1
Journal of PhytoPathology and Disease Management
Print ISSN: 3009-6111 Online ISSN: 3009-6170
Volume 12, Issue 1, 2025, Pages 1–7
Article
Biological control of root rot and wilt diseases of cucumber using
certain bioagents and biocides under greenhouse conditions
Hesham K. Ghaleb1 | Mohamed M. El-Sheikh Aly1 | Moshref M. Shamrokh1
1Agricultural Botany Department, Faculty of Agriculture, Al-Azhar University, Assiut, Egypt
DOI:
10.5281/zenodo.18012239
ARK:
ark:/24629/PPDJ.v12i1.266
Received:
8 April 2025
Accepted:
15 July 2025
Published online:
20 July 2025
Correspondence:
Mohamed M. El-Sheikh Aly
Agricultural Botany Department,
Faculty of Agriculture, Al-Azhar
University, Assiut, Egypt. Email:
drmohamedelsheikh4@gmail.com
Abstract:
Ten fungal isolates belonging to Rhizoctonia solani (4 isolates), Fusarium semitectum, F.
solani, Macrophomina phaseolina, F. oxysporum f.sp. cucumerinum (2 isolates), and
Sclerotium rolfsii were recovered from infected cucumber roots collected from various
localities during the fall (2020) and spring (2021) growing seasons. Pathogenicity tests
revealed that all isolates were capable of infecting cucumber plants, causing root rot and
wilt diseases. Rhizoctonia solani isolate No. 3 was identified as the most destructive,
resulting in the highest disease severity. Similarly, F. oxysporum f.sp. cucumerinum
isolate No. 8 was the most virulent among the wilt pathogens. Significant variations were
observed among the tested cucumber genotypes regarding their susceptibility to pre-
and post-emergence damping-off. In vitro studies demonstrated that Trichoderma
asperellum, T. harzianum, T. album, and T34 exhibited varying degrees of antagonistic
activity against the pathogenic fungi. Additionally, different bacterial bioagents were
evaluated under laboratory and greenhouse conditions; Pseudomonas fluorescens
followed by Bacillus megaterium showed the highest efficacy in inhibiting the mycelial
growth of R. solani and F. oxysporum f.sp. cucumerinum, whereas Bacillus subtilis
exhibited moderate effects, and Paenibacillus polymyxa was the least effective.
Furthermore, commercial biocides (Bio-Arc, Plant Guard, and Rizo-N) were evaluated
during the fall (2022) and spring (2023) seasons under greenhouse conditions. Plant
Guard was the most effective treatment at all tested concentrations in reducing disease
severity caused by R. solani and F. oxysporum f.sp. cucumerinum. Moreover, Bio-Arc,
followed by Rizo-N at the highest concentration, resulted in a significant reduction in
the incidence of root rot and wilt diseases.
Keywords:
Cucumber, Biological control, Root rot, Rhizoctonia solani, Fusarium oxysporum.
Article | Ghaleb et al.
2 | Journal of Plant Pathology and Disease Management | Vol. 12, No. 1 |
1. Introduction
Cucumber (Cucumis sativus L.) is considered one of the
major summer vegetable crops in commercial elds in Egypt.
In recent decades, eorts have been concentrated on
cultivating the crop in protected systems (greenhouses)
during the autumn and winter seasons. Consequently, the
cultivated area of cucumber in Egypt is expanding at a
relatively fast rate, particularly in newly reclaimed desert
lands. However, cucumber plants are attacked by several
fungal diseases during various growth stages, causing
considerable yield losses under both eld and greenhouse
conditions. Among these, soil-borne diseases are particularly
destructive. Root rot and wilt, primarily caused by
Rhizoctonia solani and Fusarium oxysporum f.sp.
cucumerinum, are the most common diseases aecting
cucumber plants, leading to damping-o and signicant
economic damage (Al-Tuwaijri, 2015; Martinez et al., 2003).
