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American Journal of  
Life Science and Innovation (AJLSI)

Evaluation of  the Antifungal Effect of  Green Synthesized Metal Oxide Nanoparticles 
Against Plant Pathogenic Rhizoctonia Species

H. K. S. Madusanka1*, A. G. B. Aruggoda1, J. A. S. Chathurika2, S. R. Weerakoon3

Volume 3 Issue 2, Year 2024
ISSN: 2833-1397 (Online)

DOI: https://doi.org/10.54536/ajlsi.v3i2.4281
https://journals.e-palli.com/home/index.php/ajlsi

Article Information ABSTRACT

Received: November 08, 2024

Accepted: December 10, 2024

Published: December 15, 2024

The current study successfully synthesized zinc oxide (ZnO), copper oxide (CuO), and 
iron oxide (FeO) nanoparticles (NPs) using cost-effective and environmentally friendly 
procedures. The synthesized NPs were characterized by UV-Vis spectroscopy and SEM 
analysis. UV-Vis spectroscopy revealed characteristic absorption peaks at 356 nm for ZnO 
NPs, confirming their synthesis. SEM analysis showed a heterogeneous distribution of  
nanoparticle sizes, with ZnO NPs averaging 81 nm, CuO NPs averaging 108 nm, and FeO 
NPs averaging 82 nm. The antifungal activity of  the synthesized nanoparticles was evaluated 
at various concentrations using the poisoned food technique. The results indicated a dose-
dependent inhibition of  mycelial growth by ZnO and CuO NPs, with higher concentrations 
(500 and 1000 mg/L) showing significant inhibition compared to untreated Rhizoctonia 
species. Specifically, CuO NPs exhibited mycelial growth inhibition percentages of  70.64% 
and 73.43% at 500 and 1000 mg/L, respectively, while ZnO NPs showed inhibition 
percentages of  78.38% and 80.94% at the same concentrations. Statistical analysis using 
one-way ANOVA revealed significant differences among the treatment groups (p < 0.001). 
In contrast, FeO NPs did not exhibit a dose-dependent inhibition of  mycelial growth but 
showed a minor, statistically insignificant promotion. Among the tested NPs, CuO NPs at 
1000 mg/L achieved the highest inhibition, followed by ZnO NPs. The observed variations 
in mycelial inhibition by different nanoparticles at various concentrations underscore the 
complexity of  nanoparticle-pathogen interactions and highlight the need for further research 
to optimize their antifungal efficacy.

Keywords

Antifungal Activity, Green-
Synthesized Nanoparticles, Poisoned 
Food Technique, Rhizoctonia Species

INTRODUCTION 
The Rhizoctonia genus belongs to the Basidiomycete 
phylum and consists of  anamorphic fungi with different 
teleomorphs (Moliszewska et al., 2023). Rhizoctonia spp. 
is a significant plant pathogenic fungus responsible for 
diseases such as wilt and root rot. These diseases can 
manifest during the seedling stage as well as in the later 
stages of  crop growth, leading to early infections that 
often result in plant death (Jaiswal et al., 2014). R. solani 
produces numerous enzymes and phytotoxic substances 
that contribute to its pathogenicity, causing specific fungal 
disease symptoms in crops (Mousa et al., 2024). The 
Rhizoctonia genus comprises a vast and diverse group of  
soil-dwelling fungi, including binucleate Rhizoctonia. While 
many species within this genus are plant pathogens and 
typically polyphagous, they exhibit distinct pathogenic 
preferences for specific plant species. Despite their broad 
and varied host range, their affinity for certain plants is 
individually determined. Among them, Rhizoctonia solani 
is the most commonly reported and is recognized as a 
significant plant pathogen (Moliszewska et al., 2023).
The current research aims to investigate the antifungal 
potential of  green-synthesized nanoparticles, including 
ZnO, CuO, and FeO NPs, against Rhizoctonia species. The 
study will involve synthesizing the nanoparticles using 
green methods and characterizing their physicochemical 

properties. Antifungal activity will be evaluated over a 
range of  concentrations (0, 50, 100, 500, and 1000 mg/L) 
using the poison food technique to determine inhibition 
effects. 

