Role of Copper and Zinc Nanoparticles in Plant Disease Management

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Role of Copper and Zinc Nanoparticles in Plant Disease Management
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2. Welcome To
PG Seminar
Series
(Crop
Protection
Group)
2022-23

3. Role of Copper and Zinc Nanoparticles in Plant
Disease Management
Course No.: Pl. Path. 691
Speaker
Vaniya Ravikumar G.
Reg No: 1010120038
4th Sem., Ph. D (Agri.) Plant Pathology
Major Advisor
Dr. Lalit Mahatma
Associate Professor
Dept. of Plant Pathology
N. M. College of Agriculture,
NAU, Navsari.
Co - guide
Dr. P. D. Ghoghari
Associate Research Scientist
(Agril. Entomology)
Main Rice Research Centre,
NAU, Navsari.
1

4. Through the slides…
Introduction
History
Applications of nanotechnology in agriculture
Different approaches and methods for synthesizing
nanoparticles
Case studies
Conclusion
Characterization of nanoparticles
Factors affecting the synthesis of nanoparticles
Different types of nanoparticles using in plant disease
management
2

5. • Nanotechnology is an innovative and emerging discipline in the field of
science and technology. With its broad application, it is now becoming a
key part of life sciences, including approaches to target phytopathogens
for disease management.
• Agrochemicals application against phytopathogens is not sustainable
anymore because of insufficient bioavailability of active and low-impact
compounds.
• Hence, the nature of nanoparticles (NPs), nanoemulsions and
nanoformulations make them efficient nanopesticides to target in a very
efficient way, showing higher solubility, permeability and stability. (Haq
et al., 2020)
• The term “nanotechnology” can be defined as the controlled
manipulation of materials with at least one dimension less than 100 nm.
• Nanoparticles are the simplest form of structures with sizes in the
nanometer range. In principle, any collection of atoms bonded together
with a structural radius of <100 nm can be considered a nanoparticles.
INTRODUCTION
3

6. Fig: 01 Nanoscale integration of nanoparticles and biomolecules
Suryani Saallah and I. Wuled Lenggoro (2017)
Nanoscale
•1 nanometer (nm) means one billionth of a meter.
•1 Nanometer = 10-9 meter
4

7. 1959: Richard Feynman, a renowned physicist
first seeded the concepts of nanotechnology in his
talk “There’s plenty of room at the bottom” in
which he described the possibility of synthesis via
direct manipulation of atoms.
Dr. Norio Taniguchi, a Japanese scientist coined
the term “Nanotechnology” in 1974, and he
defined nanotechnology as “The processing of
separation, consolidation, and deformation of
materials by one atom or one molecule.”
1986: K. Eric Drexler independently used the
term “nanotechnology” in his book “Engines of
creation. The coming era of Nanotechnology”.
He is the cofounder of The foresight Institute, to
help increase public awareness and understanding
of nanotechnology concepts and implications.
5

8. • Agriculture is a basic process of cultivating plants and livestock required
for the survival of human being. It significantly affects our natural
resources and environment.
• Agricultural production through sustainable means could provide long-
term food supply for future growing population without harming our
natural environment and depletion of natural resources.
• Nanotechnology has an eminent potential to improve the agricultural
problem and issues, particularly, reducing the economic losses occurring
due to rapidly occurring climatic changes.
• With the advent of nanotechnology, it has been now possible to enhance
the efficiencies of traditionally used agricultural inputs and supplements,
in a more sophisticated, reliable, and easily accessible way, for improving
food productivity and nutritional security.
• The application of nanomaterials in agricultural sector such as crop
protection, genetic engineering, plant breeding, genetic transformation,
precision farming, and food processing industries.
Applications of nanotechnology in agriculture
6

9. Fig: 02 Applications of nanotechnology in agriculture
Controlled released nanofertilizers improve crop growth, yield and productivity. Nano-based target
delivery approach (gene transfer) is used for crop improvement. Nanopesticides can be used for
efficient crop protection. Uses of nanosensors and computerized controls greatly contribute to precision
farming. Nanomaterials can also be used to remote plant stress tolerance and soil enhancement.
Shang et al. (2019) 7

10. Fig: 03 Uses of nanoparticles in plant protection
Nanoparticles can be used for multiple plant protection purposes, such as pathogen detection
(nanodiagnostics), pest control (against microbial pathogens, fungi, bacteria and insects), weed
control, pesticide remediation, induced resistance and so on.
Shang et al. (2019) 8