Sabbagh et al. (2017) emphasized that damping-o and root
rot are serious threats to cucumber at both seedling and adult
stages under protected cultivation. Furthermore, Aljawasim
et al. (2020) reported that several fungal pathogens,
including R. solani, F. oxysporum, F. solani, Sclerotium rolfsii,
and Macrophomina phaseolina, are responsible for damping-
o and root rot in cucumber and watermelon, causing severe
losses in seed germination and plant survival. Although
chemical fungicides have provided satisfactory control of
these diseases, they are considered major contributors to
environmental pollution and pose health risks. Therefore, to
avoid the hazards associated with fungicides, alternative
control methods have been investigated. Biological control
using antagonistic microorganisms has proven to be a
successful, eective, and eco-friendly strategy to manage
various plant diseases and reduce crop damage (Fasusi et al.,
2021; Wang et al., 2018). In recent years, Trichoderma species,
benecial bacteria, and commercial biocides have been
extensively used to enhance plant growth and combat
diseases (Awad and Fayyadh, 2018; Mahmoud, 2015). For
instance, Thabet (2023) evaluated the inhibitory eect of
fungal bioagents (T. harzianum, T. asperellum, T. album, and
T34) and bacterial isolates against the linear growth of R.
solani, F. oxysporum f.sp. cucumerinum, and Verticillium
albo-atrum in vitro. Additionally, several commercial
biocides have been tested for their ecacy in controlling
cucumber root rot and wilt diseases under greenhouse
conditions, oering a promising alternative to chemical
treatments (Thabet, 2023). The present study aims to isolate
and identify the causal pathogens of root rot and wilt diseases
in cucumber. Furthermore, the investigation intends to
evaluate the ecacy of certain antagonistic bioagents (fungal
and bacterial) and selected commercial biocides in
controlling these diseases under greenhouse conditions,
providing a sustainable approach for disease management.
2. Materials and Methods
This study was conducted during the growing seasons of 2021
and 2022 under laboratory and greenhouse conditions at the
Department of Agricultural Botany, Faculty of Agriculture,
Al-Azhar University (Assiut Branch), Egypt.
2.1 Isolation, Purification, and Identification of Pathogens
Samples were collected from the roots of cucumber and
watermelon plants exhibiting typical symptoms of root rot
and wilt. These samples were obtained from various locations
in Al-Buhaira and Menoua governorates, Egypt during the
autumn growing season of 2019. Diseased roots were washed
thoroughly with tap water, cut into small pieces (0.3–0.5 cm),
surface-sterilized with 70% ethyl alcohol for 2–3 minutes,
and dried between sterile lter papers. The pieces were then
transferred onto Potato Dextrose Agar (PDA) medium
supplemented with streptomycin to prevent bacterial growth
and incubated at 25–27°C for 5-7 days. Hyphal tips or single
spores were transferred to PDA slants to obtain pure cultures
(Cowan et al., 1999). The isolated fungi were identied based
on morphological and microscopic characteristics according
to taxonomic keys (Barnett and Hunter, 1986; Nelson et al.,
1983; Sneh et al., 1991). Identication was conrmed by the
Department of Agricultural Botany, Al-Azhar University,
Assiut, Egypt.
2.2 Pathogenicity Tests
Pathogenicity tests were conducted under greenhouse
conditions during the spring season of 2020 using the
cucumber hybrid 'Hayel'. Plastic pots (25 cm diameter) were
lled with 4 kg of soil that had been previously sterilized with
a 5% formalin solution and covered with a plastic sheet for 7
days. The soil was then aerated for four weeks to remove
formaldehyde residues. The experiment included pots lled
with sterilized soil and inoculated with pathogenic fungi, as
well as control pots. Five seeds were sown in each pot, with
Article | Ghaleb et al.
3 | Journal of Plant Pathology and Disease Management | Vol. 12, No. 1 |
four replicates per treatment. Disease incidence (pre- and
post-emergence damping-o) was recorded at 15 and 30 days
after sowing, while plant survival was recorded after 45 days.
Disease severity was assessed after 60 days using a 0–5 scale
described by Liu et al. (1995).
2.3 Susceptibility of cucumber hybrids
Six cucumber hybrids (Mashoor, HCU 096, Jannt, Crystal,
Go, and Hayel) were evaluated for their susceptibility to
Rhizoctonia solani and Fusarium oxysporum f.sp.
cucumerinum during the fall 2020 season. The experimental
design and disease assessment (pre- and post-emergence
damping-o, survival percentage) were carried out as
described in the pathogenicity test. The percentage of
infection was calculated according to the formula by El-
Helaly et al. (1970).
