Cytotoxic and Genotoxic Effect of Biofabricated Zinc Nanoparticles on Mitotic Chromosomes of Drimia Polyantha

IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES

Available online at www.ijpcbs.com Research Article

CYTOTOXIC AND GENOTOXIC EFFECT OF BIOFABRICATED ZINC NANOPARTICLES ON MITOTIC CHROMOSOMES OF DRIMIA POLYANTHA (ROXB.) JESSOP Azharuddin Daphedar and Tarikere C Taranath* Environmental Biology Laboratory, P. G. Department of Studies in Botany, Karnataka University, Dharwad, Karnataka 580 003, India.

ABSTRACT Green synthesis of nanoparticles and their diverse applications are grown in various fields of nanoscience. In the present investigation, genotoxic effect of biogenic zinc nanoparticles on the root tip chromosomes of Drimia polyantha was evaluated. Biogenic zinc nanoparticles were characterized by UV-visible, XRD, FT-IR, AFM and HR-TEM analysis. The UV spectrum shows characteristic absorption peak at 374 nm. FTIR data suggested involvement of different functional groups in leaf extract were responsible for the reduction, capping and stabilization of ZnNPs. XRD results confirmed the crystalline nature of zinc nanoparticles with face centered cubic structure. AFM and HR-TEM images reveal the size and shape of nanoparticles. The size of ZnNPs ranges

between 5 to 80 nm and are spherical in nature. The effect of zinc nanoparticles on mitotic index (MI) and the chromosomal aberrations (CAs) was determined by the exposing root meristimetic cells of D. polyantha to different concentrations viz. 4, 8, 12 and 16 μg/ml of zinc nanoparticles suspension at the interval of 6, 12, 18 and 24 hrs. The investigation, revealed that the significant decrease (19.43±1.58) in mitotic index and increase in chromosomal aberrations (36.09±3.29), dose and duration dependent, when compared to control. The treated meristimetic cells showed various types of chromosomal abnormalities including bridges, sticky, laggard, diagonal, C- metaphase, multipolar anaphase, disturbed metaphase and micronucleus.

INTRODUCTION based nanoparticles are relatively new and Nanotechnology emerges from the physical, largely explored in the area of research. The chemical, biological and engineering sciences “green” route for nanoparticle synthesis makes where new techniques are being developed use of environmental friendly, non-toxic and from single atoms and molecules for multiple safe reagents. As compared to physico-chemical applications in different field of science and synthesis, biological synthesis of nanoparticles technology. Among other metal nanoparticles are environmentally benign and economically zinc have received special focus in view of their green as they are simple, inexpensive and easily distinctive, fascinating and extensive properties scale up larger scale production 7-9. Recently, the like semiconductor, chemical stability, catalytic, bio-fabrication of zinc nanoparticles synthesized piezoelectric, and ultraviolet (UV)-shielding1-2. by leaf extracts of Justica adathoda 10, Limonia The researchers have attempted to synthesize acidissima leaf mediated synthesis of zinc oxide nanoparticles within the size ranges between 1- nanoparticles 11 and Biosynthesis of silver 100 nm. Moreover, the bio-nanotechnology nanoparticles by leaf extract of Albizia saman 12 exists, from a new branch of nanotechnology, has been well documented. which integrates principles of biology with Recently, there are few reports on phytotoxicity physical and chemical procedures to generate of ZnO nanoparticles on Arabidopsis and Rye tiny-sized particles with specific functions3-6. grass (Lolium perenne) roots shows The physico-chemical based protocols for the morphological changes at high concentration of synthesis of metallic nanoparticles are often ZnO nanoparticle, leads to shrinkage of root tips expensive, toxic and utilize lethal chemicals. Bio- due to plasmolysis and contraction of epidermal

