role of nanotechnology for crop protection in horticultural crops

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WELCOME
BIG EVENTS
HAPPEN IN
SMALL
WORLD

ROLE OF NANOTECHNOLOGY FOR CROP
PROTECTION IN HORTICULTURE
Speaker
Girija Kumari, Ch.
ID No. 10468
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Seminar Incharge
Dr. T. M. Rao

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Nanotechnology/Nanoscience
Nanotechnology
 The design, characterization, production and application of
structures, devices and systems by controlling shape and size
at the nanoscale
British Standards Institution (BSI 2005)

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Nano scale
The term ‘nanotechnology’ is based on the prefix ‘nano’- Greek word
meaning ‘dwarf’
Word ‘nano’ means 10ˉ⁹ or one billionth part of a metre
1 nanometre= one billionth (10ˉ⁹) of metre
 Size range between 1 and 100 nm
The term ‘nanotechnology’ is based on the prefix ‘nano’- Greek word
meaning ‘dwarf’
Word ‘nano’ means 10ˉ⁹ or one billionth part of a metre
1 nanometre= one billionth (10ˉ⁹) of metre
 Size range between 1 and 100 nm

Richard Feynman, Physicist
“ The father of nanotechnology”
Richard Feynman, Physicist
“ The father of nanotechnology”
“There’s Plenty of Room at the Bottom”
- at American Physical Society meeting at the California Institute of
Technology on Dec-29, 1959.
“There’s Plenty of Room at the Bottom”
- at American Physical Society meeting at the California Institute of
Technology on Dec-29, 1959.
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Richard Feynman
“Adapability to manipulate, control, assemble, produce and
manufacture things at atomic precision”

Norio Taniguchi, Professor
- coined the term “Nanotechnology” (1974)
Norio Taniguchi, Professor
- coined the term “Nanotechnology” (1974)
“Nano-technology’’ - Processing, separation, consolidation and
deformation of materials by one atom or by one molecule.
“Nano-technology’’ - Processing, separation, consolidation and
deformation of materials by one atom or by one molecule.
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Nanotechnology received its greatest momentum with the
invention of the scanning tunneling microscope (STM)
It was invented by Gerd K. Binning and Heinrich Rohrer in
1985
What STM does?
It allows imaging solid surfaces with atomic scale resolution. It operates
based on tunneling current, which starts to flow when a sharp tip is
mounted on a piezoelectric scanner approaches a conducting surface at
a distance of about 1 nm. This scanning is recorded and displayed as
an image of the surface can be resolved an displayed using STM

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Scanning Tunneling Microscopy

Time Line of Nanotechnology
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~ 2000 Years
Ago
Sulfide nanocrystals used by Greeks and Romans to dye hair
~ 1000 Years
Ago
Gold nanoparticles of different sizes used to produce different colors in
stained glass windows
1959 “There is plenty of room at the bottom” by R. Feynman
1974 “Nanotechnology” - Taniguchi uses the term nanotechnology for the first
time
1981 IBM develops Scanning Tunneling Microscope
1985 “Buckyball” - Scientists at Rice University and University of Sussex
discover C60
1986 • “Engines of Creation” - First book on nanotechnology by K. Eric
Drexler.
• Atomic Force Microscope invented by Binnig, Quate and Gerbe
1989 IBM logo made with individual atoms
1991 Carbon nanotube discovered by S. Iijima
1999 “Nanomedicine” – 1st
nanomedicine book by R. Freitas
2000 “National Nanotechnology Initiative” launched
(British Standards Institution, 2005)

Physics
Biology
Material
Science
Medicine Engineering
Bionanoscience
Chemistry
Bionanoscience / Technology
 Exploitation of biomaterials, devices or methodologies on the Nanoscale Exploitation of biomaterials, devices or methodologies on the Nanoscale
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Nanoparticles
 Nanoparticles are particles between 1 and 100 nanometers in
size
 In nanotechnology a particle is defined as a small object that
behaves as a whole unit with respect to its transport and
properties
 Arranged or assembled into ordered layers, or mine layers
 Possess distinct physical, biological and chemical properties
associated with their atomic strength
(Bhattacharyya et al., 2010)
 Nanoparticles are particles between 1 and 100 nanometers in
size
 In nanotechnology a particle is defined as a small object that
behaves as a whole unit with respect to its transport and
properties
 Arranged or assembled into ordered layers, or mine layers
 Possess distinct physical, biological and chemical properties
associated with their atomic strength
(Bhattacharyya et al., 2010)

