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RESULTS
RESULTS
The effect of cadmium and chromium were elucidated in Pisum sativum and
Zea mays to observe its consequences on plant growth, physiological, biochemical
and biomass characteristics of Pisum sativum (Arkel) and Zea mays (4212). The pea
and maize plants were grown as per protocol described in material and methods.As
per the results obtained in petridish experiments and certain pot experiments,which
were conducted in wire house. The seeds were subjected to various Cadmium and
Chromium concentrations on plants growth, development and seed yield.
4.1 Effect of Cadmium on Seed Germination and Metal Tolerance Index in Pea
The pea represented gradual loss in germination upon treating with different
cadmium solutions. The differential germination percentage was recorded in pea in
caseof control and also upon treating with various cadmium solutions (Fig. 21). The
variation in germination percentage was found to be 83% under normal condition.
However, 83% germination reduced to the level of 75, 69, 56, 39 and 28% in case of
different treatment of cadmium i.e. 1, 2, 4, 8 and 16 ppm respectively. The metal
tolerance was reduced significantly loss in the range of loss in 31-81% as compared to
control (Fig. 22A,B; Appendix-I).
85
RESULTS
A
Control
B
Control
5 Days after Cadmium application
1 ppm
2 ppm
4 ppm
8 ppm
16 ppm
15 Days after Cadmium application
1 ppm
2 ppm
4 ppm
8 ppm
16 ppm
Fig. 21. Influence of differential cadmium levels on shoots and root
biodynamic in Pisum satvum var. Arkel seedlings. A and B
indicate acquisition of root and shoot canopies as affected by
cadmium treatment levels (1, 2, 4, 8 and 16 ppm) coupled with
cadmium treatment durations as indicated.
86
RESULTS
100
A
90
80
Germination (%)
70
60
50
40
30
20
10
0
120
B
Metal tolerance index (%)
100
80
60
40
20
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig. 22. Effect of different levels of cadmium, i.e. 1, 2, 4 8 and 16 ppm on
seed germination% (A) metal tolerance index% (B) shown by
Pisum sativum
var. Arkel.
Measurements
till ppm)
15 days.
Fig.2. Effect of differential
levels
of Cadmium
(1, 2,were
4, 8,taken
and 16
on seed
Values are mean (n=3) with S.E. (±).
germination % (A) metal tolerance index % (B) shown by Pisum sativum var
Arkel. Measurements were taken till 15 days. Values are mean (n=3) with S.E. (±).
87
RESULTS
4.2 Acquisition of Shoot Length, Root Length and Lateral Roots vs. Cadmium
Levels
The germinated seedlings were used to record the biodynamics in relation to
appearance/ acquisition of radical length, as influenced by different cadmium
concentrations. The data indicate that seedlings unaffected (control) by cadmium did
acquire radical length maximum throughout till 7 days after germination. The root and
shoot lengths were recorded in pea as influenced by cadmium contaminated irrigation
water (1, 2, 4, 8 and 16 ppm). Almost similar trends have been observed as shown in
Fig. 23A,B (Appendix-IB). Both shoot and root have been found to be downregulated (31-89 and 58-89% in relation to levels of cadmium. The lower level of
cadmium application (1 ppm) cause effectively to down-regulate root and shoot
growth. Upon, increase in cadmium levels i.e., 1, 2, 4, 8 and 16 ppm, the root and
shoot both have been found severely down-regulated in case measurements were
made 15 days after the application of cadmium as compared to control. The lower
level of treatment (1 ppm) followed parallel trends with much accuracy. Nearly, 31%
loss in shoot length could occurred in case seeds were treated with 1 ppm of cadmium
till the termination of the observation (15 days). The 2, 4, 8 and 16 ppm cadmium
application could down-regulate the shoot length ca. 43, 56 and 78and 89% as
compared to control (Fig. 23A,B; Appendix-IB).
Similarly, the down-regulation in root length was found ca. 58, 70, 74, 82 and
89% (1, 2, 4, 8 and 16 ppm) cadmium treated respectively in case cadmium treatment
was allowed for 15 days. The number of leaves and lateral roots shown similar trends
as influenced by treatment levels and durations both. The loss in leaf number and
lateral roots (21-79 and 42-84%) occurred in case treated with 1, 2, 4 and 16 ppm
cadmium for 15 days, in abled seedlings for growth. The effect of cadmium treatment
of total number of leaves on per seedlings basis. The higher levels of cadmium (16
ppm) could cause loss in leaf reduction to the tune of ca. 79% as compared to control
as shown in Fig. 23C,D (Appendix-IIA,B).
88
RESULTS
12
7
B
A
6
Root length (cm)
Shoot length (cm)
10
8
6
4
4
3
2
2
1
0
0
10
12
C
D
9
10
Number of lateral roots
8
8
Leaf number
5
6
4
7
6
5
4
3
2
2
1
0
0
0
1
2
4
8
0
16
1
2
4
8
16
Cadmium treatment (ppm)
Cadmium treatment (ppm )
Fig. 23. Effect of cadmium on shoot length (A), root length (B), number of leaves (C)
and number
of lateral
rootslength
(D) in
sativum
Arkel.
The seedlings
Fig.3. Effect
of Cadmium
on shoot
(A)Pisum
,root length
(B) var.
number
of leaves
(C) and
allow
to (D)
grow
till 15sativum
days after
application
of various
concentrations
numberwere
of lateral
roots
in Pisum
var Arkel.The
seedlings
were allowed
to grow
(1,
2,
4,
8
and
16
ppm).
Values
are
mean
(n=3)
with
S.E.
(±).
till 15 days after application of various concentrations ( 1,2,4,8,and 16 ppm) .Values are
means (n=3) with S.E.(±).
89
RESULTS
4.3 Biomass and Moisture Content vs. Cadmium Levels
The loss in shoot fresh mass and dry mass of per seedlings. The loss in shoot
biomass found in the range of 40% in case of lower level (1 ppm) of cadmium
applied. The higher levels (16 ppm) of loss (87%) in shoot fresh mass was also
recorded. The losses in shoot dry mass was found in the range of 80% in case treated
with higher level (16 ppm) of cadmium (Fig. 24A; Appendix-IIIA). The stability
levels as influenced by cadmium were revealed by recording loss in root fresh and dry
mass, indicates that loss in root fresh mass was shown by pea seedlings. The loss has
been found in the range of 59% in case of lower level (1 ppm) of cadmium applied.
The higher levels (16 ppm) of loss (91%) in root fresh mass was also recorded (Fig.
24B; Appendix-IIIB). The losses in root dry mass was found in the range of 83% in
case treated with higher level (16 ppm) of cadmium in due course of time (15 days
after treatment). The moisture content was also reduced. The loss has been found in
the range of 2% in case lower level (1 ppm) of cadmium applied (Fig. 24C;
Appendix-IVA).
4.4 Photosynthetic Pigments of Pisum sativum vs. Cadmium
The photosynthetic pigments i.e., chlorophylls a and b was found to be
affected. The chl b was found more significantly affected as compared to chl a. About
31% chlorophyll a and 46% chlorophyll b (1 ppm) were found down-regulated in pea
seedlings, which could reach about 76% and 93% (16 ppm) after 15 days of cadmium.
Similarly total chlorophyll was also found to be down-regulated in pea, which could
also reach about 79% (16 ppm) after 15 days of cadmium. The loss in carotenoids was
also observed. The values of carotenoids have shown down regulation ca. 29-65% (1,
2, 4, 8 and 16 ppm) depending upon treatment levels within 15 days as shown by pea
seedlings was recorded (Fig. 25A-D; Appendix-IVB).
90
RESULTS
0.70
0.06
Fw
Dw
A
0.50
0.04
0.40
0.03
0.30
0.02
0.20
0.10
0.01
0.00
0.00
0.60
0.03
FW
Fresh wt of root (g)
Dry wt of shoot (g)
0.05
DW
B
0.50
0.03
0.40
0.02
0.30
0.02
0.20
0.01
0.10
0.01
0.00
0.00
Dry wt of root (g)
Fresh wt of shoot (g)
0.60
94
C
Moisture
content
(%)
Moisture
content
93
92
91
90
89
88
87
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig.4. EffectFig.
of 24.
different
( 1,2,4,8,and
Effect Cadmium
of differentconcentrations
cadmium concentrations
(1, 2, 16
4, 8ppm)
and of shoot
ppm)
of shoot
andweight
dry weight
(A),moisture
root fresh
fresh and dry weight16 (A)
,root
fresh fresh
and dry
(B) and
content (C) in
and
dry
weight
(B)
and
(%)
moisture
content
(C)
in
Pisum sativum var Arkel.The seedlings were allowed to grow till 15 days.Values are
sativum var. Arkel. The seedlings were allowed to
means (n=3-5) withPisum
S.E.(±)
grow till 15 days. Values are mean (n=3-5) with S.E. (±).
