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Marigold ( Tagetes erecta ): The Potential Value in the Phytoremediation of

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DOI: 10.1016/S1002-0160(17)60351-5

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Pedosphere 27(3): 559–568, 2017
doi:10.1016/S1002-0160(17)60351-5
ISSN 1002-0160/CN 32-1315/P
⃝c 2017 Soil Science Society of China
Published by Elsevier B.V. and Science Press

Marigold (Tagetes erecta): The Potential Value in the

Phytoremediation of Chromium

Lı́via C. COELHO1,∗ , Ana Rosa R. BASTOS1 , Paulo J. PINHO1 , Guilherme A. SOUZA1 , Janice G. CARVALHO1 ,
Viviane A. T. COELHO1 , Luiz Carlos A. OLIVEIRA2 , Rimena R. DOMINGUES1 and Valdemar FAQUIN1
1 Soil Science Department, Federal University of Lavras, Lavras 37200000 (Brazil)
2 Chemistry Department, Federal University of Minas Gerais, Belo Horizonte 31270901 (Brazil)
(Received December 30, 2016; revised March 9, 2017)

ABSTRACT
Environmental pollution with chromium (Cr) is harmful to humans, animals and plants, while in plants it causes diminished growth,
anatomical alterations and death. In the present study, the potential value of marigold (Tagetes erecta) in the phytoremediation of
Cr has been investigated. The randomized experimental design involved the exposure of plants to nutrient solutions containing 0.00,
0.04, 0.08, 0.12, 0.16 or 0.24 mmol L−1 Cr(III). Chromium toxicity was observed at Cr(III) concentrations ≥ 0.12 mmol L−1 as
demonstrated by diminished growth of the aerial parts and reduced density of the root system. Increasing Cr(III) concentrations in
the nutrient solution resulted in a higher bioaccumulation of total Cr in the tissues, although translocation from roots to aerial parts
was not efficient (maximum value of 25% at 0.12 mmol L−1 Cr(III)). The Cr bioaccumulation was up to 11-fold greater in roots than
in the aerial parts. Tagetes erecta exhibited leaf plasticity when exposed to Cr, indicating the existence of a tolerance mechanism
to Cr in this species. Chromium caused a reduction in xilem vases, resulting in a plastic effect in T. erecta leaves that increased
the metal tolerance in culture solution. Tagetes spp. are potential Cr hyperaccumulators; at Cr(III) concentrations up to 0.12 mmol
L−1 , the plants accumulated levels above that proposed for hyperaccumulators and still maintained a considerable growth and even
flourished. However, this study was conducted in nutrient solution, and studies on species confirmation as Cr hyperaccumulator should
be conducted in soils for further clarification.
Key Words: anatomical alterations, heavy metal, hyperaccumulor, leaf anatomy, toxicity

Citation: Coelho L C, Bastos A R R, Pinho P J, Souza G A, Carvalho J G, Coelho V A T, Oliveira L C A, Domingues R R, Faquin
V. 2017. Marigold (Tagetes erecta): The potential value in the phytoremediation of chromium. Pedosphere. 27(3): 559–568.

INTRODUCTION lue for drinking-water (WHO, 2011).

Chromium (Cr) is a heavy metal that is found
widely in the trivalent or hexavalent forms in soils,
rocks, volcanic ashes and groundwaters (Shanker et
al., 2005). It is employed extensively in the production
of steel, pigment, paint and paper, in galvanization
processes, in the refining of oil and in the wood and
leather industries (Kabata-Pendias and Mukherjee,
2007; Gupta et al., 2010). The release of Cr residues
into the atmosphere and wastewaters can cause si-
gnificant damage to living organisms, owing primarily
to the conversion of Cr(III) to Cr(VI). Although the
trivalent form of Cr is essential for human and ani-
mal nutrition and plays a key role in glucose and lipid
metabolism, the hexavalent form is toxic and muta-
genic owing to its strong oxidizing capacity and its
ability to cross biological membranes (Ferreira, 2002;
Kolomaznı́k et al., 2008). Thus, a concentration of 0.05
mg L−1 of total Cr was established as the guideline va-
Although an essential biological role for Cr in
plants has yet to be established, some studies have
indicated that it may stimulate plant growth at low
concentrations (ca. 1 µmol L−1 ) (Hossner et al., 1998).
Furthermore, there appears to be considerable inter-
and intra-species variations regarding the absorption
of Cr and its subsequent retention in roots and/or
translocation to other tissues via the xylem. A number
of factors influence the storage and translocation of Cr
in plants, including stage of development, duration of
exposure to the metal and the presence of other chemi-
cal elements. Root tissues play an essential role in the
differential tolerance of a species to heavy metals, sin-
ce they are able to regulate absorption from the rhi-
zosphere and the subsequent sequestration (in order
to preserve the integrity and primary functions of the
plant) and/or translocation to aerial parts (Soares et
al., 2001).
Chromium accumulation differs greatly between si-

