Upload
others
View
34
Download
2
Embed Size (px)
Citation preview
Available online at www.worldscientificnews.com
WSN 49(2) (2016) 204-222 EISSN 2392-2192
Biological synthesis of Titanium Dioxide nanoparticles by Curcuma longa plant extract and
study its biological properties
Dr. Raghad DH. Abdul Jalill1, Rasha S. Nuaman1, Ahmed N. Abd2,* 1Department of Biology, College of Science, AL-Mustainsiriyah University, Baghdad, Iraq
2Department of Physics, College of Science, AL-Mustainsiriyah University, Baghdad, Iraq
*E-mail address: [email protected]
ABSTRACT
The objective of this study was biosynthesis of titanium dioxide nanoparticles (TiO2 NPs) using
Curcuma longa aqueous extract and characterize them. Study their effect on the growth, sporulation,
pathogenicity of Fusarium graminearum and some wheat plants parameters compared with standards
industrials nanoparticles. C. longa aquatic extract was used to biosynthesis TiO2 NPs by two methods.
At first method, TiO2 was found in both colloidal solutions (CS) and nanopawder while it found in jest
nanopawder in second method. All biosynthetic nanoparticles were in nano size. It was: 91.37 nm,
76.36 nm of (CS) and nanopawder for first methods respectively while it was 92.6 nm of nanopawder
in second method. All nanoparticles have good optical properties. Crystal’s shape of nanopawder were
in three form: anatase, rutilr and brookite and it was anatase in colloidal solutionat first method while
it was pure anatase innanopawder when at second method. The average crystallite size of was
calculate by Scherer's equation, it was 43.088 nm and 22.881 nm for nanopawder and colloidal
solution respectively at first method. It was 45.808 nm for nanopawder at second method. All
concentrations of nanoparticles were reduced fungal and spores. These decreasing were more effective
using biosynthetic compare with industrial synthetic nanoparticles. There were reductions in damping-
off caused by F. graminearum by biosynthetic NBs in both varieties of plant (Al-Rasheed and
Tamuze-2). These was better than the effect of industrial synthetic nanoparticles. The resistance to
damping off and the growth of plant in Al-Rasheed variety was more sensitive compared with
Tamuze-2 variety especially at higher concentrations. There were decrease in all plant's parameters at
most concentrations of TiO2 biological synthetic compare with industrial synthetic nanoparticles in Al-
Rasheed variety, while there were inductions in some plant's parameters by biosynthetic nanoparticles
World Scientific News 49(2) (2016) 204-222
-205-
compared with industrial synthetic in Tamuze-2 variety. Finally, C. longa can be used to biosynthesis
TiO2 NPs with good biological properties.
Keyword: biosynthesis; Curcuma longa; damping-off; nanoparticles; plant; TiO2
1. INTRODUCTION
Nanotechnology is the term used to cover the design, construction and utilization of
functional structures with at least one characteristic dimension measured in nanometers, (1).
Such materials can be designed to exhibit novel and significantly improved physical, chemical
and biological properties, phenomena and processes as a result of the limited size of their
constituent particles or molecules, (1). Generally metal nanoparticles can be prepared and
stabilized by physical, chemical and biological methods with little modifications for different
metals, (2), (3), (4), (5), (6). The properties of nanoparticles and its applications are different
and dependent on their size, distribution and morphology, (7), or even on synthesis methods.
The main disadvantages of physical methods are the quality of the product, which is less as
compared to nanoparticles produced by chemical methods. Usually these methods require
costly vacuum systems or equipment to prepare nanoparticles,(8).
Increasing sensibility towards green chemistry and biological processes has led to
develop an environment-friendly process for the synthesis of non-toxic nanoparticles. A vast
array of biological resources available in nature including living plants, (9), plant products,
plant crud extracts, algae, fungi, yeast, (10), bacteria, (11) and viruses could all be employed
for synthesis of nanoparticles. Biological methods are regarded as safe, cost-effective,
biocompatible, non-toxic sustainable and environment friendly processes, (12). In addition,
most bioprocesses occur under normal air pressure and temperature, resulting in vast energy
savings, high-yield and low cost, (13).
Recent reports of plants towards production of nanoparticles is said to have advantages
such as easily available, safe to handle and broad range of biomolecules which mediate
synthesis of nanoparticles (Salam et al., 2012) (4). Some mineral nanoparticles have been
synthesized inside live specimens of the plants Brassica juncea, Medicago sativa and
Heliannus annus, (9). Ag-NPs were successfully synthesized under room temperature at
different volumes of C. longa extract with an average size of 10.46, 8.18, and 4.90 nm for 5,
10, and 20 mL of aqueous C. longa tuber powder extract, with spherical-shaped structures
(14). Other plant mediated process to developed its extract for phytosynthesis of titanium
dioxide nanoparticles: Nyctanthes, Annona squamosapeel extract, (15), Catharanthus roseus
leaf aqueous extract (16) and Ecliptaprostrata (17), Azadirachta indica (18).
TiO2 nanoparticles become a new generation of advanced materials due to their brilliant
and interesting optical, dielectric, and photo-catalytic characteristics from size quantization. It
is one of the most widely used nanostructures in various Areas, (19). Some researches were
focused on its effect on bacteria, algae, plankton, fish, mice, and rats, (20), (21) but its effect
on fungi or plants' growth was unclear. Some studies suggest that interactions of TiO2 NPs
with other chemicals or physical factors may result in an increase in toxicity or adverse
effects, (22), such as photo-catalytic activity of titanium dioxide nanoparticles that prevents
the fungal colonization of wood samples over long time when compared to untreated ones,
(23). TiO2 nanoparticles have some effect on plant growth.