91
RESULTS
0.600
0.18
A
0.16
Chl b (mg g-1 FM)
Chl a (mg g-1 FM)
0.500
0.400
0.300
0.200
B
0.14
0.12
0.1
0.08
0.06
0.04
0.100
0.02
0.000
0
0.8
0.45
0.7
0.4
0.6
0.35
Carotenoid (mg g-1 FM)
Total Chl (mg g-1 FM)
C
0.5
0.4
0.3
0.2
D
0.3
0.25
0.2
0.15
0.1
0.1
0.05
0
0
0
1
2
4
8
16
0
Cadmium treatment (ppm)
1
2
4
8
16
Cadmium treatment (ppm)
Fig. 25. Effect of different cadmium concentrations (1, 2, 4, 8 and 16 ppm) on
photosynthetic pigments in Pisum sativum var. Arkel. Chl a (A), chl b (B),
total chl (C) and carotenoid (D). Cadmium treatment maintained for a
Fig .5.Effect of
different
Cadmium
concentrations
(1, 2, with
4 ,8,and
ppm) on photosynthetic
period
of 15
days. Values
are mean (n=3)
S.E.16(±).
pigments in Pisum sativum var Arkel. Chl a A) ,Chl b (B) Total Chl (C) and Carotenoid
(D).Cadmium treatment maintained for a period of 15 days.Values are means (n=3) with
S.E.(±).
92
RESULTS
4.5 Peroxidase, Catalase and Lipid Peroxidase vs. Cadmium
The effect of cadmium treatment in pea seedlings was also correlated with
certain stress inducible enzymes such as peroxidase and catalase. These enzymes
genes generally gets switched on during adverse experiences by plants as shown in
Fig. 26A,B (Appendix-VA) clearly opted increasing trends in pea seedling. Almost
there intrinsic abilities in relation to increase in peroxidase (24-76%) and (18-100%)
catalase are correlated with cadmium (1 ppm-16 ppm) levels. Both these enzymes are
stress mitigating biomolecules therefore; biologically both of them have behaved as
per biological rule in supporting the biological system. Similarly lipid peroxidase
activity could enhanced ca. 258% higher in the case of leave in case treated with 16
ppm cadmium contaminated water for a period of 15 days (Fig. 26C; Appendix-VA).
4.6 Amylase vs. Cadmium
The amylase activity was significantly reduced at higher concentration of
cadmium exposure. The loss in total amylase i.e. 13% occurred in case treated with 1
ppm cadmium but higher treatment 16 ppm i.e. 92% caused loss. Similarly α amylase
(17%) and β amylase (6%) occurred in case treated with 1 ppm cadmium but higher
treatment 16 ppm i.e. 91% caused loss and 96% for a period of 15 days as shown in
Fig. 27A-C (Appendix-VB).
4.7 Total Sugar vs. Cadmium
The total sugar shown similar trends as influenced by treatment levels. The
loss in sugar (20%) occurred in case treated with 1 ppm cadmium but higher treatment
16 ppm (58%) for 15 days of irrigation of cadmium contaminated water as shown in
Fig. 28A (Appendix-VIB).
4.8 Protein vs. Cadmium
The total protein showed similar trends as influenced by cadmium treatment
levels. Protein content as revealed in favoured that the cadmium treatment might have
down regulated protein synthesis which may be associated with 70S and 80S
ribosome’s both. The leaf has shown a negative impact in acquiring normal protein
synthesis. The leaves have shown about 12% loss at 1 ppm while higher treatment of
16 ppm caused 79% loss (Fig. 28B; Appendix-VIA).
93
RESULTS
80
Peroxidase ( ∆ OD g-1 FM )
A
70
60
50
40
30
20
10
Catalase
-1 -1
(µmol
decomposed
2O2 g FM)
Catalase ( µmolHH
2O2 g FM )
0
100
90
B
80
70
60
50
40
30
20
10
Lipid peroxidase ( µmol MDA g
-1
FM)
0
80
C
70
60
50
40
30
20
10
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig.6.Effect of
Cadmium
contaminated
watercontaminated
on enzymes activities
of peroxidase
Fig.
26. Effect
of cadmium
water on
enzymes (A)
catalase(B) and lipid peroxidase
(C)
in
seedlings
growth
.
Pisum
sativum
Arkel
activities of peroxidase (A) catalase (B) andvar
lipid
seedlings were exposedperoxidase
to 0,1,2,4,8,and
Cd forgrowth.
a period of
15 days.Values
(C) 16
in ppm
seedlings
Pisum
sativum are
means (n=3) with S.E.(±).
var. Arkel seedlings were exposed to 0, 1, 2, 4, 8 and
16 ppm Cd for a period of 15 days. Values are mean
(n=3) with S.E.(±)
94
RESULTS
Total amylase
-1
(starch hydrolysed mg g FM)
10
A
9
8
7
6
5
4
3
2
1
0
α amylase
(starch hydrolysed mg g-1 FM)
7
B
6
5
4
3
2
1
0
3.5
β amylase
(starch hydrolysed mg g-1 FM)
C
3
2.5
2
1.5
1
0.5
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig .7. Effect of Cadmium on total amylase (A) α amylase (B) and β amylase (C) in Pisum
27. Effect
of cadmium
totalatamylase
(A) after
 amylase
(B)  of
sativum Fig.
var Arkel
.Measurements
wereontaken
specific time
the application
amylase
(C) in Pisum
sativum var.
Measurements
different concentrations
(0,1,2,4,8,and
16 ppm).Each
dataArkel.
point represents
mean (n=3) with
were
taken
at
specific
time
after
the
application
of
S.E (±).
different concentrations (1, 2, 4, 8 and 16 ppm). Each data
point represents mean (n=3) with S.E. (±).
95
RESULTS
6
A
Sugar (µg g-1 FM)
5
4
3
2
1
0
0
1
2
4
8
16
180
B
Protein (µg g-1 FM)
160
140
120
100
80
60
40
20
0
0
1
2
4
8
16
Cadmium treatment (ppm)
28. Effect
of cadmium
sugar
(A) andcontent
protein(B)
content
(B) sativum
Fig. Fig.
8. Effect
of Cadmium
on sugaron(A)
and protein
in Pisum
in
Pisum
sativum
var.
Arkel
leaf.
The
different
var Arkel leaf .The different Cadmium contaminated water ( 1, 2, 4, 8,and 16 ppm)
contaminated
waterare(1,means
2, 4,(n=3-5)
8 and replicates
16 ppm) with
was maintainedcadmium
for a period
of 15 days.Values
was
maintained
for
a
period
of
15
days.
Values
are
S.E.(±)
means (n=3-5) replicates with S.E. (±).
96
RESULTS
15 Days after treatment
45 Days after treatment
60 Days after treatment
Control
1 ppm
2 ppm
4 ppm
A
B
C
8 ppm 16 ppm
Fig. 29A. Influence of differential cadmium levels in plant
morphology in Pisum sativum var. Arkel. A-C indicate
acquisition of shoot appearance. The pots were irrigated
with cadmium treatment levels (1, 2, 4, 8 and 16 ppm)
once in a week followed by normal irrigation to the
level of field capacity.
97
RESULTS
A
15 Days after treatment
B
45 Days after treatment
C
60 Days after treatment
D
Control 1 ppm 2 ppm 4 ppm
8 ppm
16 ppm
60 Days after treatment
Fig. 29B. Influence of differential cadmium levels on root
morphology in Pisum sativum var. Arkel. A-C
indicate acquisition of root appearance and Dindicate pod formation. The pots were irrigated with
cadmium treatment levels, i.e. 1, 2, 4, 8 and 16 ppm,
once a week followed by normal irrigation to the
level of field capacity.
