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American-Eurasian J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
ISSN 1818-6769
© IDOSI Publications, 2008
Corresponding Author: Hossein Mirseyed Hosseini, Soil Sci. Dept., Soil and Water Facult. Univ. of Tehran, Karaj, Iran
93
Effect of Sunflower and Amaranthus Culture and Application of
Inoculants on Phytoremediation of the Soils Contaminated with Cadmium
Babak Moteshare Zadeh, Gholam Reza Savaghebi-Firozabadi,
12
Hossein Ali Alikhani and Hossein Mirseyed Hosseini
33
Ph.D. Student in Soil Science Engineering, Associate Professor and Assistant Professors,
1 23
University College of Agriculture & Natural Resource, University of Tehran, Karaj, Iran
Abstract: Cadmium is a toxic element in plant and human nutrition which is added to the soil from different
sources especially consumption of phosphate fertilizers with high cadmium concentration, application of
industrial wastewaters, mine extraction operations and metal melting and leads to contamination of environment.
Phytoremediation is an effective, economical and biocompatible method for remediation of contaminated soils.
This research was executed in order to study effect of sunflower and amaranthus culture and inoculants of two
native bacteria resistant to cadmium on phytoremediation of this metal. In a calcareous soil of Karaj region
(Fine Loamy, Mixed Super Active Thermic Xeric Haplocambids) and in green house conditions, effect of
culture of the two plants, sunflower and amaranthus and three levels of control inoculants (BO),
Bacillus mycoides M1 (B1), Micrococcus roseus M2 (B2) and four levels of control cadmium concentration
(0, 50, 100 and 200 mg/kg) was studied in a factorial experimental design with random blocks basic design with
three replications. Concentration of cadmium, iron and zinc were measured in shoot and root as a function of
dry material and photosynthetic chemical efficiency. The analysis of variance analysis showed that application
of inoculant significantly (P< 0.01) increased phytoremediation efficiency and effect of amaranthus in cadmium
phytoextraction was higher than that of sunflower. It seems that effect of sunflower on phytoremediation is
generally through phytoestabilisation and effect of amaranthus is through phytoextraction process. Treatments
of cadmium increased concentration of this element in plant and decreased photosynthetic quenching (Fv/Fm).
It is recommended to do more studies in this field and under field conditions.
Key words: Pytoremediation Soil pollution Cadmium Sunflower Amaranthus
INTRODUCTION heavy and toxic metal for the plant and prevents the
With growth and development of industries, melting with agricultural products [2]. Concentration of cadmium
metals and consumption of chemical fertilizers, in uncontaminated soils is usually less than 0.5 mg/kg,
contamination of heavy metals have been turned into a but it can reach 3 mg/kg depending on the type of parent
serious environmental problem [1,2]. Contaminant heavy materials. Higher values result from activities of human
metals including cadmium, chromium, copper, mercury, being such as mining, production and consumption of
lead and nickel are defined as elements with metal phosphate fertilizers and activated sludge. Concentration
properties and atomic number higher than 20 and density of cadmium in uncontaminated plant is between 0.05
higher than 6 g/cm3 [2,3]. When heavy metal ions are in and 2 mg/kg [7,8]. Annually, extent of entrance and
higher levels in the environment, they are transmitted accumulation of cadmium in agricultural soils increases
through phytoextraction by roots to shoots leading to and 40-70% of cadmium in agricultural products result
disorder in metabolism of plant and fertility of soil and directly from phosphate fertilizers and rock phosphate is
decrease in function of products [4,5]. In addition, a high considered as major source of heavy metal increase [9].