World Scientific News 49(2) (2016) 204-222
-206-
The results of (24) approve that TiO2 nanoparticles (NPs) (50 nm of anatase shape) were
increased promoter indicator of Latyfia variety of wheat but it has negative effect on
germination percentage, germination rate and Shoot and root length. Additionally, exposed
wheatgrass seeds to high concentrations of nano TiO2 particles (80 ppm) led to diminished
germination rate, (25). In the same time, (26) showed that the use of TiO2 NPs on the
germination of wheat at the proper and lower concentrations have additive effect and in high-
concentration have reduction effect.
The objective of this study was biosynthesis of TiO2 nanoparticles by C. longa plant
extract and characterized the products Moreover, we study its biological properties and if it is
different compared with the same size of industrial synthesis nanoparticles including their
effect on fungal growth and germinationof two varieties of wheat (T. aestivum).
2. MATERIALS AND METHODS
2. 1. Biological synthesis of Titanium Dioxide nanoparticles by C.longa plant extract
Preparation
Plant extraction: Fifteen gram of powdered material of C.longa was extracted by a
Soxhlet extractor with 300 ml of distal water at 40 °C for 3-4 h. The extract was filtered by
Whatman number 4 filter paper. The residue was removed. The filtrate was used forbiological
synthesis of nanoparticles directly as soon as possible.
Titanium Dioxide bulk particles (TiO2 BPs): it procured from Sigma Aldrich, China.
Its molar mass, density and size were: 79.87 g/mol, 4.2 g/cm3 and (400-800 nm), respectively.
Distil water was used to prepare solutionswith different concentrations.
Synthesis: This was done by two methods. In first method, 50 ml of filtrate was mixed
with 2.5 ml of titanium dioxide bulk particles (50 mg/ml), in flask. So, the final
concentrations of it was 2.38 mg/ml. It placed in magnetic steer hot plate with 50 C and 1000
rpm /second for 5 hr. The same above method had done in second method but the filtrate was
mixed with 5 ml of TiO2 bulk particles (TiO2 BPs), (50 mg/ml). The final concentrations of it
was 4.55 mg/ml. The period time was 8 hr. The solutions allowed to cool at room
temperature. The solution was repeated centrifugation at 15,000 rpm for 10 min. The
supernatant (colloidal solutions) kept for Characterization. The precipitate formed was
washed with double distilled water and then centrifuged at 1500 rpm for 10 min. This was
repeated three time. The obtained precipitate (nanopawder) was dried at room temperature for
24 h. and characterized as described fallowing.
Characterization
The exact configuration of the fabricated particles, phase purity, structure, average
particle size, morphology of crystals and distribution were measured using the fallowing
technique. Standard industrials NPs were, also, analyzed.
Atomic force microscopy (AFM): size, surface topography and granularity volume
distribution of biosynthesized nanoparticles characterized using Atomic Absorption
Spectroscopy (AA‐680, Shimadzu‐Japan), (characterized by Dr. Abdul Kareem Al-Samaraii
Lab. Baghdad/Iraq,(27).
World Scientific News 49(2) (2016) 204-222
-207-
UV–visible analysis: nanoparticles were recorded by a Schimadzu 1601
spectrophotometer in 200-800 nm range operated at a resolution of 1 nm (28). The absorption
coefficient (α) calculate from the optical spectrum using the formula, (29), (30): α = 2.3026
A/t. Where: (A) and (t) are the measured absorbance and thickness of the sample,
respectively. The optical band gap energy (Eg) evaluated from the absorption spectrum, and
the optical absorption coefficient (α) near the absorption edge gave according to (31):
where: h, ν, B, and Eg are Plank's constant, frequency of incident photons, constant, and
optical bandgap energy respectively. Energy gap of TiO2 NPs (Eg) was estimated by plotting
hν versus (αhν)1/2
according Tauc plot, (32).
X-ray diffraction. X-ray diffraction (XRD) was used to confirm the crystal structure
(crystal phases and crystallite size of each Phase ( of TiO2 nanoparticles. XRD analysis was
performed using an X-ray diffractometer with Cu-Kα crystal radiation (λ = 1.54 Å) scanning
at a rate of (5/min-1
) for (2θ) range of (20º-70º). The diffraction peaks were identified by
comparison with (JCPDS-84-1286), (JCPDS 21-1272), according 2θ. The full width at half
maximum (FWHM) in the XRD was used to determine the crystallite size using Scherer's
equation (33).
The strain value and the dislocation density value can be evaluated by using the
relations in the fallowing equations:
( )
( )
SEM analysis: TiO2 biosynthetic nanoparticles were analysis by scanning electron
microscope (SEM), Vega Tescan (USA), in the Center of Nanotechnology and Advanced
Materials/ University of Technology/ Iraq. Biological synthesis nano-powder (which
synthesis according above first method) used to compare its effect, in all following
experiments, with the effect of industrial synthetic NPs, which procured from Sigma Aldrich
(USA). These nanoparticles were white color, anatase shape withsize 50 nanometer, assay
99.0%. Sterilized distilled water was used to prepare different concentrations of industrial and
biological synthesis NPs.
Antifungal and anti-pathogenic activity of nanoparticles
A pure isolate of F. graminearum fungi were get from Department of Biology, College
of Science, AL-Mustainsiriyah University, Baghdad, Iraq. The antifungal activity of all
nanoparticles were evaluated against F. graminearum. PDA medium prepared with different
concentrations of nanoparticles. There was negative control of distill water in all experiment.
All experiment has run in three replicate. Petri dishes were inoculated in the center with 4 mm
of fungal plugs. Incubated at 28 ±2 °C for 8-10 days. The radials growth of the colonies
measured. The percentage of inhibition of mycelial growth was calculated. The spore
World Scientific News 49(2) (2016) 204-222
-208-
suspensions, (10 ml of sterile distilled water per petri old dishes), was collected,centrifuged
then calculated by hemocytometer. The percentage of spores’ inhibition rate was calculated.