Nanocarbon
Fullerene
Tubes
Cones
Carbon black
Horns
Rods
Foams
Nanodiamonds

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Carbon nanotubes
Iijima in 1991
Appeared to be made up of a perfect network of hexagonal graphite
rolled up to form a tube
Exhibit unusual photochemical, electronic , thermal and mechanical
properties

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Nanoparticles in disease and pest management
(Crop Protection)
Biopolymer nanoparticles eg. Chitosan
Metallic nanoparticles eg. Silver nanoparticles
Silica nanoparticles
Copper nanoparticles
Zinc nanoparticles
Nanocomposites eg. Chitosan Silver NP

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Chitosan
Has various applications in biology due to its biodegradable and
nontoxic properties
chitosan and chitosan nanoparticles are found to be more effective
against plant pathogens like Fusarium solani
The chitosan therefore could be formulated and applied as a natural
antifungal agent in nanoparticles form to enhance its antifungal activity
(Ing et al., 2012)

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Mode of action of chitosan against fungi can be explained by
following mechanisms:
The positive charge of chitosan interacts with negatively charged
phospholipid components of fungi membrane, which in turn alter cell
permeability of plasma membrane and causes the leakage of cellular
contents, which consequently leads to death of the cell (García-Rincón
et al., 2010)
Chitosan chelates with metal ions, which has been implicated as a
possible mode of antimicrobial action (Rabea et al., 2003). On binding
to trace elements, it interrupts normal growth of fungi by making the
essential nutrients unavailable for its development (Roller and Covill,
1999).
It is suggested that chitosan could penetrate fungal cell wall and bind
to its DNA and inhibit the synthesis of mRNA and, in turn, affect the
production of essential proteins and enzymes (Sudarshan et al. ,
1992; Kong et al., 2010)

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Metallic nanoparticles
Metallic nanoparticles ( Ag, Zn, Cu, Si) possess unique chemical and
physical properties, small size, huge surface to volume ratio, structural
stability and strong affinity to their targets (Kumar et al., 2010)
These can be used as new antimicrobial agents and an alternative to
synthetic fungicide to delay or inhibit the growth of many pathogens
species because of its multiple mode of inhibition

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Silver nanoparticles
Silver has been used as an antimicrobial agent since ancient
civilizations; it has been used extensively due to its broadspectrum and
multiple modes of antimicrobial activity (Wei et al., 2009)
Silver exhibits higher toxicity to microorganism and lower toxicity to
mammalian cells
The application of silver nanoparticles as antimicrobial agents is
because of its economical production and multiple modes of inhibitory
action to microorganisms (Clement and Jarrett, 1994)
Its specific antimicrobial mechanisms are still unclear

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Ag acts as plant-growth stimulator and reduces unwanted
microorganisms in soils and hydroponics systems (Sharma et al., 2012)
Silver in ionic or nanoparticle forms has a high antimicrobial activity and
is therefore widely used for various sterilization purposes (Park et al. ,
2006)
Some studies found that inhibition of fungal pathogens with silver
nanoparticles is concentration dependent and also on type of silver
nanoparticles used
Cont........

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Silica nanoparticles
Silicon (Si) increases disease resistance and stress resistance in plants
(Brecht et al., 2004)
It also stimulates the physiological activity and growth of plants (Carver
et al., 1998)
Torney et al. (2007) used honeycomb mesoporous silica nanoparticle
(MSN) system with 3nm pores to deliver DNA and chemicals into plant
cells and intact leaves

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Copper nanoparticles
Copper-based fungicides produce highly reactive hydroxyl radicals which
can damage lipids, proteins, DNA, and other biomolecules
It plays an important role in disease prevention and treatment of large
variety of plants (Borkow and Gabbay, 2005)
Because of its bio-compatibility, these nanohydrogels are included as a
new generation of copper-based bio-pesticides and it could also be
developed into an efficient delivery system for copper based fungicides for
plant protection (Brunel et al., 2013)
Complexation of copper with chitosan nanogel was shown to have strong
synergistic effect between chitos an and copper in inhibiting the growth of
phytopathogenic fungus Fusarium graminearum

