Curr Microbiol (2014) 69:192–201
DOI 10.1007/s00284-014-0573-0
Inhibitory Effect of Selenium Against Penicillium expansum
and Its Possible Mechanisms of Action
Zhi-lin Wu • Xue-bin Yin • Zhi-qing Lin •
Gary S. Bañuelos • Lin-xi Yuan • Ying Liu •
Miao Li
Received: 14 December 2013 / Accepted: 8 February 2014 / Published online: 30 March 2014
Ó Springer Science+Business Media New York 2014
Abstract Some organic and inorganic salts could inhibit
the growth of many pathogens. Selenium (Se), as an essential
micronutrient, was effective in improving the plant resis-
tance and antioxidant capacity at a low concentration. Pen-
icillium expansum is one of the most important postharvest
fungal pathogens, which can cause blue mold rot in various
fruits and vegetables. In this study, the inhibitory effect of Se
against P. expansum was evaluated. The result showed that
Se strongly inhibited spore germination, germ tube elonga-
tion, and mycelial spread of P. expansum in the culture
medium. The inhibitory effect was positively related to the
concentration of Se used. Fluorescence microscopy obser-
vation of P. expansum conidia stained with propidium iodide
(PI) indicated that the membrane integrity decreased to 37 %
after the conidia were treated with Se (20 mg/l) for 9 h. With
Z. Wu
M. Li (&)
Key Laboratory of Agri-Food Safety of Anhui Province,
School of Plant Protection, Anhui Agricultural University,
Hefei 230036, China
e-mail: miaoli@ustc.edu.cn
Z. Wu
e-mail: 13732639644@163.com
Z. Wu
X. Yin
L. Yuan
Y. Liu (&)
Advanced Lab for Selenium and Human Health, Suzhou Institute
for Advanced Study, University of Science and Technology of
China, Suzhou 215123, China
e-mail: louie@ustc.edu.cn
Z. Lin
Environmental Sciences Program & Department of Biological
Sciences, College of Arts and Sciences, Southern Illinois
University Edwardsville, Edwardsville, IL 62026, USA
G. S. Bañuelos
Agricultural Research Service, United States Department of
Agriculture, Parlier, CA 93648, USA
123
the use of an oxidant-sensitive probe 2,7-dichlorofluorescin
(DCHF-DA), we found that Se at 15 mg/l could induce the
generation of intracellular reactive oxygen species (ROS).
Furthermore, methane dicarboxylic aldehyde (MDA) con-
tent, hydrogen peroxide (H2O2), and superoxide anion (O2-)
production rate in P. expansum spores exposed to Se
increased markedly. Compared with the control, the activi-
ties of superoxide dismutase (SOD) and the content of glu-
tathione (GSH) were reduced, confirming that damage of Se
to cellular oxygen-eliminating system is the main reason.
These results suggest that Se might serve as a potential
alternative to synthetic fungicides for the control of the
postharvest disease of fruit and vegetables caused by P.
expansum.
Introduction
Penicillium expansum is a widely spread fungal pathogen
that causes blue mold rot in a variety of fruits, including
apples, pears, peaches, and cherries [31]. This pathogen not
only induces blue mold rot that causes the loss and shortens
the shelf-life of harvested fruit, but also produces patulin in
infected apple fruit, a secondary metabolite that is toxic to
humans and animal [32]. Patulin is produced by various
filamentous fungi with P. expansum being generally
regarded as one of the main producers. Because of the
potential carcinogenic effects of Patulin, Europe and the
United States have established a maximum limit for patulin
contamination in apple-based products [26]. Management
of decay caused by P. expansum has become important for
