Ajps - 2014112818074195 (2014)
Ajps - 2014112818074195 (2014)
Ajps - 2014112818074195 (2014)
Abstract
Arsenic is a great hazard to the environment since it is a major source of soil and water contami-
nation leading to phytotoxicity. Arsenic caused reduction in seed germination and growth of
plants including mung bean (Vigna radiata) causing loss of crop productivity. This investigation
evaluates effect of arsenic toxicity in mung bean with an attempt to ameliorate it by the applica-
tion of curcumin. Arsenic induced reduction in fresh, dry mass, length of root and shoot and pig-
ment content. The result of the present study suggests that curcumin at lower concentration ame-
liorates arsenic induced toxicity in Vigna radiata.
Keywords
Arsenic Stress, Curcumin, Vigna radiata
1. Introduction
Arsenic (As) contamination in environment, from both anthropogenic and natural sources occurs is a global
problem. As is a metalloid having properties of both metals and non-metals, and can undergo different ranges of
chemical interactions in plants. Historically, the extensive use of arsenic-based pesticides has led to considerable
contamination of domestic and agricultural land, through their use as lawn herbicides, and insecticides for rice
plants, cotton, etc. [1]. Arsenic is a non-essential element and, in general, inorganic ones are phytotoxic. Arsenic
is toxic to a wide range of organisms, including plants [2]. Studies on arsenic toxicity have shown that plant
species not resistant to arsenic suffer considerable stress upon exposure, with symptoms ranging from inhibition
of root growth to death [3]-[6].
Exposure of plants to arsenic causes reduction in photosynthetic rate [7], for example: in maize (Zea mays)
How to cite this paper: Upadhyaya, H., Shome, S., Roy, D. and Bhattacharya, M.K. (2014) Arsenic Induced Changes in Growth
and Physiological Responses in Vigna radiata Seedling: Effect of Curcumin Interaction. American Journal of Plant Sciences, 5,
3609-3618. http://dx.doi.org/10.4236/ajps.2014.524377
H. Upadhyaya et al.
plant, the rate of CO2 fixation in young plants treated with arsenic decreased by 20% and functional activity of
PSII was reduced significantly [8]. The decreased photosynthetic rate under stress condition could be due to
both stomatal and mesophyll limitations. The mesophyll factors could be of a different nature, such as distur-
bances in the chlorophyll, light and biochemical reactions from the Calvin Cycle [7]. In rice seedlings arsenic
toxicity causes changes in carbohydrate metabolism leading to the accumulation of soluble sugars by altering
enzyme activity. Sucrose synthase possibly plays a positive role in synthesis of sucrose under As-toxicity [9].
Arsenic is known to induce oxidative stress in plants [10], resulting in a range of responses in plants, includ-
ing readjustment of transport and metabolic processes and growth inhibition in plants. As a result, the electron
transport processes are impeded developing toxic systems. Several toxic reactive oxygen species (ROS) are
generated in the cell wall region as well as inside the cell during the process, which affects membrane permea-
bility, enzyme activity, metabolic pool, plant biomass, leaf chlorosis and necrosis [11], probably due to the con-
version of arsenate to arsenite, a process that usually occurs in plants by synthesizing both enzymatic and
non-enzymatic antioxidants. Arsenate acts as a phosphate analogue and is transported across the plasma mem-
brane via phosphate transport systems. Inside the cytoplasm it reacts with phosphate, replacing ATP to form
ADP-As, precisely because of disruption of energy flow in cells [12]. On the other hand, Bertolero et al. [13]
reported that arsenate reduced to arsenite in plant tissue, does not normally have enough cytoplasmic concentra-
tions to exert toxicity. Arsenite reacts withSH group of enzymes and proteins due to suppression of cellular
function and death [14]. The conversion of arsenate to arsenite, in vitro observation, is brought about by gluta-
thione [15], with the concentration of high cellular plant tissues [16].