LITERATURE REVIEW
Rhizoctonia spp cause significant damage to agricultural 
crops cuasing several diseases. Sheath blight, caused 
by Rhizoctonia solani, is a significant challenge for rice 
farmers, leading to yield losses ranging from 25% to 
50% (Chowdhury et al., 2019). Rhizoctonia species are 
responsible for several diseases across different crops: 
soybeans suffer from aerial blight and web blight, 
tomatoes are susceptible to soil rot and fruit rot, peppers 
are affected by damping-off, and eggplants are prone to 
brown spot (Tu et al., 1996).
Disease management for Rhizoctonia spp is done by 
Cultural Practices and Fungicides. Cultural practices 
includes Select cultivars that are resistant, tolerant, or 
less susceptible to Rhizoctonia diseases,  purchase certified 
disease-free seed or disease free transplants, crop rotation 
etc. Although cultural approache is most essential control 
strategy, chemical management is still an important 
tool to decrease R. solani (Tsror, 2010).  As chemical 
management, applying Pentachloronitrobenzene-based 
fungicides , Tebuconazole,  carbendazim,  iprodione, and 

1 Department of  Agricultural and Plantation Engineering, The Open University of   Sri Lanka, Nawala, Nugegoda, Sri Lanka
2 Department of  Urban Bio-resources, University of  Sri Jayawardanapura, Ganagodawila, Nugegoda, Sri Lanka
3 Department of  Botany, The Open University of  Sri Lanka, Nawala, Nugegoda, Sri Lanka
* Corresponding author’s e-mail: hksma@ou.ac.lk



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chlorothaloniland systemic fungicides such as carboxin, 
triadimefon, and thiophanate-methylis effective against 
Rhizoctonia (Karkee & Mandal, 2020; Chaube & Pundhir, 
2005).
The growing concern over alternative approaches to 
chemical management of  pathogenic fungi is closely 
linked to soil and environmental health. This includes 
issues such as increased pesticide residues in soil and 
water bodies, reduction in beneficial soil organisms, 
changes in soil physical and chemical properties, and 
pesticide resistance in pathogens (Sharma et al., 2019). 
Chemical control is the most widely used method to 
manage soilborne fungal pathogens. However, the 
negative environmental and health effects, disruption 
of  natural ecosystem balance, and the emergence of  
fungal resistance to chemical pesticides highlight the 
need for sustainable and cost-effective alternatives. 
Research has demonstrated that nanoparticles show 
promise as effective antimicrobial agents and biosensors 
for detecting plant pathogens, particularly against soil-
borne varieties, offering a comparatively environmentally 
friendly solution (Dutta et al., 2023). Nanoparticles smaller 
than 100 nm possess a higher surface area to volume ratio 
and enhanced reactivity, which enhances their suitability 
for widespread application in both human and plant 
pathology (Jeevanandam et al., 2018).

MATERIALS AND METHODS
Biologic Material and Chemical Reagents
Fresh samples of  S. molesta and Mimosa pigra were collected 
from Uhana, Ampara, Sri Lanka. High-purity Zinc Nitrate 
hexahydrate (Zn(NO3)2·6H2O) was procured from Loba 
Chemie Pvt. Ltd. Extra pure analytical reagent grade 
cupric sulfate (CuSO4·5H2O, minimum purity 99.5%) 
was acquired from Sisco Research Laboratories Pvt. Ltd. 
Sodium hydroxide pellets (NaOH, minimum purity 98%) 
were sourced from Loba Chemie Pvt. Ltd. Anhydrous 
ferric chloride of  analytical research grade (FeCl3) was 
obtained from HiMedia Laboratories Pvt. Ltd. Potato 
Dextrose Agar (PDA) was purchased from HiMedia 
Laboratories Pvt. Ltd. Fusarium species were provided by 
the Plant Virus Indexing Centre in Homagama, Sri Lanka.