11. Fig: 04 Nanoparticles as protectants or carriers to provide crop protection
Worrall et al. (2018) 9

12. Fig: 05 Different approaches and methods for synthesizing nanoparticles
Patra and Hyun Baek (2015) 10

13. UV- vis spectroscopy to follow up the reaction process
Fourier transform infrared (FTIR)
spectroscopy
for detecting types of chemical bonds in a
molecule and analyzing the characteristic
functional groups present in the synthesized
nanoparticles
X-ray diffraction (XRD), transmission
electron microscopy (TEM), and
scanning electron microscopy (SEM).
Information about particle size, crystal
structure, and surface morphology is
obtained
Atomic force microscopy (AFM) or
scanning force microscopy (SFM)
used to image and manipulate atoms and
structures on a variety of surfaces
Energy-dispersive X-ray (EDX)
spectroscopy
used for the elemental analysis or chemical
characterization of nanoparticles
Dynamic light scattering (DLS)
analysis
used to determine the size distribution
profile of nanoparticles
Fig: 06 Characterization of nanoparticles
11

14. Fig: 07 Factor affecting the synthesis of nanoparticles 12

15. • Most nanoparticles information relating to plant disease has involved
the use of metalloids, metallic oxides, or nonmetals as either
bactericide/fungicides or nanofertilizers to affect disease resistance.
• Which include the metalloids B and Si; the metallic oxides of Ag, Al,
Au, Ce, Cu, Fe, Mg, Mn, Ni, Ti, and Zn; and the nonmetal S.
• Given that the vast majority of plant pathology literature focuses on the
nanoparticles Ag, Cu, and Zn, they are discussed first and in greater
detail.
 Nano-Silver: Nanoparticles of Ag were the first to be investigated in
plant disease management given their historically known antimicrobial
activity. Among all the nano-products, maximum are impregnated with
nano-sized silver. Silver nanoparticles (AgNP) are used in a wide range
of applications, including pharmaceuticals, cosmetics, medical devices,
clothing and water purification due to their antimicrobial properties.
Different types of nanoparticles using in plant disease
management
13

16. Fig: 08 Illustrative examples of nanoparticles studies with micronutrients and non-
nutrients on diseases of citrus and maize
Elmer et al. (2018) 14

17.  Nano-Copper: The well-known antimicrobial properties of Cu and
its long history in controlling plant diseases make nano-Cu a logical
choice for plant disease management.
• The first studies that examined nano-Cu as a bactericide/fungicide in
the field were done by Giannousi et al. (2013) who engineered nano-
CuO, Cu2O, and Cu/Cu2O composites and compared foliar
applications to registered commercial Cu-based fungicides,
including Kocide 2000 35 WG, Kocide 30 WG, Cuprofix Disperss
and Ridomil Gold Plus, for their ability to suppress Phytophthora
infestans in tomato.
• They monitored the disease for 10 days once symptoms appeared
and reported that the most effective product for suppressing leaf
lesions was the CuO nanoparticles at 150–340 μg mL-1, followed by
the nano-Cu/Cu2O composite.
• Importantly, the nano-Cu products were equal or superior to the
commercial Cu-based products, delivered less Cu/ha and did not
cause overt phytotoxicity.
15

18.  Nano-Zinc: Similar to Ag and Cu, the antimicrobial activity of
Zn nanoparticles to plant pathogens has been examined by several
laboratories.
• Most in vitro assays have found that nano-Zn inhibits bacteria, a
range of fungal pathogens, including Alternaria alternata,
Botrytis cinerea, Fusarium oxysporum, Mucor plumbeus,
Penicillium expansum, Rhizoctonia solani, Rhizopus stolonifer,
and Sclerotinia sclerotiorum, as well as the nematode
Meloidogyne incognita.
• More importantly, field and greenhouse studies have
demonstrated disease suppression with nano-Zn.
• More attention has been given to managing bacterial diseases
with nano-Zn than diseases caused by other pathogens.
16

19. Fig: 09 Schematic illustration of CuNPs application for protection of plants and its growth
promotion. CuNPs provide protection to crop plants
Mahendra et al. (2018) 17

20. Fig: 10 Probable entry points for nanoparticles in plants
Mahendra et al. (2018) 18

21. Fig: 11 Schematic illustration for possible mechanism of CuNPs on microbes:
CuNPs act on microbial cell wall and disturbs its components, which leads to membrane damage. Membrane
damage decreases the electrochemical potential, which affects membrane integrity. In addition, CuNPs target
DNA, interferes with protein synthesis, and cause damage leading to death of microbial cell.
Mahendra et al. (2018) 19

22. Fig: 12 Mechanisms of Zn oxide nanoparticles antimicrobial activity
Anita Tanwar (2021) 20

23. 01
Iliger et al. (2021)
Kashmir, India. 21

24. Concentration
/
Treatments
Mycelial growth diameter (mm)
25
ppm
50
ppm
100
ppm
150
ppm
250
ppm
500
ppm
1000
ppm
CuNP-Eucalyptus 63.50 50.42 47.50 38.25 32.83 11.25 4.75
CuNP-Mint 61.42 45.25 38.58 30.92 27.83 4.75 0.25
Carbendazim 50 WP* 20.75 20.75 20.75 20.75 20.75 20.75 20.75
Copper oxychloride 50 WP** 11.25 11.25 11.25 11.25 11.25 11.25 11.25
Control 76.00 76.00 76.00 76.00 76.00 76.00 76.00
Mean 46.58 40.73 38.82 35.43 33.73 24.80 22.60
CD p≤ 0.05
* 500 ppm.
** 2500 ppm.
Table: 01 Effect of nanoparticles on mycelial growth of Colletotrichum capsici after
10 days of incubation
22

