Several bacteria were isolated from straw mushroom (Volvariella volvacea) cultivation medium. There are three potential isolates previously characterized and has growth inhibition effect against V. volvacea. This screening result lead to the further study about the inhibition activity against phytopathogenic fungi. The aim of this research is to investigate the antifungal activity of three bacterial isolates against three phytopathogenic fungi and identification of the bacteria. The method used in this study are antifungal assay using co-culture method and disk difussion assay using the filtrate of each bacteria. The profile of antifungal compound was identified using ethyl acetate extract followed by evaporation and gas chromatography (GC-MS) analysis. Identification of each isolates was performed using 16S rDNA amplification and sequencing. Three phytopathogenic fungi i.e Cercospora lactucae (InaCC F168), Colletotrichum gloeosporides (InaCC F304) and Fusarium oxysporum f.sp. cubense (F817) were co-cultured with bacterial isolates C2.2, C3.8, and D3.3. The C3.8 isolate has highest average inhibition activity either using isolate and filtrate. The result relatively consistent against three phytopathogenic fungi. The metabolite profile of C3.8 isolate showed the Bis(2-ethylhexyl) phthalate as the main compound with 97% similarity. Bis(2-ethylhexyl) phthalate has potential effect as antibacterial and antifungal compound. According to EzBioCloud and GeneBank databases, the C2.2 isolate identified as Bacillus tequilensis, C3.8 as Bacillus siamensis and D3.3 as Bacillus subtilis subsp. Subtilis. This study also shows the potential of Bacillus siamensis C3.8 as biocontrol against phytopathogenic fungi.
Abstract. Setiawati S, Nuryastuti T, Sholikhak EN, Lisdiyanti P, Pratiwi SUT, Sulistiyanti TR, Ratnakomala S, Jumina, Mustofa. 2021. The potency of actinomycetes extracts isolated from Pramuka Island, Jakarta, Indonesia as antimicrobial agents. Biodiversitas 22: 1104-1111. Actinomycetes are one of the Gram-positive bacteria which are widely distributed and produce many secondary metabolites including those known as antibiotics, antifungals, anticancer, and antimalarials agents. The secondary metabolites of actinomycetes are abundant, which include many active compounds that have been identified because of the large diversity in the actinomycetes phylum. This study aimed to identify and screen collected by Indonesian Culture Collection (InaCC) from Bojong Gede and Pramuka Island, Jakarta, Indonesia as antibacterial and antifungal agents. Primary screening was done on 16 actinomycetes isolates by well-agar diffusion method. Antimicrobial activity was tested by using micro broth dilution methods to determine minimum inhibitory concentration (MIC). Molecular identification into level genera and species was determined by 16S rRNA gene sequencing. Out of 16 actinomycetes isolates used, 4 isolates have activity against Candida albicans ATCC 10231, Staphylococcus aureus ATCC 6538, Bacillus subtilis BTCC B-612, and Escherichia coli BTCC B-614, specifically InaCC A758, InaCC A759, InaCC A760 and InaCC A765 isolates. InaCC A758 have highest antimicrobial activity against mentioned microbial with MIC value at 50 μg/mL, 6.25 μg/mL, 31.25 μg/mL and 3.125 μg/mL, respectively. Three genera were found from the samples: i.e. Streptomyces (80%), Microbispora (13%) and Nocardia (6%). Based on 16S rRNA gene identification, the active isolates of actinomycetes InaCC A758, InaCC A759, InaCC A760 and InaCC A765 were similar to Streptomyces badius, Streptomyces olivaceus, Streptomyces sanyensis, and Nocardia otitidiscaviarum, respectively. The secondary metabolites of actinomycetes extracts from Pramuka Island can be potentially developed as antifungal and antibacterial agents.