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Zinc nanoparticles
Zinc oxide nanoparticles (ZnO NPs) could be used as an effective
fungicide in agricultural and food safety applications
Mechanism of action of zinc nitrate derived nano-ZnO on important
fungal pathogen Aspergillus fumigatus showed hydroxyl and superoxide
radicals mediated fungal cellwall deformity and death due to high energy
transfer (Prasun Patra and Goswami,2012)
ZnO nanoparticles can cause deformation of fungal hyphae and prevent
the conidiophores and conidial development which ultimately leads to the
death of fungal hyphae

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Nanoparticles for pest management
Globally insect pests cause a huge crop loss of 14% and plant pathogens cause
an estimated loss up to 13% with a value of US $2,000 billion per year (Pimentel,
2009)
Nano pesticide formulations increase the solubility of poorly soluble active
ingredient and helps in releasing the active ingredient slowly
Nanoparticles are loaded with pesticides and released slowly based on
environmental trigger (Lauterwasser, 2005)
Rotenone, a water-insoluble botanical insecticide used to control aphids, thrips ,
acari from decades , however its effective utilization has limited due to its poor water
solubility, stability, degradation and isomerization when exposed to sunlight
Nanosilica showed 100% mortality against insect pests whereas nanosulfur
inhibited the sporulation and growth of fungi (Goswami et al., 2010)

(Royal Society and Royal Academy of Engineering, 2004)
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METHODS OF NANOPARTICLE PRODUCTION

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TOP DOWN APPROACH

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BOTTOM UP APPORAOCH

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1 m
1m
1 m
Each edge is 1 m Each edge is 0.1 m, but there are 1000
cube
Volume (in cubic mts ) = 1m x 1m x 1m =1m³
Surface area (in sq. m) = (1m x 1m ) x 6 sides
= 6 sq. m
Volume = (0.1 x 0.1 x 0.1) x 1000 cubes = 1m³
Surface area = (0.1 x 0.1) x 6 sides x 1000 = 60 sq.m

Mode of action of nano particles
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Dendrimers
Quantumdots
Nanosensors
FullerenesCarbon Nanotubes
Nano Chips
C60 Cadmium selinade
3D
macromolecules
Sequence nanoscale
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Tools of Nanotechnology
C60

Applications In Agriculture
31 (Mahendra et al., 2012)

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Applications of nanotechnology in crop protection and plant nutrition

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CASE STUDY : 1
Antifungal effectiveness of nanosilver
colloid against rose
powdery mildew in greenhouses
Kim et al., 2008
Solid State Phenomena Vol. 135 , pp 15-18

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Introduction
Powdery mildew, caused by Sphaerotheca pannosa var. rosae, is one
of the most common and widespread fungal diseases of greenhouse and
outdoor roses which reduces flower production and causes weakening of
the plants
It mainly appear first on the under surface of young leaves in early
summer and the infection spreads to stems, shoots and buds
Silver have long been known to have strong antimicrobial activity and
AgNP because of their specific surface area have high activity than their
bulk siver metal
Nanosilver colloid that is a well-dispersed and stabilized silver
nanoparticle solution will be more adhesive on bacteria and fungus and so
have enhanced antibacterial activity

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Objective of the study
To examine the effectiveness of nanosilver colloid as
new fungicide against rose powdery mildew in
greenhouses

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Material and Methods
31.5 g of 99.8 % AgNO3, was dissolved in 3.7 l distilled water and 40g
PVP as stabilizer was added
1g of 98.0 % NaBH4, as reducing agent was dissolved in distilled
water of 0.2 l and this solution was slowly dropped in silver ion/PVP
solution under sonication
After adding 28.5 g quaternary ammonium chloride (Cluster Instruments
Co., 80 %) as another stabilizer was dissolved and vigorously stirred for 1
h
The particle size of nanosilver and UV-visible spectrum of nanosilver
colloidal solutions was characterized by Transmission electron
microscopy (TEM) and UV spectrometer, respectively

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The antifungal effects of nanosilver solution carried out at a commercial
greenhouse (Sung-Ju Farm), located at SungJu (Gyeongsangbuk-Do,
Korea), an important area for cut rose production
Rose plants, belonging to the 'Suncity' cultivar, were grown according to
the cultural practices normally adopted by local growers
The nanosilver solution of 500 kg with concentration of 10 ppm was
sprayed at large area of 3306 m2
infected by the rose powdery mildew
Cont....
The antifungal effects were
observed by an optical microscope

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Results and discussion
TEM images of the
nanosilver
in colloidal solution of 1000
ppm
Average size was 1.5 nm with size
distribution of 1-5 nm
 In addition the images showed that
nanosilver particles were densely and well
dispersed in the colloidal solution

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Absorption spectra of a nanosilver colloidal solution after
dilution; (a) only stabilizers (b) 1 ppm, (c) 5 ppm.