2.4 In Vitro Studies
2.4.1 Antagonistic activity of Trichoderma spp.
The inhibitory eect of Trichoderma harzianum, T.
asperellum, T. album, and T34 (obtained from the Biological
Control Unit, ARC, Giza) was evaluated against R. solani and
F. oxysporum f.sp. cucumerinum using dual culture
technique. A mycelial disc (6 mm) of the pathogen was
placed on one side of a PDA plate, and a disc of the antagonist
was placed on the opposite side. Plates inoculated with the
pathogen alone served as controls. Four replicates were used
for each treatment, and plates were incubated at 25±2°C. The
percentage of growth inhibition was calculated using the
following formula:
Inhibition(%)= 𝐶 − 𝑇
𝑇 ×100
Where: C = Radial growth of the pathogen in the control. T =
Radial growth of the pathogen in the treatment.
2.4.2 Antagonistic activity of bacteria
Four bacterial isolates (Bacillus subtilis, B. megaterium,
Paenibacillus polymyxa, and Pseudomonas uorescens),
obtained from MERCIN (Faculty of Agriculture, Ain Shams
University, Egypt), were tested. Bacterial isolates were
streaked 2 cm from the edge of PDA plates, and a 6 mm
pathogen disc was placed in the center (Abou-Aly, 2008;
Landa et al., 1997). Plates were incubated at 25°C for 5 days.
The inhibition zone was measured, and the percentage of
inhibition was calculated as described above.
2.5 Ecacy of commercial biocides under greenhouse
conditions
Three commercial biocides (Bio-Arc, Rizo-N, and Plant
Guard) were evaluated during the fall (2022) and spring
(2023) seasons. Sterilized soil was infested with R. solani or F.
oxysporum f.sp. cucumerinum at a rate of 1% (w/w). The
biocides were applied to the infested soil at three rates (1, 2,
and 3 g or cm³/kg soil) before sowing. Control pots contained
infested soil without biocides. Each treatment consisted of 5
replicates (one pot per replicate) with 5 seeds of the hybrid
'Hayel' per pot. Pre- and post-emergence damping-o, plant
survival, and disease severity were recorded as previously
described.
2.6 Statistical analysis
The obtained data were subjected to statistical analysis of
variance (ANOVA). The Least Signicant Dierence (L.S.D.)
test was used to compare treatment means at a probability
level of 0.05, as described by Gomez and Gomez (1984).
3. Results and Discussion
3.1 Isolation and pathogenicity tests
Ten fungal isolates were recovered from infected cucumber
roots collected from dierent localities. The isolated fungi
included Rhizoctonia solani (4 isolates), Fusarium
oxysporum f.sp. cucumerinum (2 isolates), F. semitectum, F.
solani, Macrophomina phaseolina, and Sclerotium rolfsii.
Pathogenicity tests revealed that all fungal isolates were
capable of infecting cucumber plants ('Hayel' hybrid),
causing varying degrees of root rot and wilt symptoms. Data
presented in Table (1) indicate that R. solani isolate No. 3 was
the most destructive pathogen, recording the highest disease
severity (88.12%), as well as high pre- and post-emergence
damping-o. Among the wilt pathogens, F. oxysporum f.sp.
cucumerinum isolate No. 8 was the most virulent, causing
77.15% disease severity. Signicant dierences were observed
among the isolates compared to the control. M. phaseolina
(isolate No. 7), F. semitectum (isolate No. 5), and S. rolfsii
(isolate No. 10) exhibited lower disease severity, recording
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4 | Journal of Plant Pathology and Disease Management | Vol. 12, No. 1 |
38.35%, 41.65%, and 42.25%, respectively. These results are in
agreement with Martinez et al. (2003) and Al-Tuwaijri (2015),
who identied R. solani and F. oxysporum as the most
common and aggressive pathogens on cucumber.
Consequently, the most aggressive isolates (R. solani No. 3
and F. oxysporum No. 8) were selected for further studies.
Table 1: Disease severity of cucumber plants caused by the isolated fungi under greenhouse conditions during the fall
2022 growing season.
Isolate No.