98
RESULTS
4.9 Shoot Length and Root Length of Pisum sativum vs. Cadmium Durations and
Levels
The long-term (upto ca. 60 days) cadmium irrigation consequences were
planned to reveal seed to seed status of cadmium treatment. These studies were made
in earthen pots. The morphological approaches for pea cultivar have also been affected
their morphological appearances and differential growth behavior. Higher the treatment,
lower the plant height/shoot length was observed in comparison to pea cultivar grown
by providing normal irrigation levels (Fig. 29A,B).
The shoot length growth behavior was recorded in due course of time i.e., 15-60
days at the interval of 15 days. The lower level of cadmium treatment (1 ppm) could
cause loss in shoot length ca. 15-5% within 30 days which could get further enhanced to
the level of 63-50% in case treated with four fold higher cadmium solution (16 ppm).
Similarly the root length growth behavior was recorded in due course of time i.e., 15-60
days at the interval of 15 days. The lower level of cadmium treatment (1 ppm) could
cause loss in root length ca. 21-24% within 30 days which could get further enhanced to
the level of 80-56% in case treated with four fold higher cadmium solution (16 ppm) as
shown in Fig. 30A,B (Appendix-VIIA,C).
4.10 Number Leaves of Pisum sativum vs. Cadmium Irrigation
The effect of cadmium treatment irrigation was correlated with retention of
total number of leaves on per plant basis. The total number of leaves have shown
down regulation in retaining their number almost 15-38% depending upon the
treatment levels within 15 days as shown by pea. The enhancement in days after
treatment have been found correlated in an increasing order in response to loss in total
number of leaves. The pea cultivar has shown ca. 20% loss (1 ppm) in total leaves
after 60 days in comparison to 38, 46, 49 and 58% in case treated with 2, 4, 8 and 16
ppm levels of the cadmium after 60 days after treatment (Fig. 30C; Appendix-VIIB).
99
RESULTS
60
Shoot length (cm)
A
15
30
45
60
50
40
30
20
10
0
50
45
B
15
30
45
60
Root length (cm)
40
35
30
25
20
15
10
5
0
50
45
C
15
30
45
60
Leaf number
40
35
30
25
20
15
10
5
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig.10. Fig.
Effect30.
of different
on shoot
length(A)
,root length
Effect ofCadmium
differentirrigation
cadmium
irrigation
on shoot
length(B) and
leaf number (C) in(A),
Pisum
sativum
var
Arkel.
The
plants
grown
under
the
application
root length (B) and number of leaves (C) in Pisum
of different cadmium
concentrations
(1,2,4,8,and
ppm) grown
for a period
of 60the
sativum
var. Arkel.
The 16
plants
under
days.Values are means
(n=3)
with
S.E.(±).
application of different concentrations (1, 2, 4, 8 and 16
ppm) for a period of 60 days. Values are mean (n=3)
with S.E. (±).
100
RESULTS
4.11 Pisum sativum of Biomass vs. Cadmium Irrigation
The impact of cadmium treatment on plant biomass was also evaluated in
relation to levels and durations. The data shown in Fig. 31 also in favored inability in
parallel with the higher levels of cadmium in relation to retention of shoot fresh mass
and shoot dry mass. The loss in shoot fresh and dry mass as shown from 22-20 and
21-17% in comparison to 66-54 and 61-77% in case used lower (1 ppm) and higher
(16 ppm) levels of cadmium in response to treatment duration for a period of 60 days
(Fig. 31B; Appendix-XA,B). The stability levels as influenced by cadmium were
revealed by recording loss in root fresh biomass and dry mass. The loss in root fresh
and dry mass has been found in the range of 25-11 and 13-21% in case of lower level
(1 ppm) of cadmium applied for irrigation in due course of time (15-60 days after
treatment). The higher levels of loss (78-57 and 69-78%) in root biomass were also
recorded (Fig. 31C; Appendix-IXA,B). The similar trends were found in leaf fresh
and dry weight as shown in Fig. 31A (Appendix-VIIIA,B).
Total fresh and dry biomass were also reduced with increasing concentration
of cadmium levels in pea plants. The data shown in Fig. 32 indicates that loss in total
fresh mass was shown by Pisum sativum cultivars. The loss has been found in the
range of 28-17% in case of lower level (1 ppm) of cadmium applied for irrigation in
due course of time (15-60 days after treatment). The higher levels of loss in total fresh
biomass were also recorded. The losses in total dry mass were found in the range of
57-73% in case irrigated with higher level (16 ppm) of cadmium due course of time
(15-60 days after treatment), as shown in Fig. 32A,B (Appendix-XIA,B).
4.13 Photosynthetic Pigments of Pisum sativum vs. Cadmium
The morphological appearance was found affected due to applied cadmium
levels therefore photosynthetic pigments viz., total chlorophyll, chlorophyll a and
chlorophyll b content were recorded (Fig. 33). The values of chlorophyll
concentration found pea has shown gradual loss in retaining photosynthetic pigment
i.e., chlorophylls. The loss in photosynthetic pigment is correlated with cadmium
levels and durations. About 14% total chlorophyll was found down-regulated in pea
after 15 days of cadmium in lower treatment (1 ppm), which could reach about (25%)
in case cadmium treatments continued till 60 days. The higher levels (16 ppm) of
cadmium and duration both have extended loss in higher (80%) levels of the total
chlorophyll content (Fig. 33C; Appendix XIIIA). Similarly in case of chlorophyll a
101
RESULTS
and chlorophyll b (Fig. 33A,B; Appendix-XIIA,B). The loss in carotenoids was also
observed. The values of carotenoids have shown down regulation ca. 17-56% (1 ppm)
within 15-60 days (Fig. 33D; Appendix-XIIIB). The enhancement in days after
treatments have been found correlated in an increasing order in response to loss in
total carotenoid (26-78%).
15
45
10
6
30
60
8
6
30
60
a
4
3
2
2
1
0
0
6
15
45
12
B
30
60
Shoot dry weight (g)
Shoot fresh weight (g)
5
4
14
10
8
6
5
15
45
30
60
15
30
45
60
b
4
3
2
4
2
1
0
0
8
4.5
15
45
7
30
60
C
4
c
3.5
6
Root dry weight (g)
Root fresh weight (g)
15
45
A
Leaf dry weight (g)
Leaf fresh weight (g)
12
5
4
3
2
1
3
2.5
2
1.5
1
0.5
0
0
0
1
2
4
8
16
0
1
2
4
8
16
Cadmium treatment (ppm)
Cadmium treatment (ppm)
Fig.11.Changes
in fresh
weight
of leaf
shoot (B-b)
and(B-b)
root (C-c
Pisum
Fig. 31.
Changes in
freshand
anddrydry
weight
of(A-a)
leaf ,(A-a)
, shoot
and) in
root
(C-c )
sativum
var
Arkel
plant
treated
with
various
levels
of
Cadmuim
(1,2,4,8,and
16
ppm).The
in Pisum sativum var. Arkel plant treated with various levels of cadmium
cadmium water irrigation applied till 45 days (once in a week).Afterward , the response of
2, growth
4, 8 and
ppm). The cadmium
irrigation
applied
plant(1,
shoot
was16
observed.Measuremenst
werewater
conducted
on three
replicatetill 45 days
(once
in
a
week).
Afterward,
the
response
of
plant
shoot
growth was
plants.Values are means (n=3) with S.E.( ).
observed. Measurements were conducted on three replicate plants. Values
are means (n=3) with S.E. (±).
102
RESULTS
35
A
15
30
45
15
30
45
60
Total fresh weight (g)
30
25
20
15
10
5
0
16
B
60
Total dry weight (g)
14
12
10
8
6
4
2
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig.32.
Effect of
cadmium
irrigation
weight
(A)
Fig.12. Effect
of Cadmium
irigation
water on
total freshwater
weighton
(A)fresh
and total
dry weight
and
weight
Pisum
sativumofvar.
Arkel.
(B) in Pisum sativum
vs. total
Arkel.dry
Plants
grown (B)
underinthe
the application
different
theof 60
application
different
concentrations (1, 2,Plants
4, 8, and grown
16 ppm) under
for a period
days .Values of
are means
(n=3)
with S.E.(±).
concentrations (1, 2, 4, 8 and 16 ppm) for a period of
60 days. Values are mean (n=3) with S.E. (±).
