ANTICANCER COMPOUNDS FROM MEDICINAL PLANTS


BIOLOGICA NYSSANA  4 (1-2)  December 2013: 71-74  Sućur, J.,et al.  Oxidativ e stress in soy bean seedings treated with…  

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Original Article 

 

Oxidative stress in soybean seedlings treated with Thymus 
serpyllum aqueous extract 
 
 

Jovana Šućur, Milan Popović, Dejan Prvulović, Đorđe Malenčić, Biljana Kiprovski
 

  

 

Faculty of Agriculture, University of Novi Sad, Trg Dositeja Obradovića 8, 21000 Novi Sad, 
Serbia 
 

* E-mail: bkiprovski@polj.uns.ac.rs 
 

 

Abstract:  

Šućur, J., Popović, M., Prvulović, D., Malenčić, Đ., Kiprovski, B.: Oxidative stress in soybean seedlings 

treated with Thymus serpyllum aqueous extract . Biologica Nyssana, 4 (1-2), December 2013: 71-74. 

The effect of different concentrations (0.05, 0.1 and 0.2%) of Thymus serpyllum L. aqueous extracts on lipid 

peroxidation process (LP), as well as reduced glutathione content (GSH) in leaves and roots of soybean 

seedlings were examined 24 and 72 h after the treatment. Our results showed that only highest concentration 

of the extract used (0.2%) enhanced process of lipid peroxidation, while concentration of 0.1% stimulated 

GSH accumulation in soybean seedlings. 

Key words : allelopathy, antioxidants, biopesticides, Glycine max L., Thymus serpyllum L. 

 

 

Introduction 
 

The genus Thymus L. (Lamiaceae) consists of 
about 215 species of herbaceous perennial and 
subshrubs. Thymus species are considered as 
medicinal plants due to their pharmacological and 
biological properties (H u s s a i n  et al., 2013). It is 
commonly used as tonic, herbal tea and flavouring 
agent. Thymus oils and extracts are widely used in 
pharmaceutical, cosmetic and perfume industry also 
for flavouring and bio-preservation of several food 
products (V e r m a  et al., 2011). In the 
Mediterranean region, which can be described as the 
centre of this genus, annual aromatic herb, Thymus 
serpyllum L. inhabits cultivated and uncultivated 
lands including wastelands (V e r m a  et al., 2011). 
Research on allelopathy of T. serpyllum on weeds 
shown that water extract of T. serpyllum promoted 
test plants' growth at low concentration but inhibited 
them at high, and inhibition became stronger as 
concentration increased (Z h a n g , 2009). 

The use of allelochemicals as weed control 
agents, becomes widely investigated, however the 
impact of these bioherbicide on cultivated plants is 
less known. Since ROS (Reactive Oxygen Species)-
generation resulting in oxidative damage has been 
proposed as one of the modes of action of 
allelochemicals, the aim of this study was to 
examine the impact of Thymus serpyllum L. aqueous 
extract on soybean antioxidant properties so as to 
assess its possible side effects when applied as 
bioherbicide in soybean organic production. 
 

Material and methods 
 
The wild, aromatic plant, Thymus serpyllum 

L., was collected in northern Serbia (Vojvodina 
province) in May of 2010. Voucher specimens of 
collected plant was confirmed and deposited at the 
Herbarium of The Department of Biology, Faculty 
of Natural Sciences, University of Novi Sad. Seeds 
of soybean (Glycine max L.) cv. Sava were obtained 

11
th
 SFSES •  13-16 June 2013, Vlasina lake 

 
4 (1-2) • December 2013: 71-74 

 



BIOLOGICA NYSSANA  4 (1-2)  December 2013: 71-74   Sućur, J.,et al.  Oxidativ e stress in soy bean seedings treated with… 

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from the Institute of Field and Vegetable Crops, 
Novi Sad, Serbia.  

The aqueous extract of T. serpyllum was 
prepared with air-dried plant material (10 g) in 
boiling distilled water (100 ml). After 24 h, the 
extract was filtered through filter paper and kept at 4 
°C until application.  

The experiment was performed at the 
Laboratory of Biochemistry, Faculty of Agriculture, 
Novi Sad and conducted under controlled conditions 
(temperature 25 °C, 60-70% relative humidity, 16 h 
photoperiod). Soybean seedlings were grown for 14 
days in plastic pots (500 ml) containing sterile sand, 
after which they were transferred on plastic pots 
(700 ml) containing the Hoagland’s solution and 
3.5, 7 and 14 ml of T. serpyllum aqueous extract, 
while 3.5, 7 and 14 ml of the Hoagland’s solution 
were added in pots of control. 

Seedlings were harvested for determining the 
investigated biochemical parameters 24 and 48 h 
after the treatments. 