53 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

and cortical cells13-15 noticed that zinc oxide Phytosynthesis of zinc nanoparticles nanoparticles inhibit the growth of bacterial The 1mM fresh silver nitrate (AgNO3) solution cells due to the release of zinc ions from zinc was prepared and 10 ml of G. floribunda leaf nanoparticles, and induced toxicity via reactive extract was added to 90 ml of 1mM silver nitrate oxygen species (ROS) involving hydrogen solution and incubated at 60˚C for 1 hr. Further, peroxide which lead to DNA damage 16. Ahmed the temperature of reaction mixture was et al., 17 reported that ZnO-NPs treated Allium brought to room temperature and pH was cepa roots induce cytotoxicity and oxidative adjusted to (pH 8.0, pH 9.0 and pH 10.0), the DNA damage during mitotic cell division with color of the reaction mixture was changed from Zn2+ ions in plants. light yellow to deep yellow indicating the Getonia floribunda (Roxb.) belongs to family formation of zinc nanoparticles (Fig. 2). Combertaceae commonly known as life saver by the forest dwellers. The various parts of the plant are used to treat various ailments such as intestinal worms, colic, leprosy, malarial fever, dysentery, ulcers and vomiting 18. Moreover, it possesses antibacterial activity, anticancer and antioxidant property 19-20. In the present investigation, we have reported the biosynthesis and characterization of zinc nanoparticles using leaf extract of G. floribunda and their potential toxicity to mitotic cell division of D. polyantha (Blatt. & McCann) Stearn. family Asparagaceae. The effect of zinc nanoparticles on organism and environment are in nascent stage yet to be Fig. 1: a) G. floribunda leaf explored on different dimensions of their b) Formation of ZnNPs toxicity. However, there is no systematic work on this important aspect. Therefore, evaluation Characterization of zinc nanoparticles of the toxicological effects of nanoparticles on The biosynthesized zinc nanoparticles were plants and their mitotic activity is essential. analyzed by UV-Vis spectrophotometer (Jasco V- Hence the present investigation is carried out on 670 UV-Vis NIR spectrophotometer) operated cytotoxic/genotoxic impacts of zinc wavelength ranges between 300-700 nm with nanoparticles on D. polyantha. resolution at 1nm. The biomolecules present in

G. floribunda leaf extract and involved in MATERIALS AND METHODS bioreduction and capping of zinc nanoparticles Plant materials were investigated by FTIR analysis. The reaction All chemicals used in the present study are solution was centrifuged for 40 min at 3500- obtained from spectrochem Pvt Ltd. and S. D. 4000 rpm, to obtain pellet of zinc nanoparticles. Fine-Chem Ltd, (Mumbai, India). The plant Thereafter, purified suspension was freeze dried Getonia floribunda (synonym Calycopteris to obtain dry powder. The powder of ZnNPs floribunda) leaves (Fig. 1) were collected from mixed with potassium bromide (KBr) to obtain a the Botanical garden of Karnatak University pellet, the spectra was recorded in the range campus, Dharwad. Karnataka, India. 400-4000 cm-1. The purified dried powder of

ZnNPs was used for X-Ray diffractometer (XRD) Preparation of leaf extract analysis to confirm the crystalline nature of the About 20 g fresh leaves were thoroughly particles. XRD was performed in the 2θ range of washed in running tap water followed by double 30 to 80 degrees. Morphology, size and distilled water to remove adhered dust distribution of the nanoparticles were analyzed impurities. Washed leaves were shade dried, by using atomic force microscopy (AFM) and chopped into small pieces and added to 100 ml high resolution transmission electron of Milli-Q water taken in 250 ml of Erlenmeyer microscope (HR-TEM) images. flask and kept in water bath at 60˚C for 1 hrs.

The extract was filtered through Whatman filter Drimia polyantha assay paper No. 1. The filtrate was stored in Healthy and uniformly sized bulbs of D. refrigerator at 4˚C for further experimental polyantha (2n=20) were collected from Shivaji analysis. University campus, Kholapur, Maharashtra.