concentration of heavy metals in soil reduces biological Tests showed that triple superphosphate fertilizers, zinc
activity and fertility of soils leading to yield reduction sulphate and rock phosphate consumed in US contained
[6]. As an unnecessary element, cadmium is considered a 150.3, 295.7 and 44.4 mg/kg of cadmium respectively and
growth of root and shoots of the plants and is associated
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
94
consumption of these fertilizers increased extent of remediation were related to removal of nickel from the soil
cadmium available in soil and agricultural products [10]. with indian mustard and canola [21]. Zinc phytoextraction
With regard to emergence of heavy metals contamination with oat, barley and Indian mustard were also studied
in environment, remediation of contaminated places is a and the results showed that adding EDTA to the soil
serious challenge. These compounds are not degraded significantly led to accumulation of zinc in indian
and removed and their remediation depends on their mustard. It was also shown that barley had potential of
removal from the environment incurring high expenses. phytoremediation though this potential was not more than
In addition, during metals removal stage, one should Indian mustard [29]. In another research, zinc and
use chemicals or physicochemical material preventing cadmium absorption in Zea Maize and Thlaspi
fertilization of soil and having negative effect on caerulescens was studied and necessity of
ecosystem and biodiversity [11]. Phytoremediation is an phytoremediation expansion was emphasized with regard
effective, cheap and biocompatible method with to its economic value [30]. In a study in spain, effect of
considerable dynamic capability [12-14]. Phytoremediation sunflower on absorption of microelements and heavy
is a technology for use of plants for extraction, sequester metals including lead, arsenic, cadmium and zinc on role
or detoxification of contaminants through physical, of sunflower as suitable option for phytoremediation was
chemical and biological methods [15-17]. Some plants emphasized, in addition it was indicated that this plant has
which are called “hyperaccumulator” physiologically low potential for phytoextraction, other researchers
have suitable and usable potential for contaminated reported that sunflower was a suitable plant for zinc
soils phytoremediation purposes. So far, 400 plant contaminated soils remediation [31,32]. In addition to
species from 45 families have been reported to be agricultural plants with high biomass, identification of
hyperaccumulator [18]. Among these the most native plants around mines is one of pioneering
hyperaccumulator plants are for nickel and the least are solutions in phytoremediation. In a research, 17 plant
for cadmium with only one species known to be effective species including gramine, tree, shrub and grass which
[19]. Hyperaccumulators absorb metals in their shoots contained different and high concentration of lead,
more than 100 times more than natural plants do [20]. cadmium, copper and zinc were identified and reported
Phytoremediation of heavy metals and other inorganic around the mine [33]. In Slovenia, Thlapsi praecox Wulf
material can be in the following forms: plant containing zinc with concentration of 14590 mg/kg,
Phytoextraction: Is extraction and concentration of metals with concentration of 3500 mg/kg was identified and
from soil in shoot of the plant. introduced as Hyperaccumulator of zinc and cadmium
Rhizofiltration: Is the use of roots of plants for removal sensetively of plants root to high densities of metals led
of metals through water flow. to many researches on possibility to use microorganisms
Phytostabilisation: Is the use of plants for decrease in economization of this method [2,35]. Researches showed
expansion of metal contamination in the environment and that some of the effective and plant growth promoting
finally. rhizobacteria (PGPR) imposing mechanisms can increase
Phytovolatilization: Which is extraction and release of be increased with bacteria by production of siderophore
elements to atmosphere in the form of gas compounds is releasing iron and allowing movement of other heavy
important for mercury and arsenic [21]. The plants used in metals in soil [36]. Some of the bacteria producing
phytoremediation should have the ability for high metals Acc-Deaminase which prevents from ethylene stress,
toxicity resistant in their biomass and tolerate high decrease effects of heavy metals in plant tissues. Thus,
concentration of these in their own shoot [2]. In order to sinergisty of plant and bacterium can increase
expand application of phytoremediation with regard to low efficiency of use of phytoremediation [21]. Generally, with
biomass of hyperaccumulators, in different researches, regard to low growth speed and production of low
agricultural plants with high biomass such as sunflower, biomass in hyperaccumulators and necessity of
maize, canola, chick pea, wheat, cabbage, oat, barley and economization of metals phytoremediation on one hand
Indian mustard have been used as the replacement and expansion of heavy metals contamination in
options [22-28]. The first findings for use of plants in soil agricultural products on the other hand emphasize the
cadmium with concentration of 5960 mg/kg and lead
[34]. Production of low biomass in hyperaccumulators and
in order to expand use of phytoremediation and
absorption of metal in the soil [2]. Metal absorption may
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
95
need for study on translation of these metals to food
chain. Thus, study on agricultural plants with high
biomass (sunflower) and study and identification of
native plant species and nonagricultural plants
(amaranthus) and hyperaccumulators of heavy metals are
very important in each region. In the present research, by
emphasizing on the purposes like study on the bacteria
called PGPR group in native bacteria separated from the
soils around the lead and zinc mine in Haft Emarat, Arak,
effects of application of inoculants of two strains resistant
to metals, on function and absorption of mineral nutrition
and cadmium in the root and shoot of sunflower and
amaranthus were studied and in order to achieve the
above objectives, two laboratory and greenhouse
experiments were executed.
