Bioaccumulation and Biochemical Effects of

Mercury in the Plant Bacopa monnieri (L)

Sarita Sinha,* Manisha Gupta, and Prakash Chandra
Aquatic Botany Laboratory, National Botanical Research Institute, Lucknow, 226001 India

Plants of Bacopa rnonnieri were treated with six different concentrations of Hg (0.01, 0.5, 1.O, 2.0,
3.0, and 5.0 p g mL-l) for 4, 7, and 14 d under laboratory conditions. The metal accumulation in the
root tissues was about five times more than in the shoots. At all the concentrations of Hg, cysteine,
total -SH,reduced glutathione, and ascorbic acid contents increased in the roots of 5. monnieri up
to 7 d of exposure except the ascorbic acid content, which decreased above 2 p g mL- Hg. The
rnalondialdehyde content decreased at all the concentrations of Hg; however, protein and sugar
contents decreased above 0.1 and 2.0 p g mL- Hg, respectively, after 4 d of exposure in the roots
of the plant. The chlorophyll content in leaves decreased significantly with an increase in metal
concentrations and durations of exposure. The results suggest that an increase in cysteine, total
-SH, reduced glutathione, and ascorbic acid content by Hg treatment at the initial exposure period
are part of the overall expression of Hg tolerance in the plant, and the decrease in chlorophyll and
protein content is a consequence of Hg toxicity at higher metal concentrations and increased period
of exposure. 0 1996 by John Wiley & Sons, Inc.

INTRODUCTlON To combat metal toxicity, plant cells have sub-

Mercury is one of the major toxic pollutants in water
bodies situated adjacent to chloralkali plants. Indus-
tries such as those for the manufacture of paper and
pulp, paint, battery, and pesticide also contribute sig-
nificantly to mercury pollution of aquatic environ-
ments. Such pollution is a serious problem at Ganjam,
Orissa (India), due to indiscriminate discharge of Hg-
containing effluent from the chloralkali plant to the
adjacent low-lying agricultural land (Panda et al.,
1990, 1992).
Mercury is transformed from less toxic inorganic
species to highly toxic organic species such as methyl
mercury through biological and abiological processes.
Methyl mercury biomagnifies through trophic levels of
the food chain and causes serious health hazards (Panel
Report on Mercury, 1978).
* To whom correspondence should be addressed.
stances like glutathione (GSH), ascorbate, carot-
enoids, and enzymes like superoxide dismutase, ascor-
bate peroxidase, and glutathione reductase that
participate in scavenging active 0, species such as
0; and H,O, generated in chloroplasts during photo-
synthesis (Halliwell, 1982). Glutathione is the major
free thiol compound in plants and is the substrate for
dehydroascorbate reductase, which converts dehy-
droascorbate to ascorbate. Glutathione and ascorbate
play a prominent role in defense against the free radi-
cals in plants under conditions of oxidative stress (De
Vos et al., 1992; Ranieri et al., 1993). The potential for
the production of activated oxygen species is greatly
enhanced by heavy metals such as Cd, Cu, Hg, and
Ni, which are responsible for peroxidative damages
to fatty acids, nucleic acids, proteins, and chlorophyll
(Thompson et al., 1987). The metals are toxic to plants
if their accumulation levels exceed the detoxification
capacity of the plant tissues. Thus, a potentially deci-

Environmental Toxicology and Water Quality: An International Journal, Vol. 1 1 (1996)105-1 12

