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Toxicity of arsenic on germination and seedling

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ARTICLE JANUARY 2007

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Bangladesh Agricultural University

Bangladesh Agricultural University

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J. Bangladesh Soc. Agric. Sci. Technol., 4(1&2): 153 - 156, 2007

ISSN 1811-6221

Toxicity of arsenic on germination and seedling growth of rice

M.A. Hossain, M.N. Uddin and A.K.M.G. Sarwar
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh
Abstract
The study examined the effect of arsenic (As) on the germination and seedling growth of rice. Eight different
concentrations of As viz., 0.0 (control), 0.5, 1.0, 2.0, 4.0, 6.0, 8.0 and 10.0 mg As L-1 were tested on 5 rice varieties
viz., BR11, BRRI dhan30, BRRI dhan3l, BRRI dhan34 and BRRI dhan37. Arsenic contaminated water adversely
affected the seed germination and various growth parameters e.g. seedling height, number of leaves seedling-1,
length of primary leaf, length of coleoptile, number of roots seedling-1, average root length, and total dry matter of
10-day old seedlings. Germination and seedling growth of the rice varieties suffered at different magnitude in
response to the added As supplied as solution during germination. Water containing 10 mg As L-1 had full
inhibitory effect on the germination of seeds in all studied rice varieties. Whereas 8 mg As L-1 caused germination
reduction by 5.5% (93.93% reduction over control) compared to 90.6% in control. The total dry matter yield of 10day old seedlings varied between 1.02 to 11.66 mg seedling-1 in the 0 to 8.0 mg As L-1. Therefore, appropriate
precautionary measures should be taken during raising of rice seedling in the arsenic affected areas of Bangladesh.
Tolerance to As at the germination and seedling stage might be used as a selection criterion for As tolerant
varieties.
Keywords: Arsenic, rice, germination, seedling growth
Introduction
Arsenic is one of the most serious global environmental toxicants. Groundwater contamination by arsenic (As) has
been reported from many parts of Bangladesh and the country is currently facing the challenge of high As
concentrations in shallow aquifers (Nickson et al., 1998). In some areas of Bangladesh, the ground water As
contaminations reach 2 mg As L-1 which is higher than the recommended maximum concentration of As (0.1 mg
As L-1) in irrigation water (Tondel et al., 1999). Background levels of As in soils can be between 4 - 8 mg As
kg-1 but in the areas irrigated with As contaminated water, the soil level can reach up to 83 mg As kg-1 (Ullah,
1998).
Rice (Oryza sativa L.) is the staple food and the main agricultural crop of Bangladesh. In Bangladesh, 70% of
the total cropped area and 82% of the irrigated area are used for rice cultivation (BBS 2002). Groundwater is the
primary source of irrigation water especially in the dry seasons. The production of rice is dependent
involuntarily on As contaminated irrigation water, which explain the importance of As issue in rice. All
activities related with rice cultivation such as germination, raising of seedlings, transplanting and growing rice
in main field are mostly done by ground water irrigation. Plants can develop the toxicity symptoms while they
are exposed to excess As either in soil or in solution culture such as, inhibition of seed germination, reduction in
root and shoot growths, decrease in plant height and tiller number, and reduction of grain yield (Abedin et al.,
2002; Jahiruddin et al., 2004; Liu et al., 2005; Uddin et al., 2005). However, only a little information about the
effect of As on seed germination and seedling growth of rice is available (Tsutsumi, 1983). Tsutsumi (1983)
reported that As depressed the radicle growth in rice and arsenite is more toxic than arsenate. Therefore, a
laboratory experiment was carried out to evaluate the effects of different concentration of As in irrigation water
on the germination and seedling growth of rice.
Materials and Methods
The laboratory experiment was conducted at the Department of Crop Botany, Bangladesh Agricultural
University, Mymensingh. Eight different concentrations of arsenic viz., 0 (control), 0.5, 1.0, 2.0, 4.0, 6.0, 8.0
and 10.0 mg As L-1 were evaluated on seed germination and seedling growth of 5 rice varieties viz., BR11,
BRRI dhan30, BRRI dhan3l, BRRI dhan34 and BRRI dhan37. Arsenic was supplied as a solution of
Na2HAsO4.7H2O. The highest As treatment (10.0 mg As L-1) of this study was much higher than the reported
maximum level of As (2.0 mg As L-1) in irrigation water of Bangladesh. All the seeds of the respective varieties
were surface sterilized by soaking them in 1% NaOCl for one minute followed by rinsing with deionized water.
Germination of seeds was tested on moist filter paper (Whatman No.1). Two filter papers were placed on each
of 15.0 cm diameter Petri dishes. Each Petri dish was then moistened with 10 mL of respective As solution.
Fifty sterilized seeds of each variety were laid on filter paper bed in each Petri dish. Each dish was then covered