25. Fig: 13 Mycelial growth Inhibition (%) of Colletotrichum capsici by copper
nanoparticles
Eucalyptus
Mint
23

26. Concentration
/
Treatments
Lesion size (mm)
Nanoparticles applied before
inoculation
Nanoparticles applied after
inoculation
250
ppm
500
ppm
1000
ppm
Mean 250
ppm
500
ppm
1000
ppm
Mean
CuNP-Eucalyptus 20.20 12.70 5.26 12.72 21.70 17.20 7.50 15.63
CuNP-Mint 6.30 0.00 0.00 2.10 14.80 10.50 0.00 8.43
Carbendazim 50 WP* 13.50 13.50 13.50 13.50 22.30 22.30 22.30 22.30
Copper oxychloride 50
WP**
15.50 15.50 15.50 15.50 22.70 22.70 22.70 22.70
Control 65.30 65.30 65.30 65.30 65.30 65.30 65.30 65.30
Mean 24.16 21.40 19.91 21.82 29.36 27.70 23.56 26.87
CD p≤ 0.05
* 500 ppm.
** 2500 ppm.
Table: 02 Effect of copper nanoparticles on lesion size caused by Colletotrichum
capsici chilli under Detached fruit method
24

27. Fig: 14 Effect of copper nanoparticles on lesion number caused by Colletotrichum
capsici under detached fruit method 25

28. 02
Oussou-Azo et al. (2020)
Kagoshima, Japan. 26

29. Fig: 15 Effects of copper forms (Cu-NPs, CuO-NPs, CuO) at various concentrations of
0 (CT), 50, 100, 200, 500, and 1000 mg/ml on Colletotrichum gloeosporioides
hyphal diameter growth 27

30. 03
Malandrakis et al. (2019)
Athens, Greece. 28

31. Fungal strain
EC50
A (μg/ mL ) (mean ± SD B)
Cu-NPs CuO-NPs ZnO-NPs Cu(OH)2
B. cinerea 310.23 ± 2.34 C >1000 670.29 ± 16.32 493.47 ± 21.65
A. alternata 296.56 ± 8.72 >1000 235.48 ± 15.09 210.07 ± 3.15
M. fructicola 219.46 ± 10.12 >1000 628.22 ± 24.05 861.04 ± 44.18
C. gloeosporioides 161.78 ± 14.42 >1000 551.05 ± 18.22 563.44 ± 9.82
F. solani 261.16 ± 12.54 >1000 554.41 ± 30.05 765.61 ± 10.62
F. oxysporum f. sp
Radicis lycopersici
328.12 ± 20.30 >1000 847.44 ± 24.18 856.96 ± 73.82
V. dahliae 191.17 ± 11.33 >1000 386.48 ± 23.14 786.32 ± 55.08
A=EC50 = Effective concentration causing 50% reduction in mycelial growth rate.
B =Standard deviation.
C=Within rows, values followed by the same letter do not differ significantly according to
Tukey's HSD test (α = 0.05).
Table: 03 Effect of NPs on mycelial growth of seven plant pathogenic fungal
strains
29

32. Fungal strain
EC50
A (μg/ mL ) (mean ± SD B)
Cu-NPs CuO-NPs ZnO-NPs Cu(OH)2
B. cinerea 3.05 ± 2.30C 422.11 ± 15.39 16.29 ± 4.00 10.35 ± 0.10
A. alternata 7.69 ± 1.00 678.20 ± 26.56 17.54 ± 3.61 255.82 ± 1.20
M. fructicola 6.45 ± 0.27 215.02 ± 11.08 20.32 ± 2.10 355.51 ± 2.81
C. gloeosporioides 17.44 ± 3.18 178.20 ± 23.12 10.36 ± 1.27 90.65 ± 12.34
F. solani 18.84 ± 2.44 >2000 6.81 ± 0.29 878.24± 30.17
F. oxysporum f. sp
Radicis lycopersici
29.04 ± 4.32 >2000 164.50 ± 24.31 25.23 ± 3.45
V. dahliae 13.32 ± 0.81 252.12 ± 10.33 4.66 ± 0.14 15.25 ± 1.17
A=EC50 = Effective concentration causing 50% reduction in mycelial growth rate.
B =Standard deviation.
C=Within rows, values followed by the same letter do not differ significantly according to
Tukey's HSD test (α = 0.05).
Table: 04 Effect of NPs on spore germination in terms of colony formation of seven
plant pathogenic fungal strains
30