ensuring the quality and safety of various fruits [6]. Cur-
rently the disease caused by P. expansum is mainly con-
trolled by the intensive use of synthetic fungicides.
However, the use of fungicides is becoming increasingly
Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum
restricted because of concerns for environment and health,
as well as the development of fungicide resistance by
pathogens [29].
Selenium (Se) had been considered as a toxic element
until it was found also to be essential in 1957 [40]. Se
deficiency has been found to directly damage human
health, and more than 40 types of diseases have been found
to be associated with Se deficiency, such as Keshan dis-
ease, Kashin–Beck disease, cancer, cardiovascular disease,
liver disease, and cataracts [12, 45]. Se is of great interest
in biochemistry [17], chemistry [7, 21–24], medicine, and
medicine-related fields [11, 13, 25]. It is also used for
bioisosteric replacement of oxygen and sulfur in bioactive
molecules to obtain more bioactivity or safety [30]. Singh
(2012) had synthesized a new series of compounds called
Novel 3-(substituted/unsubstituted phenylselenonyl)-1-
ribosyl/deoxyribosyl-1H-1, 2, 4-triazole containing the 1,
2, 4-triazoles, Se atom, and sugar moiety, which was safer
more effective than traditional fungicides [43]. Selenium
sulfide has been proved to possess an efficient ability to
inhibit Pityrosporum, a yeast thought to play a pathogenic
role in seborrheic dermatitis and dandruff [5]. A mixture
fungicide of Se and dithane was used by A. A. RAK to
assess the efficiency of fungicide action against certain
fungi such as Aspergillus funiculosus and Alternaria tenuis.
Among of them, A. funiculosus showed no growth in the
presence of mixture of 2.5 ppm Se and 20 ppm dithane, or
at 10 ppm of each [38]. Se as an essential nutrient at such
very low concentrations, as well as the application of very
low concentrations of the fungicide, would certainly reduce
the hazardous effect of such pollutant in the environment.
However, whether Se alone has fungicidal activity
toward a plant pathogenic ascomycete and that the mech-
anism of action still remains essentially unexplored. In this
study, bacteriostasis experiment in vitro was designed to
explore the inhibition effect on P. expansum. The effects of
Se on the control of blue mold diseases in apple plants were
investigated, and the underlying mechanisms by which Se
caused fungal death, and affected intracellular reactive
oxygen species (ROS) and the integrity of the plasma
membrane of P. expansum was assessed. These results may
provide a novel strategy for controlling of blue mold on
postharvest fruits and vegetables.
Materials and Methods
Pathogen and Chemicals
The fungal pathogen P. expansum link (CGMCC3.3703)
was used in this study. Penicillium expansum was obtained
from Key Laboratory of Plant Resources in North China,
Institute of Botany, Chinese Academy of Sciences. The
193
fungus was inoculated and re-isolated from apple fruit to
maintain pathogenicity. The isolates were routinely grown
on potato dextrose agar (PDA) plates for 2 weeks at 25°C.
The conidia were obtained from the surface of the agar and
suspended in 5 ml of sterile distilled water containing
0.1 % v/v Tween 20. Spore suspensions were filtered
through four layers of sterile cheesecloth to remove
mycelia fragments. A hemacytometer was used to calculate
the number of spores and the spore concentration was