Anti-pathogenic activity: two variety of dry wheats' seeds (T. aestivum) (Al Rasheed
variety and Tamuze-2 variety) were taken from Ministry of Science and Technology- Seed
Technology Center. Germination percentage of them were 100% for both varieties. Mycelium
agar plug technique was used for pathogenicity test, (36). Seeds immersed in a 1% sodium
hypochlorite solution for three minute then washed with sterilized distilled water three
times.Washed seeds were soaked with various concentrations of each of industrial and
biological synthetic nanoparticles for 72 hours. Five seeds per petri dishes, were inoculated
with (0.8 cm) diameter of old culturesmycelium on the center of petri dishes.Untreated seeds
exposed to fungi and untreated seeds without exposed to fungi were used as control negative
and positive respectively. Five replicate were get for each treatment. Each replicate contains
ten seeds. The seeds were germinated in 28 C for ten days. The symptoms were: seedlings
fail to emerge (pre-emergence damping off) or seedlings collapse, submerged in a mass of
whitish fungal growth. The number of dead seeds or dead seedling were determined after
seven days to calculate total percentage of damping- off as fallowing: Damping – off % = [(S-
s)/S]*100. Where is: S = average of germinated seed in control plates, s = average of
germinated seed in platestreated with NPs.
2. 2. The effect of nanoparticles on germination parameters of wheat.
The seeds of two variety of wheat (T. aestivum) (Al Rasheed variety and Tamuze-2
variety) were soaked in different concentrations of nanoparticles suspensions for 72 hours.
There was negative control (distill water) for all experiment. All seeds were incubating at (27
±1 C, 12 h. light: 12 h. dark). All experiment has run in three replicate. The number of new
germinated seeds was recorded daily. A seed was considered germinated when the radicle
showed at least 2 mm in length. The fallowing parameters were calculating: germination
percentage, (37), germination rate, (38) and mean germination time, (37).
3. RESULTS
3. 1. Biological synthesis of TiO2 NPs by C.longa plant extract
First method: In the first method, TiO2 was found in both supernatant and precipitate. We
called them colloidal solutions and nanopawder respectively. Size range were (80 - 110) and
(50 - 110) nm with average diameter: 91.37 nm, 76.36 nm of colloidal solutions and in
nanopawder (precipitate) respectively, (Figure 4).Root mean square (RMS) and roughness
average (RA) of colloidal were: 0.872 nm and 0.745 nm. While root mean square (RMS) and
roughness average (RA) of nanopawder were: 0.773 nm and 0.674 nm, respectively. Figure
(1) showed AFM topographic images of biosynthesis TiO2 nanoparticles by first method.
The XRD pattern of this biosynthesis nanopawder showed the presence of seven peaks.
Strong diffraction peaks were: (25.3201° (101), 48.0516° (200) and 37.818° (004) indicating
that nanoparticles structure of these peaks were anatase crystalline. Other peaks were: 27.5°
(110) and 54.4° (211) corresponded to anatase form and one brookite form (crystallographic
plane = 121) with 2 theta = 30.8°. The average crystallite size of TiO2 nanoparticles was
calculate by Scherer's equation, it was 43.088 nm. The XRD patterns of biosynthesis colloidal
World Scientific News 49(2) (2016) 204-222
-209-
solutions shown in Figure (3) indicated that it was in the form of anatase TiO2. Optimum
peaks at 25.37° corresponded to the 101 planes of anatase form. Table (1) showed a summary
of X-ray characterization of TiO2 biosynthesis nanoparticles. Scherer's equation was used to
calculate the average crystallite size of TiO2 nanoparticles. It was 22.881 nm. The XRD
patterns of pure stander industrial synthetic nanoparticles were shown in Figure (3). The
crystal shape of them was pure anatase form. Strong diffraction peaks were: 25.3425°,
48.0686°, 37.8455°, 53.86°, 62.78°, 68.78° and 75.2° corresponded to: (101), (200), (004),
(111), (002) (116) and (215) crystallographic planes. The average crystallite size of TiO2
nanoparticles (according to Scherer's equation) was 16.473 nm. Figure (4) showed SEM
image of TiO2 biosynthesis nanoparticles (nanopawder) using first method. Particle size (>50
nm).
A
C
Fig. 1. (A) Granularity volume distribution chart of TiO2 NPs synthesis by C. longa plant
extract using first method. (B) and (C): AFM topographic images of colloidal solutions and
nanopawder respectively.
B
World Scientific News 49(2) (2016) 204-222
-210-
Industrial synthetic NPs
Biosynthesis NPs (nanopawder)
World Scientific News 49(2) (2016) 204-222
-211-
0
20
40
60
80
100
120
140
160
180
10
12
,42
14
,84
17
,26
19
,68
22
,12
4,5
22
6,9
42
9,3
63
1,7
83
4,2
36
,62
39
,04
41
,46
43
,88
46
,34
8,7
25
1,1
45
3,5
65
5,9
85
8,4
60
,82
63
,24
65
,66
68
,08
70
,57
2,9
27
5,3
47
7,7
6
Inte
nct
y (a
. u
)
2 Theta (deg.)
A (101)
Fig. 3. X-ray pattern of TiO2 biosynthesis NPs (nanopawder) using first method. Identified
phases of anatase (A), rutile (R) and brookite (B).
Table 1. Summary of X-ray characterization of TiO2 biosynthesis NPs (nanopawder and
colloidal solutions) compared with industrial synthetic NPs.