33. 04
Pariona et al. (2019)
Veracruz, Mexico. 31

34. Fig: 16 Antifungal activity of Cu-NPs against: (a) Fusarium solani, (b) Neofusicoccum sp.
and (c) Fusarium oxysporum. (The columns indicate different concentrations of
Cu-NPs: (I) 0 (controls), (II) 0.1, (III) 0.25, (IV) 0.5, (V) 0.75 and (VI) 1.0 mg/ mL
of Cu-NPs). 32

35. Fig: 17 Inhibition of radial growth (IRG) (%) of Cu-NPs against Fusarium solani,
Neofusicoccum sp., and Fusarium oxysporum. (*) indicates significant
differences (P < 0.05) in comparison to their controls (treatments without
Cu-NPs) 33

36. 05
Banik, S. and Alejandro P. L (2017)
Nagaland, India. 34

37. Fig: 18 Radial growth of Alternaria alternata in presence of CuNPs and CoC with
different concentration. (Bars represent data with the same letter within
the same days after inoculation (DAI) are not significantly different (HSD,
p<0.05). 35
Concentration mg/ L

38. Fig: 19 Radial growth of Trichoderma harzianum in presence of CuNPs and CoC.
(Bars represent data with the same letter within the same days after
inoculation (DAI) are not significantly different (HSD, p<0.05). 36
Concentration mg/ L

39. Fig: 20 Growth inhibition of Phytophthora cinnamomi at 7 days after inoculation
(DAI) by CuNPs and copper oxychloride (CoC)
Fig: 21 Growth of Trichoderma harzianum and Cu accumulation in mycelia in
presence of CuNPs and copper oxychloride (CoC) 37

40. Fig: 22 Effect of copper nanoparticles on Pseudomonas syringae in vitro condition.
(A) Number of colonies under different treatments and concentrations
(mg/L) of CuNPs and copper oxychloride (CoC). Data with the same letter
are not significantly different (HSD, p<0.05).
A
38
Concentration mg/ L

41. Fig: 23 Effect of copper nanoparticles and copper oxychloride (CoC) on Pseudomonas
syringae in vitro condition. (B) Growth of Pseudomonas syringae colonies in
Petri dishes. Data with the same letter are not significantly different (HSD,
p<0.05).
B
39

42. Fig: 24 Effect of copper nanoparticles on Rhizobium tropici in vitro condition. (A)
Number of colonies under different treatments and concentrations (mg/L)
of CuNPs and copper oxychloride (CoC). Data with the same letter are not
significantly different (HSD, p<0.05).
A
40
Concentration mg/ L

43. Fig: 25 Effect of copper nanoparticles and copper oxychloride (CoC) on Rhizobium
tropici in vitro condition. (B) Growth of Rhizobium tropici colonies in Petri
dishes. Data with the same letter are not significantly different (HSD,
p<0.05).
B
41

44. 06
Saharan et al. (2015)
Rajasthan, India. 42

45. Treatment
(%)
Seedling growth (A)
Germination
(%)
Seedling length
(cm)
Fresh weight
(gm)
Dry weight
(gm)
SVI
(Seed vigour
index)
Control (B) 96 ± 0.2 13.3 ± 1.8 0.37 ± 0.01 0.015 ± 0.001 1270.5 ± 7.5
Cu– chitosan
NPs 0.08
100 ± 0.0 17.1 ± 0.9 0.44 ± 0.01 0.018 ± 0.001 1701.3 ± 10.0
Cu– chitosan
NPs 0.10
100 ± 0.0 16.9 ± 0.2 0.45 ± 0.01 0.019 ± 0.002 1699.5 ± 9.1
Cu- chitosan
NPs 0.12
100 ± 0.0 15.7 ± 0.3 0.43 ± 0.02 0.017 ± 0.001 1578.7 ± 8.7
Chitosan
(0.1%) (C)
100 ± 0.0 15.0 ± 0.2 0.42 ± 0.01 0.016 ± 0.001 1500.0 ± 5.7
CuSO4 (0.1%)
(D)
60.6 ± 0.0 10.2 ± 0.5 0.23 ± 0.01 0.010 ± 0.001 618.1 ± 7.7
A: Each value is mean of triplicate and each replicate consisted of 10 seedlings. Mean ± SE followed by same
letter in column of each treatment are not significant different at p = 0.05 as determined by Tukey–Kramer
HSD.
B: Control with water.
C: Dissolved in 0.1% acetic acid.
D: Dissolved in water.
Table: 05 Effect of Cu–chitosan NPs on seedling growth of tomato (data recorded
after 10 days of growth)
43

