Biological control by using antagonistic microorganisms is an alternative that is currently being carefully studied and used as a control of plant diseases. Endophytic microorganisms also play an important role in controlling plant diseases which are induction-resistant. Endophytic antagonist fungi have high activity in producing enzymes that can be used to control pathogens. The purpose of this study was to determine the effectiveness of rubber plant endophytic fungi as biological agents against fungi
Keywords: Endophytic fungusbiological controlHevea brasiliensisColletotrichummetabolitepathogen
In Indonesia, the deciduous disease of leaves of
The last few years the excavation of microbial resources found in plant tissues has received much attention. Endophytic fungus is also defined as a fungus that consumes all or part of its life cycle between cells and within healthy tissue cells of host plants, usually not causing symptoms of disease. Endophytic fungi are an important component of plant microecosystems and have been found in every plant species that has been studied and it is estimated that there are more than one million endophytic fungi present in nature (Dreyfuss & Chapela, 1994).
Endophytic mushroom groups are able to produce antibiotic compounds that are active against bacteria and pathogenic fungi in humans, animals and plants. Symbiosis between mushrooms and healthy plants in certain tissues are able to produce mycotoxins, enzymes and antibiotics. The association of several endophytic fungi with host plants can protect the latter from several virulent pathogens, both bacteria and fungi (Purwanto, 2008).
Syamsafitri and Hasanuddin (2013) have explored endophytic fungi from different rubber clones resulted in 13 endophytic isolates. The identified endophytic fungi were derived from the genus
Efforts to achieve productivity of rubber plants are currently not optimal. One of the causes is the disruption and declining of secondary leaves caused by the fungus
Are there differences in the effectiveness of some endophytic fungi from rubber clones as biological agents in the control of C. gloeosporioides?
Are there differences in the effectiveness of the extraction of endophytic fungi metabolites and spores as biological agents in the control of C. gloeosporioides?
Purpose of the Study
To determine the effectiveness of several endophytic fungi from rubber clones as biological agents against fungi
This research was conducted on an experimental farm at Sungai Putih Rubber Research Center, Galang Subdistrict, Deli Serdang Regency, North Sumatra Province with a site altitude of ± 25 metres above sea level (MASL) from February to March 2019. Samples included endophytic pure isolate clones (RRIM-931, RRIM-901, PB-330 and PB-260) from Bandar Betsy plantation, C.gloesporioides isolates from Sungai Putih plantation and Hevea brasiliensis PB 260 clone. The research method was factorial randomized block design (RBD) with 3 replications and combinations with 5 replications.
Endophytic factor (E) consisted of 4 treatments which were E1 = RRIM-931 rubber clone from code isolate Bb 1 (endophyte), E2 = rubber clone RRIM-901 from code isolate Bb 2 (endophyte), E3 = PB-330 rubber clone from code isolate Bb 3 (endophyte), and E4 = PB-260 rubber clone from code isolate Bb 4 (endophyte).
Application methods (M) consisted of 4 types which were M0 = control (water treatment) M1 = spores 10⁴, M2 = metabolites, and M3 = metabolites + spores. The number of treatment combinations was 16 (4x4), with each treatment being repeated three times.
Making Endophytic Metabolites
Five isolates were taken from each endophytic type and were placed inside a bottle containing 100 ml of GDP media. This process was carried out in a laminar water flow, followed by a week in shaker (Sungai Putih Rubber Research Center, 2018).
Endophye Spore and Metabolite Application
An application of 180 ml of distilled water to the upper and lower surfaces of rubber leaves was carried out in the afternoon. The surface of the leaves were then applied with
The applications of spores and metabolites with a density of 10⁴ were sprayed using a hand sprayer on the leaves of the entire rubber plant. Incubation using
The latent period saw the onset of
Observation of disease occurence was carried out by calculating deciduous plants of C. gloeosporioides leaves in each trial plot. Observations were made on the 2nd, 4th, 6th, 8th, 10th and 12th day after inoculation (HSI). Data obtained was calculated by the following formula :
KP = n x 100 %
--- z where :
KP : disease incidence, n : number of leaves symptomatic of leaf fall disease, z : number of leaf samples (Inte Agrios, 2005).
Measurements for the severity of the disease according to the number of days after conidial inoculation were done using the scale of the attack on the leaves. Measurements of the pathogenic disease intensity or scale of attack at the Sungai Putih Rubber Research Institute in the farm with PB 260 clones were followed according to the formula by Towsendt and Hueberger (as cited in Unterstenhover, 1963) :
∑ (ni x vi)
I = -------------- X 100 %
N x Z
I : disease severity, ni : number of first leaves, vi : scale of attack, N : number of leaves observed, Z : highest value
Symptoms of leaf spots were set on 7 scales ( 0 – 6 ), with a spotting diameter of 2 mm.