0 1996 by John Wiley & Sons, Inc. CCC 1053-4725l96102105-08
105
106 SINHA ET AL.
sive factor in determining the outcome of oxidative
stress is the speed with which plants can activate their
antioxidant reserves (Howe and Merchant, 1992; Ra-
nieri et al., 1993). Correlation studies have indicated
that this response is an important aspect of stress tol-
erance.
The plant Bacopa monnieri is a fast proliferating
species in wetlands. It has been shown to accumulate
toxic metals in our earlier studies (Sinha and Chandra,
1990; Gupta et al., 1995a; Rai et al., 1995; Vajpayee et
al., 1995). Lenka et al. (1992) reported that this plant
grows prolifically at Ganjam, Orissa (India), where Hg
concentrations in sediments and water are 41.3 mg kg-'
and 4 pg L-', respectively. The study reported here
was undertaken, therefore, to assess the accumulation
of Hg and some underlying biochemical responses such
as cysteine, total -SH, reduced glutathione, chloro-
phyll, protein, ascorbate, sugar, and malondialdehyde
content of B . monnieri.
MATERIALS AND METHODS
Rooted shoots of B . monnieri (10-15 nodes long) were
taken from the plants growing in hydroponic tubs at
National Botanical Research Institute, Lucknow. They
were acclimatized in 5% Hoagland's solution for six
weeks. Different concentrations of Hg-namely, 0.01,
0.5, 1.0, 2.0, 3.0, and 5.0 pg mL-'-were prepared
using HgC1, in 5% Hoagland's solution. Plants of B .
monnieri were treated with different concentrations of
Hg. The metal solution in all the treatment flasks was
changed after 7 days. Three sets of flasks were used
for each treatment and for the control. Plants without
Hg Conc. in Root (ug g-1 dw)
300 I
0 01 0 05 10 20 50
Hg @g mt-1)
Fig. 1. Mercury concentrations in roots of 6.monnieri.
Values are means of triplicate determinations. F value
(conc.) = (*) 10.32; Fvalue (expo.) = (**) 6.08. * p < 0.01 ;
(**) p < 0.05.
ig Conc. in Shoot (ug g-1 dw)
5c
* 44 y = 0.46+2.89x ( ~ 0 . 9 5 )
4c
+ 74 y = 2.04+5.70r ( r = 0 . 9 8 )
+++ ,44 y = 1.26+9.87r ( r S . 9 7 )
30
20
10
0
0.01 0.05 1.0 2.0 30 5.0
Hg (us mi-1)
Fig. 2. Mercury concentrations in shoots of 6.monnieri.
Values are means of triplicate determinations. F value
(conc.) (*) = 28.73; F value (expo.) = (**) 2.75. (*) p <
0.01 ; (**) nonsignificant.
metal served as the control. The experiment was per-
formed under standard physiological conditions pro-
viding 16 h light (1 15 pmol m-2 s-')/8 h dark photope-
riod using fluorescent tube lighting (Philips) at 25 k
2C. Plants were harvested after 4, 7, and 14 d. The
blotted-dry fresh roots and leaves were used for bio-
chemical studies.
Harvested plants were washed thoroughly with dis-
tilled water, dried, and digested at room temperature
in concentrated HNO, : HC104(4 : 1, v/v) until dissolu-
tion, followed by gentle heating on an aluminum block.
The concentration of mercury both in root and shoot
tissues was measured using a cold vapor Atomic Ab-
sorption Spectrophotometer 2380.
Chlorophyll in the leaves (100 mg) was extracted in
80% chilled acetone and estimated by the method of
Arnon (1949) using Spectronic 1201 Milton Roy Spec-
1
trophotometer. Protein content was measured by the
method of Lowry et al. (1951) using bovine serum aibu-
min as the standard protein.
For the measurement of lipid peroxidation in roots
(700 mg) and leaves (700 mg), the thiobarbituric acid
(TBA) test, which determines malondialdehyde (MDA)
as an end product of lipid peroxidation (Heath and
Packer, 1968), was used.
Free cysteine content both in the leaves (700 mg)
and roots (100 mg) was measured by the method of
Gaitonde (1967). Ascorbic acid content was determined
colorimetrically both in the root (100 mg) and leaf
(100 mg) tissues following the method of Keller and
Schwagner (1977). Total sugar (monosaccharides) was
estimated in roots (100 mg) and leaves (100 mg) by the
phenol H2S0, method (Dubios et al., 1958). Total -SH
content in roots was measured spectrophotometrically
 EFFECTS OF Hg IN 6 . MONNlERl 107