46. Fig: 26 Effect of Cu– chitosan NPs on tomato seedling growth at various concentration 44

47. Treatment
(%)
% Inhibition (mycelial growth) (A) % Inhibition (spore germination)
Alternaria solani Fusarium
oxysporum
Alternaria solani Fusarium
oxysporum
Control (B) 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0 0.0 ± 0.0
Cu– chitosan NPs
0.08
52.7 ± 1.1 60.3 ± 1.2 46.2 ± 3.9 56.6 ± 3.6
Cu– chitosan NPs
0.10
68.7 ± 1.3 73.3 ± 0.3 60.1 ± 1.8 79.9 ± 2.2
Cu- chitosan NPs
0.12
70.5 ± 0.5 73.5 ± 3.1 61.5 ± 3.1 83.0 ± 1.5
Chitosan (0.1%)
(C)
23.1 ± 0.2 20.5 ± 1.2 18.0 ± 0.8 24.1 ± 1.2
CuSO4 (0.1%) (D) 25.0 ± 0.7 22.4 ± 0.1 21.3 ± 0.9 27.0 ± 1.1
A: Each value is mean of triplicate from two experiments. Mean ± SE followed by same letter in column of
each treatment are not significant different at p = 0.05 as determined by Tukey–Kramer HSD, % inhibition
rate was calculated compared to the mycelia growth and spore germination in control (0%).
B:Control with water.
C:Dissolved in 0.1% acetic acid.
D: Dissolved in water.
Table: 06 Effect of Cu–chitosan NPs on in vitro mycelial growth and spore
germination of A. solani and F. oxysporum
45

48. Treatment
(%)
Early blight Fusarium wilt
Disease severity
(%) A
PEDC (%) A Disease severity
(%) A
PEDC (%) A
Control (B) 55.0 ± 2.8 00.0 ± 0.0 66.6 ± 3.3 00.0 ± 0.0
Positive control
(C)
18.0 ± 0.0 73.7 ± 0.3 18.0 ± 0.0 73.7 ± 0.5
Cu– chitosan
NPs 0.08
15.3 ± 1.7 72.3 ± 1.8 50.0 ± 0.0 38.8 ± 5.5
Cu– chitosan
NPs 0.10
8.6 ± 0.6 84.2 ± 0.6 41.6 ± 3.3 49.9 ± 9.6
Cu- chitosan
NPs 0.12
6.6 ± 1.7 87.7 ± 3.3 33.3 ± 3.3 61.1 ± 5.5
A: Each value is mean of triplicate from two experiments. Mean ± SE followed by same letter in
column of each treatment are not significant different at p = 0.05 as determined by Tukey–
Kramer HSD.
B:Control with water.
C:Mancozeb (0.2%).
PEDC = percentage efficacy of disease control was calculated as compared to control.
Table: 07 Effect of Cu–chitosan NPs on control of early blight and Fusarium wilt
of tomato in pot conditions
46

49. 07
Kanhed et al. (2014)
Maharashtra, India. 47

50. Sr. no. Pathogenic fungi
Zone of inhibition (mm)
CuNPs IPA Bavistin (A) CuNPs +
Bavistin (B)
01 Phoma distructiva 22 ± 1 00 12 ± 0.5 22 ± 0.5
02 Curvularia lunata 21 ± 0.5 00 00 21 ± 0.5
03 Alternaria
alternata
18 ± 1.1 00 00 18 ± 1
04 Fusarium
oxysporum
24 ± 0.5 00 10± 0.5 26 ± 0.5
All experiments were done in triplicate.
Table: 08 Antifungal activity of CuNPs against four plant pathogenic fungi Phoma
distructiva, Curvularia lunata, Alternaria alternata, Fusarium oxysporum
48

51. Fig: 27 Antifungal activity of CuNPs against 1. Phoma distructiva, 2. Curvularia
lunata, 3. Alternaria alternata and 4. Fusarium oxysporum, where (A)
copper nanoparticles, (B) IPA, (C) Antifungal agent (Bavistin) and (D)
Copper nanoparticles + Bavistin 49

52. 08
Ouda. S. M. (2014)
Cairo, Egypt. 50

53. Incubation
periods
(Days)
Control
Fungal growth diameter (mm) (Alternaria alternata )
Copper nanoparticles Silver/ copper nanoparticles
1 5 10 15 1 5 10 15
2 28.5 ± 0.5 27.5 ±
0.5
26.0 ±
0.0
27.0 ±
0.2
27.0 ±
0.1
28.5 ±
0.5
28.5 ±
0.5
28.0 ±
1.0
21.5
± 0.5
3 38.5 ± 0.5 38.0 ±
0.0
38.0 ±
0.5
37.0 ±
0.0
38.5 ±
0.5
33.0 ±
0.0
38.5 ±
0.5
38.5 ±
0.0
30.0
± 0.5
5 64.5 ± 0.5 64.5 ±
0.5
64.5 ±
0.5
62.5 ±
0.5
62.5 ±
0.5
68.0 ±
1.0
67.5 ±
0.5
68.5 ±
0.5
43.0
± 1.0
6 80.0 ± 0.0 80.0 ±
0.0
80.0 ±
0.0
70.5 ±
0.0
69.0 ±
0.0
79.0 ±
0.0
78.0 ±
0.0
78.0 ±
0.0
49.5
± 0.0
Table: 09 Antifungal activities of copper and silver/ copper nanoparticles against
Alternaria alternata at different concentrations (mg/L) and for different
incubation periods
51