Methods Sample (hkℓ)
planes
Crystal
shape
2 theta
(DEG)
FWHM
(deg)
D
(nm)
x10-4
(lines2m-4
)
x 1014
(lines/m2)
Firstbiosynthesis
method
nanopawder
101 anatase 25.320 0.198 40.926 33.866 5.971
200 anatase 48.052 0.175 49.474 28.015 4.086
004 anatase 37.818 0.215 38.864 35.663 6.6207
colloidal 101 anatase 25.37 0.354 22.881 60.574 19.1
second
biosynthesis
method
nanopawder
101 Anatase 25.330 0.179 45.222 30.649 4.89
200 Anatase 48.075 0.168 51.602 26.86 3.756
004 Anatase 37.814 0.206 40.601 34.137 6.066
industrial
synthetic nanopawder
101 anatase 25.343 0.4957 16.34 84.825 37.456
200 anatase 48.067 0.529 16.368 84.678 37.327
004 anatase 37.846 0.5 16.713 82.93 35.801
(hkℓ) planes: crystallographic plane; FWHM: Full width at half maximum; D: dimension of
Crystal in nm; x10-4
: strain value; x 1014
: dislocation density.
Biosynthesis NPs (colloidal
solutions)
World Scientific News 49(2) (2016) 204-222
-212-
Fig. 4. SEM image of TiO2 biosynthesis NPs (nanopawder) using first method.
Particle size (>50 nm).
Second method: TiO2 was found in precipitate only as nanopawder and absent in supernatant
(colloidal solution). Size of nanoparticles wererange between (10 - 140) nm with average
diameter 92.6 nm of nanopawder. The percentage volume of particles with size 30 nm was
29.145. Figure 5 (B), showed AFM topographic images of TiO2 nanoparticles synthesis by
C.longa plant extract (supernatant) using second method.
A
World Scientific News 49(2) (2016) 204-222
-213-
Fig. 5. (A) Granularity volume distribution chart of TiO2 NPs powder synthesis by
C. longa plant extract using second method. (B): AFM topographic images.
The XRD patterns shown in Figure (7) indicated that the crystal shape of biosynthesis
nanopawder was pure anatase form. Optimum peaks were: 25.3302°, 48.0745 °, 37.8144°,
53.86° and 62.82° corresponded to: (101), (200), (004), (105) and (002) crystallographic
planes, Figure (7). Table (1) showed a summary of X-ray characterization of TiO2
biosynthesis nanoparticles. The average crystallite size of TiO2 nanoparticles was calculate
by Scherer's equation, it was 45.808 nm.
Fig. 7. X-ray pattern of TiO2 biosynthesis NPs (nanopawder) using second method. Identified
phases is pure anatase (A).
0
100
200
300
400
500
600
700
10
12
,52
15
,04
17
,56
20
,08
22
,6
25
,12
27
,64
30
,16
32
,68
35
,2
37
,72
40
,24
42
,76
45
,28
47
,8
50
,32
52
,84
55
,36
57
,88
60
,4
62
,92
65
,44
67
,96
70
,48
73
75
,52
78
,04
Inte
nct
y (a
. u)
2 Theta (deg.)
A (101)
A (004)
A (200)
A (105)
A (002)
B
World Scientific News 49(2) (2016) 204-222
-214-
SEM analysis: Figure (8) showed SEM image of TiO2 biosynthesis nanoparticles (nano-
pawder) using second method. Particle size (<90 nm).
Fig. 8. SEM image of TiO2 biosynthesis nanoparticles (nanopawder) using first method.
Particle size (<90 nm).
There are some advantages of using plants for the synthesis of nanoparticle such as:
they are easily, available, safe to handle and possess a broad variability of metabolites that
may aid in reduction. A number of plants are being currently investigated for their role in the
of nanoparticle, (39). According to (40) plant extracts are believed to act as reducing and
stabilizing agents in the nanoparticle synthesis. The nature of plant extract affects the kind of
nanoparticles synthesized in a highly critical manner with the source of plant extract being the
most vital factor and different concentrations of biochemical reducing agents affecting the
morphology of synthesized nanoparticles, (41).
In current study C. longa successful to synthesis pure anatase nano-size TiO2 NPs. This
ability of this plant may be due to presence of terpenoids, flavonoids and proteins was
considered to be responsible for the formation and stabilization of titanium nanoparticles,
(42). The different concentrations of C. longa plant extractmay be responsible for different
sizes nanoparticles that synthesis in current result. This result similar to(14) who reported that
Ag-NPs were synthesized with an average size of 10.46 ±5.58, 8.18 ±3.53, and 4.90 ±1.42 nm
for 5, 10, and 20 mL of different concentrations of plantaqueous extract, with spherical-
shaped structures. There are no reports about biosynthesis of TiO2 NPs by C. longa but there
were some reports of other plants which able to synthesis TiO2 NPs, for example: Nyctanthes
success to synthesis titanium dioxide nanoparticles with size of nanoparticles ranging from
100 to 150nm, (43). TiO2 NPs were synthesized from Annona squamosapeel extract (Roopan
et al., 2012) (16), Catharanthus roseus leaf aqueous extract (16), Ecliptaprostrataextract (44),
World Scientific News 49(2) (2016) 204-222
-215-
and Azadirachtaindica extract(45). (42) reported that TiO2 NPs biologically synthesis using
leaf extract of Azadirachta indica, with mixture of rutile and anatase phases, size ranged from
15 to 42 nm. (46) reported that Euphorbia prostrata extract can synthesis (TiO2) NPs with
spherical shape and an average size 83.22 nm. (47) successes to biosynthesis of tetragonal
TiO2 NPs with (32 nm) average in particles size from Aloe vera extract.
3. 2. Antifungal and anti-pathogenic activity of nanoparticles
Table (2) showed that the highest fungal and spore Inhibition rate found in (200 mg/ml)
of industrial syntheticnanoparticles, while the lowest fungal Inhibition rate found in (20, 0.2)
mg/ml. The highest fungal and spore Inhibition rate were found at (20 mg/ml) of biological
synthetic nanoparticles and the lowest of them were found at (0.2 mg/ml).There were decrease
in fungal and spore inhibition rate at all concentrations of biological synthetic compare with
industrial syntheticnanoparticles except (20 mg\ml) concentration, it has the same spores’
inhibition rate in each treatment (40.816 %). In both varieties, Rasheed and Tamuze-2, there
were decrease in percentage of damping-off at all concentrations of biological synthetic
compare with industrial synthetic nanoparticles. The response to nanoparticles of Al-Rasheed
variety was more sensitive to resistance damping off compared with Tamuze-2 variety.