103
RESULTS
4.13 Catalase, Peroxidase, and Lipid Peroxidase vs. Cadmium
The effect of cadmium irrigation in pea plant was also correlated with certain
stress inducible enzymes such as peroxidase and catalase. These enzymes genes
generally gets switched on during adverse experiences by plants. The data shown in
Fig. 34 clearly opted increasing trends in pea plant. Almost their intrinsic abilities in
relation to increase in peroxidase and catalase activity (%) are correlated with
cadmium levels, trends as observed with the increase in peroxidase activities
(Fig. 34A,B; Appendix-XIVA,B). Both these enzymes are stress mitigating
biomolecules therefore; biologically both of them have behaved as per biological rule
in supporting the biological system. Similarly lipid peroxidase was also increased
with increasing concentration of cadmium (Fig. 34C; Appendix-XVA).
4.14 Total Sugar and Proline vs. Cadmium
The total sugar shown similar trends as influenced by treatment levels. The
range of loss in sugar (6-12%) occurred in case treated with 1 ppm cadmium within
15-60 days but higher treatment 16 ppm (31-36%) for 15-60 days in plant growth as
shown in Fig. 35B (Appendix-XVIA). But in case of proline activity could enhanced
in range of ca. 70-18% within 15-60 days while enhanced the days and treatment (16
ppm), proline activity expressed ca. 156-90% within 15-60 days after treatment (Fig.
35A; Appendix-XVB).
104
RESULTS
2
1.6
15
30
45
A
60
15
1.8
30
45
60
B
1.4
Chl b (mg g-1 FM)
Chl a (mg g-1 FM)
1.6
1.4
1.2
1
0.8
1.2
1
0.8
0.6
0.6
0.4
0.4
0.2
0.2
0
0
0.8
3.5
15
30
45
C
60
15
Carotenoid (mg g-1 FM)
Total Chl (mg g-1 FM)
30
45
D
60
0.7
3
2.5
2
1.5
1
0.6
0.5
0.4
0.3
0.2
0.5
0.1
0
0
0
1
2
4
8
0
16
1
2
4
8
16
Cadmium treatment (ppm)
Cadmium treatment (ppm)
Fig. 33. Effect of differential levels of cadmium irrigation on chl a (A), chl b (B),
total chl (C) and carotenoid (D) in Pisum sativum var. Arkel. plants for a
period of 60 days. The values represent mean of three replicates (n=3) with
Fig.13.Effect
S.E. (±).of differential levels of cadmium irrigation on Chl a (A), Chl b (B), total
Chl (C) and Carotenoid (D) in Pisum sativum var Arkel .plant for a period of 60 days .The
values represent mean of three replicates (n=3) with S.E.(.(±)
105
Catalase ( µ mol decomposed H2O2 g-1 FM)
RESULTS
400
350
15
A
30
45
A
60
300
250
200
150
100
50
0
Peroxidase (∆ OD g-1 FM)
18
16
15
B
30
60
90
B
14
12
10
8
6
4
2
Lipid peroxidation (µmol MDA g-1 FM )
0
140
120
15
C
30
60
90
C
100
80
60
40
20
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig .14 Effect of of Cadmium irrigation water on enzymes activities of peroxidase (A)
Fig. 34. Effect of cadmium irrigation water on enzymes
catalase (B) and lipid peroxidase (C) in Pisum sativum var Arkel. Plants were exposed
of peroxidase
and
to 1, 2, 4, 8 andactivities
16 ppm cadmium
for a period(A)
of 60catalase
days.Data(B)
shown
are lipid
mean values
peroxidase
(C) indone
Pisum
sativum
var.
Arkel.
Plants
S.E.(±) of independent
experiments
in three
replicates
(n=3)
.
were exposed to 1, 2, 4, 8 and 16 ppm cadmium for a
period of 60 days. Data shown are mean values
S.E.(±) of independent experiments done in three
replicates (n=3).
106
RESULTS
80
15
A
30
45
60
70
-1
Proline
g FM
-1 FM)
Proline
(mg( gmg
60
50
40
30
20
10
0
20
15
30
45
60
18
B
Sugar ( µg g-1 FM )
16
14
12
10
8
6
4
2
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig. 35. Effect of differential cadmium concentrations (1, 2, 4,
8 and 16 ppm) proline activity (A) and sugar activity
(B) in Pisum sativum var. Arkel. The different
Fig .35. Effect of differential Cadmium concentrations (0, 1,2,4,8,and 16 ppm)
concentrations were analysed for a different time
proline activity (A) and sugar activity (B) in Pisum sativum var. Arkel. The
intervalswere
(15,analysed
30, 45forand
60 days).
Vertical
different concentrations
a different
time intervals
(15,bars
30 45 and
represent
means
(n=3)
with
S.E.
(±).
60 days).Vartical bars represent means (n=3) with S.E.(±).
107
RESULTS
4.16 Cadmium vs. Flower, Pod and Seeds
The differential cadmium levels (1, 2, 4, 8 and 16 ppm) have down regulated
acquisition of total number of flowers emerged, pods formed and also seed numbers.
The impact of cadmium irrigation on flower fresh and dry biomass was also evaluated
in relation to levels and duration of the cadmium as shown in Fig. 36 also favored
inability in parallel with the higher levels of cadmium in relation to retention of
flower fresh mass and dry mass (Fig. 36A; Appendix-XVIB). The loss in flower
fresh and dry mass ranged from 16-70 and 22-65% in response to treatment (1-16
ppm) and duration (45 days). The cadmium has also reduced pod formation process
which has resulted eventually in the form of loss of total number of seeds at maturity
(Fig. 36B). Pod formation process followed by seed setting during treatments under
our experimental conditions after 60 days of growth. The loss in seed number ranged
20-74% in response to treatment and duration (Appendix-XVIIA). The loss in seed
fresh and dry mass was observed. The fresh mass ranged from 17-82% and dry mass
16-85% respectively (Fig. 36C; Appendix-XVIIB).
108
RESULTS
2.5
0.25
DW
Flower fresh weight (g)
2
0.2
1.5
0.15
1
0.1
0.5
0.05
A
0
Flower dry weight (g)
FW
0
60
Seed number
50
40
30
20
10
B
25
Seed fresh weight (g)
FW
Dw
20
15
10
5
C
0
0
20
18
16
14
12
10
8
6
4
2
0
Seed dry weight (g)
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig. 36. Effect of differential cadmium concentrations (1, 2,
4, 8 and 16 ppm) on fresh and dry weight of
flower (A), seed number (B) and seed fresh and
dry weight (C) in Pisum sativum var. Arkel for a
period of 45 days (flower) and 60 days (seed).
Values
are means
(n=3) with S.E.(±)
Fig .16 .Effect of differential
Cadmium
concentrations
(1,2,4,8,and 16 ppm) on fresh and dry
weight of flower (A), seed number (B) seed fresh and dry weight (C) and no of pod (D) in
Pisum sativum var Arkel for a period of 45 days (flower ) and 60 days (seed). Values are means
(n=3 with S.E. (±).
109
RESULTS
4.17 Harvest Index vs.Cadmium
The harvest index characteristic was recorded to understand ultimate impact of
cadmium irrigation on plant performance and plant productivity in relation to
economic yield. The harvest index values as shown in Fig. 37 (Appendix-XVIII)
indicate continuous loss in the plant biomass gain compared to Cadmium treatment. It
was also observed i.e. 1.3, 6.5, 11.7, 21.6 and 48.7% loss in case plants allowed to
grow as influenced by cadmium treatments (1, 2, 4, 8 and 16 ppm) maintained in
earthen pots for a duration for a period of 60 days.
1
0.9
Harvest index (g g-1)
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
1
2
4
8
16
Cadmium treatment (ppm)
Fig. 37. Effect of cadmium concentrations (1, 2, 4, 8,
and 16 concentrations
ppm) on harvest
index
maturity
Fig.17. Effect of Cadmium
(1, 2, 4, 8,
and 16atppm
) on harvest
after6060
in sativum
Pisum (var
sativum
Arkel
index at maturity after
daysdays
in Pisum
Arkel var.
) seedlings.Values
are means of threeplants.
replications
with are
S.E. means
(±).