adjusted to 5.0 9 105 spores/ml before conducting sub-
sequent experiments.
Se in the form of sodium selenite was purchased from
Sigma Chemical Co. (St. Louis, MO). Potato dextrose
broth (PDB), propidium iodide (PI), 2,7-dichlorodihydro-
fluorescein diacetate (DCHF-DA) were purchased from
Sigma. All other chemicals used in this study were of
guarantee reagent.
Inhibitory Effect of Selenium on Spore Germination
and Germ Tube Elongation of P. expansum
Inhibition of spore germination and germ tube elongation
of P. expansum by sodium selenite was measured as pre-
viously reported [37]. In brief, aliquots of a spore sus-
pension of P. expansum were added to wells of a 24-well
microtitration plate containing PDB medium to obtain a
final concentration of 5 9 105 spores/ml. The culture
medium was supplemented with different concentrations of
sodium selenite at 0, 5, 10, 15, and 20 mg/l with normal
pH. The microtitration plate was incubated at 25°C on a
rotary shaker at 200 rpm. About 300 spores were examined
microscopically for germination rate and germ tube length
after 9 h of incubation. Spores were considered germinated
if the germ tube was equal to or greater than the diameter
of the spore. Germ tube length was determined with an
ocular micrometer and germinated spores were expressed
as percentage of the total number of evaluated spores.
There were three replicates for each treatment, and the
experiment was repeated twice.
Effect of Selenium on Mycelial Growth of P. expansum
The effect of sodium selenite on mycelial growth of
P. expansum was assayed in PDA following the method of
Droby et al. [14]. A 5-mm diameter plug of mycelial agar
was obtained from the growing edge of 7-day-old cultures
of the fungi and placed in the center of a 9-cm-diameter
Petri dish containing PDA medium with different concen-
trations (0, 5, 10, 15, and 20 mg/l) of sodium selenite
solutions. Sodium selenite solutions were filtered through a
0.45 lm Millipore filter before being added to the auto-
claved PDA medium that had cooled to about 60°C. Radial
growth of P. expansum was observed after incubation at
123
194
25°C for 3, 4, and 5 days. Results are reported as the
diameter of the fungal colony (mycelium) in the Petri dish
minus the diameter of the agar plugs (5 mm). Each treat-
ment contained three replicates and the entire experiment
was repeated twice.
Measurement of Reactive Oxygen Species
The oxidant-sensitive probe DCHF-DA was used to assess
the intracellular ROS levels in P. expansum according to
the methods described previously [9, 10]. Spores of
P. expansum were cultured in PDB medium supplemented
with 0–15 mg/l sodium selenite as described above and
collected after 6 and 9 h of incubation. The spores were
washed with 10 mM potassium phosphate buffer (pH 7.0)
and incubated for 1 h in the same buffer containing 10 lM
DCHF-DA (dissolved in dimethylsulfoxide). After washing
twice with potassium phosphate buffer, spores were
examined under a Zeiss Axioskop microscope (Carl Zeiss,
Oberkochen, Germany) using a fluorescein 2,7-dichlor-
odihydro-specific filter.
Membrane Integrity Assay and Microscopy
Spores of P. expansum were treated with sodium selenite at
0–20 mg/l in PDB medium. After 6 and 9 h of incubation
at 25°C, spores were collected and stained with 10 lg/ml
PI for 5 min at 30°C. Because PI is membrane impermeant,
only cells that have lost membrane integrity will show red
staining. The spores were centrifuged and washed twice
with 10 mM potassium phosphate buffer (pH 7.0) to