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Arsenic toxicity in rice

by a lid, and incubated at room temperature (28 - 32C). Each treatment was replicated three times following
randomized complete design (CRD). The seeds were allowed to germinate and grow for 10 days. During this
period the Petri dishes were moistened by respective solutions of As when needed. Lids of all the dishes were
removed during plumule growth. Data were recorded from 10-day old seedlings on germination, seedling
height, number of leaves seedling-1, length of primary leaf, length of coleoptile, number of roots seedling-1,
average root length, and total dry matter seedling-1. Experimental data were analyzed statistically by using
MStat-C computer package following ANOVA technique and the means were separated by Duncan's New
Multiple Range Test.
Results
Seed germination
Arsenic contaminated water had a marked effect on germination of the rice seeds (Table 1 and Fig. 1A). The
results showed that the % seed germination over control decreased significantly with the increasing As
concentration. The germination varied between 5.5 to 90.6% in 0 to 8.0 mg As L-1 treatment. Germination of
seeds were totally failed in 10 mg As L-1 solution of all the varieties studied and seeds of BRRI dhan3l and
BRRI dhan37 did not germinate even at 8 mg As L-1 solution.. The highest (90.6 %) germination was recorded
in control treatment and lowest (5.5%) in 8 mg As L-I solution. Germination decreased by 15.45, 25.39, 35.54,
56.51, 75.5 and 93.93 % over control in 0.5, 1.0, 2.0, 4.0, 6.0 and 8.0 mg As L-1 treatment, respectively. BRRI
dhan34 showed the higher and BRRI dhan30 showed the lower % germination in most As treatment (Fig. 1A).
Seedling growth parameters
The means of different growth parameters viz., seedling height, number of leaf seedlling-1, length of primary
leaf, length of coleoptile, number of root seedling-1, root length and total dry matter seedling-1 were significantly
influenced by different As treatments (Table 1). The maximum seedling height (8.88 cm) was recorded in
control treatment and the minimum (1.08 cm) in 8 mg As L-1 treatment. There were significant varietals
differences in seedling height (Fig. 1B). BR11 produced taller seedlings at lower concentrations of As solution
(0 to 4 mg As L-1), while BRRI dhan30 produced the taller seedlings at higher concentrations (6 to 8 mg As L-1)
and BRRI dhan34 produced shorter seedlings at all concentrations of As. The number of leaves seedling-1 varied
from 1.0 to 3.0 in the 0 to 8.0 mg As L-1 treatment, however, no significant differences were experienced up to
1.0 mg As L-1 treatment. The treatment of As between 2.0 to 6.0 mg As L-1 produced statistically similar number
of leaves seedling-1. The length of primary leaf varied between 0.28 to 1.33 cm in the As treatment. The length
of primary leaf progressively shortened with increasing concentration of As from 2.0 to 8.0 mg As L-1. The
length of coleoptile was also affected by As almost in the similar way as primary leaf. The highest number of
root seedling-1 (4.6) was recorded in control (0 mg As L-1) which was statistically identical with that of 0.5 mg
As L-1. The lowest number of root seedling-1 (0.6) was traced in 8.0 mg As L-1. Up to 4.0 mg As L-1, the variety
BRRI dhan31 produced the highest number of root seedling-1 (Fig. 1C). On the contrary, BRRI dhan30
produced the lowest number of roots in all As treatments. The treatment receiving no As produced the highest
root length (2.44 cm), while the lowest root length (0.21 cm) was observed in 8.0 mg As L-1. The total dry
matter seedling-1 was found to be strongly affected by the concentration of As in water. Total dry matter
decreased significantly with increasing As concentration (Table 1). The range of total dry matter seedling-1 due
to As contamination varied from 1.02 to 11.66 mg seedling-1. Arsenic contaminated water accounted for the
reduction of dry matter seedling-1 by 19.04, 36.88, 44.94, 66.04, 76.67 and 91.25 % in 0.5, 1.0, 2.0, 4.0, 6.0 and
8.0 mg As L-1 treatments, respectively over control. Total dry matter production also varied among the varieties
(Fig. 1D). BR11 and BRRI dhan31 produced the highest and the lowest total dry matter contents seedling-1,
respectively. Neither BRRI dhan31 nor BRRI dhan37 produced any dry mass in 10 mg As L-1 as germination of
these two varieties failed in this treatment.
Discussion
The germination of rice seed decreased significantly with the higher concentrations of As in water and the
adverse effect was different among the varieties. The lower percentage of seed germination corroborates with
the result of Liu et al. (2005) who reported reduced germination in wheat seed with increasing concentration of
As. Sodium arsenate (Na2HAsO4.7H2O) was found as a potent inhibitor of mungbean seed germination and
seedling growth. Germination of mungbean was totally inhibited at or above 50 M Na2HAsO4.7H2O, while
inhibition of seedling elongation started at a lower concentration of 5 M As(V) and was drastically reduced at
20 M As(V) (Swarnakar & Mukherji, 2005). In our study we found that water with 10 mg As L-1 had full lethal
effect on germination of seeds in all rice varieties and 8 mg As L-1 also showed lethal action on some rice

Hossain et al.