Values
of three replications
with S.E.(±)
4.17 Effect of Chromium on Zea maysseed Germination and Metal Tolerance
Index
The maize represented gradual loss in germination under treatment with
different chromium solutions. The differential germination percentage was recorded in
control and also upon treating with various chromium solutions. The differential
germination percentage was recorded in maize variety in case of control and also upon
treating with various chromium solutions. The germination percentage was found in the
range of 87% under normal condition free from chromium. However, 87% germination
reduced to the level of 77, 63, 57, 43 and 37% in case differentially treated with
110
RESULTS
chromium levels i.e., 1, 2, 4, 8 and 16 ppm respectively. The metal tolerance was
reduced significantly in the range of loss in 17-53% as compared to control as shown in
Fig. 39A (Appendix-XIXA).
4.18 Acquisition of Shoot Length, Root Length vs. Chromium Levels
The germinated seedlings were used to record the biodynamics in relation to
appearance/ acquisition of radical length, as influenced by different chromium levels.
The data indicate that seedlings unaffected (control) by chromium did acquire radical
length maximum throughout till 7days after germination. The root and shoot lengths
were recorded in maize as influenced by chromium (1, 2, 4, 8, and 16 ppm). Almost
similar trends have been observed in both. The root and shoot growth both have been
found down-regulated in relation to levels of chromium. Upon, increase in chromium
levels i.e., 1, 2, 4, 8 and 16 ppm, the root and shoot both have been found severely
down-regulated in case measurements were made 15 days after the application of
chromium. The higher level of chromium i.e., 16 ppm did not allow neither
germinated nor any further associated development processes (Fig. 40A,B;
Appendix-XIXB).
Our data indicate down-regulated shoot length acquisition in comparison to
control under the influence of chromium treatment. The lower level of treatment (1
ppm) followed parallel trends with much accuracy. Nearly, 34% loss in shoot length
could occurred in case seeds were treated with 1 ppm of chromium till the termination
of the observation (15 days). The 2, 4 and 8 ppm chromium application could downregulate the shoot length ca. 39, 48 and 67% compared to control. Further, ca. 79%
loss shoot length could get down-regulated in case chromium level enhanced (16
ppm) after 15 days application of chromium treatment (Fig. 40A). Similarly, the
down-regulation in root length was found to ca. 28, 37, 51, 87 and 92% (1, 2, 4, 8 and
16 ppm), concentrations treated respectively in case chromium treatment allowed for
15 days as compared to control (Fig. 40B).
4.19 Number of Leaves of Zea mays vs. Chromium
The effect of chromium treatment of total number of leaves on per seedlings
basis. The higher levels of chromium (16 ppm) could cause loss in leaf reduction to
the tune of ca. 50% compared to control as shown in Fig. 40C (Appendix-XX).
111
RESULTS
A
Control
15 Days after treatment
1 ppm
B
Control
2 ppm
4 ppm
8 ppm
16 ppm
15 Days after treatment
1 ppm
2 ppm
4 ppm 8 ppm 16 ppm
Fig. 38. Influence of differential chromium levels on shoot and root
bio-dynamics in Zea mays var. 4212 seedlings. A and B
indicate acquisition of root and shoot canopies as affected
by chromium treatment levels, i.e. 1, 2, 4, 8 and 16 ppm
coupled with Chromium treatment durations as indicated.
112
RESULTS
100
A
90
Germination %
80
70
60
50
40
30
20
10
0
0
1
2
4
8
16
120
Metal tolerance index (%)
B
100
80
60
40
20
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig. 39. Effect of chromium contaminated water on germination
% (A) metal tolerance index % (B) in Zea mays L.
Fig 2. Effect of Chromium contaminated water on germination % (A) metal tolerance
seedlings to grow till 15 days after concentrations (1, 2,
index % (B) in Zea4,mays
L. 16
seedlings
to growaretillmean
15 days
after
concentrations
(1, 2, 4,
8, and
ppm). Values
(n=3)
with
S.E.
8, and 16 ppm) .Values
(±). are mean (n=3) with S.E. (±).
113
RESULTS
14
Shoot length (cm)
A
12
10
8
6
4
2
0
14
Root length (cm)
B
12
10
8
6
4
2
0
3
C
Leaf number
2.5
2
1.5
1
0.5
0
0
1
2
4
8
Chromium treatment (ppm)
16
Fig. 40.
Effect of chromium
contaminated
water solution
Fg .3.Effect of Chromium
contaminated
water solution
in shoot length
(A) root length (B) and
in
shoot
length
(A)
root
length
and leaf ( 1, 2 ,4, 8
leaf number (C) in Zea mays L. seedlings at different Chromium(B)
concentrations
number
(C) inofZea
mays Each
L. seedlings
at represents
different mean (n=3)
and 16 ppm) was maintained
for a period
15 days.
data point
chromium concentrations (1, 2, 4, 8 and 16 ppm)
with S.E.(±).
was maintained for a period of 15 days. Each data
point represents mean (n=3) with S.E. (±).
114
RESULTS
4.20 Biomass and Moisture Content vs. Chromium Levels
The stability levels as influenced by chromium were revealed by recording
loss in root fresh and dry mass. The data shown in Fig. 41B (Appendix-XXIB)
indicates that loss in root fresh mass was shown by maize seedlings. The loss has been
found in the range of 49% in case lower level (1 ppm) of chromium applied. The
higher levels (16 ppm) of loss (92%) in root fresh mass was also recorded. The losses
in root dry mass was found in the range of 89% in case of treated with higher level
(16 ppm) of chromium in due course of time (15 days after treatment).Similarly loss
in shoot fresh biomass and dry mass. The data shown in Fig. 41A (Appendix-XXIA).
The loss has been found in the range of 44% in case lower level (1 ppm) of chromium
applied. The higher levels (16 ppm) of loss (88%) in shoot fresh mass was also
recorded. The losses in shoot dry mass was found in the range of 38% in case of
treatment with higher level (16 ppm) of chromium toxicity. The moisture content was
also reduced. The loss has been found in the range of 2% in case lower level (1 ppm)
of chromium applied. The higher levels (16 ppm) of loss (15%) in moisture content
was also recorded shown in Fig. 41C (Appendix-XXIIA).
4.21 Photosynthetic Pigments of Zea mays vs. Chromium
The photosynthetic pigment as shown in Fig. 42 (Appendix-XXIIB) i.e.,
chlorophyll a and b found affected. The chl b was found more significantly affected
compared to chl a. About 14% chlorophyll a and 20% chlorophyll b (1 ppm) were
found down-regulated in maize seedlings, which could reach about 48 and 77% (16
ppm) after 15 days of chromium (Fig. 42A,B). Similarly total chlorophyll was also
found down-regulated in maize, which could also reach about 59% (16 ppm) after 15
days of chromium treatment (Fig. 42 C). The loss in carotenoids was also observed.
The values of carotenoids have shown down regulation ca. 16-56% (1-16 ppm)
depending upon treatment levels within 15 days as shown by maize seedlings was
recorded (Fig. 42D; Appendix-XXIIB).
115
RESULTS
0.10
FW
DW
0.09
0.8
0.08
0.7
0.07
0.6
0.06
0.5
0.05
0.4
0.04
0.3
0.03
0.2
0.02
0.1
A
0.01
0
0.00
0.6
0.04
FW
DW
0.5
0.04
Root fresh wt (g)
0.03
0.4
0.03
0.3
0.02
0.02
0.2
Root dry wt (g)
Shoot fresh wt (g)
0.9
Shoot dry wt (g)
1
0.01
0.1
0.01
B
0
0.00
100
90
Moisture content
80
70
60
50
40
30
20
10
C
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.4.Effect of Chromium
water solution
on shootwater
fresh weight and dry
Fig. 41. contaminated
Effect of chromium
contaminated
weight (A) Root fresh weight
and
dry
weight
(B)
and
moisture
%
(C) in Zea mays L .
solution on shoot fresh weight and dry
seedlings at different Chromium
treatments
(1, 2,
4 ,8 and
was maintained for a
weight (A)
Root fresh
weight
and16
dryppm)
weight
period of 15 days.Each data
mean
withmays
S.E (±).
