Biol Trace Elem Res (2012) 146:101–106
DOI 10.1007/s12011-011-9222-7
Impact of Bulk and Nanosized Titanium Dioxide (TiO2)
on Wheat Seed Germination and Seedling Growth
Hassan Feizi & Parviz Rezvani Moghaddam &
Nasser Shahtahmassebi & Amir Fotovat
Received: 30 August 2011 / Accepted: 27 September 2011 / Published online: 7 October 2011
# Springer Science+Business Media, LLC 2011
Abstract The impacts of different concentrations of bulk
and nanosized TiO2 on seed germination and seedling
growth of wheat were studied in a randomized completely
design with four replications in the College of Agriculture,
Ferdowsi University of Mashhad, Iran, in 2011. The
experimental treatments included five concentrations of
bulk (1, 2, 10, 100, and 500 ppm), five concentrations of
nanosized TiO2 (1, 2, 10, 100, and 500 ppm), and control
(without any TiO2). Results indicated that among the wheat
germination indices, only mean germination time was
affected by treatments. The lowest and the highest mean
germination time (0.89 vs. 1.35days) were obtained in
10 ppm concentration of nanosized TiO2 and control
treatments, respectively. In addition, shoot length, seedling
length, and root dry matters were affected by bulk and
nanosized TiO2 concentrations, significantly. Shoot and
seedling lengths at 2 and 10 ppm concentrations of
H. Feizi (*) : P. Rezvani Moghaddam
Department of Agronomy, Faculty of Agriculture,
Ferdowsi University of Mashhad,
P.O. Box 91775-1163, Mashhad, Iran
e-mail: hasanfeizi@yahoo.com
P. Rezvani Moghaddam
e-mail: rezvani@um.ac.ir
N. Shahtahmassebi
Department of Physics, Faculty of Basic Sciences and Nano
Research Center, Ferdowsi University of Mashhad,
P.O. Box 91775-1163, Mashhad, Iran
e-mail: nasser@um.ac.ir
A. Fotovat
Department of Soil Science, Faculty of Agriculture, Ferdowsi
University of Mashhad,
P.O. Box 91775-1163, Mashhad, Iran
e-mail: afotovat@yahoo.com
nanosized TiO2 were higher than those of the untreated
control and bulk TiO2 at 2 and 10 ppm concentrations.
Employing nanosized TiO2 in suitable concentration could
promote the seed germination of wheat in comparison to
bulk TiO2 but in high concentrations had inhibitory or any
effect on wheat.
Keywords Nanosized TiO2 . Germination . Seedling .
Wheat
Introduction
The use of nanoparticles in commercial products and
industrial applications has increased greatly in recent years.
However, understanding of the interaction mechanisms at
the molecular level between nanoparticles and biological
systems is largely unknown [1]. In recent times, the rapidly
increasing applications of engineered nanoparticles, with
sizes smaller than 100 nm in various areas of the economy,
such as textiles, electronics, pharmaceutics, cosmetics, and
environmental remediation, have received a lot of attention
and concern [2]. Nanotechnology allows wide advances in
agricultural research, such as reproductive science and
technology, transfer of agricultural and food wastes to
energy and other useful by-products through enzymatic
nanobioprocessing, disease prevention, and treatment in
plants using various nanocides [2].
The ninth most abundant element and the second most
abundant transition metal in the earth's crust is titanium
element (about 6.320 ppm). The most important effects of
Ti compounds on plants are enhancement of the yield of
various crops (about 10–20%); an improvement of some
essential element contents in plant tissues; an increase in
the peroxidase, catalase, and nitrate reductase activities in
102 Feizi et al.