Chlorophyll (mg g - l f w ) and correlation analysis were performed wherever nec-

n essary (Gomez and Gomez, 1984).
1.5

1
c5
4 7 14
Treatment duration (d)
Fig. 3. Mercury concentrations (pg mL-') affecting total
chlorophyll content of 8. rnonnieri leaves. Values are
means of triplicate determinations. Standard deviations
are represented by vertical bars. The t test (compared to
the control): a = p < 0.05; b = p < 0.02; c = p < 0.01.
with Ellman's reagent (Ellman, 1959). Reduced gluta-
thione and oxidized glutathione in roots were measured
by the method of Hissin and Hilf (1976) using Fluores-
cence Spectrophotometer (Hitachi Model No. 6.50-60).
Analysis of variance was performed for calculating
the statistical significance of the results using com-
pletely randomized block design. Student's t test was
applied to see the statistical significance of the results
as compared to the control. Simple linear regression
RESULTS
Accumulation of Hg in the roots (Fig. 1) of B . monnieri
was concentration ( p < 0.01) and duration of exposure
( p < 0.05) dependent, while in shoots (Fig. 2) the up-
take was only significant ( p < 0.01) with respect to the
concentrations of Hg. Accumulation in the roots was
about five times more than in shoots and the maximum
accumulation of 273 and 48.7 p g g-' dry weight (dw)
were observed in roots and shoots, respectively, at 5
pg mL-' Hg after 14 d of exposure.
Total chlorophyll content decreased significantly
with increase in metal concentrations, showing reduc-
tions of 23, 3.5, and 3.5%, as compared to their respec-
tive controls at 5 pg mL-' Hg after 4, 7, and 14 d of
exposures, respectively (Fig. 3).
Protein content (Table I) of roots decreased with an
increase in metal concentration (>0.01 pg mL-' Hg)
and duration of exposure. Significant reductions of 20,
30, and 47% (compared to their respective controls)
were observed after 4 , 7 , and 14 d of exposures, respec-
tively, at 5 pg mL-' Hg. The protein content of the
leaves (Table I) decreased with an increase in metal
concentration (>0.01 p g mL-' Hg) and duration of
exposure. Significant reductions of 38 and 61% (com-
pared to their respective controls) were recorded at 5
pg mL-' Hg after 7 and 14 d of exposure, respectively.

TABLE I. The effect of mercury on protein content ( p g g-' fwa) of roots and leaves 01
6. monnierib

Hg Concentration
( m mL-') 4 Days 7 Days 14 Days
0.0 18.60 t 0.74 18.57 ? 0.03 19.10 ? 0.14
(10.22 ? 0.50) (10.86 ? 0.11) (11.19 +- 0.30)
0.01 19.80 k 0.89 19.20 ? 0.14b 18.93 ? 0.25a
(10.62 t 0.26) (10.43 ? 0.93) (10.41 t 0.81)
0.50 18.56 -t 1.0 18.22 ? 0.38b 18.23 t 0.29a
(10.16 t 0.67) (10.27 ? 1.17) (10.16 t 0.85)
1 .o 16.87 -+ 0.67 16.67 t I.0la 16.17 ? 1.05a
(10.03 5 0.98) (10.00 ? 0.65) (8.50 ? 1.20)
2.0 17.00 ? 0.37 14.89 -+ 0 . 4 8 ~ 13.10 t 0 . 6 2 ~
(9.69 -t 0.32) (8.15 t 0.65)b (5.81 t 0 . 2 2 ) ~
3.0 15.17 t 1.05a 14.54 t 0.81b 11.95 ? 0 . 8 6 ~
(9.69 t 0.92) *
(7.50 0 . 5 6 ) ~ (5.98 t 0.87)b
5.0 14.97 t I . l l a 13.03 ? 0 . 8 9 ~ *
10.03 O.lld
(9.67 ? 0.87) (6.70 +- 0 . 3 0 ) ~ *
(4.35 0 . 3 4 ) ~

Fresh weight
a
Each value is the mean of triplicate determinations 2SD. Values in parentheses are protein content
in leaves. The t test (compared to the control): a = p < 0.05; b = p < 0.02; c = p < 0.01 ; d = p < 0.001.
108 SINHA ET AL.

