Sensitivity of Colletotrichum Capsici Isolated from Chili Pepper
(Capsicum annum) against Synthetic Fungicides
Sartini and Saipul Sihotang
Biology Department, Medan Area University, Medan, Indonesia
Keywords: Synthetic fungicide, Colletotrichum capsici, Capsicum annum.
Abstract: The sensitivity of Colletotrichum capsici isolated from chili pepper (Capsicum annum) againts synthetic
fungicide (benomyl and hexaconazole) was studied. The aim was to determine the effect of the fungicide to
the growth of C. capsici. Completely factorial randomized design was used, the first factor was C. capsici
and the second was benomil and hexaconazole at doses 0, 250, 500, 750 and 1000 ppm. Results showed that
both benomil and hexaconazole inhibit the growth of Colletotrichum capsici, particularly doses at 750 and
1000 ppm showed the highest mycelial inhibition.
1 INTRODUCTION
Chili pepper (Capsicum annuum L.) is one of the
horticultural products and become the priority
strategic program in of the Ministry of Agriculture,
Indonesia Republic in 2015-2019. Based on Badan
Pusat Statistik (2017), national chili pepper yield in
2016 decreased sharply 8.47 tons/ha with an area of
123,404 ha. According to Than et al. (2008) and
Kim et al. (2008) reported that Colletotrichum
capsici was the main pathogen on chilli pepper and
cause a negative impact on reducing economic value
and chili production, The infections caused by C.
capsici reaches 50% (Prathiba et al. (2013).
The use of fungicides continuously on farm to
control pathogenic fungi might have negative impact
on reducing the sensitivity of the pathogenic genes.
The chemical compounds of fungicide such as Cu-
hydroxide, diphenoconazole, mankozeb, maneb,
chlorotalonil, and propineb that continuously used to
control pathogenic fungi lead to increase fungal
resistance (Moorman and Lease, 1992; Suganda
2001; Ziogas et al. 2005). Kumar et al. (2007)
reported that the sensitivity of C. gloeosporioides
isolated on mangoes was caused by the presence of
chemical compounds in fungicide. Colletotrichum
gloeosporioides commonly found on chili pepper.
The use of fungicide to control the pathogenic
fungi are effective in inhibiting fungal growth,
however, the side effect of the chemical compounds
to the fungi required to be studied. The aims of the
recent study was to determine the effect of synthetic
fungicide containing benomyl and hexaconazole as
an active compounds to control the mycelial growth
of C. capsici isolated from chili pepper.
2 MATERIALS AND METHOD
2.1 Preparation of Fungal Isolates
The research was conducted from March to
September 2019 in Microbiology and Biotechnology
Laboratory, Faculty of Biology, Medan Area
University. Colletotrichum capsici used in this study
was isolated from chili pepper as culture collection
of Microbiology and Biotechnology Laboratory,
Medan Area University. The fungal isolate was
subcultured in potato dextrose agar (PDA) and
incubated for 5 days (29
o
C).
2.2 Determination Sensitivity of C.
capsici on Fungicide Compounds
The sensitivity of C. capsici was determined by
calculating the relative resistance level (I) to the
diameter colony. The concentration of fungicide
used based on the manufacturer's i.e hexaconazole
(1000 ppm) and benomil (1000 ppm). The relative
resistance of the compounds used at doses 250, 500,
750 and 1000 ppm. Three replications were used for
each treatment. Each dose of the compound was
490
Sartini, . and Sihotang, S.
Sensitivity of Colletotrichum Capsici Isolated from Chili Pepper (Capsicum annum) against Synthetic Fungicides.
DOI: 10.5220/0010205400002775
In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 490-493
ISBN: 978-989-758-556-2
Copyright
c
2022 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
made by mixing the fungicide in PDA. Each
compound was homogenate by adding 10 ml sterile
distilled water before it was added to the PDA at 40-
45°C and poured into a sterile petri dish (9 cm
diameter). Fungal mycelia containing PDA medium
(0.5 cm diameter) was placed into the middle of the
petri dish containing PDA plate. The plates were
incubated for 7 days (29°C). The percent reduction
of colony was determine according to Kumar et al.
(2007) as follows:
I
𝑥100%
Where:
I = Percent reduction in growth of fungal,
C = Radial growth (mm) of control,
T = Radial growth (mm) of treatment
2.3 Determination the Resistance of C.
capsici to Benomyl and
Hexaconazole
The resistance of C. capsici to benomyl and
hexaconazole was determined by the method of
repeated subculture which starts from a relative
inhibitory concentration> 90% (very sensitive)
through poisoning of growing media. Isolates from
concentration treatments with I > 90% were sub-
cultured back to new media with the same
concentration. Measurement of the diameter to
determine the (I) is done by the method as in the
previous experiment. Subculture experiments were
stopped when there was a change in the level of
sensitivity of each isolate to the fungicide active
ingredient being tested.
2.4 Determination of C. capsici
Sensitivity Rate for Fungicide
Compounds
The sensitivity level of C. capsici isolates to
fungicide active ingredients was determined based
on the level of relative inhibition (I) based on Kumar
et al (2007) as follows: Very sensitive (I) >90%,
Sensitive : 75% < I ≤ 90%, Moderate resistance:
60% < I ≤ 75%, Resistant: 40% < I ≤ 60%, and Very
resistant: I ≤ 40%.