54. Incubation
periods
(Days)
Control
Fungal growth diameter (mm) (Botrytis cinerea )
Copper nanoparticles Silver/ copper nanoparticles
1 5 10 15 1 5 10 15
2 34.0 ± 1.0 29.0 ±
0.5
25.5 ±
0.5
25.0 ±
0.0
25.0 ±
0.0
34.0 ±
0.0
34.5 ±
0.5
33.0 ±
1.0
25.0
± 0.0
3 52.0 ± 1.0 45.0 ±
2.0
45.5 ±
0.5
45.0 ±
1.0
45.0 ±
0.0
52.0 ±
0.5
52.0 ±
0.5
53.5 ±
0.5
34.0
± 1.0
5 79.0 ± 0.0 79.0 ±
0.0
79.0 ±
0.0
79.0 ±
0.0
75.0 ±
0.5
79.5 ±
0.5
78.5 ±
0.5
78.5 ±
0.5
56.0
± 1.0
6 85.0 ± 0.0 85.0 ±
0.0
82.0 ±
0.0
82.0 ±
0.0
81.0 ±
0.0
85.0 ±
0.0
85.0 ±
0.0
85.0 ±
0.0
62.0
± 1.0
Table: 10 Antifungal activities of copper and silver/ copper nanoparticles against
Botrytis cinerea at different concentrations (mg/L) and for different
incubation periods
52

55. 09
Merino et al. (2021)
Saltillo, Mexico. 53

56. Table: 11 Percent inhibition means, sporulation inhibition per cent, ± standard deviation of
Fusarium oxysporum in PDA medium at different concentrations of ZnO and
ZnO NPs at 8 days of evaluation
54

57. Fig: 28 Antifungal activity of ZnO NPs on Fusarium oxysporum in vitro test: (a) 0
ppm, (b) 3,000 ppm, (c) 2000 ppm, (d) 1,600 ppm, (e) 1,200 ppm, (f) 800
ppm, (g) 400 ppm, (h) 200 ppm and (i) 100 ppm
D
A C
B
G H
F
I
E
55

58. Fig: 29 Control of Fusarium oxysporum with ZnO NPs and ZnO in Solanum
lycopersicum tomato plants. (a) ZnO NPs at 3,000 ppm, (b) ZnO NPs at
1,500 ppm, (c) ZnO NPs at 100 ppm, (d) ZnO at 3,000 ppm, (e) ZnO at
1,500 ppm, (f) ZnO at 100 ppm
A
B
C
D
E
F
56

59. 10
Carvalho et al. (2019)
Florida, USA. 57

60. Fig: 30 In vitro activity of Cu/Zn hybrid nanoparticles on X. perforans population at 15 min,
1 h, 4 h and 24 h. The Cu/Zn hybrid nano particle treatments were 1000, 500, 200
and 100 μg/ mL. Micron-sized commercial copper at the same concentrations; and
sterile tap water served as the control. (A) Copper-tolerant X. perforans GEV485.
(B) Copper-sensitive X. perforans 91–118. 58

61. Fig: 31 Effect of Cu/Zn hybrid nanoparticles on development of tomato bacterial spot in
planta in growth chamber experiments. (The area under disease progress curve
(AUDPC) was calculated using Horsfall-Barratt scale collected every day since the
third day after inoculation until twentieth day after inoculation.). 59

62. Fig: 32 Percentage of leaves showing evident bacterial spot symptoms by the total
number of leaves in the plant after application of Cu/Zn nanoparticles,
Kocide 3000, Kocide 3000 amended by Mancozeb and untreated samples.
Data collected after the last in planta experiment (20 days after
inoculation). 60

63. 11
Elsharkawy et al. (2018)
Kafrelsheikh, Egypt. 61

64. Fig: 33 Disease symptoms caused by Pseudomonas syringae pv. tomato infection in
water-treated tomato plants (control) and tomato treated with ZnO
nanoparticles by foliar spray after 1 week of Pseudomonas syringae pv.
tomato challenge inoculation. 62

65. Fig: 34 Induced suppression of disease symptoms and the number of Pseudomonas
syringae bacteria in tomato plants in response to treatments with ASM,
BTH, streptomycin, and ZnO nanoparticles after 1 day of Pseudomonas
syringae inoculation. Columns represent mean values. 63

66. Table: 12 Effects of ASM, BTH, streptomycin, and ZnO NPs on defense enzymes
(POD) and (PPO) in tomato at 1 day after Psuedomonas syringae
challenge inoculation
64