remove residual dye. Then, the spores were observed on a
Zeiss Axioskop microscope (Carl Zeiss, Oberko-chen,
Germany) equipped with individual fluorescein rhodamine
filterset (Zeiss no. 15: excitation BP 546/12 nm, emission
LP 590 nm). Images were collected using an Axiocam
MRc digital camera (Carl Zeiss).
Measurement of Cellular Leakage
The leakage of cytoplasmic contents from mycelium of
P. expansum was determined according to the method of
Lewis and Papavizas [26] with some modifications. Peni-
cillium expansum was grown in conical flasks (250 ml)
containing 100 ml PDB medium at 25°C on a rotary shaker
at 200 rpm and mycelia harvested after 3 days of incuba-
tion. After being pooled and washed with sterile distilled
water, the mycelia were re-suspended in 100 ml sterile
distilled water containing 0–20 mg/l sodium selenite, and
incubated on a rotary shaker at 25°C for 1, 2, 3, 4, and 5 h.
Mycelia were then filtered from the solutions through a
0.2 lm pore size membrane and the aqueous solutions
were used for determination of soluble proteins and
123
Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum
carbohydrates. The Bradford assay [4] was performed on
the filtrates to quantify proteins by the various treatments.
Soluble carbohydrate was determined with anthrone
reagent using glucose as the standard [26].
Measurement of Cellular Enzymatic Activity
Penicillium expansum was grown in conical flasks (50 ml)
containing 20 ml PDB medium with 0–15 mg/l sodium
selenite at 25°C on a rotary shaker at 200 rpm and mycelia
were harvested after 24, 48, and 72 h of incubation. The
leakage of cytoplasmic contents from mycelium of
P. expansum was determined according to the method of
Chander [8] with some modifications. 0.5 g mycelia were
filtered from the solutions and ground into homogenate
with phosphate buffer (pH 6.8) and a small quantity of
quartz sand in liquid nitrogen. The homogenate was cen-
trifuged at 12,000 rpm for 20 min and the supernatant was
used for determination of all kinds of enzymatic activity.
The Bradford assay [4] was performed on the supernatant
to quantify release of proteins by the various treatments.
The content of glutathione (GSH), methane dicarboxylic
aldehyde (MDA), and superoxide dismutase (SOD) was
determined with the method of Liu et al. [28]. Hydrogen
peroxide (H2O2) production rate was determined with the
method of Mondal [35]. Superoxide anion (O2-) produc-
tion rate was determined with the method of Xu [46].
Absorbance of the supernatant under different wavelengths
was measured by a ultraviolet spectrophotometer
(UV-2450, Shimadzu, Japan).
Statistical Analysis
All statistical analyses were performed with the SPSS
software version 13.0 (SPSS Inc., Chicago, IL, USA).
Analysis of variance (ANOVA) was carried out to deter-
mine the effects of the treatments. When the treatment
effects were statistically significantly (p \ 0.05), the
Duncan’s multiple range test was used for means separa-
tions. Data presented in this article were pooled across
three independent repeated experiments.
Results
Effects of Se on Spore Germination and Germ Tube
Elongation of P. expansum
The inhibitory effect of Se on spore germination and germ
tube elongation was positively related to the concentration
of Se used (Fig. 1). Spore germination of P. expansum was
significantly (p \ 0.05) inhibited by Se at 10 mg/l
(Fig. 1a). In contrast, the inhibitory effect of Se on germ
Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum 195