TABLE II. The effect of mercury on MDA content (nmol g-' fw) in the roots and
leaves of 6. monnieri"

Hg Concentration
(pg rnL-') 4 Days 7 Days 14 Days

0.0 6.84 t 0.37 6.82 i 0.32 6.69 i 0.38

(4.31 t 0.21) (4.34 i 0.25) (4.37 i 0.31)
0.01 6.56 i 0.41 6.45 i 0.30 6.40 i 0.37
(3.92 t 0.21) (3.86 5 0.20) (3.05 5 0.21)a
0.50 6.00 i 0.38 5.99 i 0.27 6.06 i 0.38
(3.90 i 0.15) (3.84 i 0.18) (2.91 i 0.18)a
1 .oo 5.99 i 0.25 5.97 i 0.35 5.90 t 0 . 4 0 ~
(3.82 5 0.19) (3.81 i 0.18) (2.71 i 0.17)b
2.0 5.81 t 0.39 4.72 i 0.26b 4.25 i 0.29b
(3.80 i 0.16) (3.49 i 0.15)a (2.61 i 0.19)b
3.O 4.65 i 0.30b 4.14 i 0 . 2 3 ~ 4.11 t 0 . 2 1 ~
(3.70 i 0.19) (3.23 i 0.12)b (2.58 i 0.13)b
5.0 4.05 i 0.37b 3.69 i 0 . 2 5 ~ 3.25 i 0 . 1 9 ~
(3.60 i 0.12)a (3.09 5 0.06)b (2.07 i 0. 12)~

Each value is the mean of triplicate determinations 2SD. Values in parentheses are MDA content in
leaves. The t test (compared to the control): a = p < 0.05; b = p < 0.02; c = p < 0.01.

The MDA content in the roots and leaves decreased
with an increase in metal concentration and duration
of exposure (Table 11). Significant decreases of 41,46,
and 51% in root, and 16, 29, and 53% (compared to
their respective controls) in leaf, MDA content were
noticed at 5 pg mL-' Hg after 4, 7, and 14 d of expo-
sure, respectively.
Cysteine content in roots (Table 111) increased with
an increase in Hg concentrations; however, decreases
of 7 and 17% (compared to the control) were noticed
at 3 and 5 p g mL-' Hg, respectively after 14 d of
exposure. In contrast to roots, cysteine content of
leaves (Table 111) decreased with an increase in Hg
concentrations. It decreased significantly to 3 1,34, and
35% (compared to their respective controls) at 5 pg
mL-' Hg after 4,7, and 14 d of exposure, respectively.

TABLE 111. The effect of mercury on cysteine content (nmol g-' fw) of roots and
leaves of 6. monnieri"