Curcumin (1, 7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione) is a polyphenol derived from the
plant Curcuma longa. Extensive research over the last decades has indicated that curcumin exhibits anti-in-
flammatory, anti-oxidant, anti-viral and anti-infectious activities [17]-[19]. Curcumin such as other polyphenols
is as strong anti-oxidant [20]. It significantly decreases lipid peroxidation, regulates antioxidant enzymes and
scavenges hyperglycemia-induced reactive oxygen species (ROS), [21]. Oxidative stress and inflammation are
closely associated with tumor growth, [22]. All such effect of curcumin has been reported from animal system
and no work has been done on the role of curcumin in plant growth. The present investigation is an attempt to
understand the role of curcumin in heavy metal stress amelioration in plant in general and arsenic stress ameli-
oration in mung bean in particular.
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3. Statistical Analysis
Each experiment was repeated thrice with each treatment sample containing ten individual plants and data pre-
sented are mean standard error (SE). The results were subjected to one way ANOVA and LSD test was used
for comparison between pairs of treatments. The data analysis was carried out using MS excel 2003 and statis-
tical package, SPSS 10.
4.2. Effect of Arsenic and Its Interaction with Curcumin on Seed Germination
The inhibitory effect of arsenic on the germination of mung seeds was evident from relatively lower germination
rate (28%) in arsenic treated seeds as compared to control (84%) (Table 1). The germination percentage of cur-
cumin treated seed was found to be 75%, 15%, 10% at 25, 50, & 100 M treatment respectively. Seed germina-
tion percentage reduced under arsenic stress but curcumin at 25 M concentration showed protective effect on
seed germination when treated alone or in combination with arsenic. The length of radicle growth also varied
among the treatments as shown in Figure 1.
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Table 1. Effect of arsenic, curcumin and their interaction on Chl a, Chl b, Chl a:b, total Chlorophyll, total Carotenoid con-
tents of growing seedings & seed germination of mung bean [Vigna radiata (L.) Wilczek]. Data presented are mean SE (n
= 3). Mean value superscript with a & b indicates significant mean difference from control & arsenic treatment at p =
0.05 in multiple comparison test by LSD respectively.
mass was observed (Figure 4). However, the total dry mass of mung bean seedlings decreased by 24% in As
treated seedlings as compared control. Azad et al. [26] also reported a decline in shoot length in Oryza sativa at
higher concentration of arsenic. Arsenate in plants is absorbed and translocated through the phosphate channels
[27] [28]. In our present study, the root growth was significantly inhibited than shoot growth. This may be attri-
buted to the poor translocation of arsenate to the shoot due to the antagonistic effect of phosphate ions present in
the nutrient solution on are senate for root to shoot translocation through the phosphate channels or the increased
level of chelation of arsenate in the roots.
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Figure 2. Effect of arsenic (50 M), curcumin (25, 50 & 100 M) and their interaction [50 M As + curcumin (25, 50 & 100
M)] on root (A) and shoot (B) length of growing seedlings of mung bean [Vigna radiata (L.) Wilczek]. Data presented are
mean SE (n = 3). Mean value superscript with a & b indicates significant mean difference from control & arsenic
treatment at p = 0.05 in multiple comparison test by LSD respectively.
Figure 3. Effect of arsenic (50 M), curcumin (25, 50 & 100 M) and their interaction [50 M As + curcumin (25, 50 & 100
M)] on root (A) and shoot (B) dry mass of growing seedlings of mung bean [Vigna radiata (L.) Wilczek]. Data presented
are mean SE (n = 3). Mean value superscript with a & b indicates significant mean difference from control & arsenic
treatment at p = 0.05 in multiple comparison test by LSD respectively.