Preparation of  the Mimosa Pigra Extract and  
Synthesis of  ZnO NPs
M. pigra leaves were thoroughly washed with tap water 
and rinsed with DDW to remove all debris. The washed 
leaves were then dehydrated at 42°C for 24 hours, ground, 
and sieved to remove large particles. Eight grams of  the 
dried leaf  powder were mixed with 100 mL of  DDW 
and stirred for 2 hours at 60°C and 400 rpm. The extract 
was centrifuged at 6000 rpm for 10 minutes, and the 
supernatant was filtered three times using Whatman No. 
1 filter paper. Three grams of  Zinc Nitrate hexahydrate 
(Zn(NO3)2·6H2O) were dissolved in 60 mL of  the plant 
extract at room temperature and stirred for 1 hour. The 
solution was then heated in a thermal bath at 60°C for 12 

hours to remove the water content. The resulting viscous 
paste was incubated at 100°C for 2 hours to form a 
crystalline powder-like material, which was subsequently 
calcined at 500°C for 2 hours.

Preparation of  the Salvinia molesta  extract and  
Synthesis of  FeO NPs
S. molesta was thoroughly washed and dehydrated at 
42°C for 24 hours. The dried plants were then ground 
and sieved to obtain a fine powder. Five grams of  the 
powdered leaves were immersed in 80 mL of  DDW and 
stirred for 1 hour at 60°C. The extract was filtered three 
times using Whatman No. 1 filter paper. This leaf  extract 
was mixed with a pre-prepared 0.1 M FeCl3 solution in 
a 2:3 ratio. The pH of  the mixture was raised to 10 by 
incrementally adding 1 M sodium hydroxide. The reaction 
mixture was left undisturbed for 24 hours to facilitate the 
synthesis of  FeO NPs. Subsequently, the mixture was 
centrifuged at 6000 rpm for 15 minutes, and the sediment 
was collected. The synthesized FeO NPs were washed 
trice with DDW and then calcined at 500°C for 2 hours. 
The resulting solid pellets were crushed using a mortar 
and pestle to obtain a fine powder-like structure.

Characterization Techniques
To verify the successful synthesis of  ZnO, CuO, and FeO 
nanoparticles, UV-Visible (UV-Vis) spectroscopy analysis 
was conducted with a resolution of  1 nm over a range of  
190 nm to 800 nm using a quartz cuvette. The morphology 
of  the synthesized nanoparticles was inspected using 
Scanning Electron Microscopy (SEM). The SEM images 
were analyzed, and particle sizes were measured. 

In Vitro Evaluation of  Antifungal Activity on 
Mycelial Development
The antifungal activity of  the nanoparticles against 
Fusarium species was assessed using the poisoned food 
technique. Potato dextrose agar (PDA) medium was 
prepared with nanoparticle suspensions at concentrations 
of  0 (negative control), 50, 100, 500, and 1000 mg/L, and 
Petri dishes (9 cm) were prepared with three replicates 
for each treatment. Agar discs (8 mm in diameter) 
containing mycelia of  Fusarium species (7 days old, 28 ± 
2°C) were then inoculated onto these plates. The plates 
were incubated at 28 ± 2°C until the fungal growth nearly 
covered the plate (9 cm). Mycelial growth was measured as 
diameter (mm). The percentage inhibition was calculated 
using the following formula (1):
Inhibition percentage (%) = C-T/C ×100             (1)
Where “C” is the diameter of  the fungal disc in the 
control, and “T” is the diameter of  the fungal disc treated 
with different nanoparticle concentrations. Analysis 
of  variance (ANOVA) was employed to determine 
significant differences between each mycelial growth 
treatment, with a 95% confidence interval (α = 0.05) 
applied. Dunnett’s test was used to identify differences 
between the means of  the inhibition percentage data 



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compared to the control group. Duncan’s Multiple Range 
Test (DMRT) was used to identify differences between 
the means of  the inhibition percentage data for each 
treatment. The analysis was conducted using R statistical 
software (RStudio 2023.09.1+494).