155

Table1. Effect of arsenic (As) contaminated irrigation water on germination and seedling growth of rice
Arsenic
Germinaadded
tion
(mg As L-1)
( %)
0.0
90.6a
0.5
76.6b
1.0
67.6c
2.0
58.4d
4.0
39.4e
6.0
22.2f
8.0
5.5g
7.70
LSD(0.05)

Reduction
in germiation (%)
0
15.45
25.39
35.54
56.51
75.50
93.93
--

Seedling
height
(cm)
8.88a
7.86b
6.86c
5.06d
3.80e
2.92f
1.08g
0.70

Leaf
number
seedling-1
3.0a
3.0a
2.8a
2.4ab
2.0b
1.8b
1.0c
0.77

Length of
primary
leaf (cm)
1.33a
1.28a
1.21ab
1.06b
0.84c
0.57d
0.28e
0.19

Length of
Root
Root
Total dry
Reduction
coleoptile number
length
matter
in total dry
seedling1(mg) matter (%)
(cm)
seedling-1 (cm)
0.48a
4.6a
2.44a
11.66a
0
0.44ab
4.1ab
1.47b
9.44b
19.04
0.40b
3.8b
1.03c
7.36c
36.88
0.36b
3.0c
0. 81cd
6.42d
44.94
0.28c
2.0d
0.56de
3.96e
66.04
0.22c
1.1 e
0.26ef
2.72f
76.67
0.11 d
0.6e
0.21f
1.02g
91.25
0.08
0.57
0.31
0.93
--

In a column, the means having common letter(s) do not differ significantly at 5% level by DMRT.

Figure 1. Effect of different concentrations of arsenic on percent seed germination (A), seedling height (B), root
numbers seedling-1 (C) and total dry matter seedling-1 (D).

varieties (Fig. 1A). Arsenic acts as a metabolic inhibitor. Once inside the cytoplasm it competes with phosphate,
for example replacing phosphate in ATP which leads to disruption of energy flow in cells (Meharg, 1994).
Arsenic is also toxic to plants as it reacts with enzymes and tissue proteins. Seed germination is energy (ATP)
depending process and during germination many enzymes are critically involved to transform complex seed
storage into simpler form and translocate them in growing region of seedlings (Meharg, 1994). So, higher As
concentration might be associated with disruption of enzymatic activity and energy synthesis of seeds which
eventually accounted for the death of seed tissues and inhibition of seed germination.
Rice seeds subjected to higher concentrations of As reduced the seedling height. Reduced shoot growth in rice
due to application of arsenate had been reported by Abedin et al. (2002) and Marin et al. (1992). Abedin et al.
(2002) reported that plant height ranged between 91.1 and 84.1 cm with lower ranges of arsenate doses (0-1.0
mg As L-1), while with higher arsenate doses (2.0 - 8.0 mg of As L-1) plant height decreased to 79.2 - 63.8 cm.
Leaf number plant-1 was less affected by As in water. Similar to germination of seeds, the adverse effect of As
on seedling height was different among the varieties (Fig. 1B).

156

Hossain et al.

Rice root development was also affected by the As containing irrigation water. The treatment receiving 0.5
mg As L-1 produced significantly lower root length compared to that of control treatment whereas no
significant differences in root number were found in the same concentration. The reduction in root biomass
was more pronounced in higher arsenate content (Abedin et al., 2002). Tsutsumi (1983) observed that the
radicle growth was almost entirely suppressed by arsenite at 10-4.0 M. It might be due to induction of
phytotoxicity by As which resulted in restricted root growth. The adverse effect of As on number of roots
plant-1 was also different among the varieties (Fig. 1C). Marin et al. (1992) also reported a differential
response in rice root dry matter production between two rice varieties, Lemont and Mercury, when treated
with different concentrations of arsenite and arsenate in hydrophonic culture. Considerable reduction in total
dry matter in the highest arsenate treatment recorded in this experiment may be primarily as a result of stunted
seedling height, reduced root number and root length, and reduced number of leaf seedling-1 (Table 1). Exposure
to arsenic, causing a reduction in shoot biomass and/ or growth, was also reported by Tsutsumi (1983) for rice.
Therefore, appropriate precautionary measures like use of surface water or As free irrigation water, tolerant
varieties etc., should be taken during raising of rice seedling in the arsenic affected areas of Bangladesh.
Tolerance to As at germination and seedling stages might be used as a selection criterion for As tolerant
varieties.
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Ministry of Planning. Govt. of the People's Republic of Bangladesh. pp. 192-202.
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