Eurycoma longifolia (pasak bumi) is known as a medicinal plant that contains biologically active compounds. Studies on endophytic bacteria associated with pasak bumi and their biocontrol activities have not been widely reported. The objective of this study is to isolate potential endophytic bacteria associated with E. longifolia possessing biocontrol activity against plant pathogenic fungi, Sclerotium rolfsii, Fusarium solani, F. oxysporum, and Colletotrichum gloeosporioides. Endophytic bacteria were isolated from the roots of E. longifolia using the plant piece method and identified based on 16S rRNA genes analysis. Antagonist test of bacterial isolates was conducted by dual confrontation method. The mechanisms of fungal growth inhibition were evaluated based on their ability to produce hydrolytic enzymes, antibiotic, and volatile organic compounds. Two isolates were obtained and identified as Stenotrophomonas maltophilia (Apb1) and Serratia marcescens (Apb2). Apb2 was able to inhibit the growth of four tested fungi and showed protease, chitinase as well as cellulase activities. The crude extract and volatile organic compound produced by Apb2 were active against F. solani growth.Keywords: biocontrol, endophytic, Eurycoma longifolia, fungi, inhibition mechanism ABSTRAKEurycoma longifolia (pasak bumi) dikenal sebagai tanaman obat yang mengandung beberapa senyawa aktif secara biologis. Penelitian mengenai bakteri endofit yang berasosiasi dengan tanaman pasak bumi berikut aktivitas biokontrolnya belum banyak dilaporkan. Penelitian ini bertujuan mengisolasi bakteri endofit potensial dari tanaman E. longifolia yang memiliki aktivitas biokontrol terhadap empat strain uji jamur patogen tanaman, yaitu Sclerotium rolfsii, Fusarium solani, F. oxysporum, dan Colletotrichum gloeosporioides. Bakteri endofit diisolasi dari akar E. longifola menggunakan metode plant piece dan diidentifikasi berdasarkan analisis gen 16S rRNA. Uji antagonis isolat bakteri dilakukan dengan metode konfrontasi ganda. Mekanisme penghambatan jamur patogen tanaman dievaluasi berdasarkan kemampuannya dalam memproduksi enzim hidrolisis, senyawa antibiotik, dan senyawa organik volatil. Dua isolat bakteri endofit berhasil diperoleh dan teridentifikasi sebagai Stenotrophomonas maltophilia  (Apb1) dan Serratia marcescens (Apb2). Isolat Apb2 mampu menghambat pertumbuhan keempat jamur yang diuji dan menunjukkan aktivitas protease, kitinase dan selulase. Ekstrak kasar dan senyawa organik volatil yang dihasilkan oleh isolat Apb2 aktif menghambat pertumbuhan F. solani.
Bacillus are commonly found in nature, especially in soil and food. It has the ability to produce bioactive compounds as well as the enzyme. This study was aimed to isolate, identify, and screen their enzyme activities. Four soil samples from Mandalika, Lombok Island, West Nusa Tenggara (NTB), were used for isolation. Bacillus was isolated using the heat-shock method and characterized through Gram staining, endospore staining, and morphological phenotype. Bacillus identification was conducted based on 16S rRNA gene sequence. The hydrolytic enzyme activities were checked qualitatively using selective media, and the enzyme tested including amylase, galactosidase, lipase, protease, and cellulase. As many as twenty-two bacteria isolates were obtained from four soil samples and represented 15 distinct species. The member of bacteria genera successfully identified, consisted of Bacillus sp., Brevibacillus sp., and Fictibacillus sp. Bacillus sp. was the most isolated. Some of the isolated bacteria have the ability to produce lipase, protease, and cellulase that potential to be used in biotechnology processes. Keywords: Bacillus, hydrolytic enzymes, identification, screening
Diversity of fruit flies (Diptera: tephritidae) attracted by me lure in CSG-BG germplasm carambolae plantation. Germplasm Garden (KPN) Cibinong Science Center-Botanical Garden (CSC-BG) is one of the gardens that conserve and utilizes Indonesian local germplasm, one of which is carambolae. The current problem in KPN is the attack of fruit flies (Bactrocera spp.). This study was conducted to determine the diversity of fruit flies in the KPN carambola plantation area. A total of eight traps were placed randomly in the plantation area. Each trap contained 1 mL of methyl eugenol pheromone solution. Identification of fruit flies was carried out based on three morphological differences, they were wings, abdomen, and hind legs. There were 317 fruit flies collected and separated into three species, namely Bactrocera carambolae, B. papayae, dan B. umbrosa, of which B. carambolae had a dominance index of 0.74 and abundance index of 85%. Data on the types of fruit flies can be used as primary data to determine the preparation of pest prevention efforts.
The contribution of endophytic bacteria to the wellbeing of plants as biocontrol agents may be due to endophytic bacteria growing in the same niche as phytopathogens. This work was conducted to study the antagonistic activity of endophytic bacteria recovered from sweet sorghum against Sclerotium rolfsii, Fusarium solani, Fusarium oxysporum, Colletotrichum gloeosporioides in vitro and evaluate the mechanisms of these fungal inhibitions. We selected 78 endophytic bacteria from the stem and root of sweet sorghum plants. They were tested for antagonist activity by direct confrontation method. Antifungal compound production and lytic enzyme activity were examined to determine their mechanisms in inhibiting fungal pathogens. Antifungal compound production was checked by detecting the presence of NRPS and PKS genes. Lytic enzyme activity of the bacteria was evaluated by their ability to produce cellulase, chitinase, and protease. Selected bacteria were identified using molecular analysis based on the 16S rRNA gene. 14 out of the 78 tested isolates showed antagonistic activity and two were able to inhibit all four tested fungal strains. Four bacteria, designated as ACIL1, ACNM4, ACNM6, and ATNM4, produced natural products via NRPS pathway, but only one bacterial extract, designated as ACNM4, showed fungal inhibition. Ten isolates were able to produce hydrolytic enzymes. Endophytic bacteria identified as Burkholderia were revealed to have potential as a biocontrol agent.
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