143

1. Introduction

Strawberry (Fragaria x ananassa Duch.), a small fruit 
crop and a hybrid of two highly variable octoploid species, 
has adapted to extremely different environmental condi-
tions (Rieger, 2005). Prerequisites for successful strawber-
ry growing are suitable climate, cultivars, soil and nutri-
tion (Almaliotis et al., 2002). Seed priming enhances seed 
performance by rapid and uniform germination in normal 
and vigorous seedlings, leading to faster and better ger-
mination in different crops (Cantliffe, 2003). Priming is 
responsible for the repair of age-related cellular and sub 
cellular damage of low vigour seeds that may accumu-
late during seed development. Priming of seed promotes 
germination by repairing the damaged proteins, RNA and 
DNA (Koehler et al., 1997).

Several treatments have been used with the aim of im-
proving strawberry achene (seed) germination: humidity 
(Guttridge and Bright, 1978); exposure to red light (Iyer et 
al., 1975); and osmotic pre-treatment (Hanke, 1993).

With the rapid growth of nanotechnology, there are 
increasing concerns about the potential adverse impact 
of engineered nanoparticles (ENPs) in the environment. 
However, our understanding of how ENPs may affect 
organisms within natural ecosystems lags far behind our 
rapidly increasing ability to engineer novel nanomateri-
als (Bernhardt et al., 2010). On the other hand, Lin and 
Xing (2007) studied the positive effects of suspensions 
of nanoparticles on seed germination and root growth 

of six different crop species [radish (Raphanus sativus), 
rape (Brassica napus), rye grass (Lolium perenne), lettuce 
(Lactuca sativa), corn (Zea mays) and cucumber (Cucumis 
sativus)]. Also, the effects of nano-TiO2

 (rutile) and non-
nano-TiO

2
 on the germination and growth of naturally aged 

spinach seeds were studied and an increase of these factors 
was observed at 0.25-4‰ nano-TiO

2
 treatment (Zheng et 

al., 2005). Feizi et al. (2011) reported that nano-TiO
2
 at 

suitable concentration could promote the seed germination 
of wheat in comparison to bulk TiO

2
 while at high concen-

trations it had an inhibitory or no effect on the crop.
Limited studies have been done on the effects of 

nanoparticles on crops and thus, we decided to investigate 
the phytotoxicity or positive effects of different concen-
trations of nano-TiO

2
 on seed germination and seedling 

growth of strawberry.

2. Materials and Methods

In order to evaluate the effect of nano priming on 
the quality of seedling production during germination in 
strawberry (Fragaria ananassa L.), this experiment was 
conducted in 2013 at Shahid Chamran University Ahvaz in 
Iran using a factorial design with eight replications. Seeds 
of strawberry (Fragaria ananassa cv. Queen Eliza) were 
from the Gene Bank of Iran, at Seed and Plant Improve-
ment Institute, Tehran (Karaj). The factors studied includ-
ed different concentrations of TiO

2 
 (Control, 3.5, 5.5, 7.5 

and 9.55%) and time (24 and 48 h). In order to prepare 
nano anatase solutions a stock solution with the highest 

Application of anatase nanoparticles (TiO2) on 
strawberry seed germination (Fragaria ananassa L.)

H.R. Hosseini*, M. Chehrazi*, E.H. Dehkourdi*, M. Hosseini**
*  Department of Horticultural Sciences, Faculty of Agricultural Sciences, University of Shadid Cham-

ran Ahvaz, Khuzestan, Iran.
** Department of Management Sciences, University of Vali Asr, Kerman, Iran.

Key words: Anatase, germination, nano, strawberry,TiO
2.

Abstract: Priming enhances germination, establishment and yields in a range of crops in many diverse environments. 
This experiment evaluated the effects of soaking strawberry seeds in different concentrations (0, 3.5, 5.5, 7.5 and 9.5 
percentage) of nano anatase on germination parameters (germination percentage, germination rate index, radicle and 
plumule length, fresh weight of seedlings and vigor index) using a factorial design with eight replications. Results showed 
that an increase in the concentration of nano anatase led to significant differences in the percentage of germination, ger-
mination rate index, root and shoot length, fresh weight and vigor index of seedlings. The best nano anatase concentration 
was found to be 7.5%.