MATERIALS AND METHODS
Soil Sampling and Analysis: First, compound soil was
sampled from depth of 0-30 cm of soil from the Campus of
Agriculture and Natural Resources of University of
Tehran located in Karaj with coordinates of latitude of
northern 35°48 35 and longitude of eastern 50° 58 18 and
' '' ' ''
1315.5 meters above sea level. The soil was classified as
Xeric Haplocambids, Fine Loamy, Mixed, Super Active
thermic. Samples were air dried and passed through 2-mm
sieve and mixed uniformly. Physical and chemical
properties and concentration of elements in samples were
measured. Measurement of total nitrogen in the soil by
Kjeldal method, available phosphorus by Olsen method,
available potassium by normal ammonium acetate,
moisture content (SP), pH and electrical conductivity
determined by Rods, [38], percentage of equivalent
calcium carbonate (calcimeteric method) [39],
percentage of organic carbon by Walkly and Black
method [40] and soil tissue in hydrometric method were
determined [41]. Available concentration of zinc, lead and
cadmium were measured by DTPA method [42] and with
Atomic Absorption Spectrometry (AAS). Results are
given in table 1.
Greenhouse Test: The soil taken from Campus farmland
was passed through 4-mm sieve after air drying and
threshing. In order to create cadmium pollution in soils of
pots, cadmium chloride was used. concentration of
cadmium in soil for greenhouse test included control
treatment 0, 50,100 and 200 mg Cd /kg and three levels of
inoculants B0 (control), (Bacillus mycoides M1) B1 and
(Micrococcus Roseus M2) B2 and the test plants were
Table 1: Physical and chemical properties of soil used in greenhouse
culture before adding cadmium
Characteristic Quntity Characteristic Quntity
Soil texture Loam Total nitrogen (%) 0.08
Clay (%) 25.00 Available Phosphate (mg/kg) 17.10
Silt (%) 36.00 Available Potassium(mg/kg) 247.00
Sand (%) 39.00 SO (meq/l) 40.60
4
pH 7.90 Fe(mg/kg)* 4.28
EC(dS/m) 4.31 Cu(mg/kg)* 4.061
CaCO % 8.90 Mn(mg/kg)* 8.244
3
OC% 0.84 Zn(mg/kg)* 0.812
% SP 35.60 Pb(mg/kg)* 2.023
CO (meq/l) 0.40 Cd(mg/kg)* 0.10
3
CEC(Cmolkg ) 26.00 Cl(mg/kg) 0.0 8 4
-1
* DTPA-Extractable
sunflower and amaranthus. In each pot, some of
necessary food elements including 1/3 of nitrogen,
phosphorus and potassium consistent with weight of
the soils were weighed and were added to each pot
equally. Pots were made of poly ethylene material and
weighted about 280 g each wit diameter of 15.5 and height
of 18 cm. In each pot 3500 g of sieved soil were added
(in cadmium treatments, consistent with type of cadmium
treatment, 50, 100 and 200 mg Cd /kg of cadmium was
added and soil was mixed uniformly and poured in
the pot). Pots were irrigated based on weight loss and up
to 70%±10 of FC moisture content with distilled water.
Transfer of germinated seeds from the bottle to pots and
their inoculation with inoculants (5×10 cfuml ) were done
8 -1
on 26 July 2007. After germination, seedlings were thinned
to two plants in each pot and grown for 70 days.
Fertilizers containing micro and macro elements were
applied to the plants based on soil test results to have
optimal conditions for plant growth and reaching suitable
limits of phytoremediation. Pots were placed in growth
room with lighting of 14 hours and temperature of 24°C to
28°C and light intensity of 20000 lux for 70 days. For
measuring photosynthetic efficiency, Handy PEA
device, model RF 232 was used and Fv/Fm parameter or
photosynthetic photochemical quenching was assessed
in different treatments[43].