(B)point
and represents
moisture %
(C)(n=3)
in Zea
L.
seedlings at different chromium treatments
(1, 2, 4, 8 and 16 ppm) was maintained for a
period of 15 days. Each data point represents
mean (n=3) with S.E.(±)
116
RESULTS
0.6
0.45
A
B
0.4
0.5
Chl b (mg g-1 FM)
Chl a (mg g -1 FM)
0.35
0.4
0.3
0.3
0.25
0.2
0.15
0.2
0.1
0.1
0.05
0
0
1.2
0.4
C
D
0.35
Carotenoid (mg g-1 FM)
Total Chl (mg g-1 FM)
1
0.8
0.6
0.4
0.3
0.25
0.2
0.15
0.1
0.2
0.05
0
0
0
1
2
4
8
0
16
1
2
4
8
16
Chromium treatment (ppm)
Chromium treatment (ppm)
Fig. 42. Effect of chromium contaminated water solution on chl a (A), chl b (B), total
chl (C)
and carotenoid
(D) inwater
Zeasolution
mays L.
at different
chromium
Fig .5. Effect
of Chromium
contaminated
onseedlings
Chl a (A) Chl
b (B) Total
Chl (C)
concentrations
(1,
2,
4,
8
and
16
ppm)
was
maintained
for
a
period
of 15
and Carotenoid (D) in Zea mays .L. seedlings at different Chromium concentrations ( 1,2,4,8
days.
Each
data point
mean
S.E. represents
(±).
and 16 ppm)
was
maintained
for represents
a period of 15
days.(n=3)
Each with
data point
mean (n=3)
with S.E.(±).
117
RESULTS
4.22 Peroxidase, Catalase and Lipid Peroxidase vs. Chromium
The effect of chromium treatment in maize seedlings was also correlated with
certain stress inducible enzymes such as peroxidase and catalase. These enzymes
genes generally gets switched on during adverse experiences by plants. The data
shown in Fig. 43A,B clearly opted increasing trends in maize seedling. Almost their
intrinsic abilities in relation to increase in peroxidase (32-244%) is correlated with
chromium (1-16 ppm) levels. The data shown in Fig. 43A,B (Appendix-XXIIIA)
supports the trends as observed with the increase in peroxidase and catalase activities.
Both these enzymes are stress mitigating biomolecules therefore; biologically both of
them have behaved as per biological rule in supporting the biological system.
Similarly lipid peroxidase activity could enhanced ca. 112% higher in leave in case of
treated with 16 ppm chromium contaminated water for a period of 15 days (Fig. 43C;
Appendix-XXIIIA).
4.23 Amylase vs. Chromium
The amylase activity was significantly reduced at higher concentration of
chromium exposure. The loss in total amylase (30%) occurred in case treated with 1
ppm chromium but higher treatment 16 ppm (65%).Similarly α amylase (34%) and β
amylase (3%) occurred in case treated with 1 ppm chromium but higher treatment 16
ppm (66%) and (63%) for 15 days, in seedlings for growth as shown in Fig. 44A-C
(Appendix XXIIIB).
4.24 Protein vs. Chromium
The total protein shown similar trends as influenced by chromium treatment
levels. Protein content as revealed in Fig. 45B (Appendix-XXIVA) favoured that the
chromium treatment might have down regulated protein synthesis may associated
with 70S and 80S ribosome’s both. The leaf has shown a negative impact in acquiring
normal protein synthesis. The leaves have shown about 23% loss (1 ppm) while
higher treatment 84% loss (16 ppm).
118
RESULTS
Peroxidase (∆ OD g -1 FM)
10
9
A
8
7
6
5
4
3
2
1
Lipid peroxidase (µmol MDA g-1 FM)
Catalase ( (µmol H2O2 decomposed g-1 FM)
0
50
B
45
40
35
30
25
20
15
10
5
0
80
70
C
60
50
40
30
20
10
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig . 6 . Effect
Chromium
contaminated
solution inwater
peroxidase
(A)incatalase
Fig.of43.
Effect of
chromium water
contaminated
solution
activity (B)and lipidperoxidase
peroxidase (C)
in Zea
mays L.activity
seedlings(B)
at different
Chromium
(A),
catalase
and lipid
concentrations (1 ,2 peroxidase
,4 ,8 and 16(C)
ppm)
was
maintained
for
a
period
of
15
days .Each
in Zea mays L. seedlings at different
data point representschromium
mean (n=3)concentrations
with S.E.(±). (1, 2, 4, 8 and 16 ppm) was
maintained for a period of 15 days. Each data point
represents mean (n=3) with S.E.(±)
119
Total amylase
-1
(starch hydrolysed mg g FM)
RESULTS
6
A
5
4
3
2
1
0
α amylaseαstarch
hydrolysed
amylase
-1
(mg
FM)
g
(stardh hydrolysed mg g-1 FM)
4.5
B
4
3.5
3
2.5
2
1.5
1
0.5
0
0.7
β amylase
(starch hydrolysed mg g-1 FM)
C
0.6
0.5
0.4
0.3
0.2
0.1
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.7 . Effect ofFig.
different
concentrations
of Chromium
contaminated
water (1,2, 4, 8 and 16
44. Effect
of different
concentrations
of chromium
ppm) on total amylase (A)
α amylase (B)
and(1,β2,amylase
Zeaon
mays
contaminated
water
4, 8 and(C)
16 in
ppm)
totalL. seedlings to
amylase
amylase
(B) of
and
β amylase experiment
(C) in
grow till 15 days .Data shown
are(A),
meanα values
± S.E
independent
done in
Zea
mays
L.
seedlings
to
grow
till
15
days.
Data
three replicates.
shown are mean values ± S.E of independent
experiment done in three replicates.
120
RESULTS
Sugar ( µg g-1 FM)
4.5
A
4
3.5
3
2.5
2
1.5
1
0.5
0
0
1
2
4
8
16
Protein content (µg g-1 FM )
180
B
160
140
120
100
80
60
40
20
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.45.
Effect of different concentrations of chromium
contaminated water (1, 2, 4, 8 and 16 ppm) on sugar
(A) and protein content (B) in Zea mays L. seedlings to
grow till 15 days. Data shown are mean values ± S.E of
Fig.8. Effect of different concentrations
of Chromium
water (1,2, 4, 8 and
independent experiment
done incontaminated
three replicates.
16 ppm) on sugar (A) protein content (B) in Zea mays L. seedlings to grow till 15
days .Data shown are mean values ± S.E of independent experiment done in three
replicates.
121
RESULTS
30 Days after treatment
A
C
1
2
4
8
16
30 Days after treatment
B
60 Days after treatment
C
90 Days after treatment
D
Control
1 ppm
16 ppm
Fig. 46A. Influence of differential chromium levels on plant
morphology in Zea mays var. 4212. A-D indicate
acquisition of shoot appearance. The pots were
irrigated with chromium treatment levels, i.e. 1, 2, 4,
8 and 16 ppm, once in aaweek followed by normal
irrigation to the level of field capacity
122
RESULTS
A
30 Days after treatment
B
60 Days after treatment
C
90 Days after treatment
D
Control
1 ppm
2 ppm
4 ppm
8 ppm
16 ppm
Fig. 46B. Influence of differential chromium levels on plant morphology
in Zea mays var. 4212. A-D indicate acquisition of root
appearance and cob formation. The pots were irrigated with
chromium treatment levels, i.e. 1, 2, 4, 8 and 16 ppm, once a
week followed by normal irrigation to the level of field
capacity
123
RESULTS
4.26 Total Sugar vs. Chromium
The total sugar shown similar trends as influenced by treatment levels. The
loss in sugar (19%) occurred in case treated with 1 ppm chromium but higher
treatment 16 ppm (81%) for 15 days, caused loss in this parameter in seedlings during
growth (Fig. 45A; Appendix-XXIVB).
4.26.1 Shoot length and root length of Zea mays vs. chromium durations and
levels
The long-term (upto ca. 90 days) chromium irrigation consequences were
planned to reveal seed to seed status of chromium treatment. These studies were made
in earthen pots. The morphological approaches for maize cultivar has also been shown
in Fig. 46A-B. The consequences of 1, 2, 4, 8 and 16 ppm chromium solutions have
affected their morphological appearances and differential growth behaviour. Higher the
treatment, lower the plant height/ shoot length was observed in comparison to maize
cultivar grown by providing normal irrigation levels. The shoot length growth
behaviour was recorded in due course of time i.e., 90 days at the interval of 15 days.
The lower level of chromium treatment (1 ppm) could cause loss in shoot length ca.