India. After collection, the bulbs were cleaned 2-

3 times with tap water followed by double

distilled water. The outer dry scales were

54 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

removed cautiously from the bulbs, without broad peaks as compared to pH-8. Our destroying root primordia. The bulbs were observation was similar to the synthesis of zinc soaked in glass coupling jar containing Milli-Q oxide nanoparticles by L. aculeata leaf extract 25. water for 48 h. at room temperature. When the newly emerged roots reached 3-4 cm in length, than they were transferred to new coupling jars 1.2 contain different concentrations of zinc nanoparticles solution (4, 8, 12 and 16 µg/ml) 1.0 and treated for 6, 12, 18 and 24 hrs. After pH 8 treatment the roots were excised and pH 9 0.8 pH 10 immediately fixed in Carnoy’s solution (3 parts ethyl alcohol and 1 part glacial acetic acid i.e. 0.6 3:1) for 24 h. Then the roots were transferred to 70% ethyl alcohol and stored in a refrigerator Absorbance 0.4 at 4˚ C for cytological studies. 0.2 Squash preparation Squashes were prepared as suggested by 0.0 Sharma 21, to evaluate the mitotic index (MI) and 300 400 500 600 chromosomal aberrations (CAs). For Wavelength (nm) examination the root tips were placed in a watch Fig. 2: UV-Vis absorption spectrum of ZnNPs glass and added 9 drops of aceto-orecin and 1 synthesized by leaf extract of G. floribunda drop of HCl and warmed over a low flame for 2- 3 minutes using spirit lamp. These were kept in FTIR analysis room temperature for 5-10 minutes. After FTIR spectra of the nanoparticles were recorded staining, the meristematic region were placed in order to identify the possible biomolecules on clean slide and squashed under a cover glass involved in the reduction and and 500 cells were scored from each slide capping/stabilization of the synthesized zinc making a total between 1800-2200 cells for each nanoparticles. Figure-3 and Table-1 shows the concentration and control under a bright field FTIR spectra of ZnNPs. The spectrum of ZnNPs 22-24 microscope at 40X magnification . The shows characteristic absorbance bands at mitotic index was calculated by using formula 3438.72, 2923.69, 2854.15, 1740.87, MI= Number of dividing cells / total number of 1629.73,1458.85, 1113.80 and 1029.93 cm-1, analyzed cells X 100 and chromosomal whereas, IR bands of G. floribunda leaf extract aberrations were calculated CAs= Number of were noticed at 3434.13, 2923.94, 2853.94, abnormal cells / total no. of dividing cells X 100. 1746.28, 1631.69, 1454.28, 1115.29 and 1058.39 cm-1 respectively. A shift in the peak Statistical analysis from 3434.13 to 3438.72 cm-1 is due to the The data were analyzed by using IBM, SPSS stretching of O-H and H-bending vibrations of Statistical version 20 windows software alcohols and phenols. Shift was observed in the followed by two-way ANOVA and Tukey test. peak at 2923.94 to 2923.69 cm-1 is because of O- H stretching of carboxylic acid 26. A shift was RESULTS AND DISCUSSION observed from 2853.94 to 2854.15 cm-1 is due to UV-Vis spectroscopy the asymmetric stretching of the C-H group. The UV-vis absorption spectrum was used to Another shift was observed from 1746.28 to monitor the biosynthesis of zinc nanoparticles 1740.87 cm-1 is because of C=O stretching using G. floribunda leaf extract (Fig. 1). The modes of the carbonyl functional group in addition of 10 ml leaf extract of G. floribunda ketones, aldehydes and carboxylic acids. The was added to 90 ml of ZnNO3 solution in a 250 shift in absorption peak at 1631.69 to 1629.73 ml Erlenmeyer flask, the reaction mixture was cm-1 may be assigned to conjugation effect of N- incubated at 80˚ C for 10-15 minutes. The pH H stretching 27. The shift in the weak band was adjusted to 8, 9 and 10 using either 0.1N 1115.29 to 1113.80 cm-1 and 1058.39 to HCl or 0.1 N NaOH. The changes in the color of 1029.93 cm-1 was due to the involvement of the solution were due to reduction of ZnNO3 to aromatic and aliphatic amines (C-N stretch) or ZnNPs, has a plasmosan resonance peak at 374 C−O stretching vibrations of phenolic nm at pH 8 are represented in Figure 1 and 2. compounds. But, In case of pH 9 and 10 the absorption peak observed at 381 and 386 nm, which shows