Isolation and Purification of Native Resistant Bacteria
from Soil: After study of status of the regions
contaminated with heavy metals, Arak lead and zinc mine
in Markazi Province located 46 Km south west of Arak at
coordinates of 33° 45 and 34° latitude and 49° 30 and 49°
''
45 longitude at 2150 M above sea level. 19 samples of soil
'
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
96
were taken from different regions around the mine from physical and chemical properties for greenhouse culture.
depth of 0-30 cm randomly and more than 130 bacteria In addition, no contamination with heavy metals was
were isolated from the soils[44] and finally, two strains of observed in soil.
bacterium resistant to heavy metals (Cadmium, Lead, Zinc
and Nickel) were isolated from the bacteria by performing Cadmium Concentration in Shoots and Roots: The results
biochemical tests in laboratory (microbiology section of obtained from plant analysis showed that concentration
Razi Serum and Vaccine Production Institute) on the basis of cadmium in shoots of amaranthus in all treatments was
of Bergeys Manual [45]. Two strains of Bacillus mycoides more than that of sunflower and there was significant
M1 and Micrococcus roseus M2 were identified and used difference between B2Cd100 and B0Cd200 statistically
in greenhouse test. (P<0.01). In sunflower, with increase in concentration of
Biological Tests: These tests were done on bacteria also increased. The highest cadmium concentration in
before performance of greenhouse tests and included: amaranthus was observed in B2Cd100, B1Cd200 and
Siderophore production power by CAS-Agar B1Cd100 treatments respectively. This increase was
method [46], ACC Deaminase enzyme production power consistent with Cd50, Cd100 and Cd200 treatments. The
quality test by Penroz and Gilck, [47], test of resistance to highest cadmium concentration in shoot of sunflower
salinity [48]. was in B1Cd200, B1Cd100 and B2Cd200 treatments
Plant Harvesting and Analysis: After 70 days from treatments Figure 1. In comparison between two plants,
culture, at the beginning of reproductive period and on 4 control treatments and Cd50 had the least amount of
Oct. 2007, shoot and root of each plant from place of cadmium. Consumption of both inoculants had better
crown were cut and after washing with distilled water and effect on increase in concentration of cadmium in shoots
measurement of wet weight, they were placed in the of amaranthus in comparison with control. Effects of
special packets and dried in oven at 70°C. After recording consumption of both inoculants in sunflower associated
the samples dry weight, they were milled with mixer or with cadmium metal in shoot were not observed the same
steel blade and concentration of cadmium, iron and zinc as those in amaranthus.
were measured by wet digestion method with nitric acid Results obtained from cadmium concentration in root
and perchloric acid (3:1) using ICP-OES, model CAP-6500 of sunflower and amaranthus showed that in all applied
for metal analysis [33]. Translocation factor indicating treatments, increased concentration of cadmium was
hyperaccumulation of heavy metals in harvested plant higher in sunflower than that in amaranthus. There was
parts was obtained by dividing metal concentration in significant difference in cadmium concentration
shoot by its concentration in the root [49]. Ratios higher between all treatments of sunflower and amaranthus
than 1 indicates higher concentration of metal in shoot except B2Cd50. The highest amount of cadmium was
and are one of the factors which indicates suitability of observed in root of sunflower, in B1Cd200, B1Cd100,
the plant for use in phytoremediation. Statistical analysis B2Cd100 treatments respectively. The obtained data
of data in the form of factorial experimental design with showed that in amaranthus, the highest cadmium
random basic design in three replications was done with concentration was observed in B1Cd100, B1Cd200 and
SAS software and comparison of means was done with B2Cd200. In B0Cd200, due to emergence of toxic effects of
LSD test at 1% level and graphs were drawn with Excel cadmium, amount of cadmium decreased in root in
software. comparison to B0Cd100 treatment. In terms of inoculants,
RESULTS AND DISCUSSION bacterium in both plants was more effective than that of
Physico-Chemical Properties of Soil: The soil used in cadmium in treatments, consumption of inoculants led to
greenhouse test table 1 was selected based on metal an increase in concentration of cadmium in root and
toxicity limits and study of sources and doubling reduced toxic effects of cadmium.