Adv. Hort. Sci., 2013 27(4): 143-146

Received for publication 10 December 2013
Accepted for publication 14 January 2014



144

concentration (9.5%) was fixed and then with dilution of 
this stock solution, various concentrations of nano anatase 
were obtained. Strawberry seeds were sterilized using so-
dium hypochlorite solution (1%) for 10 min, then washed 
several times with distilled water, and soaked in nano-TiO

2
 

solutions at different concentrations. Seeds were placed in 
disinfected Petri dishes, each dish contained 100 seeds. 
All of the Petri dishes were irrigated using distilled water. 
Seeds were allowed to germinate at 25±3°C for 14 days. 
Every day the number of seeds with visible radicle were 
counted and recorded as sprouted seeds; the length of 
seedling shoot and root were also measured.

Germination rate [Eq. (1)] and germination percent-
age [Eq. (2)] were calculated using the following formula 
(Hosseini et al., 2013):

GR= ∑ n
∑ dn

 (1)

Where GR is the germination rate, Σn is the number 
of seeds germinated on the day, and Σdn is the number of 
days from the start of experiment.

GP= ∑ n
N

 ×100 (2)

Where GP is the germination percent, Σn is the number 
of seeds germinated until the last day of experiments, and 
N is the total number of seeds.

Seed vigor indices [Eq. (3)] were calculated according 
to the following formula (Hosseini et al., 2013):

VI= (RL+PL) ×GP (3)

Where VI is the Seed Vigor Index, RLis radicle length, 
PL is the plume length, and GP the germination percentage.

Statistical analysis
The data were analyzed using SAS 9.1 software. The 

significant levels of difference for all measured traits were 
calculated, and the means were compared by the multiple-
range Duncan test at 1% level.

3. Results and Discussion

Analysis of variance showed that treatment of nano an-
atase led to significant differences in germination percent-
age, germination rate, root and shoot length, allometric in-
dex and vigor index of the seedlings. Time treatment was 
not significant for root, shoot, seedling length and seedling 
allometric coefficient. Interaction of anatase nanoparticles 
and time treatment was not significant for root and shoot 
length and seedling allometric coefficient (Table 1-3).

The interaction of anatase nanoparticles and time 
showed the lowest percentage of germination in the treat-

Table 1  Analysis of variance of strawberry seed germination indices affected by nano anatase

Sources of
variation

df
Percentage of 
germination

Germination rate Shoot length Root length Seedling length Allometric VI

Nano-TiO
2

4 2260.6 * 0.184 * 11.077 * 15.366 * 0.525 * 3.33 * 103028.5 *

Time 1 136.9 * 1.089 * 0.196 ns 0. 016 ns 0.0022 ns 0.049 ns 5712.1 *

Nano*time 4 76.9 * 0.078 * 0.179 ns 0.0053 ns 0.001 ns 0.087 ns 3868.1 *

Error 30 13.7 0.180 0.148 0.145 0.004 0.069      578.23

*= significant at 1% level.
ns= non-significant.

Table 2 -  Effect of nano anatase on seed germination of strawberry after 24 hours

Concentration Germination (%) Germination rate Shoot length (cm) Root length (cm) Fresh weight (g)

0 % 10.97 e 0.60 d 1.78 e 2.0 d 0.012 d

3.5 % 40.10 d 0.60 c 8.50 d 4.5 c 0.021 c

5.5 % 68.80 ab 0.67 b 9.10 c 4.4 c 0.029 b

7.5 % 92.46 a 1.20 a 12.24 a 6.0 a 0.034 a

9.5 % 74.64 b 1.00 ab 10.00 b 4.8 b 0.030 b

Means with different letters at each column are statistically different at 1% level.