RESULTS AND DISCUSSION
The synthesized nanoparticles (ZnO, CuO, and FeO) 
were characterized to confirm their successful synthesis 
and evaluate their properties. The UV-Vis spectroscopy 
analysis revealed distinct absorption peaks, indicative of  

Figure 1: UV-Vis spectroscopy analysis of  ZnO NPs

Figure 2: SEM images of  ZnO NPs at a scale of  200 nm

Figure 3: UV-Vis spectroscopy analysis of  CuO NPs



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the formation of  nanoparticles. The morphology and size 
of  the nanoparticles were further analyzed using SEM. 

Characterization of  ZnO NPs 
The green-synthesized ZnO NPs were characterized 

using UV-Vis spectroscopy with DMSO as a reference. 
Figure 1 shows a peak absorption at 356 nm, which 
preliminarily confirms the synthesis of  ZnO NPs. The 
consistent result was observed in a previous study of  
Green synthesis of  ZnO NPs using an aqueous extract of  

Figure 4: SEM images of  CuO NPs at a scale of  10 µm 

Figure 5: UV-vis spectroscopy analysis of  FeO NPs

Figure 6: SEM images of  FeO NPs at a scale of  200 nm



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Epipremnum aureum leaves photocatalytic degradation 
of  Congo red having 326 as the maximum absorbance 
(Razia Sultana Brishti et al., 2024). Another study of  the 
synthesis of  ZnO nanoparticles using peels of  Passiflora 
foetida observed maximum absorbance at a wavelength 
ranging from 200 to 380 nm (Khan et al., 2021). In a study 
on the green synthesis of  ZnO-NPs using Juglans regia 
green husk aqueous extract, the biosynthesized ZnO-NPs 
exhibited a prominent maximum absorbance band at 360 
nm, as indicated by the UV-Vis analysis results(Dehghani 
& Parinaz Ghadam, 2023). Collectively, these results 
affirm the successful production of  ZnO NPs in the 
current study.
SEM images in Figure 2 reveal a heterogeneous 
distribution of  shapes and sizes within the nanoscale 
range, with an average particle size of  approximately 81 
nm (35-164 nm). Consistent results have been reported 
for green-synthesized ZnO NPs with particle sizes of  30-
100 nm (Thu et al., 2022), 79 nm (Alyamani et al., 2021), 
and 75-90 nm (Shanthi Sathappan et al., 2021).

Characterization of  CuO NPs
Maximum absorbance in the UV-Vis spectroscopy range 
of  190 nm to 450 nm was observed, with a peak at 224 
nm, confirming the green synthesis of  nanoparticles 
as shown in Figure 3. Similar results were reported in 
previous studies with absorption peaks at 265 nm (Kumar 
et al., 2019), 220 nm (Renuga et al., 2020), and 250–255 

nm (Kumar et al., 2017).
The SEM images illustrated in Figure 04 reveal CuO NPs 
with diverse shapes and sizes, averaging approximately 
108 nm in diameter, and show a tendency to aggregate 
and form various structures. Consistent results were 
also obtained in previous studies, reporting particle 
sizes of  18-106 nm (Selvi et al., 2019), 197 nm (Arushi 
Saloki & Daharwal, 2023), and 20-200 nm (Manivannan 
Rangasamy et al., 2023).

Characterization of  FeO NPs 
UV-Vis spectrometry indicated the formation of  green-
synthesized nanoparticles within the range of  200 to 600 
nm, with a maximum absorbance at 300 nm, confirming 
the synthesis of  FeO NPs. Consistent results were also 
observed in previous studies, with maximum absorbance 
at 295–301 nm (Karpagavinayagam & Vedhi, 2019), 293 
nm (Andrade-Zavaleta et al., 2022), and 230 and 290 nm 
(Hussain et al., 2023).
SEM images in Figure 6 show the shapes and sizes of  
the iron oxide nanoparticles, which exhibit a variety 
of  nanoscale shapes and sizes, averaging around 82 
nm (ranging from 41 to 140 nm), and tend to clump 
together and form multiple structures. Similar results 
were also observed having particle sizes 30–100 nm 
(Karpagavinayagam & Vedhi, 2019), and 88 nm (Aigbe 
& Osibote, 2024). 