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Figure 4. Effect of arsenic (50 M), curcumin (25, 50 & 100 M) and
their interaction [50 M As + curcumin (25, 50 & 100 M)] on root (A)
and shoot (B) dry mass of growing seedlings of mung bean [Vigna ra-
diata (L.) Wilczek]. Data presented are mean SE (n = 3). Mean value
superscript with a & b indicates significant mean difference from
control & arsenic treatment at p = 0.05 in multiple comparison test by
LSD respectively.
growing mung seedlings. Chl a: Chl b ratio increased by 5.75% in arsenic treated mung seedlings. Srivastava et
al. [29] also reported a similar decline of chl a, chl b content in Hydrilla verticillata at higher doses of arsenic
treatment. This decrease in chlorophyll synthesis could be due to the generation of reactive oxygen species like
superoxide and hydroxyl radicals, and hydrogen peroxide that have the potential to damage proteins, nucleic ac-
ids and amino acids involved in the biosynthetic pathway of chlorophyll synthesis [30].
4.5. Effect of Curcumin and Its Interaction with As on Growth of Mung Bean Seeds
Curcumin causes significant changes in growth of mung seedlings. Although changes in root length of mung
bean is lower relative to control but compare to arsenic treated seedlings curcumin treated samples showed in-
crease in root length. It was evident that As induced decrease in root growth is checked when curcumin and ar-
senic is treated simultaneously. There is no significant changes in shoot length in curcumin treated plants rela-
tive to control. But in comparison to arsenic treated sample, increase in shoot length due to curcumin treatment
was observed. The interaction of curcumin with As however could not increase any shoot length. The root dry
mass was decrease with curcumin treatment, but relative to control root dry mass shows no significant changes
when 50 M As and 25 M curcumin are interacted. It also clearly indicates that the toxic effect of arsenic in
root of growing mung seedlings is minimized by increasing root biomass over 350% when the plant is supplied
with 50 M As along with 25 M curcumin. Although As have no significant effect on shoot biomass, curcumin
increases shoot biomass by 11.64% at 50 M concentration relative to control. The interactive effect of arsenic
with curcumin also showed increase in shoot biomass over 23.52%, 17.64% and 11.76% in arsenic treated plant
at 25, 50, 100 M curcumin interaction respectively. Shoot dry mass increased because of curcumin interaction
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by 23.52%, 17.64%, 11.76% at 25, 50, 100 M curcumin concentration respectively with respect to arsenic
treated plant. However the total dry mass increases by 20% in plants treated with arsenic and curcumin (50 M
As + 25 M curcumin) relative to control. It also clearly indicates that total dry mass has increased by 57.89%
due to curcumin interaction (50 M As + 25 M curcumin) in comparison with As treated plants. This findings
highlight the fact that curcumin prevents cell biomembranes damage. Lipid peroxidation is a chain reaction
triggered by the free radicals produced in plants under stress conditions, leading to the damage of the cell mem-
branes. Curcumin checks lipid peroxidation by scavenging the reactive free radicals that mediate the process of
lipid peroxidation [31] [32]. Curcumin exhibits the free radical scavenging activity by donating H-atom from the
phenolic group in acidic or neutral pH [33] [34].
5. Conclusion
From the present study, it may be suggested that the arsenic induced growth reduction in mung bean seedlings
with simultaneous reduction in photosynthetic pigments but the curcumin, alleviates the adverse effect of arsenic
on mung bean seedlings. It can also be suggested the arsenic induced germination inhibition of mung bean may
be counteracted by curcumin at lower concentration. Further detail study on the curcumin effect on physiologi-
cal events during mung seed germination and physiochemical and antioxidative responses of growing seedlings
of mung bean (Vigna radiata) can reveal the exact mechanism underlying the amelioration of arsenic toxicity by
curcumin in growing seedlings of mung bean (Vigna radiata).
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Acknowledgements
Authors gratefully acknowledge the Department of Biotechnology, Govt of India, for providing research support
under the Biotech Hub (San. No.: BT/04/NE/2009 dated 21/09/2010) and Star College programs (San. No.: BT/
HRD/11/04/2011 dated 21/03/2011) to Karimganj College, Karimganj, Assam, India.
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