.

.
C SEI L FV EF R E&
Allium CG O
cepa T LSD
. . ROOTS
N A N O P A R T I C L E S

.
 Nanoparticles having small in size (<50 nm)
is absolutely
a special aisbranch
new of material
emerging field,
can comfortably penetrate and pass which
This a short-term test, into the
and
science basically
Research
lymphatic where
has
can
system there
shown
andboth
assess are
bulk evidences
silver
cytogenetic
subsequently materials
andreach
gold
effectsto
of nanoparticles
break
the several
vital down
organ can
andtobe
of nanoparticles
negative
bodysynthesized
small suspended
effects
tissues easily
particles
and in a
on
exhibit
AMONG PLANT
AMONG has
PLANT many
TESTS,
TESTS, applications
ALLIUM
ALLIUM
test CEPA IS
CEPA
solution. which
IS ONE
ONE OF THE
OF
Keeping makes
THE MOST
MOST
in minditWIDELY
WIDELY
the
NANOTECHNOLOGY growth
USED.
USED.
their
within
ALLIUM
ALLIUM and
negative
the development
CEPAabove
CEPA
effect.
range
HAS BEEN
HAS BEEN
On
of
USED
USED
the
1–100
FOR
FOR of
otherplantlet
hand,
nmworkin
EVALUATING
EVALUATING
easily accessible
potential toxicity and
of quite
fact popular
present
nanoparticles on is
CHROMOSOMAL ABERRATIONS
ABERRATIONS SINCE
SINCE 1920S
1920S ..
but due to CHROMOSOMAL
diameter by means
itsterrestrial
diverse
among of
researchers.
dedicated
plant physical,
application in
speciestois judge chemical
various
extremely fields
the limited.
efficacy
such as DNA methods.
and biological of silver and
sequencing, gold nanoparticles
pharmaceuticals,
cosmetics, agriculture, towards
biomolecular chromosomaldetection
aberrations of Allium cepa root
and diagnostics, it has gained attention and
under laboratory condition.
synthesized and used extensively in various field.
.
 Macroscopic parameters were measured after
After
72The
When 72
goldthe
h of exposure.handroot
roots
The tipsslides
silver
The
reached
roots were
were cutcut
nanoparticles
were
to 2–3
at and
analyzed
cm
their are fixed
they
base diluted
with in to
Olympus
were ethanol
cut three
and
different
and acetic Three
concentrations
acid mixturebulbs(3:1)
CH20imicroscope were
i.e; used
for1at
mg24 for
100XhL−1each
at, 55 concentration
magnification.
°C.
mg Then
L−1 and the
All calculations
processed for were
slide done
preparation
and the number counted and the length along using Minitab
by following version17 standard
10
roots
mgthe
software.
method.
with were
L .breadth
−1
and
Healthy
The
Two dipped
of
level MATERIALS
Thewhich
replicates
breadth
were mitotic
onion
ofinto
concentration
number
of
fiveindex
bulbs
1
root tips
M
significance
all of
for
measured.
roots
each
to
dividing
per
was
were
HCl
The
prepare
were
was
cellswas
bulb
used foraseach
calculated
solution
collected
accepted
concentration
length
slide for
per number of
and thewere
from the
heated
nearby
at p <
made. at
After 60that
vegetable
0.05. °Cmicroscopic
for 4–5
market.
Cytogenecity
the
total
minanalysis.
dried
observed
followed
Three
roots
cells. was
were Theby
healthy
Along
the transfer
onion
statistically
carefully
slides
with of
thiseach bulbs
shaved
portion
to
(12–15in Sg)
distilled
analysed
off orderTEST
TMICROSCOPIC
Aand
Tto
by SYSTEM
Iand
Sgrown
T
MACROSCOPIC EXAMINATION
summarized were
water I
expressed C
kept
Student’s
expose
treatment
N
of A N A
O
mitotic
AND
P L
as
for
the
and
A
the
directly R A
few TREATMENTS
t-test.
phases,
N
total
fresh
control
T I C A
EXAMINATION
in
minutes.
L
the E SL
length
the The
were Ynanoparticles
presence
Sprepared
Ilevel
S andthe
Finally
meristematic of
by in
root
tissue.
and total breadth of the root system. The mean
cylindrical
tips
Then were
significance
the crushed
glass
was
roots tubes
withonion
accepted
of
following
frequency 2%
inaceto
normal
aceto
of at
were p
orcein
chromosomeorcein
< lighting
0.05.
grown
squash withcondition
in flatdifferent
technique.
aberrations endThe at
of
values for all parameters were calculated.
room
metal
medium temperature
rod and
containing root
the
METHODS
tips
(20
cover
(fragments,
bothwere
°C) slip
Seven roots of each bulb were fixed in a freshly
The
the
10 mg slide
testL−1)suspension
and
and 5micronuclei
the mincover
gold followed
was
(1 mg
prepared mixture of absolute ethanol and
kept
for
silver
Lby
were
was
anaphase
replaced
slip
72 inhcarefully
(1 1mg
staining
−1, wasalso
5 mg
M
alongHCl−1,for
bridges
L
with 2%
determined.
sealed
daily
L −1 and
with
5
to
about
lowered
etc.)
mg
aceto
with
10 Amg
−14–and
control.
and
L
maintain
on
clear
L−1)
orcein.
minimum Staining
of 500 was
cells continued
were for about
counted for
constant
finger nail
glacial polish.
nanoparticles alongconcentrations
acetic acid
10
The
(3:1
min
prepared
with
v/v)
and
for
then
24
it
of
controlh
was
slides
at 4 suspensions
°C.
squashed.
were
(double ready
The cover
for
of
distilled
each slide.
nanoparticles.
microscopic
water) duringstudy. 72sliph. was sealed with clear finger nail polish.
 TABLE 1c
No. of TABLE 1b
counted
No. Dividing cell Prophase Metaphase Anaphase Telophase Mitotic index Mean ± SD
of4. X-ray diffraction of (a) silver
Fig.
cells Dividing nanoparticle (b) gold nanoparticle.Mitotic
Fig.
counted1. UV–Vis spectroscopy of (a) silver
Prophase Metaphase nanoparticle and (b)
Anaphase gold nanoparticle. Mean ± SD
Telophase
cell
Fig. 3. Transmission electron microscopy ofSilver
(a) silver index
nanonanoparticle and (b) gold nanoparticle.
cells TABLE 1a
(1 mg/L) Gold nano
(1 mg/L) No.
Replica1 500 of 291 280 7 4 0 58.20%
Mitotic 57.1% ± 1.22
Replica1 counted
500 Dividing
272 cell Prophase
260 Metaphase
8 Anaphase
3 Telophase
1 Mean ± SD ± 1.7
index54% 52.4%
Replica2
Replica2
Replica3
The500500surface
cells
500
263
279
morphology
287
257
268
279
4
5
and2
4
4existance
2
0
3 of
52.60%
55.80%
1 57.40%