∗ Corresponding author. E-mail: liviacoelho 6@hotmail.com.

560
tes and plants. The hyperaccumulators must achieve
their high metal concentrations while remaining hea-
lthy enough to maintain a population (van der Ent
et al., 2013). Given the very low Cr concentrations in
plants, it is proposed to set 300 mg kg−1 for hyper-
accumulator status both on normal and metalliferous
soils. Since no specific Cr transporters have been iden-
tified in plants, it is likely that the metal is transported
by carriers of essential elements. Indeed, it has been
demonstrated that Cr competes with Fe, S and P for
binding sites in protein-transporters (Shanker et al.,
2005). Additionally, all biological processes associated
with Cr, including absorption, translocation and accu-
mulation, depend on the oxidation state of Cr. There-
fore, the absorption of Cr(VI) probably occurs via an
active mechanism, whereas that of Cr(III) is by passive
transport (Hossner et al., 1998; Shanker et al., 2005).
Marigold (Tagetes erecta) is an attractive and wide-
ly grown ornamental species of the family Asteraceae
(Compositae) (Lorenzi and Souza, 2001). Members of
the genus Tagetes are also of interest as sources of bi-
ologically active compounds (Vasudevan et al., 1997)
and natural food colorants (Barzana et al., 2002). In
this context, the pigment extracted from T. erecta is
used in the preparation of vegetable oils, pasta, bread,
juices, mustard and milk derivatives (Delgado-Vargas
and Paredes-López, 2003), and is added to chicken feed
in order to intensify the yellow coloration of broiler
flesh and egg yolks (Martinez et al., 2004).
The rapid growth and well-developed root system
of T. erecta, together with its ability to act as a pio-
neer on poor soils, suggest that this species might be
suitable for the remediation of areas degraded by me-
tal pollution. Indeed, in the last decade, phytoreme-
diation technologies have received increasing attention
from scientists and private companies for the clean-up
of soils contaminated by organic and inorganic sub-
stances, by either reduction, removal, degradation or
immobilization of the contaminants (Das and Maiti,
2007).
Thus, T. erecta could be employed with the mu-
ltiple purpose of decontaminating urban areas and
brownfield sites, while providing environment-enhan-
cing green spaces and a source of commercially valu-
able products. Within this context, the objectives of
the present study were to investigate: i) the growth of
T. erecta in nutrient solutions containing different con-
centrations of Cr(III); ii) Cr bioaccumulation in roots
and aerial parts of the plants and iii) signs of Cr toxici-
ty in the anatomy of the leaves. The accomplishment
of these objectives will improve our understanding re-
L. C. COELHO et al.
garding the possible mechanisms by which T. erecta
tolerates the presence of Cr and verify the phytoreme-
diation potential of this species.
MATERIALS AND METHODS
Experimental layout
This study was conducted in the Soil Science De-
partment at Federal University of Lavras, Lavras, Mi-
nas Gerais, Brazil (21◦ 14′ S, 45◦ 00′ W, 910 m above sea
level). Nutrient stock and Cr(III) solutions were pre-
pared using analytical grade (PA) reagents and deio-
nized water. Commercially acquired seeds of T. erecta
were germinated in 72-cell styrofoam trays containing
substrate. After 11 d, the seedlings were transferred to
plastic trays containing 36 L Hoagland and Arnon nu-
trient solution (Hoagland and Arnon, 1950), the ionic
strength of which (25%, 50% or 100%) was increased
sequentially every 7 d, when constant aeration condi-
tions were maintained.
Following the adaptation period, the plantlets were
transferred to separate 3-L plastic pots containing
appropriate treatment solutions, and were subjected
to continuous aeration with compressed air supplied
using transparent hoses. The experiment was a com-
pletely randomized design and involved six treatments
with four repetitions each. The treatments consisted
of Hoagland and Arnon nutrient solutions (100%) con-
taining 0.00 (non-treated control), 0.04, 0.08, 0.12,
0.16 or 0.24 mmol L−1 Cr(III) as Cr(NO3 )3 ·9H2 O.
The concentrations of Cr(III) employed were sele-
cted on the basis of preliminary assays (results not
shown). Throughout the experiment, the volume of nu-
trient was maintained daily by addition of deionized
water, and the nutrient was replaced by fresh solution
every 15 d. During the growing period, plants were sup-
ported by styrofoam sheets. At the start of flowering,
which occurred 10 weeks after the commencement of
treatment, plants were evaluated with respect to stem
diameter and height. Plants were then harvested, sepa-
rated into roots and aerial parts, washed under running
water, rinsed with distilled water and dried to constant
weight in a hot air oven at 65–70 ◦ C. The dried plant