concentration of each treatment in comparison with the It is concluded that amaranthus had suitable
previous treatment, contaminated with cadmium at ability for phytoremediation by phytoextraction
rates (0, 50, 100 and 200 mg Cd /kg) [50,51]. Results of the method, transmitting more cadmium from root to
analysis showed that the intended soil have suitable shoot and sunflower in response to cadmium was
cadmium in soil, concentration of this element in shoot
and B1Cd200 was different significantly from other
results showed that consumption of inoculants of Bacillus
Micrococcus and with increase in concentration of
0
20
40
60
80
100
120
C
d0
C
d
50
C
d100
Cd
200
Cd0
C
d50
Cd1
00
Cd2
00
Cd0
Cd50
Cd1
00
Cd2
00
B0 B1 B2
Treatments
Shoot Cd Concentration (mg/kg DW)
Amaranthus
Helianthus
0
20
40
60
80
100
120
140
160
180
200
Cd
0
Cd50
Cd100
Cd200
Cd
0
Cd
50
Cd
100
Cd
2
00
Cd
0
Cd
5
0
Cd
100
Cd
200
B0 B1 B2
Treatments
Root Cd Concentration (mg/kg DW)
Amaranthus
Helianthus
0
50
100
150
200
Cd0
C
d
5
0
Cd100
Cd200
Cd0
Cd50
Cd100
Cd200
Cd
0
Cd50
Cd100
Cd200
B0 B1 B2
Treatments
Shoot Fe Concentration (mg/kg DW)
Amaranthus
Helianthus
0
500
1000
1500
2000
2500
Cd0
Cd50
Cd10
0
Cd2
0
0
Cd0
Cd50
C
d1
00
Cd200
Cd0
Cd50
C
d10
0
Cd200
B0 B1 B2
Treatments
Root Fe Concentration (mg/kg DW)
Amaranthus
Helianthus
0
20
40
60
80
100
120
Cd
0
Cd
50
Cd
10
0
Cd20
0
Cd0
Cd
50
Cd100
Cd
20
0
Cd0
Cd5
0
Cd10
0
Cd
20
0
B0 B1 B2
Treatments
Shoot Zn Concentration (mg/kg DW)
Amaranthus
Helianthus
0
20
40
60
80
100
120
140
160
Cd0
Cd50
Cd
100
Cd20
0
Cd
0
Cd50
C
d10
0
Cd
20
0
Cd
0
Cd50
Cd100
Cd
200
B0B1B2
Treatments
Root Zn Concentration (mg/kg DW)
Amaranthus
Helianthus
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
97
Fig. 1: The concentration of Cd, Fe and Zn in shoot and root of A. retroflexus and H. annuus grown in different cadmium
and inoculant treatments. B0: Control, B1: Bacillus mycoides inoculant, B2:Micrococcus roseus inoculant.
Significantly different at P<0.01
considered a suitable plant for phytoremediation use them for stabilizing metal in soil and prevention
through phytostabilization method. On the basis of from expansion of contamination as result of water
research performed, plants confronting with tension of erosion and wear. With regard to results of the present
heavy metals use two ways [52,53]: 1- prevention from research and findings of other researchers in the use
entrance of metal to shoot and association in root of plants with high biomass and ability to absorb
2- Metal detoxication. On this basis, the plants metal in root in phytostablization method, use of
which prevent from entrance and transfer of metals plants like sunflower can be suitable option for
have low potential for phytoextraction, but, one can phytoremediation [11,31].
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
98
Iron Concentration in Shoots and Roots: Results of iron and in sunflower; it was found in B0Cd50, B0Cd0 and
analysis in shoot of two plants showed that with increase B1Cd0 treatments respectively and seems that decrease in
in concentration of cadmium, concentration of iron in concentration of zinc in Cd200 was due to toxicity of
shoot of both plants increased. Although, increase in cadmium in this treatment Figure 1.
concentration of iron in both plants decreased in Cd200
treatments in comparison to Cd100, this decrease was Study on Characteristics of Bacteria: In table 2, some of
due to toxic concentration of cadmium and statistically, the specifications of bacteria were identified and two
it was not significant. In amaranthus, the highest iron strains were used in greenhouse test. Two strains of
concentration in shoot was observed in B2Cd50, B2Cd200 bacteria (B1 and B2) had some characteristics of Plant
and B1Cd200 treatments respectively and in sunflower; it Growth Promoting Rhizobacteria (PGPR). Application of
was observed in B1Cd100, B2Cd0 and B1Cd200. inoculant of these bacteria in greenhouse test showed
The highest iron concentration in root of amaranthus better results in comparison to Control treatment (B0).
was observed in B1Cd100, B1Cd200 and B0Cd100 These bacteria were resistant to high concentrations of
respectively and in sunflower; it was found in B1Cd0, heavy metals and were able to produce siderophores.