Hg Concentration
(pg rnL-') 4 Days 7 Days 14 Days

0.0 115.3 i 6.6 110.2 i 6.3 118.4 i 5.9

(44.5 i 2.7) (48.0 i 4.7) (48.2 5 3.8)
0.01 124.2 2 7.8 134.7 5 5.3a 124.5 t 4.9
(35.1 i 3.5) (48.9 5 3.9) (42.7 i 2.9)
0.50 125.5 2 5.7 145.6 i 6.Sa 135.2 i 6.2
*
(33.8 2.9)a (47.5 2 5.8) (40.3 i 4.7)
I .O 130.7 t 6.7 167.8 2 7.5b *
140.5 5.9a
(34.5 t 3.1) (45.7 i 4.7) (40.5 t 4.5)
2.0 *
133.8 5.9 170.5 i 9.5b 142.7 i 8.5
(33.5 i 2.5)a (39.5 2 3.8) (35.3 i 3.9)
3.0 *
140.1 6.8a *
180.2 11.5b 110.5 i 7.3
*
(33.3 2.6)a (38.3 t 2.7) (33.5 t 2.7)a
5.0 148.8 i 7.5a 200.9 k 1 0 . 9 ~ 98.5 t 5.3a
(30.8 & 3.5)a (31.7 2 2.5)a (31.5 i 2.5)a
Each value is the mean of triplicate determinations t S D . Values in parentheses are cysteine content
in leaves. The t test (compared to the control): a = p < 0.05; b = p < 0.02; c = p < 0.01.
 EFFECTS OF Hg IN B. MONNlERl 109

160
400

100

aoo
so

0 0

a .s

1.4

0.S

Fig. 4. Mercury affecting ascorbic acid content in 5. monnieri roots (A) and leaves (B), and sugar content of
B. monnieri roots (C) and leaves (D). Values are means of triplicate determinations. Standard deviations are repre-
sented by vertical bars. The t test (compared to the control): a = p < 0.05; b = p < 0.02;c = p < 0.01.