parts were weighed separately using a precision digital
scale, reduced to a powder in a Wiley mill and analyzed
for total Cr concentration.
Quantitative determination of Cr and calculation of
plant indicators
The concentrations of total Cr in root and aerial
tissues of experimental plants (mg kg−1 ) were determi-
POTENTIAL OF MARIGOLD IN CHROMIUM PHYTOREMEDIATION
ned according to the methodology described by Mala-
volta et al. (1997). The total Cr bioaccumulation (BA,
mg) in plant tissue (roots or aerial parts) was calcula-
ted using the equation:
BA = Total Cr concentration in the tissue ×
Dry biomass of the tissue (1)
The capacity of a plant to absorb Cr from the nu-
trient solution was determined by calculating the phy-
toextraction coefficient (PEC) (Henry, 2000) according
to the equation:
Total Cr concentration in the plant
PEC = (2)
Total Cr concentration in the solution
where the total Cr concentration in the plant was ta-
ken as the sum of the concentrations of Cr in roots
and aerial parts, and the total Cr concentration in the
solution was calculated as the product of the concen-
tration of Cr(III), the volume of nutrient solution per
plastic pot and the number of changes of nutrient so-
lution during the trial period. The higher the value of
PEC, the greater the absorption of Cr.
The influence of Cr(III) concentration in the solu-
tion on plant yield was determined by calculating the
relative dry biomass yield (RBY, %) according to the
equation:
Dry biomass in the presence of Cr
RBY = × 100 (3)
Dry biomass in the absence of Cr
The translocation index (TI, %) was employed to
evaluate the capacity of T. erecta to translocate Cr
from the roots to the aerial parts (Paiva et al., 2002),
and was calculated according to the equation:
Total Cr concentration in the aerial parts
TI = ×
Total Cr concentration in the plant
100 (4)
Light microscopy
Leaves of plants that had been exposed to Cr were
examined under the light microscope in order to detect
possible anatomical alterations. Leaves were collected
from the mid-third sections of each of the experimental
plants, and subsequently fixed in formaldehyde, acetic
acid and 70% ethanol for 72 h and stored in 70% alco-
hol (Johansen, 1940). Cross sections of the leaves were
cut with a microtome, clarified in 5% sodium hypochlo-
rite solution (Kraus and Arduim, 1997) and stained
with astra blue/safranin. Slides were prepared in 20%
glycerin and observed under a light microscope. Pho-
tomicrographs were recorded using a digital camera,
561
and anatomical characteristics were analyzed with the
help of UTHSCSA Image Tool software.
Statistical analysis
Data were submitted to analysis of variance (ANO-
VA) and the mean values were compared using the
Scott-Knott test at the 5% probability level. The con-
centrations of Cr(III) in the nutrient solutions were
considered dependent variables in the regression analy-
ses. The strengths of the relationships (as described by
the regression equations) were estimated from the coef-
ficients of determination (R2 ), and the significances of
such relationships were established using the Student’s
t test at the 5% probability level. Regression models
which presented the highest R2 values were selected to
explain the variation within the data and the biological
significance of the model. All statistical analyses were
performed using SISVAR software (Ferreira, 2003).
RESULTS
Growth of T. erecta
Exposure to Cr(III) affected all of the studied
growth parameters of T. erecta, namely, stem diame-
ter, plant height and dry biomass of roots and aerial
parts (Fig. 1). Stem diameter and dry biomass of the
aerial parts diminished linearly with increasing Cr(III)
concentrations. However, plant height and dry biomass
of roots varied with Cr(III) concentration in a quadra-
tic fashion, with plant height reaching an estimated
maximum value of 97 cm at 0.005 mmol L−1 Cr(III)
and root dry biomass attaining an estimated maximum
value of 6.33 g at 0.05 mmol L−1 Cr(III).
Biomass production of T. erecta
Production of dry biomass (evaluated as RBY va-
lues) of T. erecta decreased markedly as the concentra-
tion of Cr(III) in the nutrient solution increased (Table
I), with a reduction of 82% being recorded after expo-
sure to 0.24 mmol L−1 Cr(III).
The reduction in linear growth with increasing con-
centrations of Cr(III) could be readily observed, es-
pecially in the concentration range from 0.12 to 0.24
mmol L−1 (Fig. 2a). At these concentrations, the leaves
were slender and fragile, and flowering was premature
in comparison with those of the non-treated plants.
Furthermore, the root system was dark in color, re-
latively undeveloped, and exhibited an abnormal ar-
chitecture with thicker lateral roots in comparison
with the white dense roots of the non-treated plants
(Fig. 2b).
562 L. C. COELHO et al.