A 100
a
a 80
a
Se
70
Spore germination(% of total)

80 3d
60 4d

Mycelial growth(mm)
a 5d
60 50 b
40
a b
40
b
30
b
20
20
c
c 10 c
c d
d d d
0 e e e
0
0 5 10 15 20 0 5 10 15 20
Concentration(mg/L) Concentration of selenium(mg/L)

B 60 Fig. 2 Inhibition of selenium at different concentrations on mycelial

Se growth of P. expansum in potato dextrose agar. Growth of
a P. expansum was observed after 3, 4, and 5 days of incubation at
50 25°C. Treatments followed by different letters within each sampling
interval are statistically different by the Duncan’s multiple range test
Germ tube length(µm)

40 (p \ 0.05)

b
30

germ tube elongation, and there was a significantly positive

20
c
10
0 Detection of Reactive Oxygen Species With DCHF-DA
0 5 10
Concentration(mg/L)
Fig. 1 Effect of selenium (Se) on a germination of conidia and
b elongation of germ tubes of P. expansum after 9 h incubation at
25°C. Treatments followed by different letters are statistically
different according to the Duncan’s multiple range test (p \ 0.05)
tube elongation of P. expansum was already evident when
the concentration reached 5 mg/l (Fig. 1b). This indicated
that spore germination was less sensitive to Se than germ
tube elongation. When the concentration of Se reached
15 mg/l, \10 % of P. expansum spores germinated. Both
spore germination and germ tube elongation were almost
completely inhibited by Se at 20 mg/l.
Inhibitory Effect of Se on Mycelial Growth
of P. expansum
As shown in Fig. 2, Se was effective in inhibiting the
mycelia growth of P. expansum on PDA. After incubation
for 3–4 days at 25°C, the growth of P. expansum was
completely inhibited when the concentration of Se reached
to 20 mg/l. Moreover, the inhibitory effect on mycelia
growth was more notable than on spore germination and
correlation between Se concentration and the effect on
mycelial growth.
d
e
15 20
DCHF-DA as an oxidant-sensitive fluorescent dye was
used to monitor the changes in intracellular ROS levels.
Compared with control, more cells under Se stress were
stained with the DCHF-DA, indicating that an increasing
amount of oxidizing molecules was produced in cells
exposed to Se (Fig. 3a). After 6 h of incubation, a majority
of spores in control did not show any obvious fluorescence
signal, whereas partial spores (about 25 %) were stained by
DCHF-DA in Se treatment (Fig. 3b). With increasing time,
the percentage of stained spores in Se treatment reached
65 % after 9 h of incubation, which were significantly
higher compared with the control (Fig. 3c).
Detection of Plasma Membrane Integrity With PI
The plasma membranes of P. expansum were markedly
damaged by Se (p \ 0.05) (Fig. 4a). Membrane integrity
of P. expansum spores declined with increasing incubation
time in PDB containing Se (Fig. 4b). Most (90 %)
untreated spores appeared to have intact plasma mem-
branes whereas membrane integrity of spores treated with
20 mg/l Se decreased to 72 % at 6 h and 32 % at 9 h
(Fig. 4c).

123
196 Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum

A b Fig. 3 Effect of sodium selenite on production of reactive oxygen

Brightfield DCHF-DA species in spores of P. expansum. Spores incubated for a 6 h and
b 9 h. c The percentage of spores stained with DCHF-DA. Bars
represent standard deviation of the treatment means of data. Lower
case letters indicated significant differences at p \ 0.05

Cellular Leakage and Determination of SOD Activities,

H2O2, MDA, GSH Content, and O2- Production Rate

The leakage of proteins and carbohydrates were signifi-

cantly increased under Se stress compared with the control
group (Fig. 5). To determine whether toxicity of Se to
P. expansum gave rise to the change of enzyme activity in
mycelium, the activity of cellular enzymatic were deter-
mined. As shown in Fig. 6a and b, both SOD activities and
GSH content presented the similar decreasing trend from 1
to 3 days. Moreover, upon exposure to Se, the values of
6h SOD and GSH were significantly lower than those of the
B control at each time point. The descent speed of SOD
Brightfield DCHF-DA activities and GSH content were even faster compared to
the control group. It reached minimum for SOD activities
4.5 U/mg protein and GSH content 2 lg/g, respectively.
MDA content significantly increased under Se stress
compared with that in the control, especially after 3 days
with a maximum of 11.8 lg/g (Fig. 6c). H2O2 content and
O2- production rate of cellular without Se still remained
increasing, but the rising speed were obviously lower than
that had been treated by 15 mg/l Se. They increased to 5
and 8.7 nmol/g for H2O2 and O2-, respectively (Fig. 6d, e).

Discussion

Se plays an important role in scavenging active oxygen

9h (AO) species as an essential constituent of glutathione
C 80 peroxidase. Several reports proposed a possible induction
b of toxic AO by Se compounds in vitro [41]. In this study,
Control
70
Se Se could significantly inhibit spore germination and
cells stained with DCHF-DA(%)

60 mycelia growth of P. expansum. The mechanism of action

appeared to be closely related to the generation of AO. In
50
this regard, direct plasma membrane damage and induction
40
of ROS resulted in cell apoptosis.
The results of PI staining and microscopy showed that
30 b
the plasma membrane integrity of P. expansum conidia
a clearly declined in PDB containing Se with the increase of
20
incubation time (Fig. 3). Compared with the control, the
10
a leakage of proteins and carbohydrates from mycelia was
more significant in the Se treatment (Fig. 5). These results
0
6 9 confirmed previous findings that approximately 68 % of
Time(h) the conidia were cracked and ruptured, with leakage of cell