Table 2. Comparison between the effect of industrial synthetic NPs and biological
syntheticNPs on Inhibition rate, Spores inhibition rate and pathogenicityof F. graminearum
Con.
(mg/ml)
% F.IR % S.IR % F.IR % S.IR % Path.
IS BS IS BS Al-Rasheed variety Tamuze-2 variety
IS BS IS BS
200 39.583 53.061 29.166 34.693 50 33.333 50 40
20 12.5 40.816 31.25 40.816 46.667 40 56.667 46.667
2 14.583 46.938 12.5 20.408 53.333 46.667 56.667 50
0.2 12.5 40.816 8.333 6.122 53.333 56.667 53.333 63.333
CT- 0 0 0 0 100 100 100 100
CT-2 - - - - 0 96.667 0 96.667
Con.: concentrations; F.IR: fungal inhibition rate; S.IR: spores’ inhibition rate; Path.:
pathogenicity;IS: industrial synthetic NPs; BS: biological synthetic NPs; CT-: untreated fungi
or untreated seeds exposed to fungi; CT-2: untreated seeds without exposed to fungi.
3. 3. Effect of nanoparticles on germination parameters of T. aestivum
In Al-Rasheed variety, there were decrease in all plant's parameters at all concentrations
of biological synthetic NPs compare with industrial synthetic NPs except (20-2 mg\ml)
World Scientific News 49(2) (2016) 204-222
-216-
concentrations. These concentrations of biological synthetic were induction mean germination
time compare with industrial synthetic. In compared with control, there were:
a. Germination percentages: There were reduction in seed germination using different
concentrations of industrial and biosynthetic nanoparticles.
b. Germination rate: The higher concentration of industrial synthesis nanoparticles
reduced these parameters significantly. The same reduction found using (2-200)
mg/ml of biosynthesis nanoparticles, (P < 0.05). All other treatments were not
affected.
c. Mean germination time: it was not affected by all concentrations of industrial or
biosynthesis nanoparticles, (P < 0.05), except the induction in these parameter at 2
mg/ml of biosynthesis nanoparticles, significantly. All other treatments were not
affected.
d. Mean daily germination: each concentrationsof: (20-200) mg/ml of industrial
synthesis nanoparticles and (2-200) mg/ml of biosynthesis nanoparticles were
reduced MDG.
e. Germination value: the reduction in these parameters weresignificance at all
treatment of biosynthesis nanoparticles while these significant reduction was found
at (20-200) mg/ml concentrations of industrial synthesis.
f. Promoter Indicator: the reduction in these parameters weresignificance at all
treatment of industrial synthesis while these significant reduction was found only at
higher concentrations, (Table 3).
In Tamuze-2 variety: the comparison between the effect of industrial and biosynthetic
nanoparticles were different in action. The decreasing was found in different concentrations of
only promoter indicator. While there were inductions by biosynthetic nanoparticles compared
with industrial synthetic in: germination percentage (2-20) mg/ml, germination rate (2-200)
mg/ml, mean germination time (all concentrations), mean daily germination (2-20) mg/ml and
germination value (2-20) mg/ml. All other concentrations were reduced above parameters.
a. Germination percentages: There were reduction in seed germination using different
concentrations of industrial and biosynthetic nanoparticles.
b. Germination rate: The changing in these parameters were not significant in all
treatment except the induction on it at (200 mg/ml) of biological synthetic
nanoparticles.
c. Mean germination time: There were reduction in it using different concentrations
of industrial and biosynthetic nanoparticles at (P < 0.05).
d. Mean daily germination: The same feature found in MDG.
e. Germination value: the significant reduction found at (20 mg/ml) of industrial
synthesis and (20-200) mg/ml of biosynthetic nanoparticles. All other treatments
were not significant.
f. Promoter Indicator: the reduction in these parameters weresignificance at all
treatment of biosynthesis nanoparticles while these significant reduction was found
at (20-200) mg/ml concentrations of industrial synthesis.
Overall, the result found that Al-Rasheed variety was more sensitive to nanoparticles
compared with Tamuze-2 especially at the higher concentrations.
World Scientific News 49(2) (2016) 204-222
-217-
Table 3. Comparison between the effect of industrial synthetic NPs and biological synthetic
NPs on Germination percentage, Germination rate, Mean germination time, mean daily
germination, Germination Value and Promoter Indicator of Al-Rasheed variety of
T. aestivum.
Co
n.
(mg
/ml)
Industrial synthetic nanoparticles Biological synthetic nanoparticles
V.
% G
P
GR
MG
T
MD
G
GV
PI
% G
P
% G
R
MG
T
MD
G
GV
PI
Al-
Ra
shee
d
200
66
.667
0.5
85
B
2.6
00
A
66
6.6
67
B
78
3.3
33
B
3.2
50
B
43
.333
0.1
79
B
2.3
33
A
43
3.3
33
B
50
0.0
00
B
1.0
83
B
20
66
.667
0.7
16
A
2.6
67
A
66
6.6
67
B
96
6.6
67
B
3.5
83
B
66
.667
0.5
364
B
3.1
67
A
66
6.6
67
B
96
6.6
67
B
2.3
33
A
2
86
.667
0.8
52
A
2.8
67
A
86
6.6
67
A
14
66
.66
7B
5.0
83
B
83
.333
0.7
89
B
3.8
00
B
83
3.3
33
B
13
66
.66
7B
2.8
33
A
0.2 90
.