Fig. 1 Influence of increasing concentrations of Cr(III) in nutrient solutions on stem diameter, plant height, dry biomass of aerial parts
and dry biomass of roots of Tagetes erecta.

TABLE I
Influence of increasing concentrations of Cr(III) in nutrient solutions on the relative biomass yield (RBY) and the ratio of total Cr
bioaccumulation (BA) in the roots (BAroots ) to that in the aerial parts (BAap ) of Tagetes erecta

Parameter Concentration of Cr(III) (mmol L−1 )

0.00 0.04 0.08 0.12 0.16 0.24

RBY (%) 100.0 87.1 78.6 75.1 64.2 18.0
Ratio of BAroots /BAap – 9 11 3 10 4

Concentration and bioaccumulation of total Cr in T.
erecta
Concentrations of total Cr in roots and aerial parts
of T. erecta increased linearly with increasing concen-
trations of Cr(III) in the nutrient solutions (Fig. 3), in-
dicating a direct correlation between the bioavailability
of the metal and the absorption by plants.
The variation in BA of total Cr in the aerial parts
of T. erecta presented a quadratic profile, whereas it
followed a somewhat erratic pattern that could not be
described by a simple mathematical function in the
roots (Fig. 3). For this reason, the ratio of BA in roots
and aerial parts was also extremely variable (Table I),
although total Cr BA in roots was always much greater
(up to 11 fold) than in the aerial parts. The largest
values of BA in roots and shoots were both observed
when the concentration of Cr(III) in the nutrient so-
lution was 0.16 mmol L−1 , and under these conditions
BA in the roots was 10-fold greater than that of the
aerial parts.
Phytoextraction and translocation of total Cr by T.
erecta
The value of PEC provides information regarding
the capacity of a plant to extract a specific contami-
nant from the surrounding substrate. The higher the
PEC value, the greater the level of uptake (Henry,
2000). In the case of T. erecta, the highest value of PEC
was obtained when Cr(III) was present in the medium
at a concentration of 0.16 mmol L−1 (Fig. 4). At this
concentration of Cr(III), the calculated value of total
Cr BA in the roots was maximal (Fig. 3), while the TI
was one of the lowest values recorded at different levels
POTENTIAL OF MARIGOLD IN CHROMIUM PHYTOREMEDIATION
Fig. 2 Influence of increasing concentrations of Cr(III) in nutrient solutions on linear growth and aerial parts (a) and root morphology
(b) of Tageteserecta. Cr (0), Cr (0.04), Cr (0.08), Cr (0.12), Cr (0.16) and Cr (0.24) = 0.00, 0.04, 0.08, 0.12, 0.16 and 0.24 mmol L−1
Cr(III), respectively.
of Cr(III) (Fig. 4). The TI is an indicator of the capaci-
ty of a plant species to transport nutrients and other
substances from the roots to leaves. As shown in Fig. 4,
the translocation of Cr from roots to the aerial parts
of T. erecta was typically modest, with a maximum
translocation of just 25% at 0.12 mmol L−1 Cr(III). In-
deed, most of the Cr absorbed by the roots of T. erecta
was accumulated there, with a maximum accumulation
of 92% at 0.08 mmol L−1 Cr(III).
Anatomical characteristics in leaves of T. erecta
The compact and dorsiventral mesophyll of the leaf
blade of T. erecta (a dicotyledon) which was clear-
ly differentiated into palisade and spongy parenchyma
(Fig. 5a). The former comprised a single layer of cells,
while the latter consisted of four or five layers of round