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Photographs of rose effected with powdery mildew
before treatment immediately after treatment 2 days after

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Photographs of leaves with powdery mildew
Before treatment a week after treatment

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Before treatment
Optical microscope (80 magnification) images of powdery mildew
on leaf of rose
week after treatment

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Conclusion of the study
The double-capsulized nanosilver showed high dispersity and stability
The photographic results showed that the effects of nanosilver colloidal
solution against rose powdery mildew was very high and durable for a
week
In addition, the nanosilver did not have phyto-toxicity on the plants cell
of leaves, stem and buds of rose plants
As a result, well dispersive and stabilized nanosilver could be
recommended as new fungicide for powdery mildew

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DNA-tagged nano gold: A New Tool
For The Control Of The Spodoptera
litura Fab.
(Chakravarthy et al., 2012)
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CASE STUDY : 2
African Journal of Biotechnology Vol. 11(38), pp. 9295-9301, 10 May, 2012

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Introduction
Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is an extremely
serious pest, the larvae of which can defoliate many economically important
crops cutting across over 40 families
Polyphagous, voracious feeder and very prolific pest
The use of insecticides in agricultural fields leads to an ecological
imbalance in nature and thus in some countries including India, several
insecticides have been banned (Yadav, 2010)
A very recent approach to the control of insect pests is the use of DNA-
tagged nano particles

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 To examine the potential of DNA-tagged nano particles
for the control of Spodoptera litura
Objective of the study

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Material and Methods
A solution of HAuCl4 dissolved in 20 ml of distilled water was used as
the solvent for the preparation of gold (Au) nanoparticle
The solution was continuously stirred in a bath at 110°C for an hour
and then quickly treated with C6H5Na3O7
The Au nanoparticle solution was then further reacted with an
aqueous solution of calf-thymus DNA-sodium salt to obtain DNA
tagged Au nanoparticles
To determine the virulence/lethal concentration of DNA-tagged nano
particle solution, serial dilutions of the nano particle solution ranging
from 200, 300, 400 to 500 ppm were prepared
 10 μl of the suspension was dispensed on the semi-synthetic
chickpea (Cicer arietinum) based diet filled into 5 ml glass vials

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Second instar larvae of S. litura of uniform age and size were released
onto the diet 20 min after surface treatment with DNA-tagged gold
nanoparticles, at all four concentrations viz., 200, 300, 400 and 500 ppm
A control diet was maintained where chickpea based semi-synthetic diet
was applied and used without DNA-tagged gold nanoparticles
Observations of larval settlement on the diet were taken from first day
onwards
Larval mortality was recorded from 3rd till 10th day at 24 h interval.
Each treatment was replicated thrice
Cont.....

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Results and discussion
As the concentration and days after treatment increased, the larval
mortality of 2nd instar S. litura larvae also increased. The maximum
mortality of 30.0 (33.2), 57.5 (49.6) and 75.0 (60.5) was obtained at 500
ppm on 3rd, 4th and 5th day, respectively
There were statistically significant differences between the treatments
and the days after treatment required for 50% mortality of the larvae
At the highest concentration (500 ppm) of the DNA-tagged gold nano
particle, feeding was reduced, larvae turned sluggish and were unable
to orientate towards the source

Treatment (PPM) Percentage of larval mortality days after treatment
3rd
4th
5th
200 10.0 (16.0)b 27.5 (31.4)c 35.0 (36.0)b
300 22.5 (28.2)a 42.5(40.7)b 62.5(52.3)a
400 25.0(29.7)a 55.0 (47.9)ab 72.5 (58.6)a
500 30.0 (33.2)a 57.5 (49.6) a 75.0(60.6)a
Control 0.0 (0.6)c 0.0 (0.6)d 0.0 (0.6)c
SEM ± 2.92 2.87 3.05
CD at 5 % 8.81 8.66 9.20
51 (Chakravarthy et al., 2012)
Effect Of Different Concentrations Of DNA-tagged
With Nano Particle On 2nd
Instar S. litura, Three,
Fourth And Fifth Days After Treatment

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Effect of different concentrations of DNA-tagged gold nanoparticles on
2nd instar S. litura larvae