B1Cd100 and B0Cd50 respectively. B1Cd100 treatment in Results obtained from determination of concentration of
the amaranthus had the highest concentration. In terms of iron in different organs of the tested plants showed that
effect of inoculant,, similar to results of shoot, Bacillus concentration of iron in root of sunflower was higher
inoculant had obtained better results. than that in shoot. In fact, in some part of the plant which
Zinc Concentration in Shoots and Roots: The results concentration of iron was also high. Observations of this
showed that zinc concentration in shoot of amaranthus result and findings of other researchers indicated that the
was generally higher than that in sunflower. The highest plant confronting with stress of heavy metals increases
concentration of zinc in amaranthus was observed in extent of iron absorption. This increase is a reply to metal
B2Cd200, B2Cd100 and B1Cd0 respectively and in stress and also is as result of effect of plant growth
sunflower; it was found in B2Cd0, B0Cd50 and B0Cd0 promoting rhizobacteria. Ability to produce microbial
respectively. In sunflower, increase in concentration of siderophores can be one of the important factors of iron
cadmium led to decrease in zinc concentration in the concentration increase in shoot and root of the plant.
shoot in such a way that there was significant difference Synergistic effects of bacterium and plant in these
between concentration of zinc in Cd200 and Control (Cd0) conditions are so close that this synergism in decrease of
(P<0.01). Results obtained from application of bacterium metal stress on the one hand and increase of iron
inoculant showed that there was not a significant concentration in root and shoot of the plant and
difference between two inoculants in sunflower, but in facilitation in absorption of iron and zinc on the
amaranthus, there was significant difference among some other hand leads to improvement of phytoremediation
of the treatments Figure 1. conditions. Ability to reduce ACC Deaminase Enzyme
Zinc concentration in different treatments of which prevent from synthesis of ethylene in the plant,
amaranthus had significant difference in terms of reduces ethylene tension in the plant and is one of the
concentration of cadmium and consumption of inoculant. ways of plant’s confrontation with undesirable
The highest concentration of zinc in amaranthus was conditions. Similar research with isolation of resistant
observed in B2Cd200,B2Cd100 and B1Cd100 respectively bacteria from contaminated soils and application of
there is the maximum association of cadmium (root),
Table 2: Some specification of growth promoting rhizobacteria – identification of bacteria on the basis of [45]
Resistance to Resistance to Resistance to Resistance to Posetive or Salinity
Strain of bacteria Pb (1000 mg/l) Zn (1000 mg/l) Ni (1000 mg/l) Cd (1000 mg/l) Negative Gram ACC deaminase (60 ds/m) Siderophore
Bacillus mycoides M1 + + + + + + +-
Micrococcus roseus M2 - - + - + + ++
Bacillus circulans M3 + - - - - - ++
Bacillus pumilis M4 + - - - + + +-
Bacillus coagularis M5 - + - - - + ++
Bacillus cereus M6 + - - - + + ++
Micrococcus luteus M7 + - - - + - ++
+ = proper reply to the intended characteristic - = lack of the intended characteristic
0
2
4
6
8
10
12
14
Cd
0
Cd
50
Cd
100
C
d200
Cd0
Cd
50
Cd
100
C
d200
Cd0
Cd
50
Cd
10
0
Cd
200
B0 B1 B2
Treatments
Shoot Dry Weight (g)
Amaranthus
Helianthus
0
10
20
30
40
50
60
70
80
90
100
Cd0
Cd50
Cd100
Cd200
Cd0
Cd50
Cd100
Cd200
Cd0
Cd50
Cd100
Cd200
B0 B1 B2
Treatments
Shoot Wet Weight (g)
Amaranthus
Helianthus
0
1
2
3
4
5
6
7
8
Cd0
Cd50
Cd100
Cd200
C
d0
C
d5
0
Cd
10
0
Cd200
Cd0
Cd5
0
Cd
100
Cd
20
0
B0B1B2
Treatments
Root wet Weigh t (g)
Amaranthus
Helianthus
0
1
2
3
4
5
6
7
8
Cd0
Cd50
C
d
100
Cd200
Cd0
C
d
5
0
Cd
100
Cd200
C
d
0
Cd5
0
Cd100
Cd
200
B0 B1 B2
Treatments
Root wet Weight (g)
Amaranthus
Helianthus
0.5
0.55
0.6
0.65
0.7
0.75
0.8
0.85
C
d0
C
d50
Cd100
Cd
2
00
Cd0
C
d50
C
d100
Cd
2
00
Cd
0
C
d
50
Cd1
00
C
d
2
00
B0 B1 B2
Treatments
FV/FM
Amaranthus
Helianthus
0
0.5
1
1.5
2
2.5
3
3.5
4
C
d
0
Cd
50
C
d
1
00
C
d200
Cd
0
C
d
50
C
d1
00
C
d2
0
0
Cd
0
C
d
50
C
d
10
0
C
d
2
00
B0B1B2
Treatments
TF
Amaranthus
Helianthus
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
99
Fig. 2: Shoot and root wet, dry weight, Fv/Fm and TF of A. retroflexus and H. annuus grown in different Cadmium and
inoculant treatments. B0: Control, B1: Bacillus mycoides M1 inoculant, B2: Micrococcus roseus M2 inoculant.