The tested fungi
Pre-emergence damping-off (%)
Post-emergence damping-o (%)
Survival (%)
Disease Severity (%)
1
Rhizoctonia solani (1)
15.00
35.00
50.00
65.35
2
Rhizoctonia solani (2)
12.00
22.00
66.00
73.40
3
Rhizoctonia solani (3)
22.00
46.00
32.00
88.12
4
Rhizoctonia solani (4)
18.00
38.00
44.00
72.35
5
Fusarium semitectum
12.00
24.00
64.00
41.65
6
Fusarium solani
40.00
22.00
38.00
52.10
7
Macrophomina phaseolina
12.00
24.00
64.00
38.35
8
F. oxysporum f.sp. cucumerinum
20.00
55.00
25.00
77.15
9
Fusarium oxysporum
12.00
52.00
36.00
63.75
10
Sclerotium rolfsii
16.00
36.00
48.00
42.25
-
Control (Uninfected)
0.00
0.00
100.00
0.00
L.S.D at 5%
4.18
5.36
6.65
7.15
3.2 Reaction of cucumber genotypes
The susceptibility of six cucumber genotypes to the most
aggressive isolates was evaluated under greenhouse
conditions. Data in Table (2) show that all tested genotypes
were susceptible to infection, though signicant dierences
were observed. 'Jannt' and 'Go' were the most susceptible
varieties to R. solani, recording disease severity of 65.40% and
55.10%, respectively. Conversely, 'Mashhor' and 'Hayel' were
the least susceptible to R. solani. Regarding F. oxysporum f.sp.
cucumerinum, 'Go' followed by 'Crystal' exhibited the highest
disease severity (55.20% and 48.75%, respectively), while
'Mashhor' (28.45%) and 'Jannt' (32.40%) showed the lowest
disease severity. These ndings are consistent with reports by
Al-Tuwaijri (2015) and Thabet (2023), who noted varietal
dierences in resistance to root rot and wilt pathogens.
Table 2: Response of six cucumber genotypes to root rot and wilt diseases incited by the most aggressive Rhizoctonia
solani and Fusarium oxysporum f.sp. cucumerinum isolates under greenhouse conditions.
Cucumber genotype
Rhizoctonia solani (Isolate 3)
F. oxysporum f.sp. cucumerinum (Isolate 8)
Pre %
Post %
Surv. %
D.S. %
Pre %
Post %
Surv. %
D.S. %
Hayel
25.00
20.00
55.00
35.25
10.00
15.00
75.00
45.15
HCU 096
15.00
20.00
65.00
45.75
10.00
10.00
80.00
38.75
Crystal
18.00
22.00
60.00
46.25
20.00
30.00
50.00
48.75
Go
22.00
28.00
50.00
55.10
15.00
20.00
65.00
55.20
Jannt
30.00
35.00
35.00
65.40
10.00
10.00
80.00
32.40
Mashhor
20.00
10.00
70.00
35.00
10.00
10.00
80.00
28.45
Control
0.00
0.00
100.00
0.00
0.00
0.00
100.00
0.00
L.S.D at 5%
10.50
8.37
5.15
12.11
8.75
7.15
9.75
10.65
Pre = Pre-emergence damping-off; Post = Post-emergence damping-off; Surv. = Survival; D.S. = Disease Severity.
3.3 In vitro biological control
3.3.1 Ecacy of Trichoderma spp.
The antagonistic activity of Trichoderma species against the
pathogenic fungi is presented in Table (3). Results indicated
that all tested bioagents signicantly inhibited the mycelial
growth of the pathogens compared to the control.
Trichoderma spp. grew rapidly over the mycelium of F.
oxysporum and R. solani, preventing their development. T34
(biocontrol agent) exhibited the highest inhibitory eect,
reducing the growth of R. solani by 86.33% and F. oxysporum
by 88.65%. T. asperellum followed, showing 78.35%
inhibition of R. solani. T. harzianum recorded the lowest
inhibition percentages against both pathogens. These results
align with El-Sheshtawy et al. (2009) and Malathi (2015), who
reported the ecacy of Trichoderma spp. in
hyperparasitizing soil-borne pathogens.
Article | Ghaleb et al.
5 | Journal of Plant Pathology and Disease Management | Vol. 12, No. 1 |
Table 3: Antagonistic activity of Trichoderma spp. on the mycelial growth inhibition of the pathogenic fungi in vitro.
Trichoderma Bioagents
Mycelial Growth Inhibition (%)
Against R. solani
Against F. oxysporum f.sp. cucumerinum
Trichoderma asperellum
78.35
82.66
Trichoderma harzianum
66.66
69.25
Trichoderma album
71.25
79.33
T34 (Biocontrol)
86.33
88.65
Control
0.00
0.00
L.S.D at 5%
1.39
2.15
3.3.2 Ecacy of bacterial bioagents
Data in Table (4) demonstrate that all tested bacterial isolates
signicantly reduced the linear growth of R. solani and F.
oxysporum. Pseudomonas uorescens gave the highest
inhibition of R. solani (81.75%) and F. oxysporum (86.67%),
followed by Bacillus megaterium. Bacillus subtilis showed
moderate ecacy, while Paenibacillus polymyxa was the least
eective. The use of microbial antagonists oers an eective
and eco-friendly strategy for controlling soil-borne
pathogens, as supported by Gravel et al. (2004).