plant tissues; and an enhancement of the chlorophyll different concentrations of bulk and nanosized TiO2 on seed
content in paprika (Capsicum anuum L.) and green alga germination and seedling growth of wheat.
(Chlorella pyrenoidosa) [3]. Plants interact with their
atmospheric and edaphic environments strongly and are
expected to be affected by exposure to engineered nano- Materials and Methods
particles [4]. Lu et al. [5] have shown that a combination of
nanosized SiO2 and TiO2 could increase the nitrate Description of Materials
reductase enzyme in soybean (Glycine max), increase its
abilities of absorbing and utilizing water and fertilizer, Wheat (T. aestivum L. var. Pishtaz) seeds were taken from
promote its antioxidant system, and in fact accelerate its the Agricultural Research Center of Khorasan Razavi
germination and growth. Also it has been found that TiO2 Province, Iran. Nanosized TiO2 powder was as AERO-
nanoparticles encourage spinach (Spinacia oleracea) seed XIDE® TiO2 P25 that was supplied by Evonik Degussa
germination and plant growth [6]. Navarro et al. [7] stated GmbH Company. Specific surface area of nanosized TiO2
that engineered nanoparticles could sequester nutrients on was 50 m2 g−1, average primary particle size was 21 nm,
their surfaces and thus serve as a nutrient stock to the and purity was >99.5%. The size and topography of TiO2
organisms, particularly those engineered nanoparticles nanoparticles (Figs. 1 and 2) were determined by scanning
having high specific surface area. These positive effects tunneling microscope (STM) in the Central Laboratory of
could be probably due to the antimicrobial properties of Ferdowsi University of Mashhad, Iran.
engineered nanoparticles, which can enhance strength and X-ray diffraction (XRD) pattern of TiO2 nanoparticles
resistance of plants to stress. was shown in Fig. 3. XRD measurement showed that the
Other studies were reported that nanosized TiO2 can used TiO2 nanoparticles were made by 80% anatase and
promote plant photosynthesis and nitrogen metabolism and 20% rutile. Analysis of particles in X-ray diffraction
then greatly improve growth of spinach at a suitable indicates tetragonal particles and crystalline nature of
concentration [6, 8–10]. But Yang and Watts [11] conclud- TiO2 particles.
ed that root elongation of corn (Zea mays), cucumber Bulk TiO2 was supplied by AppliChem GmbH Compa-
(Cucumis sativus), soybean (G. max), cabbage (Brassica ny with 99% purity, and particle size was determined by
oleracea), and carrot (Daucus carota) was inhibited by scanning electron microscope (SEM) in the Central Labo-
uncoated alumina nanoparticle treatment. It has been ratory of Ferdowsi University of Mashhad, Iran (Fig. 4).
reported that the germination rate and the germination and XRD measurement showed that the used bulk TiO2
vigor indexes of spinach at 250–4,000 ppm nanosized TiO2 particles were made by 100% anatase. Analysis of particles
treatments were improved in comparison with bulk-TiO2 in X-ray diffraction indicates tetragonal particles and the
treatment [6]. They added that during the spinach growth crystalline nature of bulk TiO2 particles (Fig. 5).
period, the plant dry weight, chlorophyll formation,
ribulose bisphosphate carboxylase/oxygenase activity, and
the photosynthetic rate were increased. These results
suggested that the physiological effects were related to the
nanometer-size particles. Gao et al. [12] revealed that
treated S. oleracea by nano-anatase TiO2, Rubisco carbox-
ylase activity was 2.67 times that of control Rubisco. Lee et
al. [13] studied phytotoxicity and bioavailability of copper
nanoparticles to the plants Phaseolus radiatus and Triticum
aestivum, employing plant agar test as growth substrate.
They concluded that the growth rates of both plants were
constrained when exposure to nanoparticles and the
seedling lengths were negatively related to the exposure
concentration of nanoparticles. Contents of nanoparticles in
plant tissues increased with increasing nanoparticle con-
centration in growth media; therefore, bioaccumulation was
concentration dependent.
A few studies have been done on the effects of
nanoparticles on crops particularly on wheat which is one
of the most important crops in the world. In this work, we
decided to find out the phytotoxicity or positive effects of Fig. 1 Image of nanosized TiO2 by STM
Impact of Bulk and Nanosized TiO2 on Wheat Seed
Fig. 2 Topographic image of nanosized TiO2 by STM
Experimental Design and Data Observation
In order to study the effect of different concentrations of
bulk and nanosized TiO2 on wheat germination, a random-
ized completely design with four replications was
employed. The experimental treatments included five