Significantly different at P<0.01
inoculant showed that in zinc and cadmium contamination iron resource for the plant. This case is the best reason for
conditions, motion of these metals in Rhizosphere prevention from formation of chlorosis and yellowness of
increased and concentration of iron increased in the leaves in confrontation with high densities of heavy
plant. Bacteria had abilities of siderophores production, metals. Results showed that the role of effective bacterium
ACC Deaminase Enzyme and IAA (Oxine) and their was to facilitate provision of iron necessary for the plant
consumption increased phytoremediation in the for growth in case of presence of heavy metal. On the
contaminated soils [1,21]. Bacterial complex of Iron- other hand, bacteria applying mechanisms such as
siderophore can be used by the plant and be a suitable decrease in ethylene stress, decrease in pH of
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
100
Rhizosphere and development of roots growth provide wet weight was observed in B0Cd200, B1Cd200 and
better opportunities for settlement and promotion of B0Cd50 treatments. In amaranthus, the highest wet weight
plant growth [2,21]. of root was observed in B2Cd0, B0Cd50 and B0Cd100
Shoots Wet Weight: Since presence of heavy metals B2Cd200, B2Cd100 andB1Cd100 treatments. The obtained
leads to decrease in growth and function of the products results showed that application of inoclant in sunflower
[2,4,5], in this expriment, similar results were also obtained. was effective to concentration at Cd100 and in higher
In different treatments with increase in concentration of concentration, it had no considerable effect. There was
cadmium in the media, shoot wet weight and the highest not significant difference between two inoculants
extent of this decrease was found in Cd200 treatment for (P<0.01). In amaranthus, similar results were obtained
both plants Figure 2. The highest shoot wet weight of Figure 2.
shoot of the plant in sunflower was found in B0Cd0,
B1Cd0 and B2Cd0 and the lowest was found in B0Cd200, Roots Dry Weight: Results of root dry weight in
B0Cd100 and B2Cd200 treatments. which were not different treatments showed that although increase in
different statistically (P<0.01). In addition amount of concentration of cadmium led to decrease in root dry
biomass of sunflower was more than that of amaranthus. weight, this decrease was more intense in control
In amaranthus, the highest shoot wet weight was found treatment (without inoculant). Between two plants, root
in B2Cd0, B0Cd50 and B0Cd100 and the lowest shoot wet dry weight of sunflower was more than that of amaranthus
weight was found in B2Cd200, B2Cd100 and B1Cd200 and the highest dry weight in sunflower was observed in
treatments. In terms of effect of inoculant in shoot wet B0Cd0, B1Cd0 and B2Cd100 treatments and the lowest dry
weight, there was statistically significant difference weight was observed in B0Cd200, B0Cd100 and B1Cd200
between two plants (P<0.01) and in amaranthus, Bacillus treatments. In amaranthus, the highest dry weight of root
incoulant showed better results except in B2Cd0. in was observed in B0Cd100, B0Cd0 and B0Cd50 treatments
sunflower, Bacillus and Micrococcus inoculants and the lowest dry weight was observed in B2Cd200,
showed better results than control in wet weight B2Cd100 and B1Cd100 treatments. There was significant
increase Figure 2. difference among different treatments in terms of
Shoots Dry Weight: With increase in concentration of cadmium, there was not significant difference between
metal in media, shoot dry weight decreased in both two inoculants statistically and application of inoculant
plants and the lowest amount was observed in Cd200. in showed better results than that of control treatment
sunflower which had higher biomass than amaranthus, the Figure 2. Study on data obtained from wet weight and dry
highest shoot dry weight was observed in B1Cd0, B0Cd0 weight of shoot and root showed that heavy metal stress
and B2Cd0 treatments and the lowest shoot dry weight led to decrease yield of wet and dry material. This stress
was observed in B1Cd200, B0Cd100 and B0Cd50 was highest in concentrations of 200 mg/kg. Results of