55 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

Fig. 3: FTIR spectra of ZnNPs synthesized by G. floribunda and leaf extract

Table 1: FTIR absorption peaks and their functional groups of ZnNPs synthesized by leaf extract of G. floribunda Absorption peak Absorption peak cm-1of Sl.No. cm-1of leaf Functional groups Silver nanoparticles extract O-H stretch, H-bending vibrations of alcohols 1 3434.13 3438.72 and phenols 2 2923.94 2923.69 O-H stretch Carboxylic acid, Alkyl C-H Stretch 3 2853.94 2854.15 Asymmetric stretching of the C-H group C=O stretching modes of the carbonyl 4 1746.28 1740.87 functional group in ketones, aldehydes and carboxylic acids 5 1631.69 1629.73 N–H stretching 6 1454.28 1458.85 C-H bond 7 1115.29 1113.80 Aromatic amines Aliphatic amines (C–N stretch) or C−O 8 1058.39 1029.93 stretching vibrations of phenolic compounds

XRD analysis X-ray diffraction (XRD) analysis of the ZnNPs synthesized by G. floribunda leaf extract has 1400 (111) been carried out (Fig. 4). The presence of peaks 1200 located at 2θ angles corresponds to (111), (200), (220) and (311) shows high intense peak 1000 can be indexed as 38.45˚, 44.12˚, 64.26˚ and 800

78.48˚ planes of face cubic centered (fcc) of zinc, Intensity 600 200 respectively (Figure-4). Thus, the XRD spectrum 220 confirmed crystalline nature of ZnNPs. The 400 311 average size of zinc nanoparticles can be 200 calculated by using (111) Braggs reflection 0 D=Kʎ/cosθ. Where K is the scherrer constant 30 40 50 60 70 80 90 (K=0.94) λ is the wavelength of the X-ray, β is Angle (2) the FWHM (full width and half maximum) of the peak and θ is the half of the Bragg angle. The Fig. 4: X-ray Diffraction pattern of ZnNPs size of the nanoparticle found to be 40.15 nm.

56 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

AFM and HR-TEM analysis The two and three dimensional view of sample The morphology and size of zinc nanoparticles represents structure of the nanoparticles with a as revealed by AFM and HR-TEM found to be height of 12.4 nm (Fig. 5b). The distance distinct nanostructures. The particles are between from each other nanoparticle is 16.3 to monodispersed and spherical in shape with size 48.5 nm (Fig. 5c). range between 15 and 85 nm (Fig. 5a and Fig. 6).

Fig. 5: Atomic force microscopy images of ZnNPs synthesized by leaf extract of G. floribunda a) 2D image b) 3D image c) particle distribution of ZnNPs

Fig. 6: HR-TEM images of biosynthesized ZnNPs synthesized by leaf extract of G. floribunda a) 50 nm image b) 10 nm image of HR-TEM