Biochemical analyses were carried out 
spectrophotometrically using an UV/VIS 
spectrophotometer model 6105 (Jenway, UK). Lipid 
peroxidation (LP) was measured at 532 nm using 
the thiobarbituric acid (TBA) test. The total amount 
of TBARS (TBA-reactive substances) is given as 
nmol malondialdehyde (MDA) equivalents mg

-1 

proteins (M a n d a l  et al., 2008). Reduced 
glutathione (GSH) was determined according to 
S e d l a k  a n d  L i n d s a y  (1968) and expressed as 
µmol GSH mg

-1
 proteins.  

Values of the biochemical parameters were 

expressed as means  standard error of 
determinations made in triplicates and tested by 
ANOVA followed by comparison of the means by 
Duncan’s multiple range test (P<0.05). Data were 
analysed using STATISTICA for Windows version 
11.0. 

 

Results and discussion 
 

The toxicity of many allelochemicals can 
largely be attributed to the formation of 
semiquinone radicals that donate electrons to 
molecular oxygen, forming reactive oxygen species 
(ROS), such as superoxide anions (O2

.-
), and more 

reactive hydroxyl (
.
OH) or hydroperoxyl (HO2

-
) 

radicals (H a m m o n d k o s a k  & J o n e s , 1996). 
This plant-plant allelopathic interaction generates a 
cascade of signaling events-including Ca

2+
 influx 

and proton efflux that actives the NADPH-oxidase 
complex (generating 

.
O2

-
) and pH-sensible cell wall 

peroxidases (producing H2O2), which produces an 
oxidative burst (D o s  S a n t o s  et al., 2008). 

Enhancement of ROS production during the 
oxidative burst is one of the earliest reactions 
elicited in response to various abiotic and biotic 
stimuli. Some allelochemicals rapidly depolarize the 
cell membrane, increasing membrane permeability, 
inducing lipid peroxidation and causing a 
generalized cellular disruption that ultimately leads 
to cell death (D e v i  &  P r a s a d , 1996; Z e n g  et 
al., 2001; Y u  et al., 2003). The peroxidation of 
unsaturated lipids of biological membranes is the 
most prominent symptom of oxidative stress in 
animals and plants (M a n d a l  et al., 2008). 
Malondialdehyde (MDA) content, an end-product of 
lipid peroxidation process, is used as oxidant bio-
marker. It is suggested that different concentrations 
of allelochemicals induce production of MDA, 
however due to their antagonistic and synergistic 
effect MDA content is not always in reciprocity 
with these chemicals (D i n g  et al., 2007; 
H a d d a d c h i  &  G e r i v a n i , 2009). 

Our results showed that 24 h after the 
treatment with T. serpyllum aqueous extracts no 
significant difference in the LP intensity in soybean 
leaves and roots between plants form control and 
treatments were recorded. The significant increase 
was recorded 72 h after the treatment only in leaves 
of soybean plants treated with the highest 
concentration of T. serpyllum aqueous extract (Fig. 1). 

Allelochemicals might directly inhibit 
oxidizing enzymes in some way, leaving the plant 
vulnerable to oxidative damage, similar to the 
reports of Q i a n  et al. (2009). It has been 
documented that GSH can remove ROS and also 
protect proteins from the oxidation of protein thiol 
groups into denaturation (N o c t o r , 2002). T. 
serpyllum aqueous extract with concentration of 
0.1% induced production of GSH in leaves of 
soybean plants 24 h, and in leaves and roots 72 h 
after treatment (Fig. 2). Also, 24 h after the 
treatment with T. serpyllum aqueous extract with 
concentration of 0.2%, roots of soybean had highest 
level of GSH content (Fig. 2).  

 

Conclusion 
 

In conclusion, our results showed that low 
concentration of Thymus serpyllum L. aqueous 
extract did not induce lipid peroxidation in soybean 
seedlings, while the highest concentration used 
(0.2%) enhanced lipid peroxidation process 72 h 
after the treatment. Furthermore, concentration of 
0.1% of T. serpyllum aqueous extract stimulated 
GSH accumulation in soybean seedlings which 
propose its possible stimulative effect on antioxidant 
system. 



BIOLOGICA NYSSANA  4 (1-2)  December 2013: 1-7  Balian, D., Mujagić-Pašić, A..  Morphological v ariability  of  the f ruit… 

 

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Fig. 1. Lipid peroxidation level in leaves and roots of soybean seedlings 24 and 72 h after treatment with 
different concentrations (%) of T. serpyllum aqueous extracts (v/v) and in control (C). Values marked with 
different letter differ significantly at P<0.05 (Duncan’s test).  

 

 
Fig. 2. Reduced glutathione content in leaves and roots of soybean seedlings 12 and 48 h after treatment with 
different concentrations (%) of T. serpyllum aqueous extracts (v/v) and in control (C). Values marked with 
different letter differ significantly at P<0.05 (Duncan’s test). 
 
 

Acknowledgements . This study was carried out within a 

project of the Ministry of Education, Science and 

Technological Development, Republic of Serbia, Grant 

N
o
 TR-31022. 

 

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