and thin-walled cells that were loosely packed and
characterized by small intercellular spaces. The epider-
mis of the adaxial and abaxial surfaces exhibited a sin-
gle layer of cylindrical cells. Vascular bundles of vari-
able dimensions were integrated into the spongy me-
sophyll. The collenchyma, which provides mechanical
support to the leaves, was readily discernable beneath
563
the epidermis and close to the main vein (Fig. 5b), to-
gether with xylem (upper surface) and phloem (lower
surface) vessels.
Various structural modifications of T. erecta were
observed at different concentrations of Cr(III), which
could contribute to tolerance towards Cr. The thick-
ness of the spongy parenchyma increased when Cr(III)
concentrations were in the range from 0.04 to 0.12
mmol L−1 (Table II). However, the thickness of the
palisade parenchyma was significantly reduced and the
number of xylem diminished in the presence of 0.16
mmol L−1 Cr(III) in comparison with untreated plants
(Fig. 5c).
DISCUSSION
Growth of T. erecta
At high concentrations, Cr(III) causes oxidative
stress, alterations in chloroplast structure and degene-
ration of photosynthetic pigments. Furthermore, Cr
also reduces the catalytic activity of antioxidant en-
zymes (Taiz and Zeiger, 2004) and cellular metabolism
in shoots (Shanker et al., 2005). These factors may have
564
Fig. 3 Concentration and bioaccumulation (BA) of total Cr in the aerial parts and roots of Tagetes erecta exposed to nutrient solutions
containing different concentrations of Cr(III).
Fig. 4 Variations in Cr phytoextraction coefficient (PEC) and
translocation index (TI) of Tagetes erecta exposed to nutrient
solutions containing different concentrations of Cr(III). Vertical
bars are standard errors of the means (n = 4).
contributed to the observed reduction in linear growth
L. C. COELHO et al.
of plants exposed to Cr(III) at concentrations ≥ 0.12
mmol L−1 (Fig. 2a). In contrast, very low concentra-
tions of Cr(III) may stimulate plant growth (Hossner
et al., 1998), and could have exerted a beneficial effect
on T. erecta exposed to Cr(III) with concentrations <
0.12 mmol L−1 .
According to Castilhos et al. (2001a, b), the dry
biomasses of leaf and root tissues of soybean plants
were not significantly affected by exposure to Cr(VI)
at 5 mg L−1 (ca. 0.096 mmol L−1 ) or to Cr(III) at 20
mg L−1 (ca. 0.38 mmol L−1 ) in comparison with plants
grown in the absence of the heavy metal. However, dry
biomass was significantly reduced when soybean plants
were exposed to Cr with concentrations above the sta-
ted levels. Tolerance to low concentrations of the heavy
metal has also been reported for hydroponically-grown
tomato (Moral et al., 1995) and lettuce (Figliolia et al.,
1992) plants in which growth was diminished only in
the presence of Cr(III) at 100 mg L−1 (ca. 1.92 mmol
L−1 ).
Biomass production of T. erecta
Production of dry biomass of T. erecta decreased
markedly with increasing concentrations of Cr(III) in
the nutrient solutions (Table I). Similar findings have
been reported for eelgrass (Vallisneria spiralis), com-
mon purslane (Portulaca oleracea), cauliflower (Bras-
sica oleracea var. botrytis), cabbage (Brassica oleracea
POTENTIAL OF MARIGOLD IN CHROMIUM PHYTOREMEDIATION 565