123
Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum 197

A A
Brightfield PI 25
Control
Se

Soluble carbohydrate(µg/mL)
20

15

10

0
0 1 2 3 4 5
Time(h)

B 35

6h
30 Control
B Se
Brightfield PI

Solubleprotein(µg/mL)
25

20

15

10

0
0 1 2 3 4 5
Time(h)

Fig. 5 Effect of Se at 20 mg/l on leakage of carbohydrates (a) and

protein (b) of P. expansum mycelia cultured in sterile double-distilled
water at 25°C. Values for both soluble proteins and carbohydrates
were expressed as lg/ml of aqueous solutions
9h
C 110
contents when exposed to Se. Therefore, Se treatment
100
possibly precipitated plasma membrane disintegration and
90 leakage of intracellular proteins and sugars, which resulted
Integrity of menbrane(%)

80 in inhibiting the growth of fungal phytopathogen. In

addition, Se can inhibit fungal pathogens (Table 1) at
70
minimum concentrations, as reported in the literature. This
60 finding suggests that Se might serve as a potential alter-
50 native to synthetic fungicides for the control of the post-
harvest disease of fruit and vegetables caused by
40
P. expansum.
control Se
30 Both the production and clean-up of free radicals exists
20 are dependent upon stability of the dynamic balance
0 1 3 6 9 in vivo, the excessive free radical can damage the organ-
Time(h) ism. To withstand biotic stress and abiotic stress, a perfect
AO scavenging system has been formed in the long term of
Fig. 4 Effect of sodium selenite on plasma membrane integrity of
P. expansum conidia. Spores incubated for a 6 h and b 9 h. evolution process, which mops up the reactive oxygen
c Percentage of plasma membrane integrity of P. expansum spores compounds and protects cell from the infection of

123
198 Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum

A 18 B Control
a a Se
12 a
16 a a Control a
Se a
SOD activity(U·mg-1 protein)

14 a 10

GSH content(µg/g)
12
8
b
10 b
a
a 6
8
b
6 4 b
b
4
b
2
2

0 0
0 1 2 3 0 1 2 3
Time after treatment(d) Time after treatment(d)

C b D 10

Superoxide anion producing rate(nmol/h·g)

12 b
Control
b Control
Se Se
10 8
b a b
a
MDA content(µg/g)

8 a a a b
6 a
a
6 a
4
a a
4

2
2

0 0
0 1 2 3 0 1 2 3
Time after treatment(d) Time after treatment(d)

E b
5
Control
Hydrogen peroxide content(nmol/g)

Se
4

b
3
b
a
2 a

a
1
a a

0
0 1 2 3
Time after treatment(h)

Fig. 6 Effects of Se at 15 mg/l on activities of SOD (a), GSH (b), Columns with different letters at each time point are significantly
and MDA (c) content, O2- (d) and H2O2 (e) production rate of different according to Duncan’s multiple range test at p \ 0.05
P. expansum spores after 1, 2, and 3 days of incubation at 25°C.