Effect of ZnNPs on mitotic chromosomes of D. evidence of mitodispersive effect of ZnNPs on polyantha cell division, which cause inhibition of DNA The present investigation was undertaken to synthesis or blocking G2 phase to prevent the determine the effect of biogenic zinc cell from entering mitosis 29-30. This is similar to nanoparticles synthesized by leaf extract of G. previous studies wherein the aqueous extract of floribunda on root tip meristimetic cells of D. different medicinal plants caused inhibition of polyantha. D. polyantha is a wild bulbous plant mitotic cell division in Allium cepa root tips 31-35. used for cytotoxic and genotoxic assay. The root The tiny sized nanoparticles have greater tip cells were treated with viz. 4, 8, 12 and 16 tendency to enter subcellular organelles and µg/ml concentrations of ZnNPs solution for 6, may interact directly or indirectly with the 12, 18 and 24 hrs. The results revealed that the genetic material (DNA) via nuclear pores mitotic index values found to be decreased with causing clastogenic effects on cell cycle leading increasing concentrations of zinc nanoparticles to DNA damage and formation of micronucleus up to 16 µg/ml for 24 hrs exposure duration, was apparent through oxidative stress 36-37. when compared to control (Table 2 and Figure Similar results have been reported by several 7). Statistically, the data found to be significant workers by using different plant model systems at (p˂0. 05). The decrease in mitotic index was such as Zea mays, Nigella sativa, Triticum dose and duration dependent in root tip cells of aestivum, Allium cepa, Vicia faba, and Drimia D. polyantha. Coelcho et al. 28 suggested that a indica which support our studies 38,39,10,40,17,12. reduction in the number of dividing cells is an

57 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

Fig. 7: Chromosomal abnormalities observed in root tip meristematic cells of D. polyantha exposed to different concentrations of ZnNPs. a) sticky metaphase b) laggard at metaphase c) single bridge at anaphase d) multipolar anaphase e) multibridge at anaphase f) spindle disturbance at anaphase g) C-metaphase h) micronucleus

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Table 2: The effect of ZnNPs on mitotic index (MI) of root tip meristematic cells of D. polyantha

No. of Concentr Total No. No. of non dividi Duration ation of cells dividing P% M% A% T% MI% ng (hrs) (µg/ml) examined cells cells Control 2417 1413 1004 92.74±2.05 2.89±0.76 2.56±0.70 1.77±0.07 58.25±7.08 4 2184 1130 1054 94.28±1.49 2.84±1.01 1.72±0.07 0.42±0.01 51.71±5.47 8 2158 1044 1114 93.49±1.44 3.39±0.73 1.36±0.09 0.39±0.02 48.43±2.68 6 12 2246 1048 1198 92.30±1.04 3.70±0.20 1.52±0.06 0.39±0.02 46.63±3.12 16 2204 1003 1201 90.72±1.64 4.90±0.77 1.09±0.04 0.69±0.01 45.54±2.58 4 2093 926 1167 90.47±0.82 3.90±1.07 1.82±0.01 0.53±0.01 44.23±1.22 8 2207 975 1232 89.62±1.33 4.93±0.44 1.54±0.03 0.50±0.06 44.15±1.11 12 12 2160 911 1249 87.45±2.40 5.46±0.57 1.99±0.07 0.54±0.09 42.15±0.99 16 2202 930 1272 87.40±0.64 5.28±1.27 2.58±0.85 0.62±0.06 42.18±1.28 4 1915 709 1206 84.36±3.04 4.67±1.18 3.40±0.06 0.69±0.02 37.01±0.87 8 2080 745 1335 82.43±4.10 5.63±0.36 3.08±0.52 0.53±0.04 35.81±0.42 18 12 2115 690 1425 82.65±3.40 5.04±0.79 3.38±0.48 0.84±0.07 32.60±1.43 16 1935 655 1280 78.75±1.91 5.65±0.67 3.79±0.82 1.04±0.06 34.01±3.79 4 1992 543 1449 73.04±3.73 4.64±0.57 2.65±0.15 1.12±0.05 27.24±3.20 8 1813 477 1336 68.32±3.17 4.84±0.90 2.73±0.05 1.67±0.03 26.39±2.45 24 12 1861 412 1449 59.69±1.16 4.68±0.52 2.88±0.56 1.98±0.03 22.09±0.32 16 1748 341 1407 49.20±1.35 3.50±0.63 1.71±0.07 2.65±0.08 19.43±1.58 Where, P- prophase; M-metaphase; A-anaphase; T-telophase; MI- mitotic index Stastically significant at p˂0.05, analyzed by Two-way ANOVA (Tukey test)