concentrations (1, 2, 10, 100, and 500 ppm) of bulk and
five concentrations (1, 2, 10, 100, and 500 ppm) of
nanosized TiO2 and untreated control (without any TiO2
types). The experiment was conducted in laboratory
conditions with natural light and an average temperature
of 20 ± 1°C at the College of Agriculture, Ferdowsi
University of Mashhad, Iran, in 2011.
One hundred seeds of similar size were randomly
selected and placed on moistened paper as four groups of
25 seeds in Petri dishes, and then 10 ml of each
concentration treatment was added to each Petri dish. For
the control, only distilled water was added to Petri dishes.
Germination tests were performed according to the rule
issued by the International Seed Testing Association [14].
All concentrations of TiO2 and the control were run at the
Fig. 3 XRD pattern of nano
TiO2 particles
103
Fig. 4 Image of bulk TiO2 particles by SEM
same time and consequently under equal light and
temperature conditions. The number of germinated seeds
was noted daily for 8 days. Seeds were considered as
germinated when their radicle showed at least 2 mm length
[14]. Mean germination time was calculated based on
Matthews and Khajeh-Hosseini [15] (Eq. 1):
P
F:X
MGT ¼ P ð1Þ
F
Where F is the number of seeds newly germinated at the
time of X, and X is the number of days from sowing.
Germination rate was determined based on Maguire [16]
(Eq. 2):
Germination rate ¼ ða=1Þ þ ðb  a=2Þ þ ðc  b=3Þ
þ . . . :: þ ðn  n  1=N Þ ð2Þ
104
Fig. 5 XRD pattern of bulk
TiO2 particles
where a, b, c,…, n are number of germinated seeds after 1,
2, 3,…, Ndays from the starting of their imbibitions.
Seedling vigor was computed based on Vashisth and
Nagarajan [17] (Eqs. 3 and 4):
Vigor index I ¼ germination%  seedling lengthðroot þ shootÞ
ð3Þ
Vigor index II ¼ germination%  seedling weightðroot þ shootÞ
ð4Þ
Statistical Analysis
Analysis of variance was performed on a randomized
completely design. The data were analyzed using MSTAT-
C software. The significant levels of difference for all
measured traits were calculated, and the means compared
by multiple range Duncan test at 5% level.
Results and Discussion
In terms of wheat germination indices, only mean germi-
nation time was affected by studied treatments. In addition,
among the seedling growth indices, shoot length, seedling
length, and root dry matter were affected significantly by
bulk and nanosized TiO2 concentrations (Tables 1 and 2).
Although the highest germination percent (98%) was
shown in both 2 ppm bulk and nanosized TiO2 concen-
trations, the treatments had no significant effect on seed
germination percentage (Table 1). In general, lower mean
germination time is state earlier germination of seed. The
lowest mean germination time (0.89 day) was found in
Feizi et al.
10 ppm concentration nanosized TiO2, and the highest
(1.35 day) was shown in the control treatment. Therefore,
10 ppm concentration nanosized TiO2 treatment reduced
mean germination time by 34% in comparison to untreated
control, while 10 ppm concentration bulk TiO2 did not
reduce mean germination time (1.35 day) in comparison
with control (Table 1). Zheng et al. [6] demonstrated that
nanosized TiO2 helped the water absorption by the spinach
seeds, and as result of it, the germination of the seeds
accelerated.
The effect of studied treatments on root length was not
significant, but they had a significant effect on shoot and
seedling lengths. Shoot and seedling lengths at 2 and
10 ppm concentrations of nanosized TiO2 were higher than
those of the untreated control (8% and 7.3%, respectively)
and bulk TiO2 (10.2% and 7%, respectively) treatments.
Increasing concentrations of nanosized TiO2 after 10 ppm
decreased shoot and seedling lengths. The lowest shoot
length was achieved at 100 ppm nanosized TiO2 and 2 ppm
bulk TiO2 treatments (Table 1). Lee et al. [13] analyzed
toxicity and bioavailability of copper nanoparticles to the
bean (Phaseolus radiatus) and wheat (T. aestivum) plants.
They stated that the exposure of bean and wheat plants to
copper nanoparticles decreased growth and seedling
lengths; and mentioned criteria were negatively related to
the exposure concentration of nanoparticles. The results of
Zheng et al. [6] showed that the growth of spinach plants
was greatly improved at 250–4,000 ppm nano TiO2
concentrations, but there was no improvement at higher
concentrations.
The results indicated that nanosized TiO2 in an appro-
priate concentration could promote the seed germination
and seedling growth of wheat in comparison to bulk TiO2.
Ruffini and Cermonini [4] suggested that nanoparticles can
explain their actions depending on both the chemical
Impact of Bulk and Nanosized TiO2 on Wheat Seed 105