Replica3
both500
silver264and 253
gold Control
was
7
assessed
3 1
by SEM-
50.60%
(5 mg/L)
(5 mg/L)
Replica1
Replica1 500
500
EDX 340
258
study
317
247
(Figs.
12
5
2a and
6
5
2b).
5
1
68%
51.60%
68% ± 3
47.3%
53% ± 1.4 ±
Replica1
Replica2 500
500 249
328 238
318 87 22 0 2 49.80%
65% 2.19
Replica2 500 265 251 7 5 2 53%
Replica2
Replica3 500
500 233
355 221
341 7
115 2 44 1 21 46.60%
71%
Replica2 500 272 261 54.40%
Replica3
The500
exact size228 and shape 220
of silver and5 2
gold nanoparticles 1 45.60%
were presented
(10 mg/L) Fig.XRD
Theinsharp 3.andstudy
Fig.
From 2a.ofTEM
2b.
the
distinct both
(a) FESEM silver
it was
peaks ofand and(b)
observed
silver andgold
EDX ofhighlighted
that
gold silver
gold
average sizethe
nanoparticle.
nanoparticles crystal
ofwere
silver and gold
observed
(10 mg/L) structure of bothbetween
the silver and
nanoparticles
from UV–vis ranges
spectral signature 25at 440gold
and 40
nmnm nanoparticle
andand 17–24
550 nm,(Fig. 4).
respectively.
nm respectively.
Replica1
Replica1 500
500 213
285 206
277 4 32 11 42.60%
57% 41.4%
55.8% ± 1.2
± 1.44