There was a significant ( p < 0.01) increase (89% as
compared to the control) in ascorbic acid content of
roots (Fig. 4A) at 5 pg mL-' Hg after 4 d of exposure.
The ascorbic acid content increased up to 2 pg mL-'
Hg as compared to the control after 7 d of exposure.
However, a continuous decrease in ascorbic acid con-
tent was observed at 14 d at all the metal concentra-
tions; the maximum reduction (52% as compared to
the control) was observed at 5 pg mL-' Hg after 14 d
of exposure. A similar trend was observed in the leaves
(Fig. 4B).
Sugar content of roots was increased up to 1 pg
mL-' Hg at 4 d followed by a decrease with an increase
in Hg concentration and duration of exposure (Fig. 4C).
However, sugar content in leaves (Fig. 4D) increased at
all the concentrations of Hg up to 4 d and up to 1
pg mL-' Hg at 7 d of exposure (compared to their
respective controls).
Total thiol (-SH) content (Table IV) of the roots and
leaves increased with an increase in metal concentra-
tion. The maximum increases of 138 and 90% ( p < 0.02)
were observed in the roots and leaves of B . monnieri,
respectively, at 5 pg mL-' Hg (compared to their re-
spective controls) after 7 d of exposure. The GSH mea-
surement results in the roots of the plant showed that
there was a significant increase in GSH levels beyond
1.0 pg mL-' Hg after 7 d of exposure. The maximum
increase of 142% ( p < 0.01) was found at 5 pg mL-'
Hg as compared to the control. However, oxidized
glutathione remained unaffected (data not shown).
DISCUSSION
Bacopa monnieri is known to accumulate toxic metals
under laboratory (Sinha and Chandra, 1990; Gupta et
al., 1995a) and field conditions (Vajpayee et al., 1995;
110 SlNHA ET AL.
TABLE IV. The effect of mercury on total -SH (roots
and leaves) and glutathione (roots) contents in the
plants of 6. monnieri after 7 d of exposurea
Hg Concentration Total -SH GSH
(pg rnL-') (prnoi g-' fw) (nrnoi g-' fw)
0.0 2.02 t 0.15
(1.02 ? 0.05) 240 ;tr 22
0.5 2.11 ? 0.10
(1.11 ? 0.04) 280 ;tr 24
1.o 2.62 2 0.16a
(1.42 ? 0.06)b 333 i: 30a
2.0 3.42 ? 0 . 1 7 ~
(1.64 ? 0 . 0 8 ) ~ 390 5 28b
5.0 4.81 ? 0 . 2 0 ~
(1.94 t 0 . 0 9 ) ~ 580 5 41c
a Each value is the mean of triplicate determinations 2SD. Values
in parentheses are total SH content in leaves. The t test (compared
to the control): a = p < 0.05; b = p < 0.02; c = p < 0.01.
Rai et al., 1995). Emergent macrophytes like B . mon-
nieri, Cyperus rotundus, Eichhornia crassipes, and
Marsilea spp. growing in the vicinity of the chloroalkali
plant at Ganjam, Orissa (India), are reported to accu-
mulate 8.9-25.37 pg g-' Hg in roots and 1.17-13 pg
g-' in shoots when the Hg concentration was 4 pg L-'
in the water (Lenka et al., 1992). In all the plants,
accumulation was more in the roots than shoots. Our
results also show high accumulation of Hg in roots. Sen
and Mondal(1987), while working on Salvinia molesta,
reported 0.3 mg g-' Hg in the whole plant at the 5 pg
mL-' level after 3 d of exposure. In our study, maxi-
mum accumulation of Hg (273 p g g-') was found in
roots at 5 pg mL-'. High Hg content in the roots may be
due to complexation of Hg with the sulphydryl groups,
resulting into less translocation to the shoot system.
A decrease in chlorophyll content in the presence
of Hg has been reported in a number of aquatic plants
(Mhatre and Chaphekar, 1985; Sen and Mondal, 1987).
Mercury (50-250 pM) is known to inhibit the biosyn-
thesis of chlorophyll in Bajra (Pennisetum typhoideum)
by reacting with the sulphydryl requiring enzyme (Pra-
sad and Prasad, 1987). Further, Hg might be involved
in replacing Mg from the chlorophyll moiety leading
to the reduced level of chlorophyll. Sen and Mondal
(1987) reported reduction (26% of the control at 2 d)
in the chlorophyll content of Salvinia natans at and
above 5 pg mL-' Hg, while lower concentrations (0.5
and 1.0 pg mL-') had no effect. Our results conform
with the findings of these authors, showing a decrease
of 23% of the control at 5 pg mL-' Hg after 4 d of
exposure, while there was a slight decrease at lower
concentrations of Hg.
Bacopa monnieri showed a decrease in protein con-
tent both of the rodt and shoot tissues after exposure
to Hg at 5 pg mL-I. Salvinia natans also showed a
decrease (10% of the control at 2 d) in protein content
at 5 pg mL-' Hg, but lower concentrations of Hg had
no effect (Sen and Mondal, 1987). These authors re-
ported that decrease in protein content may be due to
the formation of a protein complex with Hg2+changing
the conformation and solubility of the proteins.
Polyunsaturated fatty acids (PUFA) are particularly
susceptible to lipid peroxidation (Kappus, 1985). Lipid
peroxidation was assessed in terms of malondialde-
hyde, a product of lipid peroxidation that reacts with
TBA. Depletion of PUFA relative to saturated fatty
acids has been reported under stress conditions in cu-
cumber (Kramer et al., 1991). Jones et al. (1987) also
reported that the organomercurial p-chloromercuriben-
zonate had a concentration-dependent decrease on the
fatty acid composition of Asterionella glacialis. The
results of our study conform with the findings of these
authors showing a concentration-dependent decrease
in PUFA content with an increase in Hg concentration
(data not shown). Thus, the decrease in MDA content
both in root and leaf tissues of B. monnieri may be due
to the decrease in PUFA content relative to saturated
fatty acids.
The result of our study contradicts the previous re-
ports on lipid peroxidation (Cakmak and Horst, 1991;
De Vos et al., 1989, 1991; Somashekariah et al., 1992).
These authors reported that heavy metals are related
to an increased production of an enzyme system pro-
ducing superoxide anion radical (O;-) causing alter-
ation of plasma membrane permeability.
Cysteine constitutes one of the three amino acids of
GSH and its increased production may lead to en-
hanced GSH synthesis (Reddy and Prasad, 1992). The
results of the present study conform with the findings
of these authors showing an increase in cysteine and
GSH content of Hg-treated roots of B. monnieri. Howe
and Merchanth (1992) reported significant accumula-
tion of total -SH and GSH contents in Hg-treated cells
of Chlamydomonas reinhardtii. The principal thiol-
containing compound was GSH in Hg2+treated cells.
The fast mercury-induced accumulation of GSH and
the high stability of Hg (GSH), mercaptide complexes
suggest that GSH functions as an effective scavenger
of Hg2+ ions. An increased accumulation of GSH in
Hg-treated roots of B . monnieri may be the plausible
system of Hg tolerance in the plants of B. monnieri. Our
results show a higher concentration of the sulphydryl
group in Hg-treated roots of B. monnieri than in the leaf
tissues. Most of the mercury may remain in innocuous
form in the roots of B. monnieri, where it may bind to
sulphydryl groups.
Nussbaum et al. (1988) reported that cysteine syn-
thesis increased in the roots of Phaseolus vulgaris at