1.2
31
A
3.2
33
A
90
0.0
0A
19
83
.33
3A
4.1
67
B
83
.333
1.3
14
A
3.0
67
A
83
3.3
33
A
18
00
.00
0B
2.9
17
A
CT-
10
0.
1.1
95
A
2.8
67
A
10
00
.00
A
28
33
.33
3A
6.9
17
A
10
0.0
00
1.1
95
A
2.7
00
A
10
00
.00
0A
28
33
.33
3A
3.6
67
A
Ta
mu
ze-
200
76
.67
1.7
8A
1.8
7B
76
6.6
7B
15
00
.00
A
3.4
2B
60
2.2
66
B
2.3
7B
60
0B
12
00
B
1.6
7B
20
56
.67
0.7
3A
1.8
7B
56
6.6
7B
80
0.0
0B
B
2.9
2B
73
.33
1.2
07
A
2.5
7B
73
3.3
3B
10
00
B
2.2
5B
2
76
.67
1.4
A
1.8
0B
76
6.6
7B
14
16
.67
A
4.1
7A
80
1.4
46
A
2.6
0B
80
0B
14
66
.67
A
2.5
8B
World Scientific News 49(2) (2016) 204-222
-218-
0.2
86
.67
2.1
3A
1.8
7B
86
6.6
7B
18
66
.67
A
4.5
8A
76
.67
1.4
40
A
2.3
7B
76
6.6
7B
16
66
.67
A
2.5
8B
CT-
96
.67
1.6
5A
3.1
0A
96
6.6
7A
19
33
.33
A
4.7
5A
96
.67
1.6
5A
3.1
0A
96
6.6
7A
19
33
.33
A
4.7
5A
Data shows: means; V: variety of plant; Con.: concentration; CT - : untreated; GP:
Germination percentage; GR: Germination rate; MGT: Mean germination time; MDG: mean
daily germination; GV: Germination Value; PI: Promoter Indicator; Similar letters are not
significance at (P < 0.05) vertically.
This study agrees with the positive and negative effect results of (24) who approve that
TiO2 industrials nanoparticles (NPs) (50 nm of anatase shape) were increased promoter
indicator of Latyfia variety of wheat but it has negative effect on germination percentage,
germination rate and Shoot and root length. It, also, increased total number of chemical
compounds that identified in leaves plant compared with control. Additionally, exposed
wheatgrass seeds to high concentrations of nano TiO2 particles (80 ppm) led to diminished
germination rate, (25). In the same time, (26) showed that the use of TiO2 NPs on the
germination of wheat at the proper and lower concentrations have additive effect and in high-
concentration have reduction effect.This positive and negative effects of NPs on plant growth
may depends on the concentration, size, particles shape and physical and chemical properties
of NPs, plant species, (48) as well as the methods of NPs synthesis. Some studies found that
physiological and chemical methods for NPs synthesis were more toxic to living cells and
vice versa. In current studies, there were decrease in all plant's parameters at most
concentrations of TiO2 biological synthetic compare with industrial synthetic nanoparticles in
Al-Rasheed variety, while there were inductions in some plant's parameters by biosynthetic
nanoparticles compared with industrial synthetic in Tamuze-2 variety. (49) studied the effect
of biologically synthesized Ag NPs on hydroponically grown Bacopa monnieri growth
metabolism, and found that biosynthesized AgNPs showed a significant effect on seed
germination and induced the synthesis of protein and carbohydrate and decreased the total
phenol contents and catalase and peroxidase activities.
4. CONCLUSIONS
C. longa can be used to biosynthesis TiO2 NPs by two methods. All biosynthetic
particles were in nano size with good optical properties. Crystals shape were in three form
anatase, rutilr and brookite when first method was used while it was pure anatase when
second methods was used. This Nanoparticles were reduced fungal growth, spores and
pathogenicity of F. graminearum. Al-Rasheed variety of wheat plant was more sensitive to
resistance damping off compared with Tamuze-2 variety. But its growth was more sensitive to
industrial and biological synthetic NPs compared with Tamuze-2 especially at higher
concentrations. More study of the stability of TiO2 NPs produced by C. longa and if it
World Scientific News 49(2) (2016) 204-222
-219-
agglomerates to be in micro-size are benefit. Study its activity against other organism
compared with bulk and industrial nanoparticles will gave good knowledge.
References
[1] Kelsall, R W.; Hamley, I W. and Geoghegan. M. (2005). Nanoscale Science and
Technology. John Wiley & Sons Ltd, 473p.
[2] Zhou, R.; Wu, X.; Hao, X.; Zhou, F.; Li, H. Li, H. and W. Rao. (2008). Influences of
surfactants on the preparation of copper nanoparticles by electron beam irradiation,
Nuclear Instruments and Methods in Physics Research B: Beam Interactions with
Materials and Atoms, 266(4): 599-603.
[3] Soomro, R.A.; Hussain, S. T.; Sherazi, Sirajuddin,; Memon1, N.; Shah, M. R.; Kalwar,
N.H.; Hallam, K. R.. and Shah, A. (2014). Synthesis of air stable copper nanoparticles
and their use in catalysis. Adv. Mat. Lett. 5(4): 191-198.
[4] Salam, HA.; Rajiv, P.; Kamaraj, M.; Jagadeeswaran, P.; Gunalan, S. and Sivaraj, R.
(2012). Plants: Green Route for Nanoparticle Synthesis. I. Res. J. Biological Sci., 1(5):
85-90.
[5] Sarkar, M.B.; Datta, J.; Mondal, D. and Mukhopadhyay, S. (2013). Synthesis and
morphology of silicon nanoparticles by deposition time varying LPCVD method to
demonstrate the variation of height, density and size. International Journal of Research
in Engineering and Technology. 2(8): 400-404.
[6] khashan, K.S. (2013). Synthesis, Structural and Optical Properties of Cds Nanoparticles
Prepared by Chemical Method. Eng. & Tech. Journal 31(1): 39-48.