Fig. 5 Photomicrographs of cross sections of leaves of Tagetes erecta showing: leaf blade (a) and main vein in the absence of Cr(III) (b)
and main vein under exposure to 0.16 mmol L−1 Cr(III) in nutrient solution (c). AdE = adaxial epidermis; AbE = abaxial epidermis; PP
= palisade parenchyma; SP = spongy parenchyma; Xy = xylem; Ph = phloem; VB = vascular bundles; AC = angular collenchyma.

TABLE II
Anatomical characteristics of leaves of Tagetes erecta exposed to different concentrations of Cr(III) in nutrient solution

Concentration Thickness No. of

of Cr(III) vessels in
Palisade parenchyma Spongy parenchyma Adaxial epidermis Abaxial epidermis Leaf blade Phloem xylem
µm
0.00 108.5ba) 135.8a 11.7a 9.0a 281.8b 92.4a 42.6b
0.04 107.8b 147.3b 11.9a 8.2a 289.1b 102.1a 41.6b
0.08 105.7b 150.9b 14.1b 8.5a 298.3b 97.1a 35.8b
0.12 119.2b 192.5c 14.4b 9.4a 364.4c 105.1a 28.3a
0.16 79.9a 123.9a 12.9b 9.1a 240.7a 89.9a 27.3a
0.24 103.6b 159.6b 9.9a 9.3a 289.0b 94.6a 30.5a
CVb) (%) 7.79 8.34 10.65 10.9 8.56 13.03 20.11
a) Means followed by the same letter in a column are not significantly different at the 5% probability level according to Scott-Knott
test.
b) Coefficient of variation.