Figure 7: Rhizoctonia species mycelial growth inhibition test treated with NPs synthesized by the green route: FeO 
NPs; (T1) 0 mg/L, (T2) 50 mg/L, (T3) 100 mg/L, (T4) 500 mg/L, (T5) 1000 mg/L

Figure 8: Mycelium growth inhibition % of  Rhizoctonia species by different nanoparticles



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Evaluation of  the Antifungal Effect of  the NPs
The antifungal effect on mycelial growth of  the 
synthesized nanoparticles was evaluated by amending 
the culture medium with different NP concentrations 
and recording the fungus colony diameter over 7-9 days 
until the fungus nearly covered the plate (9 cm), with one 

nanoparticle tested at a time. All tested NPs varied in their 
extent of  inhibition of  Rhizoctonia species colony growth 
at different concentrations, with statistically significant 
differences calculated among the nanoparticles. As 
illustrated in Figure 7, the inhibition of  Rhizoctonia species 
colony growth was observed. 

Table 1: Inhibition of  colony growth of  Rhizoctonia Species over control by poisoned food technique treated with 
ZnO NPs
Concentration (mg/L) Diameter (mm) Colony growth inhibition % DMRT test result
0 80.03 ± 0.59 a
50 77.7 ± 4.64 2.91% a
100 78.9 ± 3.31 1.41% a
500 44.77 ± 7.17 44.06% b
1000 44.23 ± 8.21 44.73% b

Each value is mean of  3 replicates ± SD

ZnO NPs affected the mycelium growth at different 
concentrations, as tabulated in Table 1. The one-way 
ANOVA analysis demonstrated significant differences 
among the treatment groups, with a p-value (Pr > F) of  
less than 0.001 and an F value of  35.14. This indicates 
that the variation in mycelial growth among the different 
nanoparticle concentrations is statistically significant. The 
control group (0 mg/L) exhibited a Rhizoctonia mycelium 
growth diameter of  80.03 ± 0.59 mm. For the 50 and 100 
mg/L concentrations, the mycelium growth diameters 
were 77.7 ± 4.64 mm and 78.9 ± 3.31 mm, respectively. 
According to Dunnett’s test, there was no significant 
difference between the 0, 50, and 100 mg/L groups, 
despite inhibition percentages of  2.91% and 1.41% for 
the 50 and 100 mg/L concentrations, respectively.
In contrast, the FeO NPs at higher concentrations 
(500 and 1000 mg/L) exhibited significant inhibition 
of  mycelium growth compared to the control. The 
mycelium growth diameters for the 500 and 1000 mg/L 
concentrations were 44.77 ± 7.17 mm and 44.23 ± 8.21 

mm, respectively. The inhibition percentages for these 
concentrations were substantially higher, at 44.06% for 
the 500 mg/L concentration and 44.73% for the 1000 
mg/L concentration. These results indicate that higher 
concentrations of  ZnO NPs are more effective in 
inhibiting Rhizoctonia mycelium growth.
Previous studies have also demonstrated the dose-
dependent effect of  ZnO NPs on the inhibition of  
Rhizoctonia mycelium growth. For example, Jannat et 
al. (2022) reported inhibition percentages of  17.1%, 
31%, 40%, 48.7%, and 51.1% for R. solani at ZnO NP 
concentrations of  0.05, 0.15, 0.25, 0.35, and 0.45 mg/mL, 
respectively. In another study, green-synthesized ZnO 
NPs exhibited significant growth inhibition of  R. solani, 
with inhibition percentages of  70.2 ± 0.5%, 61.4 ± 0.5%, 
48.5 ± 0.5%, 1.2 ± 0.5%, and 0.6 ± 0.5% at concentrations 
of  1.0, 0.75, 0.5, 0.25, and 0.1 mg/mL, respectively (Ali et 
al., 2022). Those previous findings also support the dose-
dependent manner of  ZnO NPs inhibition of  Rhizoctonia 
mycelium growth.