In another study, genotoxic effect can be The occurrence of various forms of analyzed by observing chromosomal chromosomal aberrations was detected in all abnormalities. Chromosomal aberrations are phases of mitosis. The chromosomes appear structural changes in chromosomes or sticky due to physiological stress as a result of chromatid breaks resulting dicenteric asphyxiation. Extreme treatment conditions for chromosomes. There were no chromosomal prolonged duration cause cellular asphyxiation aberrations were observed in the control. The causing respiratory inhibition leading to cell maximum proportion of chromosomal death. The chromosome stickiness and spindle aberrations was recorded in 16 µg/ml of ZnNPs malfunction was evident in the absence of suspension (14.79±12.27%) for 24 hrs of cellular energy ATP. In addition, laggard exposure duration. The results were statistically chromosomes are formed by the failure of significant at (p<0.05), when compared to spindle fiber or acentric chromosome formation. control. The observed chromosomal aberrations The occurrence of C- metaphase indicates the induced may be due to the presence of some risk of aneuploidy, causing partial disturbance naturally occurring phytochemical compounds in the spindle apparatus 45. Chromosomal in the leaf extract such as terpinoids, sterols, bridges were also frequent in ZnNPs suspension, alkaloids and flavanoids 18 have been implicated which occur in anaphase-telophase stages of in chromosomal damage at higher mitosis. Bridges may occur during the concentrations 41-44. When these higher translocation of unequal sister chromatid concentrations of ZnNPs enter into the root tip exchange and cause structural chromosome cells, changes the chromosomal behavior such mutation 32, 34. The Lagging chromosomes are as bridges, sticky, laggard, diagonal, C- formed by separation of daughter nuclei with metaphase, multipolar anaphase, disturbed unequal number of chromosomes and metaphase and micronucleus (Table 3 and Fig. subsequent formation of micro nuclei or 7). daughter cells with unequal sizes at intervals.

59 IJPCBS 2018, 8(1), 53-63 Tarikere C Taranath et al. ISSN: 2249-9504

Table 3: Chromosomal aberrations observed in root tip meristematic cells of D. polyantha