Scale 0 = no spots on the leaves were observed
Scale 1 = 1 to 8 spots on the leaves were observed
Scale 2 = 9 to 12 spots on the leaves were observed
Scale 3 = 13 to 16 spots on the leaves were observed
Scale 4 = 17 to 20 spots on the leaves were observed
Scale 5 = 21 to 24 spots on the leaves were observed
Scale 6 = more than 24 spots on the leaves were observed or the leaves fall.
The results of observations of the time of the appearance of symptoms of a disease attack (latent period) is the fastest is 3th day after inoculation, and the longest is 4 days. This shows that at 3th day after inoculation the penetration of the fungus Colletotrichum into the leaf tissue (Table
The initial appearance of the symptoms of
The incubation period as the initial process of the emergence of symptoms of disease in plants is influenced by environmental factors that are suitable for the development of pathogens, but instead makes plant growth inhibited. In addition to environmental factors, this latent period is also influenced by genetic factors, both from pathogens and host plants to the disease. This is evident from the data generated where the types of endophytic fungi that are applied, each of which has a response to a different disease (
Disease Incidence (%)
Disease Inctnsity (%)
The results of analysis of variance showed that the test of endophytic fungi did not significantly affect the intensity of
The average percentage intensity of
The effectiveness of endophytic fungi from the Bandar Betsy plantation had a significant effect on the intensity of the disease in the rubber plants in the farm (Figure
The absence of an interaction in observations 2, 4, 6, 8, 10 and 12 HSI showed that the effect of the application method was very significant to the intensity of the disease in the endophytic application method from the Bandar Betsy Clone RRIM 931 Garden with spore and metabolite treatments.
Deciduous disease of C.gloeosporiodes in PB 260 clone rubber plants was tested using spores and endophytic fungal metabolites from Bandar Betsy garden with isolates BB1, BB2, BB3 and BB4 clones with observations at 2 to 12 HSI. Each treatment from 4 isolates had controls with codes KE1, KE2, KE3 and KE4.
Results of the study showed that the severity and resistance of PB 260 rubber clones to C.gloeosporioides deciduous disease with endophytic treatment and the application method in all treatments were at 0 - 5%. This percentage was categorised as mild damage-resistant, thus the treatment of endophytic and effective application method as biological control in suppressing the development of C.gloeosporioides deciduous disease. Interaction between endophytic fungi from RRIM-901 rubber clone from Bb 2 isolate code (endophytes) and spore and metabolite application method showed the duration of
days after inoculation (HSI) and the presence of an influence caused by endophytic fungi in suppressing the development of fungal deciduous leaf C. gloeosporioides.
The effectiveness of endophytic fungi from the Betsy Bandar garden with the code of isolates BB1, BB2, BB3 and BB4 showed effective isolates suppressing the development of C.gloeoporiodes. BB1 with treatment of endophyte 1 Spores and E1M1 metabolites with an average disease intensity of 2.57 % is said to be able to suppress the fungal development. In the opinion of Debbab, Muller, and Mosaddak (2009), endophytic fungi are microbes that live in the internal tissues of almost all healthy plants without causing direct negative effects on their host plants. Endophytic fungi are found in the leaves, flowers, twigs and roots of plants. They synergize with their host plants through a symbiotic relationship between mutualism and some endophytic fungi are considered useful for plants by producing special substances such as enzymes that can prevent host plants from attacking pathogens such as fungi and pests. Verma, Patel, Pratap, Gangwar, and Nath (2014) said that 80% of endophytic fungi tested showed ability as antibacterial, anti-fungal, anti-allergic or as an herbicide.
The treatment of the application method has a significant effect on the 4, 6, 8, 10 and 12 HSI treatments, while the interaction of the two treatments did not significantly affect the development of leaf deciduous C.gloeoporioides. Intensity The lowest disease was obtained from endophytic treatment at E1M2 2.56% and the highest value at E2M0 treatment. 4.93%. Effective endophytic fungi suppress and prevent the development of C.gloeosporioides deciduous disease in entres.
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30 March 2020
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Syamsafitri, S., Bakti, D., Lisnawita, L., & Tistama, R. (2020). Effectiveness Of Endophytic Fungi As Biological Control Agent On Rubber Plants. In & N. Baba Rahim (Ed.), Multidisciplinary Research as Agent of Change for Industrial Revolution 4.0, vol 81. (pp. 54-63). European Publisher. https://doi.org/10.15405/epsbs.2020.03.03.8