
that living creatures including humans, fish, and birds, must
be prevented from eating the plants with significant accu-
mulation of these substances. Nonetheless, it is still easier to
isolate, cut down and remove plants growing in the con-
taminated soils (e.g., EtBr in this study), than to use strong
acids and permanganates to chemically neutralize a danger-
ous process that can further contaminate the environment
and pose additional risks to humans. Though this alterna-
tive method does not totally eliminate eventual environmen-
tal contamination, it by far produces an extremely insignifi-
cant amount of by-products compared with other processes.
Phytoremediation works in many favorable instances. It
doesn't leave behind its own pollution; it doesn't require an
unsightly treatment facility; it has no chemical add-ons; and
the tools needed (usually quite ordinary plants) are readily
available and inexpensive (Betts 1997). Also it is a safe, effi-
cient and sound clean-up technology to manage and protect
the environment. Because the sun acts as the engine, the
pollutants themselves can supply some of the fuel, and green
plants work like a well-oiled machine.
5 Recommendations and Perspectives
A more detailed and complete investigation of the phyto-
remediation potential of the different plants should be con-
ducted in actual field experiments. Exposure of plants to
EtBr should be prolonged until they reach maturity to es-
tablish the maximum response and maximum absorbing
qualities of the plants to the toxicity and mutagenecity of
EtBr. Also, time point sampling and analysis should be done
to determine at what stage of the plant's development the up-
take of EtBr should take place. Different plant parts should be
analyzed individually to determine the movement and trans-
location of EtBr from soil to plant tissues. Since this study has
established that some plants can thrive and dwell in EtBr treated
soil, an increased amount of EtBr should be explored in future
studies. It is suggested that a larger, more comprehensive ex-
ploration of phytoremediation application in the management
of toxic and hazardous wastes emanating from biotechnology
research activities should be considered especially the use of
Env Sci Pollut Res 14 (7) 2007
Ethidium Bromide
vetiver grass, a very promising perennial grass. Finally, this
study may have had paved the way for a larger and more
Percent comprehensive exploration of phytoremediation application
in the management of toxic and hazardous wastes emanat-
in Soil
ing from biotechnology research activities.
8.12b a
10.66a References
5.58b Betts K (1997): Phytoremediation project. Environmental Science and Tech-
1.52c nology 31 (8) 347–355
Clark RB (1982): Plant response to mineral element toxicity and deficiency.
1.52c pp 71–142
Clarkson DT, Hanson JB (1980): The mineral nutrition of higher plants.
1.52c
Annu Rev Plant Physiology 31, 239–298
Clemens S, Palmgren MG, Kramer U (2002): A long way ahead: Under-
standing and engineering plant metal accumulation. Trends in Plant Sci-
ence 7 (7) 309–314
Dhanker OM, Parkash II, Yujing R, Barry P, Shijin S, David S, Julie F,
Sashti NA, Meagher RB (2002): Engineering tolerance and hyper-
accumulation of arsenic in plants by combining arsenate reductase
and y-glutamylcysteine synthetase expression. Nature Biotechnology
20, 1140–1145
Fritz GG (1983): Introductory plant physiology, 2nd edition. Prentice-Hall
Inc., Englewood Cliffs, New Jersey, p 625
Hale MG, Dorcutt DM (1987): Physiology of plants under stress. Wiley
and Sons Inc., Canada
Indiana University Chemical Fact Sheet on Ethidium Bromide (1996): Indi-
ana, USA
Ona LF, Alberto AP, Prudente JA, Sigua GC (2006): Levels of lead in urban
soils from selected cities in the rice-based region of the Philippines. Env
Sci Pollut Res 13 (3) 177–183
Pressman E, Bar-tal A, Shaked R, Osenfeld K (1997): The development of
tomato root system in relation to the carbohydrate status of the whole
plant. Annals of Botany 80, 533–538
Raskin RD, Salt DE (1997): Phytoremediation of metals: Using plants to
remove pollutants from the environment. Curr Opin Biotechnology 8
(2) 2–6
Roongtanakiat N (2001): Uptake potential of some heavy metals by veti-
vergrass. Kasetsart J Nat Sci 35, 46–50
SAS (Statistical Analysis System) (2000): SAS/STAT User's Guide. Release
6.03. SAS Institute, Cary, North Carolina, 494 pp
Sigua GC (2005): Current and future outlook of spoil and sludge materials
in agriculture and environment. J Soils Sediments 5 (1) 50–52
Sigua GC, Adjei MB, Rechcigl JE (2005): Cumulative and residual effects of
repeated sewage sludge applications: Forage productivity and soil qual-
ity implications in South Florida, USA. Env Sci Pollut Res 12 (2) 80–88
Sigua GC, Holtkamp ML, Coleman SW (2004a): Assessing the efficacy of
dredged materials from Lake Panasoffkee, Florida: Implication to envi-
ronment and agriculture. Part 1 – Soil and Environmental Quality As-
pect. Env Sci Pollut Res. 11 (5) 321–326
Sigua GC, Holtkamp ML, Coleman SW (2004b): Assessing the efficacy of
dredged materials from Lake Panasoffkee, Florida: Implication to envi-
ronment and agriculture. Part 2 – Pasture Establishment and Forage
Quality. Env Sci Pollut Res 11 (6) 394–399
United States Environmental Protection Agency (2001): A citizen's guide
to phytoremediation. Accessed 12/11/2002, <http://www.cluin.org/prod-
ucts/citguide/phyto2.htm>
Truong PN (1999): Vetivergrass system for environmental protection. Tech-
nical Bulletin no.1, Pacific Rim Vetiver Network, Bangkok, Thailand
Wall Street (1996): Sunflowers bloom in tests to remove radioactive metals
from soil and water. Wall Street Journal 6 (2) 23–28
Watson L, Dallwitz MJ (1992): Grass genera of the world: Descriptions,
illustrations, identification, and information retrieval; including syn-
onyms, morphology, anatomy, physiology, phytochemistry, cytology,
classification, pathogens, world and local distribution, and references
Whitener K (1993): Phytoremediation. Environmental Science & Technol-
ogy Magazine 93 (8) 31–38
Zollinger M, Morais II (1979): Studies of the effect of chloramphenicol,
ethidium bromide and camptothecin on the reproduction of rous sar-
coma virus in infected chick embryo cells. Journal of General Virology
44, 323–331
Received: January 24th, 2007
Accepted: Februay 28th, 2007
OnlineFirst: Februay 28th, 2007
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