Table 3 - Effect of nano anatase on seed germination of strawberry after 48 hours

Concentration Germination (%) Germination rate Shoot length (cm) Root length (cm) Fresh weight (g)

0 % 0 e 0 e 0 e 0 d 0 d

3.5 % 23.2 d 0.17 d 3.5 d 2.5 c 0.011 c

5.5 % 40.6 b 0.26 b 5.3 c 2.9 c 0.017 b

7.5 % 87.6 a 0.56 a 10.3 a 5 a 0.030 a

9.5 % 46.3 bc 0.35ab 7.1 b 3.3 b 0.025 b

Means with different letters at each column are statistically different at 1% level.



145

ment without nanoparticles and the highest at a concentra-
tion of 7.5% of nanoparticles for 48 h (Fig. 1).

Zheng et al. (2007) reported that the significant effect 
of nano-sized TiO

2 
on germination is probably due to the 

small particle size which allows penetration into the seed 
during the treatment period, exerting its enhancing func-
tions during growth.

Results showed that the lowest germination rate result-
ed from treatment without nanoparticles while the highest 
rates were obtained at a concentration of 7.5% of nanopar-
ticles in 24 and 48 h. The key to increase seed geermina-
tion is the penetration of nanomaterial into the seed (Hash-
emi and Mousavi, 2013) (Fig. 2).

Khodakovskaya et al. (2009) mentioned that the carbon 
nanotubes can effectively penetrate through the seed coat, 
thus influencing seed germination. Exposure of tomato 
seeds to carbon nanotubes (CNTs) resulted in enhanced 
seed germination and growth rate.

Among the different nanoparticle TiO
2
 concentrations, 

7.5% for both times tested (24 and 48 h) showed the maxi-
mum seedling vigor index while the control showed the 
lowest (Fig. 3).

Effects positive of nano anatase are reported that na-
no-TiO

2
 (anatase) improved plant growth by enhanced 

nitrogen metabolism (Yang, 2006) that promotes the ab-
sorption of nitrate in spinach and, accelerating conver-
sion of inorganic nitrogen into organic nitrogen, thereby 
increasing the fresh weights and dry weights. Studies 
also showed the effects of nitrogen photoreduction on the 
improved growth of treated spinach plant (Yang, 2007). 
Effects of nano-TiO

2
 (anatase) on the content of light har-

vesting complex II (LHC II) on thylakoid membranes of 
spinach was studied and it showed an increase in LHC II 
content (Mingyu, 2007 a). These promote energy transfer 
and oxygen evolution in photosystem II (PS II) of spin-
ach (Hong, 2005).

It has also been found that nano anatase TiO
2
 promoted 

antioxidant stress by decreasing the accumulation of su-
peroxide radicals, hydrogen peroxide, malonyldialdehyde 
content and enhance the activities of superoxide dismutase, 
catalase, ascorbate peroxidase, guaiacol peroxidase and 
thereby increase the evolution oxygen rate in spinach chlo-
roplasts under UVB radiation (Mingyu, 2007 b).

The results indicated that nano sized TiO
2
 in an appro-

priate concentration could promote the seed germination 
and seedling growth of strawberry.

The results show that with increasing concentration of 
nano anatase to 7.5% increased germination parameters. 
The seedlings grown with nano anatase increased length 
when compare to the control seedlings. Effect of nano ana-
tase on root, shoot and seedling strawberry may be due 
to early emergence induced by nano anatase treatment as 
compared to control seeds. Rapid embryo growth resulted 
when the obstacle to germination was removed.

Although the concentration of 9.5% caused lower 
germination parameters toward concentration of 7.5%, 
showed more desirable effects of the concentration of 
5.5% nano anatase and control.

In all treatments, the better result was obtained in time 
of 48 hour. 

In order to understand the possible benefits of apply-
ing nanomaterials in agriculture, it is important to analyze 
penetration and transport of nanoparticles in the plants.

References

Fig. 1 -  Effect of interaction nano anatase and time on germination per-
centage of strawberry seeds.

Fig. 2 -  Effect of interaction nano anatase and time on germination rate 
of strawberry seeds.

Fig. 3 -  Effect of interaction nano anatase and time on vigor index of 
strawberry seeds.



146

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