Concentration laggar Multipo laggin C- CAs CB Sticky Micro MB Duration (µg/ml) d lar g meta (%) (hrs) 0.00±0. 0.00±0. 0.00±0. 0.00±0. 0.00±0. 0.00±0. 0.00±0. 0.00±0. 0.00±0. Control 00 00 00 00 00 00 00 00 00 0.10±0. 0.26±0. 0.80±0. 0.87±0. 0.81±0. 0.00±0. 0.00±0. 0.00±0. 0.69±0. 4 07 04 03 05 02 00 00 00 06 0.17±0. 0.64±0. 0.20±0. 0.10±0. 0.10±0. 0.00±0. 0.00±0. 0.00±0. 1.34±0. 8 03 06 05 07 05 00 00 00 07 6 0.30±0. 0.79±0. 0.40±0. 0.18±0. 0.18±0. 0.00±0. 0.00±0. 0.90±0. 2.05±0. 12 01 08 08 05 03 00 00 05 05 0.39±0. 1.57±0. 0.00±0. 0.00±0. 0.20±0. 0.10±0. 0.00±0. 0.20±0. 2.57±0. 16 03 09 00 00 04 07 00 04 06 0.11±0. 1.17±0. 0.77±0. 0.22±0. 0.20±0. 0.20±0. 0.11±0. 0.19±0. 3.24±0. 4 09 01 09 03 07 05 09 03 08 0.30±0. 1.33±0. 0.50±0. 0.31±0. 0.20±0. 0.10±0. 0.00±0. 0.10±0. 3.38±0. 8 09 01 07 02 05 01 00 07 08 12 0.55±0. 1.53±0. 0.98±0. 0.44±0. 0.23±0. 0.00±0. 0.10±0. 0.11±0. 4.51±0. 12 01 05 01 08 09 00 05 09 43 0.90±0. 1.66±0. 0.00±0. 0.34±0. 0.35±0. 0.31±0. 0.96±0. 0.35±0. 4.80±0. 16 06 08 00 05 06 09 07 05 09 0.28±0. 2.17±0. 1.13±0. 0.68±0. 0.69±0. 0.25±0. 0.30±0. 0.38±0. 6.84±0. 4 04 09 25 04 02 04 05 06 55 0.52±0. 2.01±0. 1.21±0. 0.93±0. 0.00±0. 0.52±0. 0.26±0. 0.65±0. 8.29±0. 8 04 07 05 05 00 09 02 08 65 18 0.57±0. 2.52±0. 1.59±0. 0.29±0. 0.69±0. 0.29±0. 0.00±0. 0.29±0. 8.04±0. 12 07 51 06 05 08 05 00 05 26 0.46±0. 2.30±0. 1.39±0. 1.67±0. 0.78±0. 0.44±0. 0.57±0. 0.63±0. 10.73±1 16 05 57 04 08 03 05 06 04 .52 0.50±0. 5.16±2. 2.30±0. 1.37±0. 1.23±0. 0.67±0. 0.33±0. 0.50±0. 14.36±2 4 05 26 17 19 07 06 07 08 .47 1.27±0. 6.01±0. 3.41±0. 1.25±0. 2.06±0. 0.42±0. 0.42±0. 0.84±0. 18.45±1 8 08 27 04 07 24 07 02 07 .27 24 1.86±0. 6.46±0. 3.26±0. 2.56±0. 2.72±0. 0.84±0. 1.34±0. 1.65±0. 23.24±3 12 04 13 90 51 34 06 03 05 .20 2.93±0. 10.55±0 4.88±0. 4.57±0. 3.47±0. 1.50±0. 2.37±0. 1.78±0. 36.09±3 16 34 .95 59 83 50 06 05 08 .29 Where, CB- chromosomal bridge; MB-multibridge at anaphase; CAs- chromosomal aberrations Stastically significant at p˂0.05, analyzed by Two-way ANOVA (Tukey test)

Earlier reports reveled that Allium cepa root tip index and increase in chromosomal aberrations cells have been widely applied to evaluate confirming the cytotoxic and genotoxic level of cytotoxicity and genotoxicity induced by zinc nanoparticles. pesticides46, herbicides47, Aromatic hydrocarbons 48-49, Textile industry dyes29, 50, ACKNOWLEDGEMENTS heavy metals 51 and higher plants 52, 10 showed The authors are thankful to the Chairman, P. G. positive results by using these parameters for Department of Studies in Botany. Karnatak the analysis of chromosomal abnormalities and University, Dharwad, for providing the also showing a greater genotoxic effect on root necessary facility and UGC-DSA-I phase of tip cells. The results of present investigation Department for financial assistance. One of the suggest that biogenic zinc nanoparticles inhibit author (A.D) acknowledges the Directorate of cell division at higher concentrations was found Minorities, Government of Karnataka, for to be strongly concentration and duration awarding Research fellowship. The Authors also dependent. The induction of chromosomal thank USIC- KUD, MIT Manipal and IITM aberrations observed in the root tip cells of D. Chennai for instrumentation facility. polyantha, was due to the bio-uptake nanoparticles from the suspensions. REFERENCES 1. Meulenkamp EA. Synthesis and growth CONCLUSION of ZnO nanoparticles. J Phys Chem B. In the present study zinc nanoparticles 1998;102:5566-5572. synthesized by G. floribunda leaf extract, showed 2. Serpone N, Dondi D and Albini A. higher cytotoxic and genotoxic activity using Inorganic and organic UV filters: their root tip meristematic cells of D. polyantha. The role and efficacy in sunscreens and toxicity is found to be concentration and suncare product. Inorg Chim Acta. duration dependent and decrease in mitotic 2007;360:794–802.

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