var. capitata), radish (Raphanus sativus), common
bean (Phaseolus vulgaris), maize (Zea mays), oat (Ave-
na sativa), cucumber (Cucumis sativus), lettuce (Lac-
tuca sativa), millet (Panicum miliaceum) and white
mustard (Sinapis alba) (Shanker et al., 2005), and the
reductions in dry biomass were attributed to Cr to-
xicity. The exposure of plants to heavy metals appa-
rently induces the generation of reactive oxygen species
(ROS), such as superoxide anion, singlet oxygen, hy-
droxyl and hydrogen peroxide, which have the poten-
tial to cause oxidative damage to biomolecules simi-
lar to that caused by abiotic and biotic stress factors,
including drought, UV radiation and chemical pollu-
tion (Foyer et al., 1994; Sanità di Toppi and Gabbriel-
li, 1999; Benavides et al., 2005). This type of oxidative
burst may lead to a considerable reduction in growth
and productivity, and may even trigger plant death
(Foyer et al., 1994).
Concentration and bioaccumulation of total Cr in T.
erecta
Some authors have reported that higher plants
present symptoms of toxicity when the concentration
of total Cr in aerial tissue exceeds 5.2 mg kg−1 dry
mass (Hossner, 1998). In maize, e.g., growth was re-
duced by 50% when Cr attained a level of 5.9 mg kg−1
in the dry plant biomass (Chang et al., 1992). On the
other hand, Losi et al. (1994) have claimed that most
plants exhibit signs of toxicity when the concentration
of Cr in the leaves is > 18 mg kg−1 . In the present
study, signs of Cr toxicity in T. erecta, i.e., reductions
in growth and dry biomass production, were observed
when the concentrations of total Cr in the aerial tissues
were even higher than the limits reported previously.
Thus, visual signs of phytotoxicity in T. erecta were
observed only when the concentration of total Cr in
the nutrient solution was ≥ 0.12 mmol L−1 , and under
these conditions the concentration of total Cr in the
aerial parts was approximately 300 mg kg−1 , whilst
that in the roots was ≥ 10 g kg−1 . Since the roots are
the main organs involved in the absorption of water,
nutrients and pollutants, it is not surprising that the
largest concentrations of Cr are located in these tissues
(Grant et al., 1998). This finding is in agreement with
566
previous reports, demonstrating that accumulation of
the Cr absorbed by roots is highly regulated in order
to minimize toxic effects, with most of Cr being im-
mobilized in the vacuoles of root cells and only small
amounts being translocated to the leaves (Losi et al.,
1994; Hossner et al., 1998; Shanker et al., 2005).
According to Tan (2000), the uptake of heavy me-
tals by plants and the subsequent translocation to aeri-
al parts vary according to species. When plants of hy-
brid willow (Salix sp.) were exposed to solutions con-
taining Cr(III), concentrations of Cr in leaves, stems
and roots varied considerably (Yu and Gu, 2007). In-
terestingly, the concentration of Cr in the roots of
plants that had been exposed to 7.5 mg L−1 (ca. 0.14
mmol L−1 ) Cr(III) was higher than that in the roots
of plants that had been exposed to 30 mg L−1 (ca.
0.58 mmol L−1 ) Cr(III). These studies demonstrate
that when plants are exposed to toxic levels of metals,
their capacity to absorb and bioaccumulate declines
markedly, probably because of a generalized reduction
in metabolic rate. In our study, at Cr(III) concentra-
tions up to 0.12 mmol L−1 , the plants accumulated Cr
levels (ca. 800 mg kg−1 ) above that proposed by van
der Ent et al. (2013) for hyperaccumulators (300 mg
kg−1 ) and still maintained a considerable growth and
even flourished.
Phytoextraction and translocation of total Cr by T.
erecta
Evidently, a high concentration of Cr(III) in solu-
tion, together with an extended time of exposure, pro-
duced serious damages to the structure and metabo-
lism of the plant, and especially to organelles re-
sponsible for the production and conversion of ener-
gy (Figs. 1, 2, 5, Table II). Thus, the level of damage
caused by Cr was associated with the concentration of
Cr(III) in the nutrient medium and the time of expo-
sure of T. erecta to the additive. Over evolutionary
time, plants have developed complex molecular and
physiological mechanisms in order to survive in metal-
rich environments and to ensure the preservation of
the species. One of these strategies is stress escape,
which is related to external protection, while anoth-
er involves tolerance, which is associated with internal
protection. Plants growing in contaminated soils can-
not escape entirely from absorbing toxic metals, al-
though they can limit damage by accumulating the
toxins in specific tissues. In this case, tolerance rather
than escape would be the main survival strategy (Paiva
et al., 2002).
Anatomical characteristics in leaves of T. erecta
On exposure to heavy metals, plants may undergo
L. C. COELHO et al.
anatomical changes, related to their anatomical plas-
ticity or adaptative measures, in order to develop tole-
rance to toxic levels of such metals in their tissues.
Following exposure to 0.16 mmol L−1 Cr(III), the con-
centration at which total Cr BA and PEC attained
their maximum values, the thickness of the palisade
parenchyma was significantly reduced in comparison
with untreated plants. This negative outcome was asso-
ciated with Cr toxicity which, according to Panda and
Choudhury (2005), can cause modifications in the ul-
trastructure of chloroplasts and cell membranes, resul-