123
Z. Wu et al.: Inhibitory Effect of Selenium Against Penicillium expansum
Table 1 Comparison of inhibitory effects of several similar inor-
ganic salts on fungal pathogen
Inorganic Minimum Inhibitory References
salts inhibitory fungal pathogen
concentration
(g/l)
Se 0.01 Penicillium Current study
expansum
Boron 1 Botrytis cinerea Qin et al. [37]
Sulfur 0.85 Penicillium Franck et al. [16]
expansum
Silicon 2 Penicillium Qin et al. [36]
expansum
Calcium 10 Penicillium Yu et al. [47]
expansum
oxidative stress [20]. SOD and GSH are the two key
enzymes of oxidative metabolism. SOD can catalyze
intracellular O2- into O2 and H2O2, while catalase (CAT)
and peroxidases (POD) can catalyze H2O2 into innoxious
substances, such as O2 and H2O [1, 3, 15]. GSH was a
crucial antioxidant to prevent membrane lipid peroxidation
and remains stable in cell membrane systems [33]. In this
study, GSH content was significantly decreased by 15 mg/l
Se treatment. The inhibition of its activity aggravates
membrane lipid peroxidation and damages the integrity of
cell membrane systems thereby accelerating spore apop-
tosis sharply. We also found that MDA content, a param-
eter reflecting lipid peroxidation, increased in P. expansum
exposed to Se, the enhanced degree of cell membrane lipid
peroxidation suggest that the inhibitory effect of Se was
associated with oxidative damage.
To verifying whether AO scavenging system was dam-
aged by Se stress, we have made a further study on the
change of intracellular ROS of P. expansum. We demon-
strated that an obvious increase was detected on the num-
ber of spores generated with ROS after 6 and 9 h using
DCHF-DA as an oxidant-sensitive fluorescent dye. ROS
level was even higher in the Se treatment as time passed,
which was consistent with the results of H2O2 and O2-
production rate in P. expansum spores. Oxidative stress
caused by Se may have resulted from two aspects: the
increase of ROS production and the decrease of ROS
detoxifying ability. Antioxidant defense systems played an
important role in mediating the ROS level [34].
Se plays an important role in scavenging AO species as
an essential constituent of glutathione peroxidase [27].
Some researchers have proposed that Se compounds are
toxic because of their prooxidant catalytic activity to pro-
duce O2-, H2O2, and likely other cascading oxyradicals
[44]. Spallhoze has used scanning electron microscopic
(SEM) to show that a certain concentration of selenite,
199
selenocysteine, and even glutathione peroxidase in vitro
could damage rat erythrocyte (red blood) cell membranes
[19]. Electron spin resonance (ESR) spectra using dimethyl
pyrroline oxide (DMPO) as a spin trapping agent was
applied to demonstrate that Se compounds, i.e., selenite
and selenium dioxide, can react with GSH and other thiols
to form selenotrisulfides that will ultimately react to pro-
duce toxic superoxide and H2O2 [46]. Many studies have
assumed that it is possible for Se compounds to generate
AO species within cells. Although the idea that AO may be
generated in vivo is controversial, because oxygen tension
in the body is lower than that in the solution under ambient
atmosphere. However, tissues consume O2 rapidly, and O2
is transported efficiently to tissues. Therefore, it may be
possible that Se compounds, such as Se2- and RSeH, react
with O2 to generate AO species within tissues in vivo [42].
Since Se has a dual function in the organism and its higher
concentrations can be toxic, the margin between Se toxicity
and essentiality is quite narrow. Se in small amounts is an
essential micronutrient for many organisms, including
humans and other animals. Se is a constituent of enzymes,
such as glutathione peroxidase and thioredoxin reductase,
and contributes to the antioxidant protection of cells
[18, 39]. Agricultural crops can be used both to remediate
Se-contaminated soils [2, 14, 48] and to increase the daily
Se intake of consumers after soil supplementation using
inorganic or organic Se sources.
In conclusion, our results showed that Se could directly
inhibit the growth of P. expansum in vitro. This observation
suggests that Se might serve as a potential alternative to
synthetic fungicides for the control of the postharvest disease
of fruit and vegetables caused by P. expansum. However,
further study on the mechanism of Se against fungal patho-
gens at the biochemical and molecular level is needed.
Acknowledgments The authors wish to thank Drs. T.F Lai and X.Q
Shi who comes from Key Laboratory of Plant Resources in North China,
Institute of Botany, Chinese Academy of Sciences for his useful com-
ments and suggestions on the language and structure of our manuscript.
This work was partially supported by the National Natural Science
Foundation of China (50949038), the Natural Science Foundation of
Anhui province (1408085MC68), Natural Science Youth Foundation of
Jiangsu Province for Youth of China (BK 2012195, BK 2012202), and
China Postdoctoral Science Foundation (20100470108).
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