[7] Pérez, J.; Bax, L. and Escolano. C. (2005). Roadmap report on nanoparticles. Barcelona,
Spain: Willems & van den Wildenberg (W&W) (ES/NL). pp. 57.
[8] Wu, SH. and Chen, DHJ. (2004). Synthesis of high-concentration Cu nanoparticles in
aqueous CTAB solutions. J. Colloid Interface Sci., 273: 165-169.
[9] Bali, R.; Razak, N.; Lumb, A. and Harris, A.T. (2006). The synthesis of metallic
nanoparticles inside live plants. Laboratory for Sustainable Technology, School of
Chemical and Biomolecular Engineering, The University of Sydney, NSW, Australia.
4p.
[10] Castro, L.; Blázquez, M.L.; Muñoz, J.Á.; González, F.G. and Ballester, A. (2014).
Mechanism and Applications of Metal Nanoparticles Prepared by Bio-Mediated
Process. Reviews in Advanced Sciences and Engineering 3: 1-18.
[11] Jha, Z.; Behar, N.; Sharma, S N.; Chandel, G.; Sharma, D. and Pandey, M. (2011).
Nanotechnology: prospects of agricultural advancement. Nano Vision, 1(2): 88-100.
[12] Shah, M.; Fawcett, D.; Sharma, S.; Tripathy, S.K. and Poinern, G.E. J. (2015). Green
Synthesis of Metallic Nanoparticles via Biological Entities. Materials 8: 7278-7308.
World Scientific News 49(2) (2016) 204-222
-220-
[13] Makarov, VV.; Love, AJ.; Sinitsyna, OV; Makarova, SS.; Yaminsky, IV.; Taliansky,
ME, Kalinina, NO. (2014). Green” nanotechnologies: synthesis of metal nanoparticles
using plants. Acta. Naturae, 6(1): 35-44.
[14] Shameli, K.; Bin Ahmad, M.; Shabanzadeh.; P. Al-Mulla, EA.; Zamanian, A.;
Abdollahi, Y.; Jazayeri. SD.; Eili, M.; Jalilian, FZ. And Haroun, RZ. (2013). Effect of
Curcuma longa tuber powder extract on size of silver nanoparticles prepared by green
method. Res Chem Intermed, 40: 1313-1325.
[15] Roopan, SM; Bharathi, A.; Prabhakarn, A.; Rahuman, AA; Velayutham, K. and
Rajakumar, G. (2012). Efficient phyto-synthesis and structural characterization of rutile
TiO2 nanoparticles using Annona squamosapeel extract. Spectrochim Acta. A Mol.
Biomol. Spectrosc. 98: 86-90.
[16] Velayutham, K.; Rahuman, AA.; Rajakumar, G.; Santhoshkumar, T.; Marimuthu, S.
and Jayaseelan ,C. (2011). Evaluation of Catharanthus roseusleaf extract-mediated
biosynthesis of titanium dioxide nanoparticles against Hippobosca maculata and
Bovicola ovis. Parasitol Res, 111(6): 2329-2337.
[17] Gong, P.; Li, H.; He, X.; Wang, K.; Hu, J. and Zhang, S. (2007). Preparation and
antibacterial activity of Fe3O4, Ag nanoparticles. Nanotechnology, 18(28): 604-611.
[18] Siegel, RW.; Hu, E. and Roco, MC. (1999). Nanostructure Science and Technology: R
& D Status and Trends in Nanoparticles, Nanostructured Materials, and Nanodevices.
Kluwer Academic Publishers, Boston, pp. 336.
[19] Seabra, AB. And Duran, N. (2015). Nano toxicology of metal oxide nanoparticles.
Metals, 5(2): 934-975.
[20] Mahmoodzadeh, H; Nabavi, M. and Kashefi, H. (2013). Effect of nanoscale titanium
dioxide particles on the germination and growth of canola (Brassica napus). J.
Ornamental Hortic Plants, 3: 25-32.
[21] Jesline, A.; John, N. P.; Narayanan, P. M.; Vani, C. and Murugan, S. (2015).
Antimicrobial activity of zinc and titanium dioxide nanoparticles against biofilm-
producing methicillin-resistant Staphylococcus aureus. Applied Nanoscience 5(2): 157-
162.
[22] Liu, K.; Lin, X. and Zhao, J. (2013). Toxic effects of the interaction of titanium dioxide
nanoparticles with chemicals or physical factors. Int J Nanomedicine ; 8: 2509-2520.
[23] Filpo, G.D.; Palermo,A.M.; Rachiele,F. and Nicoletta, F.P. (2013). Preventing fungal
growth in wood by titanium dioxide nanoparticles. International Biodeterioration &
Biodegradation. 85: 217-222.
[24] Yousef, A M. (2015). Effect of titanium dioxide TiO2 nano and bulk particles on seeds
germination and growth of wheat (Triticum aestivum L.) and rice (Oryza sativa L.).
DMS in Biology/Botany/ College of Science/ Al-Mustansiriyah University.
[25] Azimi, R.; Feizi, H. and Hosseini, M. K. (2013). Can bulk and nanosized titanium
dioxide particles improve seed germination features of wheatgrass (Agropyronde
sertorum)? Notulae Scientia Biologicae, 5(3): 325-331.
World Scientific News 49(2) (2016) 204-222
-221-
[26] Feizi, H.; Moghaddam, R. P.; Shahtahmassebi, N. and Fotovat, A. (2012). Impact of
bulk and nanosized titanium dioxide (TiO2) on wheat seed germination and seedling
growth. Biological Trace Element Research, 146(1): 101-106.
[27] Naveen, H. K.S.; Gaurav Kumar.; Karthik L.; and Bhaskara Rao K.V. (2010).