Table 1 Effect of different concentrations of bulk and nanosized TiO2 on seed germination of wheat

Concentration (ppm) Germination (%) Germination rate MGT (day) Root length (cm) Shoot length (cm) Seedling length (cm)
(seed day−1)

Bulk TiO2
1 96 a 20.48 a 1.34 a 14.92 a 15.49 abc 30.41 abc
2 98 a 20.08 a 1.18 ab 14.44 a 14.53 bc 28.97 bc
10 90 a 17.73 a 1.35 a 13.16 a 15.10 abc 28.25 c
100 95 a 18.68 a 1.13 ab 15.51 a 15.44 abc 30.94 abc
500 97 a 22.73 a 1.22 ab 15.53 a 15.76 ab 31.29 ab
Nano TiO2
1 94 a 19.60 a 1.24 a 15.46 a 15.50 abc 31.05 abc
2 98 a 15.02 a 1.20 ab 16.04 a 16.01 a 32.06 a
10 95 a 15.00 a 0.89 b 14.69 a 16.15 a 30.85 abc
100 95 a 16.20 a 1.42 a 13.94 a 14.35 c 28.29 c
500 92 a 17.89 a 1.21 ab 15.45 a 15.37 abc 30.83 abc
Control 92 a 17.85 a 1.35 a 14.35 a 14.92 abc 29.17 bc

Means in each column followed by similar letters are not significantly different at the 5% probability level using Duncan's multiple range test

compound and on the size and/or shape of the particles.
Zheng et al. [6] reported that the significant effect of
nanosized TiO2 on spinach germination is probably
attributed to the small particle size, which allows its
penetration into the seed during the treatment period,
exerting its enhancing functions during growth.
It seems that bulk TiO2 could not penetrate into the seeds
of wheat in the same way which Zheng et al. [6] reported;
therefore, the results were not as marked as those of the
nanocounterpart treatments. Yang and Watts [11] indicated
that surface area and surface characteristics play an
important role in the phytotoxicity of nanoparticles to
seedlings. Khodakovskaya et al. [18] revealed that multi-
wall carbon nanotubes (MWCNTs) at the concentration
range of 10–40 mg L−1 significantly improved the seed
germination and growth of tomato plants. They hypothe-
sized that the positive effect of MWCNTs arose from the
capability of CNTs to penetrate the seed coat and therefore
promote water uptake. Water uptake in seed germination is
important because mature seeds are fairly dry and need a
large amount of water to start cellular metabolism and
growth. Khodakovskaya et al. [18] added that the calculated
water moisture content of seeds and the detection of CNTs
inside seeds supported the hypothesis, but the specific
penetration mechanisms through the coat and the improve-
ment of water uptake by CNTs were not reported.