Replica2
Replica2 500
500 201
271 192
261 4
6 44 01 40.20%
54.20%
Replica3
Replica3
500
500
207
281
201
272
3
5 4
3 0
0 41.40%
56.20%
 Table 3. Comparative status of MI exerted by 1 mg/L AuNPs and 1 mg/L AgNPs.
Au NP Ag NP Significant
Table 6. Comparative status of chromosomal
Mean aberration
SD of AuNPs with respect to control
t value
Present
1 mg/L study
1 mg/L reveals there TABLE was 2 no chromosomal level
54 58.2 Au NP Ag NP Au NP Ag NP t value of t value of t value of
aberration
TheEFFECT
efficacy in
ofLENGTH control
silver and (Table
gold 1a).Au 10But
nanoparticlesmg/L silver on and vsthe gold MI
Au5 ofvs Allium
From
Control
52.6 the ON statistical
Au 1 mg/L
57.4 No.52.4 AND results
Au 5 mg/L
BREADTH it
of roots 57.13 Root length was
1.71 OF observed
ALLIUM Au1
(cm)1.22 conc. CEPA
Root 3.9 that
breadth ROOTgold
conc. P(cm)
Au10 vs
< 0.03
conc.
cepa nanoparticles
was compared haveby significant
applying effect
statistical on test the (Table 3). From the
result it (Au)
50.6
occurrence
has nanoparticles
been55.8
(mean
of
found
± SD)
chromosomal
that of 10 of
mean mg
(mean Lof showing
± −1
SD)
aberrations
MI AgNPs in
7.83 highest
(mean
comparison
(1 mg L
± SD)
−1) is higher
9.63 14.26
Breadth
68 is the measurement
54 49.8 of width of the
42.6 Allium cepa root which is
than the chromosomal
Control
with the
mean control.
MI of
6 ± 0.04
AuNPs aberration
From the
(1
measured by screw gauge and root length was measured by the
Table 4. Comparative status of MI mg
exertedL
3.6
−1
byin
)
±
experimentaland
5 the
0.01
mg/L it roots
is
AuNPs data
p < 0.004of
statistically
and 5 it
mg/LAllium
1.1
was
± 8.33
P < 0.002 p < 0.005
significant
AgNPs.
(P cepa
65
<
Silver
Au seen
0.03) and 52.6
that
(Table
centimeter scale. From Table
nanoparticle
NP Ag NP it
the is
3). significant
mitotic
Almost 46.6
Mean
index
similar at
valueP <40.2
0.05
for
significant
2, it was clear SD
level
control
thatdifference
the (Table
was
t
68%
breadth
value
7). ofInthe
(PSignificant
< 0.003)
715 mg/L 5 mg/L
50.6 45.6 41.4 level
rootscomparison
between 1and
mg/L for
AgNPs
of Allium cepa gold (5 with
mg7 ± L
0.1−1
AuNP,
nanoparticles
is highly ) and AgNP
it
AuNPs
affected was
4.23 ±(5 ofthe
52.4%,
mg
0.11
by 10 L mg L showed
−147.3%
) was
nanoparticles. −1and
also
0.25 ± recorded
0.06 But in
Table 7. Comparative
49.8
541.4%
mg/L4).for 51.6 statusAu of
NP chromosomal
Ag NP aberration
Au NP ofAgAgNPs
NP with respect to control
case(Table
much lower
of length But 1 mg
at 10L±
chromosomal
there ,
−10.13
higher 5 mg
is no significantL −1, 10
concentration mg
5.63 L −1 respectively
± 0.13
aberration
changes. (10 mg L
with
−1),0.28
Similar MI (Table
±of
respect 0.03AgNP
observation to of
46.6
10 mg/L 53 7 53
± 0.01 47.33 2.19
0.53 ± 0.004 1.4 3.77
0.36 ± 0.021p < 0.033
1b).
Fig.
exhibited
was 6.ItGold
Fig. 7. means
high
Mitotic
recorded inby
nanoparticle
trend case
chromosomal of
other of
induced gold
aberrations
authors. nanoparticles,
chromosomal
statistically insignificant
A. cepa
This aberration
may be (P
root cells
t value
the
of
<
due mitotic
ininduced
root
0.001)
t value
to tip by
of
lowcells
thant value
control
Control
45.6
of Allium which
Ag
silver 1 mg/L
54.4
cepa is
showing
nanoparticle significant
Ag J5 mg/L
– Chromosomal
showing A – at Ag P
10 <
mg/L0.024
bridge,
micronucleus Kat– (Table
Ag1
normalvs
interphase, Ag56).
prophase,
B –vs So,
L – it
Ag10 vs
10 mg index
L
concentration
Goldcan
decreased
−1 of AuNPs
of (Table
the with5).
nanoparticles increasing
FinallyM –it may
which the beconcentration
did concluded
notconc.
show of
that
any
conc. AgNPs
effects
conc.