ting in the inhibition of photosynthetic reactions and,
consequently, in the diminution of growth. Indeed, the
signs of Cr toxicity were readily observable in T. erec-
ta as reductions in height, dry biomass and stem dia-
meter. Similar findings have been reported for Indian
mustard (Brassica juncea), in which the sizes of the
palisade parenchyma cells were reduced when plants
were grown in soil contaminated with Zn and Cr, while
the collapse of palisade and spongy parenchyma cells
was observed when plants were cultured under Zn and
Cd stress (Maruthi Sridhar et al., 2005).
The thickness of the spongy parenchyma of T. erec-
ta increased when concentrations of Cr(III) were in the
range of 0.04 to 0.12 mmol L−1 (Table II), demonstra-
ting plasticity within this tissue in order to minimize
the accumulation of Cr in the palisade parenchyma
and diminish possible damage to developing chloro-
plasts. Vollenweider et al. (2006) showed that although
small amounts of Cd accumulated in the leaf mesophyll
of Salix viminalis that had been exposed to the metal,
the distribution through the leaf layers indicated that
levels in the photosynthesizing parts were minimal. In-
deed, plants may store metals in non-photosynthetic
tissues as a strategy to improve tolerance when toxic
levels are present. Interestingly, the thickness of the
spongy parenchyma in T. erecta did not increase when
plants were exposed to concentrations of Cr(III) ≥ 0.16
mmol L−1 . However, under these conditions, the pali-
sade parenchyma was reduced and, in relative terms,
was still thinner than the spongy parenchyma. Hence,
the rationalization regarding the preservation of pho-
tosynthetic structures under conditions of metal stress
remains applicable. Additionally, at 0.16 mmol L−1
Cr(III), the thickness of the leaf blade diminished (Ta-
ble II), caused presumably by downsizing of the me-
sophyll cells (Zhao et al., 2000; Maruthi Sridhar et al.,
2005). This type of anatomical plasticity or adaptation
ability has been observed in other plant species as a re-
sponse to stress conditions (Melo et al., 2007), and is
believed to represent a compensation mechanism that
allows sunlight to reach photosynthetically active cells
more readily. The specific combination of responses to
POTENTIAL OF MARIGOLD IN CHROMIUM PHYTOREMEDIATION
Cr stress observed in T. erecta likely account for the
maximum accumulation of Cr detected in this plant
that had been exposed to 0.16 mmol L−1 of Cr(III).
While the leaf adaxial epidermis was significant-
ly enlarged in plants that had been exposed to 0.08–
0.16 mmol L−1 Cr(III), exposure to Cr(III) had no
influence on the thickness of the leaf abaxial epider-
mis (Table II). The epidermis of roots and leaves is
very responsive to environmental stress (light, salini-
ty, humidity, drought and flooding), and variations in
the structure of this layer have been reported for vari-
ous species (Reinhardt and Rost, 1995; Enstone et al.,
2003; Melo et al., 2007). Thickening of the epidermis
may constitute a strategy to reduce water loss through
transpiration. Since the epidermis is closely associated
with root absorption, a diminished rate of transpira-
tion would imply a reduced uptake of metal ions.
In T. erecta, the number of xylem vessels, but not
phloem vessels, diminished in direct response to in-
creasing concentrations of Cr(III) in the nutrient so-
lution (Table II, Fig. 5c). Environmental factors can
affect the dimensions and organization of the vascular
tissue, as well as the number of vessels, thus modu-
lating hydraulic conductivity (Alves and Angyalossy-
Alfonso, 2000). Additionally, exposure to heavy metals
has been shown to alter the water balance in plants
(Barceló and Poshenrieder, 1990). The metal-induced
reduction in the number of xylem vessels diminishes
water flow and, consequently, the translocation of nu-
trients and the balance of growth regulators (Sandalio
et al., 2001).
It is noteworthy that the values of all of the pa-
rameters assessed in T. erecta (with the exception of
the number of xylem vessels) were not significantly di-
fferent from those of untreated controls at 0.24 mmol
L−1 Cr(III), indicating that plants could not undergo
anatomical plasticity at this toxic concentration in or-
der to protect the development of aerial parts.
In conclusion, the T. erecta accumulated levels
above that proposed for hyperaccumulators, which can
be used in the phytoremediation of Cr-contaminated
soils. Characteristics of rapid growth, well developed
root system, acclimatization facility, pioneer in soils
of low fertility and economic importance attributed to
the species potential for employment in programs of
recovery of degraded areas by the deposition of heavy
metals. This potential with a possible phytoremedia-
tion of an urban environment should also be associated
with architecture and landscaping. However, this stu-
dy was conducted in nutrient solution, and studies on
the species confirmation as hyperaccumulator should
be conducted in soils for further clarification.
567
ACKNOWLEDGEMENT
The authors thank the Coordenação de Aper-
feiçoamento de Pessoal de Nı́vel Superior (CAPES),
the Fundação de Amparo à Pesquisa do Estado de
Minas Gerais (FAPEMIG) and the Conselho Nacional
de Desenvolvimento Cientı́fico e Tecnológico (CNPq),
Brazil for financial support and scholarships.
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