Table 2 Effect of bulk and nanosized TiO2 concentrations on seed germination of wheat

TiO2 concentration (ppm) Root dry matter (mg) Shoot dry matter (mg) Seedling dry matter (mg) Vigor index I Vigor index II

Bulk TiO2
1 6.93 b 9.68 a 16.61 a 2,188 a 1,595 a
2 8.56 a 10.27 a 18.83 a 1,932 a 1,846 a
10 7.41 b 9.19 a 16.60 a 1,889 a 1,496 a
100 6.96 b 10.24 a 17.20 a 1,954 a 1,634 a
500 7.67 ab 10.33 a 18.00 a 1,826 a 1,744 a
Nano TiO2
1 6.99 b 9.49 a 16.47 a 1,948 a 1,537 a
2 7.01 b 9.49 a 16.50 a 1,874 a 1,616 a
10 7.00 b 9.84 a 16.84 a 1,645 a 1,597 a
100 7.93 ab 9.40 a 17.32 a 1,935 a 1,598 a
500 7.39 b 10.6 a 18.00 a 1,864 a 1,655 a
Control 7.16 b 9.86 a 17.01 a 1,693 a 1,493 a

Means in each column followed by similar letters are not significantly different at the 5% probability level using Duncan's multiple range test
106
Therefore, it seems that the uptake efficiency and effects of
various nanoparticles on the growth and metabolic func-
tions vary differently among plants [19].
All studied treatments affected root dry matter signifi-
cantly. The highest root dry matter was found in 2
(8.56 mg) and 500 ppm (7.67 mg) bulk and 100 ppm
(7.93 mg) nanosized TiO2 concentration (Table 2). Exper-
imental treatments have no significant effect on shoot and
seedling dry matter, vigor index I, and vigor index II
(Table 2). Owolade et al. [20] reported that seed yield of
cowpea (Vigna unguiculata Walp) was increased when
treated (as foliar application) with nanosized TiO2. They
concluded that it may be due to the photocatalyst ability of
the nanosized TiO2 which leads to an increased photosyn-
thetic rate. Lin and Xing [21] evaluated phytotoxicity of
five types of metallic nanoparticles in six higher plant
species and indicated that seed germination was not
affected except for the inhibition of nanoscale zinc on
Lolium multiflorum and nanoscale zinc oxide on maize (Z.
mays). They indicated that inhibition of root growth varied
significantly among nanoparticles and plants, and it is
partially correlated to nanoparticle concentration.
Conclusions
Interactions of plants with dosage and size of TiO2
were studied by measuring the germination indices and
vigor index of wheat seeds. The results indicated that the
nanosized TiO2 treatments in proper concentration accel-
erates the germination of the wheat seeds and increases its
vigor. Nano TiO2 improves the mean germination time
and growth of wheat seedling in comparison to bulk TiO2
and untreated control. We found that nanosized TiO2 in
low concentration has stimulatory effect on wheat
seedling. High concentrations of nanosized TiO2 had
neutral effects on studied criteria. These effects have been
obtained inversely by the bulk TiO2 particles. These
results will help to further understand inhibitory effects of
bulk and nanosized materials and are important in terms
of the relationship between size and concentration of
materials and their effects on germination and plant
growth. Finally, future studies should be directed to the
requirements to illuminate the phytotoxicity and promot-
ing mechanisms, possible uptake and translocation of
nanoparticles by plants, and physical and chemical
properties of nanoparticles in plant metabolism and the
rhizosphere.
Acknowledgments The authors acknowledge the financial support
of the project by Ferdowsi University of Mashhad. We thank Ms.
Pesyan and Ms. Hashemian for helping provide electron microscopy
images. We wish to thank Mrs. Agheli for providing valuable help
during seed germination tests.
Feizi et al.
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