showed be
anaphase
Normal
nanoparticle
the conclude
with
anaphase,
Fig.
nanoparticles.
Table higher
5.
5. Comparative
chromatin
Almost
cell
C –
But
division
status that
bridge,
chromosomal
proper
of MIa at AuNPs
anaphase
reverse
both
exerted
bridges,
by 10
(A),of
mg/Lhigher
anaphase-telophase
trend
lower
D – normal
prophase
was
(1
AuNPs mg
(B) (10
telophase,
observed
and L 10)mg/L
−1 mg
with fragment
and
EL )6.35
in
–−1
higher
AgNPs.
1on
mg/L the growth
bridge,
vagrant
and N – disturbed
metaphase 3of
chromosome root
metaphase
(C)
± 0.003 in length as
with
anaphase-telophase,
showing no well
0.001as number0.24
unoriented
chromosomal
0.8 ± chromosomes
F – cell laggards,
aberration
5.81 of ±roots.
inat different
G
7.85
0.06–
68
5
concentrations
Aucase
NP
disturbed
mg/L of
58.2
silver
Agmetaphase
NP
(10 mg15 L
±
−1) showed
exposure
with
0.11
51.6
nanoparticles.
Mean
conditions
concentration
clumping
control. highest
The
of
57
themitotic
chromosomes,
3.05 ± than
0.011
SD
chromosomal
Hindex
nanoparticle.
AuNPs.0.028tvalue
p–<normal 0.34interphase
p<
value ± 0.016was
Significant
0.006 p < 0.024
10 mg/L cell,10I mg/L
– disturbed telophase at different exposure conditions of the level
65 1057.1%
42.6
mg/L and aberration
57.4
42.6
53%Au 6 ±for
0.10
NP
1 than
53mg LNP AgNPs
−1 and 4.23
nanoparticle.
Ag
5± mg
54.2
Au NP
of L10
0.53
−1
Ag NP
mg L0.34. ± 0.001
respectively −1
71 but
40.2
at 10
55.8mg L−1 the
54.2 41.4
value
54.4
55.8
increases 56.2
1.2
to 1.44
55.8% (Table 13.29
1c).
p < 0.001
41.4 56.2
Present study
Moreover,
Therefore results
the
it can reveal
becell that both
division
concluded silver
thatwas and gold
arrested,
nanoparticles
nanoparticles
to beshows negative effect on the roots
THANK
at metaphase stage for both the nanoparticles,

CONCLUSION
supposed a potent hazardous component
for ofthe
showingAllium cepa. Bothchromosomes,
lagging
environment theandnanoparticles could
stickiness,
entire ecological
andpenetrate
anaphase into the
withroot cell and
broken cause significant
chromosome
be done to bridge.

YOU
systems. More research should unfold
Bothchanges in intracellular components, causing
their silver
overallandfate, goldtransport,
nanoparticlesend does not
exposure
remarkableany
exhibited damage to the cell
variation in division.
root The mitotic
length but
pathways in the wider environment. Further study
index decreased
remarkable from the
changes was control (68%) tointhatroot
recorded of
is going−1on to explore more about nanoparticles
10 mg L treated
diameter and number (41.4%)of forroots
gold for
nanoparticles
both typesand of
and their−1 cytogenetic effects on plants.
5 mg L treated (53%) for Silver nanoparticles.
nanoparticles.
 Lester Paul R. Sivila
Noreen Jessica S. Dacles
Kristal Gail I. Manibo
Andrea Nicole F. Rebibis
Joyce Nikko T. Fagaragan
 .
.
C SEI L FV EF R E&
Allium CG O
cepa T LSD
. . ROOTS
N A N O P A R T I C L E S