treatments. In amaranthus, the highest dry weight of research showed that increase in concentration of heavy
shoot was observed in B2Cd0, B0Cd0 and B0Cd50 metals in soil decreased biomass of the root and shoot in
treatments and the lowest shoot dry weight was observed Zea mays, sorghum, Helianthus annuus,Cynodon
in B2Cd200, B2Cd100 and B1Cd200 treatments. dactylon and Conyza discordies[54]. In another research,
Observations showed that there was difference between amount of biomass in shoot and root of sunflower in
sunflower and amaranthus in terms of effects of inoculant contaminated soil was less than that in uncontaminated
and Bacillus inoculant in amaranthus. Micrococcus soil [31]. With regard to effects of growth promoting
inoculant in sunflower showed better results in dry weight rhizobacteria, yield in inoculant treatments (B1, B2)
increase Figure 2. showed lower decrease in comparison to control and
Roots Wet Weight: Root wet weight of sunflower in all
treatments was higher than that of amaranthus. In most Photosynthetic Photochemical Efficiency: Green plants
treatments, increase in concentration of metal led to have two types of chlorophyll a and b. These two
decrease in wet weight especially in concentration of materials are green colored under white light, because
Cd200. Effect of inoculant can be discussed in this regard. they absorb radiation located in the limit of blue and
The highest root wet weight in sunflower was observed red spectrum and reflect radiation located in green
in B2Cd100, B1Cd0 and B0Cd0 treatments and the lowest spectrum. Energy obtained from absorption is spent for
treatments and the lowest wet weight was observed in
application of inoculant in both plants, in similar levels of
intensity of cadmium stress decreased Figure 1.
Am-Euras. J. Agric. & Environ. Sci., 4 (1): 93-103, 2008
101
performance of photosynthesized chemical reactions phytoremediation, effort to recognize and study this
and its remaining is lost in the form of heat. method and ways of phytoremediation efficiency increase
Researchers measuring parameters of F0 (low level of is very important. As mentioned, phytoremediation of
photosynthesis), Fm(maximum level of photosynthesis) contaminated soils is a relatively new and developing
and their difference(Fv) obtained new ratio called technology with special benefits [12-14]. On the basis of
Fv/Fm(photochemical quenching) which shows very results obtain from greenhouse and laboratory tests, it
close correlation with optical function of pure seems that by continuation of research in the fields of
photosynthesis of healty leaves[43]. Data obtained from botany and study of characteristics of growth promoting
measurement of photochemical quenching showed rhizobacteria and synergism between plant and bacterium,
that there was statistically significant difference one can be successful in development of application of
between two plants in all treatments. On the other hand, phyotremediation in soils contaminated with heavy metals
photosynthetic quenching in sunflower was higher than with economic and applied approach. In addition that
that in amaranthus. The highest amount in sunflower was some of the plants such as sunflower can be used for
observed in B1Cd0, B2Cd0 and B0Cd0 treatments and the control and prevention from distribution of metals
lowest amount was observed in B0Cd200, B2Cd200 and contamination (phytostabilization) and phytoexraction
B1Cd200 treatments. In amaranthus, the highest method can be used by identification of native and
photosynthetic quenching was observed in control nonagricultural plants which are hyperaccumulator
treatments (Cd0) and the lowest was observed in for phytoremediation of the contaminated regions.
B1Cd200, B2Cd200 and B0Cd200 treatments Figure 2. Potential of phytoremediation depends on interactions
Translocation Factor: Comparison of translocation factor of these factors and mechanism of their effect can have
showed that this factor in amaranthus was much better important role in development of phytoremediation
than that in sunflower and amaranthus had more application.
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