lower levels of Cd. These authors also reported a de-
crease in cysteine content at higher Cd concentrations
due to depletion of metal in the solution. This agrees
with our findings where initially (7 d) cysteine content
increased followed by a decrease at 14 d. Cysteine
content in the leaves of B . monnieri showed a decreas-
ing trend throughout the experiment. This could be due
to the lower availability of metal in the leaves of B .
monnieri. De Filippis (1979a) reported that sulphur-
containing amino acids (L-methionine and L-cysteine)
are known to reduce toxicity of Hg and Zn in Chlorella.
The amelioration of toxicity by sulphur compounds
may be a result of the binding of a toxic metal with
sulphydryl compounds (De Filippis, 1979b). Singh and
Pandey (198 1) also reported the formation of strong
complexes by Cd with only cysteine in cyanobacteria
showing a protective role against metal toxicity. Simi-
larly, high cysteine content in the roots of B . monnieri
may ameliorate metal toxicity.
Like glutathione and cysteine, ascorbic acid is also
known to detoxify mercury in Chlorella vulgaris by
donating electrons to the system and protecting the
integrity of -SH groups (Rai, 1979). Ranieri et al. (1993)
reported an increase of the antioxidant levels, such as
GSH (+130%) and ascorbic acid (+60%) in Cucubita
pep0 under polluted air. The increase in antioxidant
levels reduces oxidized sites of biomolecules. These
authors also reported that these detoxicants are not
completely sufficient to protect against visible injury
but do allow a normal plant growth. Recently, Gupta
et al. (1995b) reported an increase in ascorbic acid
content in Hydrilla verticillata plants treated with Cu.
This could be true for B . monnieri, which is quite resis-
tant to 5 pg mL-' Hg and showed a significant increase
in ascorbic acid content during the initial period of
metal exposure.
CONCLUSIONS
Mercury stress increased the levels of cysteine, total
-SH, GSH, and ascorbic acid in roots of B . monnieri
at the initial period of exposure. These antioxidant
systems in plants seem to bind metal in a form that
renders metal harmless, as was observed by others
(Howe and Merchant, 1992; Ranieri et al., 1993). All
these parameters render tolerance to the plant at lower
metal concentrations and initial period of exposure.
However, the antioxidant systems would not be able
to overcome the toxicity caused by higher concentra-
tion of Hg and at increased exposure periods, resulting
into significant effects on growth parameters.
Thanks are due to Dr. P. V. Sane, Director, National
Botanical Research Institute, Lucknow, for providing facili-
EFFECTS OF Hg IN 8. MONNlERl 111
ties and encouragement (NBRI Research Publication No.
444 NS). M. Gupta is thankful to CSIR (New Delhi) for
financial assistance.
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