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: ahmed_naji_abd@yahoo.com

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 )