Table 2: Inhibition of  colony growth of  Rhizoctonia Species over control by poisoned food technique treated with 
CuO NPs
Concentration (mg/L) Diameter (mm) Colony growth inhibition % DMRT test result
0 78.24 ± 1.25 a
50 73.38 ± 2.07 6.21% a
100 75.60 ± 3.58 3.37% a
500 22.97 ± 0.67 70.64% b
1000 20.79 ± 2.75 73.43% b

Each value is mean of  3 replicates ± SD

The one-way ANOVA analysis demonstrated significant 
differences among the treatment groups of  different 
concentrations of  CuO NPs, with a p-value (Pr > F) of  
less than 0.001 and an F value of  494.5. Table 2 illustrates 
the dose-dependent effect of  CuO NPs on inhibiting 
the mycelium growth of  Rhizoctonia species. The control 
group exhibited a mycelium diameter of  78.24 ± 1.25 
mm. At concentrations of  50 mg/L and 100 mg/L, the 

mycelium diameters were 73.38 ± 2.07 mm and 75.60 
± 3.58 mm, respectively, which were statistically similar 
to the control group (Duncan’s Multiple Range Test, 
DMRT), with growth inhibitions of  6.21% and 3.37%.
In contrast, concentrations of  500 mg/L and 1000 mg/L 
resulted in mycelium diameters of  22.97 ± 0.67 mm 
and 20.79 ± 2.75 mm, respectively. These values were 
statistically similar to each other (DMRT) but significantly 



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different from the control group (Dunnett’s test), 
showing inhibitions of  70.64% and 73.43%, respectively. 
These results indicate that higher concentrations of  CuO 
NPs are more effective in inhibiting Rhizoctonia mycelium 
growth. 
Previous study also suppor the findgs as at 150 ppm the 
CuO NPs synthesized by chemical reduction method 
could inhibit the growth of  R. solani by 62% (Chowdhury 
et al., 2024). Another study of  green synthesized CuO 

NPs in the concentrations of  50,100, and 200 ppm , 
maximum inhibition was recorded in 200ppm having 
11.89% inhibition of  R. solani(Fetyan et al., 2024). Similar 
study of  CuO-NPs biosynthesized from H. bacciferum leaf  
extract and antifungal activity for R. solani by 20, 50, 75, 
and 100 µg/mL they observed inhibition of  mycelium 
growth 81.48% inhibition in 100 µg/mL concentration 
(Hamdy et al., 2024).

Table 3: Inhibition of  colony growth of  Rhizoctonia Species over control by poisoned food technique treated with 
FeO NPs
Concentration (mg/L) Diameter (mm) Colony growth inhibition %
0 80.53 ± 1.28
50 78.2 ± 3.54 2.89%
100 79.54 ± 0.63 1.23%
500 82.33 ± 2.06 -2.24%
1000 82.71 ± 0.60 -2.71%

