THE ANTAGONISM MECHANISM OF Trichoderma spp TOWARDS Fusarium solani MOLD
Utami Sri Hastuti 1,1 Indriana Rahmawati 2,1 *
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 day moth year; Accepted day moth year (will be given)
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 shows: 1) seven species of Trichoderma molds have different antagonism ability towards F.solani each other; 2) the antagonism mechanism observed by light microscope and Scanning Electron Microscopy are 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 biocontrol 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 have been suggested as being responsible for their biocontrol activity, which include: competition for space and nutrients, secretion of chitinolytic enzymes, mycoparasitism, and production of inhibitory compounds [5]. The objectives of this research are: 1) to study the
antagonism ability of some Trichoderma species toward F.solani; 2) to observe the antagonism mechanism of Trichoderma spp toward F.solani by microscopic observation.
EXPERIMENT Materials
Mold isolates of : Trichoderma aureoviride, T. viride, T. citrinoviride, T.
koningii, T.parceramosum, T. harzium , T. atroviride, and Fusarium solani, Czapek Agar late medium, alcohol 95%, lactophenol solution, lactophenol cotton blue solution.
Interaction experiment procedure
The interactions between seven species of Trichoderma and F.solani was evaluated by using the dual culture technique [6]. Each mold isolates were inoculated on Czapek Agar plate medium and incubated in 25oC during 3x24 hours. The F.solani colony were cuted with the cork borer of 5 mm in diameters aceptically and put at a point and each Trichoderma spp mycelia disc was put to confront (Fig.1). The plate cultures were incubated at 25oC during 3x24 hours, then the antagonism activities of the mold hyphae were observed microscopically. The antagonism ability of each Trichoderma spp toward F.solani were count with the formula of antagonism ability.
Mycoparasitic assays
Microscopic descriptions was 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.
R1 = the pathogen mold colony radius be far from antagonism mold colony
R2 = the pathogen mold colony radius be next to the antagonism mold colony
The antagonism ability (%) = R1−R2
R1 x 100%
Fig 1. The cut of Patogen Mold and Antagonis Mold mycelia disc Settling in Dual Culture
P = F.solani mycelia disc A= Trichoderma spp mycelia
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.
Fig 2. The Growth of Pathogen Mold and Antagonist Mold Colonies in Dual Culture Methode 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
Trichoderma atroviride have 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, 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.
Fig. 3 The Mycoparasitism of Trichoderma spp towards Fusarium solani in the Light Microscope Observation (400x)
a. F. solani hyphae, b. The Trichoderma hyphae coiled or attach on the F.solani hyphae
The scanning electron mycograph shows that the Trichoderma hyphae grew paralelly and attach on the F.solani hyphae (Fig.4). It is also found that the Trichoderma hyphae penetrate the F.solani hyphae (Fig. 4). The Trichoderma could take the nutrients in the cell, so it will caused the F.solani cell death. The Trichoderma hyphae is 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. 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 will caused the hyphae distruction and the inhibition of F.solani colony growth.
Fig. 4. The Scanning Electron Micrograph 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 ( ), The deformation of F.solani hyphae caused by T.viride activity ( )
Fig 5. The Scanning Electron Micrograph 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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