3 RESULTS AND DISCUSSION
3.1 The Sensitivity of C. Capsici to
Benomyl and Hexaconazole
The sensitivity of C. capsici was determined from
the level of relative inhibition of the fungicide on the
growth of the isolate colony diameter of C. capsici..
The statistical analysis, both single factors, species
or isolates and types of active ingredients,
interactions on the sensitivity level of C. capsici
based on their relative level of resistance, can be
seen in Table 1.
Table 1 shows the fungicide have a significant
effect on C. capsici. Benomil in the 1000 ppm
showed no growth of C. capsici on observations of
2, 4 , 6 , and 8 days. Administration of hexaconazole
in the 4 days treatment of 1000 ppm showed no
growth of C. capsici on observations at day 2, day 4,
day 6 and day 8. This indicates that dose at 1000
ppm inhibit the growth of the pathogenic fungi. It
seem that the growth of C. capsici at dose 750 ppm
was inhibited. According to Andriani and Desta
(2017) benomyl is an active ingredient of systemic
fungicide by specifically targeting to disrupt mitosis
β-tubulin and cell division.
Fungicide with specific compounds showed high
response in inhibiting the growth of the fungal
pathogen. The fungicides prevents fungal infection
by forming a barrier layer on the surface of the plant
(Peres et al. 2004).
3.2 Level of Relative Barriers of C.
capsici Pathogens to Benomyl and
Hexaconazole
The level of relative inhibition of C. capsici
pathogens against benomyl and hexaconazole shows
different results as indicated by the doses and type
of fungicide (Table 2).
Potential development of C. capsici isolates
resistance to fungicide active ingredients was
selected based on relative inhibitory values> 90%.
The test of the potential for resistance development
using (I) > 90% aims to determine the ability of
these isolates to adapt in developing resistance.
Table 2 shows that doses of 750 ppm and 1000 ppm
are the best treatment for the level of relative
pathogenic inhibition.
Sensitivity of Colletotrichum Capsici Isolated from Chili Pepper (Capsicum annum) against Synthetic Fungicides
491
Table 1: Relative levels of C. capsici to benomyl and hexaconazole.
Treatments
Incubation time (da
y
s) / reduction colon
y
diameter (%)
2 4 6 8
Control
3.78
d
4.73
c
6.40
c
7.07
c
Colletotrichum × benomyl 250 ppm
0.95
c
1.02
b
1.12 b 1.03
b
Colletotrichum × benomyl 500 ppm
0.50
b
0.52
ab
0.60
b
0.87
ab
Colletotrichum × benomyl 750 ppm
0.00
a
0.28
ab
0.32
a
0.42
a
Colletotrichum × benomyl 1000 ppm
0.00
a
0.00
a
0.00
a
0.20
a
Control
3.78
c
3.82
d
4.00
c
4.03
d
Colletotrichum × hexaconazole 250 ppm
0.95
b
0.70
c
0.85
b
0.87
c
Colletotrichum × hexaconazole 500 ppm
0.35
a
0.45
bc
0.52
b
0.72
bc
Colletotrichum × hexaconazole 750 ppm
0.00
a
0.10
ab
0.27
ab
0.37
ab
Colletotrichum × hexaconazole 1000 ppm
0.00
a
0.00
a
0.00
a
0.18
a
Note: Numbers followed by same letters not significantly different (P≤0.05) according to Duncan’s multiple range test (DMRT)
Table 2: Sensitivity of C. capsici on concentration of fungicide compounds (benomyl and hexaconazole).
Compounds of fun
g
icide (ppm)/Colletotrichum capsici sensitivit
y
Benom
y
l Hexaconazole
0 250 500 750 1000 0 250 500 750 1000
0.00
(vr)
79.19
(s)
85.37
(s)
95.52
(vs)
100
(vs)
0.00
(vr)
83.10
(s)
86.43
(s)
91.69
(vs)
100
(vs)
N
ote: vr = ver
y
sensitive; s = sensitive
Figure 1: Colletotrichum capsici in PDA contain benomyl 1000 ppm (a) and hexaconazole 1000 ppm (b) after 8 days
incubation (29°C).
The growth of colony diameter had a slightly
difference in each culture during 8 DAF (days after
application). The relative level of inhibition of the
active ingredient of benomyl fungicide on the
growth of C. capsici (Figure 1).
The level of relative inhibition to the growth
diameter benomil changes in all observations.
Development of pathogenic resistance to benomyl
active ingredients at doses of 750 and 1000 ppm
occurs more slowly. This indicates that the active
ingredients are generally non-systemic which has a
low risk for developing pathogenic fungal resistance.
Benomyl is an active ingredient with specific
mechanism that interfere with mitosis β-tubulin and
cell division Djojosumarto (2008). Fungicides with
multisite mode of action mechanism by general non-
systemic which has a low risk for developing fungal
resistance to the active ingredient (Kumar et al.
(2007). The resistance involves gene mutations.
Fungal resistance to the fungicide compounds
occurred while the pathogenic fungi are exposed
continuously to the compounds (Andriani and Desta
(2017). Colletotrichum gloeosporioides tend to be
sensitive to fungicidal compounds even at low
concentrations.
IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)
492
4 CONCLUSIONS
From the results of the study it can be concluded that
the administration of benomil at doses of 750 ppm
and 1000 ppm is the best dose in inhibiting the
growth of C. capsici which is 0.00 mm and shows a
significant effect on the relative inhibition level of
the test pathogen colony.
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