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2nd instar S. litura larva at
three days after treatment
2nd instar S. litura larva four days
after treatment
The larvae ceased active
movement, the skin and
entire body became stiff
and hard and oozing of
the body content (lysis)
The body became swollen,
pulpy and fragile. It attained
almost a ‘C’- shape and
body turned dark brown
2nd instar S. litura larva at
four days after treatment

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2nd instar S. litura larva at five
days after treatment
2nd instar S. litura larva six
The larvae showed
premature molting in and
half of the body became
discoloured (fleshy white)
and the other half turned
brown
The larvae attained
pupal shape, all the
internal contents oozed
out, and eventually
death occurred

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2nd instar S. litura larva seven
Control
The dead larvae turned black

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Conclusion of the study
This study clearly demonstrates that DNA-tagged gold nanoparticle has
a devastating effect on the larval tissue of S. Litura and would therefore be
a useful component of an integrated pest management strategy
Metal nano-particles could be a better alternative to synthetic
insecticides, in addition to being a toxicant that inhibits biological and
physiological systems of insects
 This experiments clearly established that the DNA tagged gold nano
particle should be tested at concentrations higher than 500 ppm to
determine the effective dose resulting in 50% larval mortality
Subsequent to the laboratory tests on S. litura, field tests on a small
scale need to be initiated

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Other Applications Of Nanotechnology
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According to Mariya Khodakovskaya, a plant biologist, and Alexandru Biris, a
nanotechnologist founded a technique used carbon nanotubes to encourage fast
and early germination of tomato plants
Tomato seeds were planted, some with a growth medium containing carbon
nanotubes, and some without nanotubes in the growth medium
It took only three days for more than 30% of the nanotube tomato seeds to begin
sprouting. In that time, none of the non-treated seeds had even germinated
In fact, it took 12 days for 32% of the tomato seeds without nanotube help to
germinate
After four weeks, the researchers noticed that the tomato plants that had been
treated with carbon nanotubes had two times the biomass and two times the height
of their non-treated counterparts. The current theory is that the nanotubes penetrate
the seed coat of the tomato seeds, allowing water to more rapidly penetrate the
seeds and boost their development
http://phys.org/news174066714.html#jcp
Other Applications of nanotechnology (carbon nanotubes)
Nanotechnology for seed germination

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Interestingly, the root systems were similar in all of the plants, so the nanotubes did
not change the way the roots established themselves
Another issue is that the nanotubes seem to be causing abnormally long internodes,
and that might affect the ultimate outcome regarding the viability of mature plants
The mechanism of ionic interactions with the CNT surface (Miskovic 2008)
implies that redox type changes of the nutrient ion of a given oxidation state
might take place with the MWCNTs in the medium.
There is a potential for the utilization of CNTs for optimizing water transport in arid-
zone agriculture and Horticulture and of improving crop biomass yields
http://phys.org/news174066714.html#jcp

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Nanoparticles in post-harvest disease management
Chitosan, a deacetylated derivative of chitin, is found to be very effective
in reducing postharvest decay of fruit and vegetables (Liu et al. , 2007)
Chitosan/nanosilica hybrid film, extended shelf life, reduced browning
index, retarded weight loss and inhibited the increase of malondialdehyde
amount and polyphenoloxidase activity in fresh longan fruit (Shi et al.
2013)
Pulsing of nano silver (with 2.5 nm diameters) on cut gerbera (Gerbera
jamesonii) cv. Ruikou flowers for 24 h with 5 mg/L nano solution extended
vase life and inhibited the bacteria growth in vase solution for initial 2 days
when observed in vitro under microscope (Liu et al. 2009)

 Biological toxicity
 Environmental toxicity
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Nanotoxicity

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Conclusion
Nanoparticles are biodegradable and target specific, so they can be
successfully employed in production of nanocapsules for delivery of
fungicides, pesticides, fertilizers, and other agrochemicals
Nanotechnology will revolutionize agriculture including crop protection
in the near future
Over the next two decades, the green and golden revolutions would be
accelerated by means of nanotechnology

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Future prospects
More studies are needed to explore the mode of action of NPs, their
interaction with biomolecules, and their impact on the regulation of gene
expressions in plants
Research on nanoparticles with respect to crop protection should be
geared towards introduction of faster and ecofriendly nanoformulations in
future

role of nanotechnology for crop protection in horticultural crops

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