Extracellular biosynthesis of silver nanoparticles using the filamentous fungus
Penicillium sp. Archives of Applied Science Research, 2(6): 161-167.
[28] Ba-Abbad, M M.; Kadhum, A H.; Mohamad, A B.; Takriff, M S. and Sopian, K.
(2012). Synthesis and catalytic activity of TiO2 nanoparticles for photochemical
oxidation of concentrated chlorophenols under direct solar radiation. Int. J.
Electrochem. Sci. 7: 4871-4888.
[29] Cottrell, A. (1975) Introductions to Metallurgy, Arnold, London, P.173.
[30] Longhurst, R.S. (1957). Geometrical and physical optics, Longmans green, Londan.
[31] Kumar, V.; Khan, K L A.; Singh, G.; Sharma, T P. and Hussain, M. (2007). ZnSe
sintered films: Growth and characterization. Appl . Surf. Sci., 253(7): 3543-3546
[32] Tauc, J.; (1974). Amorphous and Liquid Semiconductors, Plenum Press, London and
New York. pp. 159.
[33] Cullity, BD., Elements of X-ray Diffraction, IInd Ed, Addison Wesley, London. 1974.
[34] Wei, W.; Mao, X.; Ortiz LA.; Sadoway DR. (2011). Oriented silver oxide
nanostructures synthesized through a template-free electrochemical route, Journal of
Materials Chemistry, 21(2): 432-438.
[35] Jobst, P.J.; Stenzel, O; Schürmann, M.; Modsching, N.; Yulin, S.; Wilbrandt, S.; Gäbler,
D.; Kaiser N. and Tünnermann, A. (2013). Optical properties of unprotected and
protected sputtered silver films: Surface morphology vs. UV/VIS reflectance. Adv. Opt.
Techn. 3: 91-102.
[36] Gargouri, K L.; Gargouri, S.; Rezgui, S.; Trifi, M.; Bahri, N.; and Hajlaoui, M.R.
(2009). Pathogenicity and aggressiveness of Fusarium and Microdochium on wheat
seedlings under controlled conditions. Tunisian Journal of Plant Protection, 4: 135-144.
[37] Feizi, H.; Kamali, M.; Jafari, L. and Moghaddam, P.R. (2013). Phytotoxicity and
stimulatory impacts of nanosized and bulk titanium dioxide on fennel (Foeniculum
vulgare Mill). Chemosphere 91(4): 506-511.
[38] AL-Kaisi, W.A., Muhsen, T.A.A., Hamed, A.S.: 'Effect of mycorrhiza (Glomus
mosseae) and superphosphate on physiological characters of Hodeum vulgare', College
of Basic Education Journal, 72 (2011) 765-784.
[39] Torresdey, J. L.; Parsons, J. G.; Gomez, E.; Peralta-Videa, J.; Troiani, H. E.; Santiago,
P.and Yacaman, M. J.(2002). Formation and Growth of Au Nanoparticles inside Live
Alfalfa Plants, Nano Lett. 2(4); 397-401.
[40] Mukunthan, K. and S. Balaji, S. (2012). Cashew apple juice (Anacardium occidentale
L.) speeds up the synthesis of silver nanoparticles. International Journal of Green
Nanotechnology, 4(2): 71-79.
World Scientific News 49(2) (2016) 204-222
-222-
[41] Li, X.; Xu, H.; Chen, Z.S. and Chen, G. (2011). Biosynthesis of nanoparticles by
microorganisms and their applications, Journal of Nanomaterials, 1-16.
[42] Krishnasamyet, A; Sundaresan, M.; and Velan, P. (2015). Rapid phytosynthesis of
nano-sized titanium using leaf extract of Azadirachta indica. International Journal of
ChemTech Research 8(4): 2047-2052.
[43] Sundrarjan, M. and Gowri, S. (2011). Green synthesis of titanium dioxide nanoparticles
byNyctanthes arbor-tristis leaves extract. Chalcogenide Letters 8(8): 447-451.
[44] Rajakumar, G.; Rahuman, A.A.; Priyamvada, B. V.; Khanna, G.; Kishore, K D. and
Sujin, P.J. (2012). Ecliptaprostrata leaf aqueous extract mediated synthesis of titanium
dioxide nanoparticles. Mater Lett., 68: 115-117.
[45] Sankar, R.; Rizwana, K.; Shivashangari, KS. and Ravikumar, V. (2014). Ultra-rapid
photocatalytic activity of Azadirachta indica engineered colloidal titanium dioxide
nanoparticles. Appl.Nanosci, 5: 731-736.
[46] Abduz Zahir, A.; Singh, IC.; Bagavan, A.; Kamaraj,C.; Elango, G.; Shankar, J.; Arjaria,
N.; Roopan, M.; Abdul Rahuman, A. and Singh, N. ( 2014). Synthesis of Nanoparticles
Using Euphorbia prostrata Extract Reveals a Shift from Apoptosis to G0/G1 Arrest in
Leishmania donovani.J.Nanomed Nanotechnol. 5)4): 213.
[47] Rao, KG.; Ashok, CH.; Rao, KV.; Shilpa, CH. and Tambur, V. (2015). Green Synthesis
of TiO2 nanoparticles Using Aloe Vera Extract, (2)1A: 28-34.
[48] Ma, X.; Lee, GJ; Deng, Y. and Kolmakov, A. (2010). Interactions between engineered
nanoparticles (ENPs) and plants: phytotoxicity, uptake and accumulation. Sci Total
Environ, 408(16): 3053-3061.
[49] Krishnaraj, C.; Ramachandran, R.; Mohan, K. and Kalaichelvan, PT. (2012).
Optimization for rapid synthesis of silver nanoparticles and its effect on
phytopathogenic fungi. Spectrochim Acta Part A Mol. Biomol. Spectrosc. 93: 95-99.
( Received 02 May 2016; accepted 23 May 2016 )