3.4 Ecacy of commercial biocides under greenhouse
conditions
The ecacy of commercial biocides (Bio-Arc, Plant Guard,
and Rizo-N) in controlling root rot and wilt was evaluated
during the fall (2022) and spring (2023) seasons (Table 5).
Table 4: Effect of antagonistic bacteria on the mycelial growth inhibition of the pathogenic fungi in vitro.
Antagonistic Bacteria
Mycelial Growth Inhibition (%)
Against R. solani
Against F. oxysporum f.sp. cucumerinum
Bacillus subtilis
72.33
81.66
Bacillus megaterium
79.15
84.33
Paenibacillus polymyxa
68.66
78.10
Pseudomonas fluorescens
81.75
86.67
Control
0.00
0.00
L.S.D at 5%
2.64
1.55
Table 5: Effect of commercial biocides on controlling cucumber root rot and wilt diseases under greenhouse
conditions during the fall (2022) and spring (2023) growing seasons.
Commercial Biocides
Rate of Application
Disease Severity (%)
Fall (2022)
Spring (2023)
R. solani
F. oxysporum
R. solani
F. oxysporum
Bio-Arc
1 g
14.25
16.75
16.25
15.75
2 g
10.15
11.10
10.35
9.25
3 g
6.65
5.35
7.14
6.33
Plant Guard
1 cm³
9.62
11.44
10.65
12.33
2 cm³
6.66
8.54
7.85
7.66
3 cm³
4.35
3.90
5.25
4.75
Rizo-N
1 g
16.65
14.75
16.40
17.75
2 g
12.25
10.25
11.50
12.66
3 g
9.15
6.65
6.67
7.33
Control
--
86.10
77.25
88.90
78.33
L.S.D at 5%
1.05
1.33
1.28
1.18
All tested biocides eectively reduced disease severity
compared to the control. Plant Guard was the most eective
treatment at all tested concentrations (1, 2, and 3 cm³/kg
soil), signicantly decreasing the severity of root rot caused
by R. solani and wilt caused by F. oxysporum in both seasons.
Bio-Arc, followed by Rizo-N, also provided signicant disease
control, particularly at the highest concentration (3 g/kg
soil). These treatments resulted in the highest reduction of
wilt disease compared to lower concentrations and the
control. These ndings are in accordance with El-Blasy
(2006) and Thabet (2023), who conrmed the potential of
these biocides in managing cucumber diseases.
4. Conclusion
The present study highlights the signicant threat posed by
Rhizoctonia solani and Fusarium oxysporum f.sp.
cucumerinum to cucumber production under greenhouse
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6 | Journal of Plant Pathology and Disease Management | Vol. 12, No. 1 |
conditions, as pathogenicity tests conrmed the
susceptibility of various cucumber genotypes, particularly
'Jannt' and 'Go'. In vitro investigations demonstrated the high
antagonistic potential of Trichoderma species, especially
isolate T34, and bacterial bioagents like Pseudomonas
uorescens and Bacillus megaterium, while greenhouse
experiments provided practical evidence that commercial
biocides can eectively manage these diseases. Among the
tested treatments, Plant Guard proved to be the most
eective at all concentrations, followed by Bio-Arc and Rizo-
N at higher application rates. Consequently, this study
recommends the integration of these eco-friendly biocides
into disease management programs as a sustainable
alternative to chemical fungicides, reducing environmental
pollution while maintaining cucumber productivity in
protected cultivation systems.
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Funding Information: The authors received no external
funding for this article.
Data Availability: Data are available upon request.
Correspondence and requests for materials should be
directed to Mohamed M. El-Sheikh Aly .
Author Contributions: All authors contributed equally to
this work and share rst authorship.
Human and Animal Rights: This research did not involve
human or animal subjects.
Conicts of Interest: The authors report no known
nancial or personal relationships that could have inuenced
the work presented in this article.
How to cite this article: Ghaleb, H.K., El-Sheikh Aly,
M.M., & Shamrokh, M.M. (2025). Biological control of
root rot and wilt diseases of cucumber using certain
bioagents and biocides under greenhouse conditions.
Journal of PhytoPathology and Disease Management,
12(1), 1–8.