14% within 90 days which could get further enhanced to the level of 58% in case of
treated with four fold higher chromium solution (16 ppm) as shown in Fig. 47
(Appendix-XXVA). Similarly the root length growth behavior was recorded in due
course of time i.e., 15-90 days at the interval of 15 days. The lower level of chromium
treatment (1 ppm) could cause loss in root length ca. 6% within 90 days which could
get further enhanced to the level of 31% in case of treated with five fold higher
chromium solution (16 ppm) as shown in Fig. 47B (Appendix-XXVC).
4.27 Number Leaves of Zea mays vs. Chromium Irrigation
The total number of leaves have shown down regulation in retaining their
number almost 15-69% depending upon the treatment levels within 15 days in
maizessedlings (Fig. 47C). The enhancement in days after treatment have been found
correlated in an increasing order in response to loss in total number of leaves. The
maize cultivar has shown ca. 10% loss (1 ppm) in total leaves after 90 days in
comparison to 14, 21, 25 and 32% in case of treated with 2, 4, 8 and 16 ppm levels of
the chromium after 90 days of the treatment (Fig. 47C; Appendix-XXVB).
124
RESULTS
100
90
15
A
30
Shoot length (cm)
80
60
70
90
60
50
40
30
20
10
0
90
80
15
30
60
90
B
Root length (cm)
70
60
50
40
30
20
10
0
16
15
30
60
90
C
14
Leaf number
12
10
8
6
4
2
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.10.Fig.
Effect
different
Chromium
concentrations
(1,2,4,8,and
47.ofEffect
of different
chromium
concentrations
(1, 16
2, ppm)
4, on acquisition of
shoot length(A)8,root
(B) and
leaf number of
(C)shoot
in Zea
mays (A),
L. Plants were maintained
andlength
16 ppm)
on acquisition
length
for a period of 90
days.Vertical
bars
represent
(n=3)
with
their
S.E.(±).
root length (B) and leaf number (C) in Zea mays L.
Plants were maintained for a period of 90 days.
Vertical bars represent (n=3) with their S.E.(±)
125
RESULTS
4.29 Zea mays of Biomass vs. Chromium Irrigation
The impact of chromium treatment on plant biomass was also evaluated in
relation to levels and durations. The data shown in Fig. 48, also favored inability in
parallel with the higher levels of treatment in relation to retention of shoot fresh mass
and shoot dry mass. The loss in shoot fresh mass as shown from 23% in comparison
to 62% in case used lower (1 ppm) and higher (16 ppm) levels of Chromium in
response to treatment (90 days) duration (Fig. 48B; Appendix-XXVIIIA,B).
The stability levels as influenced by chromium were revealed by recording
loss in root fresh biomass and dry mass. The data shown in Fig. 48C (AppendixXXVIIA,B) indicates that loss in root fresh mass was shown by Zea mays cultivars.
The loss has been found in the range of 38-46% in case of lower level (1 ppm) of
chromium applied for irrigation in due course of time (15-90 days after treatment).
The higher levels of loss in root biomass was also recorded. The losses in root fresh
mass were found in the range of 84-75% in case irrigated with higher level (16 ppm)
of chromium in due course of time (15-90 days after treatment), similarly losses in
leaf fresh and dry mass shown in Fig. 48 (Appendix-XXVIA, B). Total fresh and dry
biomass were also reduced with increasing concentration of chromium levels in maize
plants. The data shown in Fig. 49A,B (Appendix-XXIXA,B) indicates that loss in
total fresh mass was shown by Zea mays cultivars. The loss has been found in the
range of 35-26% in case lower level (1 ppm) of chromium applied for irrigation in
due course of time (15-90 days after treatment). The higher levels of loss in total fresh
biomass were also recorded. The losses in total dry mass were found in the range of
91-67% in case irrigated with higher level (16 ppm) of chromium in due course of
time (15-90 days after treatment).
4.29 Photosynthetic Pigments of Zea mays vs. Chromium
The morphological appearance was found affected due to applied chromium
levels therefore photosynthetic pigments viz., total chlorophyll, chlorophyll a ,
chlorophyll b and carotenoid content were recorded (Fig. 50; Appendix XXXA,B &
XXXIA,B). The values of chlorophyll concentration indicated that maize had gradual
loss in retaining photosynthetic pigment i.e., chlorophyll. The loss in photosynthetic
pigment is correlated with chromium levels and durations. About 8% total chlorophyll
was found down-regulated in maize after 15 days of chromium in lower treatment (1
ppm), which could reach about (9%) in case chromium treatments continued till 90
126
RESULTS
days. The higher levels (16 ppm) of chromium and duration both have extended loss
at higher levels in total chlorophyll content (Fig. 50C). Just like chlorophyll a and
chlorophyll b. The loss in carotenoids was also observed. However it was lower as
compared to the chlorophyll loss (Fig. 50A-C). The values of carotenoids have shown
down regulation ca. 30-7% (1 ppm) within 15-90 days while increasing treatment loss
in 78-24% (16 ppm) (Fig. 50D; Appendix-XXXIB).
25
15
30
60
A
90
15
50
Leaf dry weight (g)
Leaf fresh weight (g)
60
40
30
20
90
15
30
60
90
b
15
30
60
90
c
10
5
0
0
25
15
30
60
90
B
60
Shoot dry weight (g)
Shoot fresh weight (g)
60
15
10
70
a
30
20
50
40
30
20
15
10
20
5
10
0
0
80
20
15
60
C
90
18
16
60
Root dry weight (g)
Root fresh weight (g)
70
30
50
40
30
20
14
12
10
8
6
4
10
2
0
0
0
1
2
4
8
16
0
Chromium treatment (ppm)
1
2
4
8
16
Chromium treatment (ppm)
Fig.
48. Effect
Effectofof
different
chromium
concentrations
(1, 2,164,ppm)
8 and
16 ppm) on
Fig.11.
different
Chromium
concentrations
(1,2,4,8,and
on acquisition
of
acquisition
fresh
and
dry (B-b)
weight
leaf(C-c)
(A-a),
(B-b)
and Chromium
root (C-c)
fresh and dry
weight ofofleaf
(A-a)
shoot
andofroot
in shoot
Zea mays
L.The
water irrigation
60 days
(once in water
a weekirrigation
).Measurement
were
on in
in Zeaapplied
mays till
L. The
chromium
applied
tillconducted
60 days (once
three replicate
plantsMeasurement
.The data points
are means
with S.E.(±).
a week).
were
conducted
on three replicate plants. The data
points are means with S.E.(±)
127
RESULTS
200
A
15
Total fresh weight (g)
180
30
60
90
160
140
120
100
80
60
40
20
0
80
B
15
30
60
90
Total dry weight (g)
70
60
50
40
30
20
10
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.12.Effect of differential levels of Chromium irrigation of water on total fresh weight (A)
and total dry weight (B) in Zea mays L. Plants were grown in earthen pots (radius= 15 cm
and length
cm ) filled
soil and compost
(3:1of).The
weekly treatments
were imposed
Fig.
49.30Effect
of with
differential
levels
chromium
irrigation
of
for a period of
60
days.The
values
represent
mean
of
three
replicates
(n=3)
with
S.E.(±).
water on total fresh weight (A) and total dry weight
(B) in Zea mays L. Plants were grown in earthen pots
(radius = 15 cm and length 30 cm) filled with soil
and compost (3:1). The weekly treatments were
imposed for a period of 90 days. The values
represent mean of three replicates (n=3) with S.E.
(±).
128
RESULTS
1.8
2.5
15
30
60
90
15
A
Chl b (mg g-1 FM)
2
Chl a (mg g-1 FM)
30
60
90
B
1.6
1.5
1.4
1.2
1
0.8
1
0.6
0.4
0.5
0.2
0
0
1.4
4.5
15
30
60
90
30
60
90
D
1.2
3.5
Carotenoid (mg g-1 FM)
Total Chl (mg g-1 FM)
15
C
4
3
2.5
2
1.5
1
0.8
0.6
0.4
1
0.2
0.5
0
0
0
1
2
4
8
16
0
1
2
4
8
16
Chromium treatment (ppm)
Chromium treatment (ppm)
Fig. 50. Effect of differential levels of chromium concentrations of irrigation
water (1, 2, 4, 8 and 16 ppm) of specific time intervals i.e. 15, 30, 60,
and 90 days after chromium treatment on chl a (A), chl b (B), total chl
(C) and carotenoid (D) in Zea mays L. The values represent mean of
three replicates (n=3) with S.E. (±).
