Antagonistic Activities of Some Trichoderma spp. against Fusarium sp. and Meloidogyne spp., the Causal Agent of Pepper Yellowing Disease

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MARCO SATELLITE WORKSHOP @ IUCA
CE-SAE 2012
Sustainability of Agriculture and Environment
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ANTAGONISTIC ACTIVITIES OF SOME TRICHODERMA SPP.
AGAINST FUSARIUM SP. AND MELOIDOGYNE SPP., THE CAUSAL
AGENT OF PEPPER YELLOWING DISEASE
CITRA MAYANG WARDHIKA* , SURYANTI, TRI JOKO
Department of Crop Protection, Faculty of Agriculture, Universitas Gadjah Mada, Yogyakarta, Indonesia
*email : dhika_maknyak@yahoo.co.id
Keywords: antagonistic activities, Trichoderma spp., Fusarium sp., Meloidogyne spp., yellowing disease
Introduction
Pepper (Piper nigrum L.) is in a spices
group called the king of spices, because it is the most
widely traded commodity. Pepper is one of
Indonesian export commodities, but in this
cultivation has a problem that pepper is infected by
yellowing disease. Yellowing disease is common in
Bangka and Kalimantan. Causes very complex such
as nematode Radopholus similis and Meloidogyne
incognita, Fusarium oxysporum, and fertility and low
soil moisture [1]. Biological control is a reduction in
the amount of inoculum or activity damage caused
by the pathogen, using one or more types of
organisms [2]. This control includes the use of
variants avirulent pathogens and microbial
antagonists that influence the presence or activity of
pathogens [3].
Materials and Methods
Fusarium sp. are isolated from pepper stems
wich is showing yellowing disease symptoms.
Trichoderma spp. isolates (JA, JB, JD, JE, JF, JI, JL,
JM, JP, JQ, JX) and Meloidogyne spp. are isolated
from the rhizosphere of pepper plantations in West
Kalimantan.
Growth Inhibition Observations. Fusarium
sp. and Trichoderma spp. were grown in opposite
direction in petridish, incubated for 2 days and
observed in 2 parameters, i.e qualitative observation
of the formation of halo. Halo is clear zone that
produced between Fusarium sp. and Trichoderma
spp. Quantitative observation is halo was not form in
this observation.
Growth inhibition observation can did with
measured by the diameter of each fungi and
calculated the percentage of inhibition using the
formula : {(a-b)/a x 100%}. a = widespread culture
of control. b = widespread culture of the treatment
[4].
Micoparasitism observation. Fusarium sp. and
Trichoderma spp. were placed on object glass in
opposite direction. Incubated for 2 days and observed
under microscope for presence or absence
micoparasitism.
Parasitism Observation. Spore suspension
was prepared by adding sterile water into fungal pure
culture on agar slant, then obtained spore suspension
density calculated using haemacytometer [5].
Parasitation test against nematode larvae stage 2 (L2)
is done by using a multiwell plates filled with 0.5 ml
suspension containing nematodes by 10 L2. Each

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well containing a suspension of L2 nematodes
inoculated with Trichoderma spp. suspension with a
density of 106 up to 200 ml, and then incubated for 1
week and observed daily for the presence of
nematodes infected by the fungi, the nematodes were
dead or lyses.
Results and Discussion
Growth Inhibition Observations. Some
Tricoderma spp. isolates were formed to inhibition
zone, such as isolate JA, JD, JI, JM, JQ, and JX. The
highest percent inhibitions is JI (92,88%) (Table 1).
Table 1. The results of growth inhibition
observation
No
Isolate
code
Inhibition
zone
%
Inhibition
1
JA
+
78,222
2
JB
-
43,75
3
JD
+
91
4
JE
-
73,305
5
JF
-
41,222
6
JI
+
92,88
7
JL
-
75
8
JM
+
88,888
9
JP
-
67,888
10
JQ
+
81,222
11
JX
+
85,305
Fig 1. Halo formed Fig 2. No halo formed
Micoparasitism observation. There is only
one fungal isolate that can form micoparasitism
(isolate J1) (Fig 3.). The process of micoparasitism
interaction between Trichoderma spp. and pathogen
is divided into 4 stages, i.e chemotropic growth,
recognition, attachment, and host cell wall
degredation [6].
Fig 3. Micoparasitism Trichoderma spp. to
Fusarium sp.
Parasitism Observation. There are 4 isolate
that can lyse nematode, namely JA, JB, JL, JM
(Table 2).
Table 2. The resultsof parasitism observation
No
Isolate
code
The percetage of deaths
nematodes
Dead (%)
Lysis (%)
1
JA
100
6,7
2
JB
100
3,3
3
JD
96,7
0
4
JE
100
0
5
JF
100
0
6
JI
100
0
7
JL
100
6,7
8
JM
93,3
3,3
9
JP
100
0
10
JQ
100
0
11
JX
100
0
The role of nematophagous fungi in controlling
nematodes are as predatory larvae and eggs,
endoparasites on larvae and producing toxins. The
fungi kills nematodes by making traps infective
larvae, became the endoparasitic on larvae, penetrate
the female larvae and eggs, and kill larvae by toxins
[7].
Fig 4. Lysis on the nematode caused by
Trichoderma spp.

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Concluding Remarks
There are 6 isolates of Trichoderma spp. (JA,
JD, JI, JM, JQ, JX), which is capable of inhibiting
Fusarium sp. (forming the halo) and there is only one
Trichoderma spp. Isolate (JI) that is mycoparasitism
against Fusarium sp. All Trichoderma spp. were
tested against Meloidogyne spp. capable of causing
death Meloidogyne spp. with a percentage between
93.3 to 100%, but there are only 4 isolates were able
to cause lysis in Meloidogyne spp., which isolates
JA, JB, JL, JM. The largest percentage lysis was
6.7%, ie the treatment isolates JA and JL.
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