Each value is mean of  3 replicates ± SD

The one-way ANOVA analysis indicated no significant 
differences in the mean radial growth of  Rhizoctonia 
mycelium treated with FeO NPs (F value: 2.81, Pr > 
F > 0.05). The control and all treatments showed no 
significant differences, though minor inhibition and 
promotion zones were observed compared to the 
control. The maximum inhibition occurred at 50 mg/L, 
while the minimum inhibition was observed at 1000 
mg/L. Similar results were found in a study on the effects 
of  FeO NPs on the mycelial growth of  rot-causing 
fungi, where treatments with 0.1, 0.25, and 0.5 mg/mL 
resulted in mycelial growth of  12.00 ± 1.00 mm, 14.33 
± 0.57 mm, and 16.33 ± 0 mm, respectively, indicating 
higher concentrations led to increased growth rather 
than inhibition. Many results found in the literature do 
not align with the observed data. For instance, a previous 
study reported significant inhibition of  mycelium growth 
at various concentrations of  FeO NPs. Specifically, FeO 
NPs showed highly significant growth reduction, with 
the maximum growth inhibition (87.9%) observed at a 
concentration of  1.0 mg/mL (Ali et al., 2020). Another 
study investigated the growth inhibition percentages 
of  different concentrations of  iron oxide nanoparticles 
against two tested A. alternata strains. They observed 
mycelium inhibition at 29.85%, 47.24%, 55.95%, 65.74%, 
and 75.89% for concentrations of  50, 100, 200, 400, and 
800 ppm, respectively (Yassin et al., 2023).
Figure 8 illustrates that different concentrations of  
nanoparticles and various types of  nanoparticles 
inhibit mycelium growth of  Rhizoctonia species in a 
dose-dependent manner. The maximum inhibition was 
observed with CuO NPs at 500 and 1000 mg/L, followed 
by ZnO NPs. The FeO NPs did not inhibit the Rhizoctonia 
mycelium at higher concentrations, and instead promoted 
growth to a minor extent, although this increase was 
not statistically significant.  The inhibition of  mycelium 
growth by nanoparticles in a dose-dependent manner can 

be attributed to various mechanisms. For instance, NPs 
may cause cell membrane damage, leading to increased 
membrane permeability. This disruption can induce 
responses within intracellular organelles, potentially 
altering DNA and RNA through mutation, affecting ion 
transport pathways, and disrupting protein functions 
(Foldbjerg et al., 2015). Smaller nanoparticles, as suggested 
by Gliga et al. (2014), possess a larger surface area relative 
to their volume, facilitating easier penetration of  fungal 
cell walls and membranes. This penetration can trigger 
responses within intracellular organelles. Nanoparticles 
are also known to generate reactive oxygen species, 
which can further alter fungal membrane permeability. 
This alteration compromises cell structure and leads to 
osmotic imbalance, affecting fungal functions (Kumari 
et al., 2019). Studies have shown that silver nanoparticles 
inhibit spore germination in B. cinerea in a concentration-
dependent manner (Bayat et al., 2021). Despite these 
insights, the mechanisms by which green-synthesized 
nanoparticles affect mycelium growth inhibition remain 
incompletely understood. Further research is necessary 
to fully assess the potential impacts of  nanomaterials on 
altering fungal growth dynamics.

CONCLUSION
The current study successfully synthesized ZnO, 
CuO, and FeO nanoparticles using a cost-effective and 
environmentally friendly procedure. The synthesized 
nanoparticles were characterized by UV-Vis spectroscopy 
and SEM analysis. The antifungal activity of  the 
synthesized nanoparticles was evaluated at different 
concentrations using the poison food technique, testing 
one nanoparticle at a time. The results indicate that both 
ZnO and CuO nanoparticles exhibit a dose-dependent 
effect on mycelium growth inhibition, with higher 
concentrations (500 and 1000 mg/L) causing significant 
inhibition compared to the untreated Rhizoctonia species. 



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In contrast, FeO nanoparticles did not exhibit dose-
dependent inhibition of  mycelium growth but caused 
minor promotion, which was not statistically significant. 
The study concludes that among ZnO, CuO, and FeO 
nanoparticles, CuO nanoparticles at a concentration 
of  1000 mg/L demonstrated the maximum inhibition, 
followed by ZnO nanoparticles. The observed variations 
in mycelium inhibition by different nanoparticles 
at various concentrations highlight the complexity 
of  nanoparticle interactions with fungal pathogens, 
underscoring the need for further research to optimize 
their antifungal efficacy.

Funding
Competitive Research Grant 2020, Grant Number 
202003, The Open University of  Sri Lanka.

Declaration of  Generative AI and AI-Assisted 
Technologies in the Writing Process
During the preparation of  this work, the authors used 
ChatGPT 3.5 to improve language and readability. After 
using this tool/service, the authors reviewed and edited 
the content as needed and take full responsibility for the 
content of  the publication.

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