67. 12
El- Argawy et al. (2017)
Egypt. 65

68. Fungi Untreated
Control
ZnO Nanoparticles (ppm)
25 ppm 50 ppm 100 ppm
Fusarium
oxysporum
f. sp. betae
0.00 32.04 44.29 53.06
Sclerotium
rolfsii
0.00 72.56 73.67 77.44
Rhizoctonia
solani
0.00 49.22 50.00 59.67
Mean 0.00 51.27 55.99 63.39
Means followed different letters are significantly different at p=0.05 of probability,
inhibition effects were determined based on five replicates for each treatment.
Table: 13 In vitro percentage of inhibition of radial growth of sugar beet root rot
pathogenic fungi on PDA by nanoparticles concentrations
66

69. Fungi
Treatments (%)
TSS Sucrose
Control Vitavax ZnO Control Vitavax ZnO
Fusarium
oxysporum
f. sp. betae
17.17 24 20.33 13.97 18.78 17.25
Sclerotium
rolfsii
16 22 21.67 13.43 17.52 17.51
Rhizoctonia
solani
17 21 21.33 13.96 17.14 17.37
Mean 16.72 22.33 21.10 13.79 17.80 17.38
Table: 14 In vivo effect of nanoparticles on TSS and sucrose content of sugar beet
roots grown in soil infested with sugar beet root rot fungi in pot
experiments, 150 days after planting
67

70. Fungi
Treatments (%)
Total phenols (mg g-1 fresh
tissue
Polyphenol oxidase activity
(Absorbance min g-1 fresh tissue)
Control Vitavax ZnO Control Vitavax ZnO
Fusarium
oxysporum
f. sp. betae
4.60 8.83 7.83 0.025 0.059 0.051
Sclerotium
rolfsii
3.83 8.75 7.75 0.026 0.062 0.052
Rhizoctonia
solani
4.73 8.49 8.68 0.030 0.059 0.073
Mean 4.39 8.69 8.09 0.027 0.060 0.059
Table: 15 Effect of ZnO nanoparticles on total phenol content and polyphenol
oxidase of sugar beet roots grown in soil infested with sugar beet root
rot fungi in pot experiments, 150 days after planting
68

71. 13
Wagner et al. (2016)
Lexington, USA. 69

72. Fig: 35 Germination frequency of Peronospora tabacina spores in the presence of
Zn treatments 70

73. Fig: 36 Germination frequency of Peronospora tabacina spores in the presence of
Zn treatments 71

74. Fig: 37 Spore germination tube length as measured for selected treatments 72

75. Fig: 38 Percentage of tobacco plants infected by P. tabacina in the presence of Zn
treatments 73

76. 14
Zabrieski et al. (2015)
Logan, USA. 74

77. Fig: 39 The effect of ZnO and CuO NPs on mycelial growth of Pythium
aphanidermatum (Pa) or Pythium ultimum (Pu) on PDA at 24 hr after
inoculation. The NPs were added at the doses shown: 50, 100, 250 and
500 mg/ mL. 75

78. 15
Yehia and Ahmed (2013)
Cairo University, Egypt. 76

79. Fig: 40 Effect of ZnO nanoparticles on the colony growth diameter of Fusarium
oxysporum 77

80. Fig: 41 Effect of ZnO nanoparticles on the colony growth diameter Penicillium
expansum 78

81. Table: 16 Effect of ZnO nanoparticles at different concentrations on production of
mycotoxins (mg g-1 dry mass) by Fusarium oxysporum and Penicillium
expansum
79

82. Synthesization and characterization of Banana
Pseudostem-chitosan mediated silver nanoparticles
and their antifungal activities
Musmade (2021)
Navsari, Gujarat.
16
80

83. Treatment
No.
Treatment
details
% Growth of inhibition on PDA
Concentration (ppm)
10 50 100 150 200 250 300
T1 BSE-
AgNPs
0.00 1.76 10.88 39.12 69.12 100.0 100.0
T2 Chi-
AgNPs
0.00 8.24 69.41 86.47 100.0 100.0 100.0
T3 BSE- Chi-
AgNPs
5.59 21.47 75.00 100.0 100.0 100.0 100.0
T4 AgNO3 0.00 100.0 100.0 100.0 100.0 100.0 100.0
T5 Control 0.00 0.00 0.00 0.00 0.00 0.00 0.00
*All the figures were average of four observations
Table: 17 In vitro efficacy of synthesized AgNPs against Sclerotium rolfsii
81

84. Fig: 42 In vitro efficacy of synthesized AgNPs against Sclerotium rolfsii 82

85. Treatment
No.
Treatment
details
% Growth of inhibition on PDA
Concentration (ppm)
10 50 100 150 200 250 300
T1 BSE-
AgNPs
0.00 0.00 6.47 36.76 71.76 91.76 100.0
T2 Chi-
AgNPs
0.00 10.30 24.41 76.47 94.71 100.0 100.0
T3 BSE- Chi-
AgNPs
0.00 22.94 49.41 90.00 100.0 100.0 100.0
T4 AgNO3 0.00 34.41 100.0 100.0 100.0 100.0 100.0
T5 Control 0.00 0.00 0.00 0.00 0.00 0.00 0.00
*All the figures were average of four observations
Table: 18 In vitro efficacy of synthesized AgNPs against Pythium aphanidermatum
83

