The Antagonism Mechanism Of Trichoderma spp. Towards Fusarium solani Mold

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DOI: 10.21776/ub.jpacr.2016.005.03.260 J. Pure App. Chem. Res., 2016, 5 (3), 178-181 17 October 2016

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The Antagonism Mechanism Of Trichoderma spp. Towards Fusarium solani Mold

Utami Sri Hastuti^1,*, Indriana Rahmawati¹

1Department of Biology, Faculty of Mathematics and Natural Sciences State University of Malang, Malang, Indonesia

email: tuti_bio_um@yahoo.com

*Corresponding email : tuti_bio_um@yahoo.com

Received 18 May 2016; Revised 14 October 2016; Accepted 17 October 2016

ABSTRACT

The antagonism ability of seven Trichoderma isolates towards F. solani have been observed and tested by dual culture technique. The antagonism mechanism observed by microscopic observation with light microscope and Scanning Electron Microscopy (SEM).

The research result showed seven species of Trichoderma molds have different antagonism ability towards F. solani each other. The antagonism mechanism observed by light microscope and Scanning Electron Microscopy were mycoparasitism, antibiosis, and competition.

Key word: antagonism mechanism, Trichoderma spp, F. solani, microscopic observation

INTRODUCTION

The biological control of Fusarium solani mold, a pathogenic soil borne mold species that cause some plant diseases should be done by the using of Trichoderma spp molds.

Trichoderma spp have been studied as potential biological control agents for controlling many plant pathogens [1]. Most of the studies on Trichoderma species have been done to their activity as biological control agent toward some phytopathogenic molds in vitro [2,3,4].

The antagonism mold, Trichoderma spp have an ability to control F. solani growth by the antagonism mechanism. The antagonism ability of Trichoderma spp towards F. solani could be different each other. The antagonism mechanism could be observed by microscopic observation with light microscope and Scanning Electron Microscopy (SEM). Different mechanism has been suggested and responsible for the biological control activity, which include competition for space and nutrients, secretion of chitinolytic enzymes, mycoparasitism and production of inhibitory compounds [5]. The research objectives study the antagonism ability of some Trichoderma species toward F. solani and observe the antagonism mechanism by means of microscopic observation.

EXPERIMENT Materials

Some molds species used for research included T. aureoviride, T. viride, T. citrinoviride, T. koningii, T. parceramosum, T. harzium, T. atroviride, and F. solani. Medium and chemicals are used directly as bought from manufacturer Czapek agar plate medium, alcohol 95%, lactophenol solution, lactophenol cotton blue solution.

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J. Pure App. Chem. Res., 2016, 5 (3), 178-181 17 October 2016

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Interaction experiment procedure

The interactions between seven species of Trichoderma and F. solani were evaluated by using the dual culture technique [6]. Each mold isolates were inoculated on Czapek Agar plate medium and incubated in 25^oC during 3x24 hours. The F. solani colony were cut with the cork borer of 5 mm in diameters aseptically and put at a point and each Trichoderma spp mycelia disc was put to confront (Fig. 1). The plate cultures were incubated at 25^oC during 3x24 hours, and then the antagonism activities of the mold hyphae were observed microscopically. The antagonism ability of each Trichoderma spp toward F. solani was counted with the formula of antagonism ability.

Figure 1. The cut of pathogen mold and antagonism mold mycelia disc settling in dual culture method. P is F. solani mycelia disc and A is Trichoderma spp mycelia disc

( )

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where R1 is the pathogen mold colony radius be far from antagonism mold colony and R2 is the pathogen mold colony radius be next to the antagonism mold colony

Mycoparasitic assays

Microscopic descriptions were done on the mycoparasitism by Trichoderma spp towards F. solani. The antagonism mechanism observed by slicing of 2x2 mm the Czapek Agar medium on the interaction zone between Trichoderma spp and F. solani colonies of the dual culture plates.

RESULT AND DISCUSSION

Each Trichoderma molds that used in this research have the antagonism ability towards F. solani mold. The mold colony growth on the agar medium shows that the Trichoderma colony grew faster than the F. solani colony based on the test result by the dual culture technique. It caused the inhibition on the growth of F. solani colony (Fig. 2). This fact indicated the nutrition and space competition between the Trichoderma and F. solani mold.

The antagonism ability of each Trichoderma species towards F. solani have also observed and measured.

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Fig 2. The Growth of Pathogen Mold and Antagonist Mold Colonies in Dual Culture Method a. T. viride () and F. solani (), b. T.harzianum () and F. solani ()

The antagonism ability of each Trichoderma species were different each other as seen in the Table 1.

Table 1. The Antagonism Ability of Trichoderma spp. towards F. solani

Isolate

Number Trichoderma species Antagonism ability

1 T.aureoviride 56.58 ± 4.46 a

2 T.viride 58.18 ± 3.73 b

3 T.citrinoviride 66.82 ± 1.91 bc

4 T.koningii 68.90 ± 3.56 cd

5 T.parceramosum 72.02 ± 0.44 d

6 T.harzianum 73.32 ± 0.52 d

7 T.atroviride 78.20 ± 0.52 e

Figure 3. (right) The

Mycoparasitism of Trichoderma spp towards Fusarium solani in the Light Microscope Observation (400x): (a) F. solani hyphae, and (b) The

Trichoderma hyphae coiled or attach on the F. solani hyphae

Trichoderma atroviride has the highest antagonism ability compare with the other Trichoderma species. This mold species suggested can produce the highest β-1,3 glucanase enzyme compared with another Trichoderma species [7]. Trichoderma species are antagonistic to some phytopathogenic fungi because they have the ability to suppress the disease they cause [8]. Trichoderma uses several biocontrol mechanisms such as mycoparasitism, antibiosis, and competition for space and nutrients between the Trichoderma and F. solani in vitro. The antagonism mechanism of Trichoderma spp towards F. solani in light microscope observation shows in Fig. 3. The scanning electron mycograph shows that the Trichoderma hyphae grew in parallel and attached on the F. solani hyphae (Fig. 4). It was also found that the Trichoderma hyphae penetrate the F. solani hyphae (Fig. 4). The Trichoderma could take the nutrients in the cell, so it causes the F. solani cell death. The Trichoderma hyphae were also coiled around the F. solani hyphae (Fig. 5). It caused morphological deformations and disorganization in the cell wall structure (Fig. 4). Probably this fact due to the secretion of antifungal substance, i.e.: enzymes by Trichoderma.

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Trichoderma produced some hydrolytic enzyme, i.e.: 1,3-glucanase, citinase, protease, and cellulose, that will initiate the pathogenic mold cell wall degradation during mycoparasitism process [9,10,11]. This mycoparasitism activity of Trichoderma spp caused the disintegration of the mycelia wall and afterward it gave rise to the hyphae destruction and the inhibition of F. solani colony growth.

Figure 4. (A) SEM image of mycoparasitism of T.viride towards F. solani, T. viride hyphae;

attach on the F. solani hyphae (), F. solani hyphae (), T. viride hyphae penetrate the F.

solani hyphae (). Deformation of F. solani hyphae is caused by T. viride activity (). (B) SEM image of mycoparasitism of T. harzianum towards F. solani. T. harzianum hyphae;

coiled on the F. solani hyphae (), F. solani hyphae () CONCLUSION

The seven species of Trichoderma molds have different antagonism ability towards F.

solani. The antagonism mechanism of Trichoderma spp towards F. solani based on the light microscope and Scanning Electron Microscopy observation is by competition nutrient and space, mycoparasitism, and antibiosis.

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