129
RESULTS
Catalase ( µmol decomposed H 2O2 g-1 FM )
90
80
A
15
30
60
90
70
60
50
40
30
20
10
0
45
Peroxidase ( ∆ OD g-1 FM)
40
B
15
30
60
90
35
30
25
20
15
10
5
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig. 51. Effect of chromium irrigation on catalase (A) and peroxidase
(B) in leaves of Zea mays L. experienced with various
chromium levels (1, 2, 4, 8 and 16 ppm). Measurements
were recorded by using different leaf sample after
experiencing differential chromium levels. The observations
were recorded after chromium treatment at the time interval
of 15, 30, 60, and 90 days. The values represent mean of
three replicates (n=3) with S.E. (±).
130
RESULTS
4.30 Catalase, Peroxidase and Lipid Peroxidase vs Chromium
The effect of chromium treatment in maize plant was also correlated with
certain stress inducible enzymes such as peroxidase and catalase. These enzymes
genes generally gets switched on during adverse experiences by plants. The data
shown in Fig. 51A,B (Appendix-XXXIIA,B) clearly opted increasing trends in maize
seedling. Almost their intrinsic abilities in relation to increase in peroxidase (%) is
correlated with chromium levels. The data shown in Fig. 51 supports the trends as
observed with the increase in peroxidase activities. Both these enzymes are stress
mitigating bio-molecules therefore; biologically both of them have behaved as per
biological rule in supporting the biological system. Similarly lipid peroxidase was
also increased with increasing concentration of chromium levels. Lipid peroxidase
activity could enhanced of range in ca. 128-22% higher in the case of leaves which
were treated with 16 ppm chromium irrigation for a period of 15-90 days (Fig. 52A;
Appendix-XXXIIIA).
4.31 Total Sugar and Proline vs. Chromium
The total sugars shown similar trends as influenced by treatment levels. The
range of loss in sugars (13-26%) occurred in 1 ppm chromium within 15-90 days but
higher treatment 16 ppm (55-63%) for 15-90 days, the data as shown in Fig. 53
(Appendix-XXXIV). However, proline activity could enhanced in range of ca. 3623% (1 ppm) within 15-90 days while enhanced the days and treatment (16 ppm),
proline activity expressed ca. 300-167% within 15-90 days after treatment as shown
in Fig. 52B (Appendix-XXXIIIB).
131
RESULTS
300
Lipid peroxidase ( µmol MDA g-1 FW)
A
15
30
60
90
30
60
90
250
200
150
100
50
0
140
B
15
Proline (mg g-1 FW)
120
100
80
60
40
20
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig. 52. Response of lipid peroxidation expressed as amount
of malodialdehyde (A) and proline activity (B) in
Fig.15.Response chromium
of Lipid peroxidation
expressed
as amount
of malodialdehyde
concentrations
of irrigation
levals.
Zea
(A) and proline activity
(B)
in
Chromium
mium
concentrations
of irrigation
mays L. plants were exposed to 1, 2, 4, 8, and
16
levals. Zea mays ppm
L.plants
were
exposed
to
1,
2,
4,
8,
and
16
ppm
Chromium
chromium for a period of 90 days as compared
control.
The values
represent
mean
of three
for a period of 90 to
days
as compared
to control.The
values
represent
mean of
replicates
(n=3)
with
S.E.
(±).
three replicates (n=3) with S.E.(±).
132
RESULTS
18
15
16
30
60
14
-1
Sugar (µ g g FW)
90
12
10
8
6
4
2
0
0
1
2
4
8
16
Chromium treatment (ppm)
in sugar
in Zeaactivity
mays L. cultivar
various
Fig. Fig.16.
53. Changes
Changes
inactivity
sugar
in treated
Zea with
mays
L.
levels of Chromium (1, 2, 4, 8 and 16 ppm).The Chromium water irrigation
cultivar
treated
with
various
levels
of
applied till 6o days (once in a week ).Measurement were analysed in three
replicate chromium
plants. The data points
S.E.(±).16 ppm). The
(1, are2,means
4, with
8 and
chromium water irrigation applied till 60
days (once in a week). Measurement were
analysed in three replicate plants. The data
points are means with S.E. (±).
9
60 days
Inflorescence fresh weight (g)
8
90 days
A
7
6
5
4
3
2
1
0
5
60 days
Inflorescence dry weight (g)
4.5
90 days
B
4
3.5
3
2.5
2
1.5
1
0.5
0
0
1
2
4
8
16
Chromium treatment (ppm)
Fig.
54. Effect
of different
chromium
concentrations
(1,ppm)
2, on freshFig.17.Effect
of different
Chromium
concentrations
( 1,2,4,8,and 16
dry weight 4,
of inflorescence
(A,
B)
in
Zea
mays
L.
at
specific
intervals
8 and 16 ppm) on fresh-dry weight ofi.e. 60 -90
days.Values are means (n=3) with S.E.(±).
inflorescence (A, B) in Zea mays L. at specific
intervals i.e. 60-90 days. Values are means (n=3)
with S.E. (±).
133
RESULTS
4
Cob number plant-1
A
3.5
3
2.5
2
1.5
1
0.5
0
Length of cob (cm)
18
B
16
14
12
10
8
6
4
2
0
200
C
Seed number plant -1
180
160
140
120
100
80
60
40
20
0
0
1
2
4
8
16
Chromium treatment (ppm)
55. Effect of
differential
levelsirrigation
of chromium
irrigation
Fig.18..EffectFig.
of differential
levels
of Chromium
of water
on total
of
water
on
total
number
of
cob/
plant
formed
number of cob/ plant formed (A) length of cob (B) and number of seeds found
(A), length
cobgrown
(B) and
number
seeds 15 cm
(C) at maturity in Zea mays
L. Plantsofwere
in earthen
potsof(radius=
found
(C)
at
maturity
in
Zea
mays
L.
plants
and length 30 cm ) filled with soil and compost (3:1 ).The weekly treatments
were
in earthen
(radius=
15ofcm
and
were imposed for a period
of grown
60 days.The
valuespots
represent
mean
three
length 30 cm) filled with soil and compost
replicates (n=3) with S.E.(.(±)
(3:1).The weekly treatments were imposed for a
period of 60 days.The values represent mean of
three replicates (n=3) with S.E. (±).
134
RESULTS
1
0.9
0.8
-1
Harvest index (g g )
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
0
1
2
4
8
16
Chromium treatment (ppm)
The changes
harvest
index
in L.
Zea
mayswith
L. treated
Fig. 19.Fig.
The56.
changes
in harvestinindex
in Zea
mays
treated
variouswith
levels of
various
levels
of
chromium
(1,
2,
4,
8
and
16
as
chromium (1, 2, 4, 8, and 16 ppm ) as compared to control at maturityppm)
( 90 days
compared
to
control
at
maturity
(90
days).
The
).The Chromium water irrigation applied till 60 days ( once in a week ). Afterward
chromium
water was
irrigation
applied
days (once
in (n=3)
a
the response of plant
shoot growth
observed.
Each till
data60
represents
mean
week).
Afterward
the
response
of
plant
shoot
growth
was
with S.E. (±).
observed. Each data represents mean (n=3) with S.E.(±).
135
RESULTS
4.32 Chromium vs. Cobs and Seeds
The differential chromium levels (1, 2, 4, 8 and 16 ppm) have down regulated
acquisition of cobs formed and also seed numbers. The chromium has also reduced
cob formation process which has resulted eventually in the form of loss of total
number of seeds at maturity (Fig. 55A,B; Appendix-XXXVIIA). The loss in length
of cob formation was detected 12 and 47% in case treated with lower (1 ppm) and
higher (16 ppm) Chromium solutions. Cob formation process followed by seed setting
during treatments under our experimental conditions after 90 days (Fig.55C;
Appendix- XXXVIA, B). The losses in inflorescence fresh and dry mass are also
observed after 60 and 90 days of treatment as shown in Fig. 54 (Appendix-XXXV
A,B).
4.33 Harvest Index vs. Chromium
The harvest index values as shown in Fig. 56 (Appendix-XXXVIIB) indicate
continuous loss in the plant biomass gain compared to chromium treatment. It was
also observed i.e. 6, 9, 19, 27 and 52% loss in case plants allowed to grow as
influenced by chromium treatments (1, 2, 4, 8 and 16 ppm) maintained in earthen pots
for a duration of 90 days.
136