86. Fig: 43 In vitro efficacy of synthesized AgNPs against Pythium aphanidermatum 84

87. Synthesis of plant mediated nanoparticles and their
antimicrobial activities
Ramya (2018)
Navsari, Gujarat.
17
85

88. Treatment details AgNO3
Concentration
Inhibition %
Sclerotium rolfsii Trichoderma viridae
T1 AgNPs
synthesized
from Garlic
3mM 18.82 2.35
5mM 64.71 19.61
T2 AgNPs
synthesized
from Datura
3mM 14.90 25.10
5mM 32.16 33.73
T3 AgNO3 Solution 3mM 11.37 61.96
5mM 29.80 61.96
T4 Garlic clove extract 66.27 56.08
T5 Datura leaves extract 27.45 35.69
S. Em ± 1.65 1.47
C.D at 5% 4.94 4.40
C.V % 8.59 6.86
Table: 19 In vitro efficacy of green synthesized AgNPs against S. rolfsii and T.
viridae
86

89. Fig: 44 In vitro efficacy of silver nanoparticles synthesized by green method against tomato
collar rot causing by Sclerotium rolfsii
(A) Garlic AgNPs, (B) Datura AgNPs, (C) AgNO3 Solution,
(1) 1 mM AgNO3 Solution, (2) 2 mM AgNO3 Solution, (3) 3 mM AgNO3 Solution, (4) 5 mM
AgNO3 Solution 87

90. Fig: 45 In vitro efficacy of silver nanoparticles synthesized by green method against
Trichoderma viridae
(A) Garlic AgNPs, (B) Datura AgNPs, (C) AgNO3 Solution,
(1) 1 mM AgNO3 Solution, (2) 2 mM AgNO3 Solution, (3) 3 mM AgNO3 Solution, (4) 5 mM
AgNO3 Solution 88

91. Treatment
details
AgNO3
Conc.
Inhibition %
Xanthomonas
oryzae pv.
oryzae
Bacillus
megaterium
Azotobacter
chroococcum
Frateuria
aurantia
Pseudomonas
fluorescens
T1 AgNPs
synthesized
from Garlic
3mM 1.38 1.14 1.10 1.11 1.14
5mM 1.65 1.10 1.11 1.13 1.13
T2 AgNPs
synthesized
from
Datura
3mM 1.29 1.10 0.71 1.17 1.13
5mM 1.44 1.13 0.71 1.21 1.13
T3 AgNO3
Solution
3mM 1.37 1.32 1.14 1.17 1.33
5mM 1.37 1.40 1.12 1.24 1.32
T4 Garlic clove extract 1.66 1.53 1.35 1.20 1.43
T5 Datura leaves extract 1.37 1.38 0.71 0.71 1.35
S. Em ± 0.01 0.01 0.01 0.02 0.02
C.D at 5% 0.04 0.03 0.04 0.05 0.05
C.V % 1.50 1.42 2.50 2.47 2.35
Table: 20 In vitro efficacy of green synthesized AgNPs against different bacteria
89

92. Fig: 46 In vitro efficacy of silver nanoparticles synthesized by green method against
different bacteria
(A) Garlic AgNPs, (B) Datura AgNPs, (C) AgNO3 Solution,
(1) Xanthomonas oryzae pv. oryzae, (2) Bacillus megaterium, (3) Azotobacter chroococcum,
(4) Frateuria aurantia, (5) Pseudomonas fluorescens
C3
90

93. • Applications of nanotechnology have a promising future in
agricultural science, and they can be a great source of innovation
to improve yields and significantly contribute to precision
agriculture farming practices.
• Engineered nanoparticles are increasingly used in disease
management strategies as bactericides/fungicides and as
nanofertilizers to enhance plant health.
• To date, most reports have centered on NPs of Ag, Cu, and Zn for
their antimicrobial activity and their ability to alter host defense.
We predict that nanoparticles will ultimately may play a major
role in suppressing disease in both the greenhouse and field.
• One major impact of using nano particles in disease management
is the large reduction in active metals entering the environment
when compared to the conventional metallic fungicides.
CONCLUSION
93

94. • One challenging obstacle facing researchers is that
nanomaterials might behave differently in different plant/disease
systems, requiring that each disease system may need separate
evaluation.
• Given the large challenges currently faced in agriculture, the
development of nano-enabled disease suppression strategies will
most certainly be an important tool in the effort to achieve and
sustain global food security.
• Nanotechnology can provide solutions for agricultural
applications and has the potential to revolutionize the existing
technologies used in disease management.
94

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