Production and Characterization of Extracellular Protease from Bacillus sp. 140-B Isolated from Pineapple Plantation in Lampung, Indonesia

Bacillus sp. being industrially important organisms produces a wide variety of extracellular enzymes including protease. Bacillus sp. 140-B isolated from rhizosphere area of pineapple plantation in Lampung Province, Indonesia was tested for production of protease on skim milk agar medium. The aims of this study were to characterize and investigate some properties of  protease activity from Bacillus sp. 140-B. Bacillus sp. 140-B showed  protease activity qualitatively by clear zone diameter of 15.0 mm and the highest protease activity was 35.02 Unit · mg–1 protein at 14 h after incubation. Some properties of protease activities from Bacillus sp. 140-B including effects of temperature, pH and several metals were observed in this experiment. The protease activity from Bacillus sp. 140-B had optimum pH of 7,0 and the optimum temperature was 60 °C. Several metals which were evaluated on protease activity showed that Mn could increase protease activity, while other metals (Ca, Co, and Hg) showed decreasing protease activity of Bacillus sp. 140-B. Keywords: Bacillus sp. 140-B; enzyme activity; enzyme properties; proteas

STUDY OF PROPOXUR DEGRADING BACTERIA ISOLATED FROM VARIOUS SAMPLING SITE OF RICE FIELD FROM NGAWI

Propoxur (2-isopropoxyphenyl-N-methylcarbamate) is a carbamateinsecticide which has negative impact on the environment. Propoxur is readily degraded by soil microorganism in most soils. Increased enviromental degradation of propoxur constitute the result of increase in activities of soil pesticide-degrading bacteria. The aims of the present study are to isolate andscreen the bacteria with high potential ability to degradepropoxur from soil samples. Isolation and selection were carried out by enrichment cultures technique.Propoxur degrading bacteria were isolated from 5 soil samples in different locations of rice field from Ngawi. The biodegradation ofpropoxur by isolated bacteria were analysed by High Performance Liquid Chromatography (HPLC). Ten consorsia were selected and purified. One isolate was able to use propoxur at 3000 ppm as sole source of carbonfor the growth. The result suggested that H2-NG isolate hydrolysedpropoxur with the yield of 2-isopropoxyphenol and methylamine. Keywords: Ngawi, Isolation, Propoxur, Biodegradation, Bacteri

Secondary Metabolites Production by Actinomycetes and their Antifungal Activity

Wilt desease of banana caused by Fusarium oxysporum f.sp. cubense (FOC) is one of the most destructive deseases of banana in the tropics. Actinomycetes are the most economically and biotechnologically valuable prokaryotes able to produce wide range of bioactive secondary metabolites. The aims of the present study are to isolate and screen the actinomycetes with high potential ability to produce secondary metabolites that have inhibitory activity against plant pathogenic fungi, Fusarium oxysporum f. sp. cubense. Two isolates from Lampung and Cianjur showed activity against fungi. The isolates designed as L.3.1 and CiIA5b. The metabolites from potent stain was produced by extraction of culture filtrate with ethyl acetate : methanol (4:1), it was tested for their antifungal activity by well diffusion method. Evidence for in vitro antibiosis of L.3.1 and CiIA5b isolates was demonstrated by the zone of fungal-growth inhibition. Production of secondary metabolites was analysis by thin layer chromatography (TLC) and bioautography assays. In this study, the metabolites from L.3.1 and CiIA5b have showed good antifungal activity. Keywords: Actinomycetes; antifungal activity; bioautography; secondary metabolites; thin layer chromatography

PCR-based Specific Detection of Bacillus in Liquid Organic Fertilizer

Rapid molecular PCR-based detection method for Bacillus species used in the production of Beyonic® liquid organic fertilizer was carried out based on nucleotide sequence data from the 16S rRNA gene. The method involved sequencing the 16S rRNA gene of several Bacillus species and identifying around 16-22 specific nucleotide bases from 5' and 3' ends in the Bacillus 16S rRNA gene sequences. One specific primer pair for Bacillus detection was determined as follow: 5' - CAT AAG ACT GGG ATA ACT CCG GG - 3' (forward) from positions of 85-107 bp, and 5’ - CCA GGC GGA GTG CTT AAT GC - 3’ (reverse) from positions of 836-854 bp. PCR assay and gel electrophoresis analysis showed that the primer pair was specific to the genus Bacillus

Potensi Isolat Azospirillum spp. Asal Lahan Pasir Sebagai Kandidat Rizobakteri Pemacu Pertumbuhan Tanaman (RPPT)

Lahan pasir besi merupakan lahan marginal yang memiliki kandungan bahan organik rendah, kapasitas pertukaran kation rendah, kemampuan menahan air rendah, dan kandungan besi (Fe) berkisr 38.59%. Spesies anggota genus Azospirillum selain dikenal mampu tumbuh di lingkungan marginal juga merupakan salah satu angota rizobakteri pemacu pertumbuhan tanaman (RPPT. Penelitian ini bertujuan untuk mengetahui potensi isolat Azospirillum spp. asal lahan pasir besi sebagai kandidat RPPT. Penelitian untuk mengetahui karakter fisiologis Azospirillum spp. Mencakup kemampuan melarutlan fosfat, produksi fitohormon IAA, siderofor, dan ACC­deaminase, serta sifat antagonisme terhadap beberapa fungi patogen dilakukan secara deskriftik. Hasil penelitian menunjukkan bahwa enam isolat Azospirillum spp. (HR 154, KR33, HR2, HR141, HR124, HR1S2) asal lahan pasir besi mampu melarutkan fosfat, Menghasilkan fitohormon IAA, siderofor dan empat isolat (HR154, HR92, HR141, HR152) mampu menghasilkan ACC­dearinase. Pelarutan fosfat yang tinggi ditunjukkan oleh isolat KR33 dan HR141 dengan indeks pelarutan masing­masing sebesar 135,38 dan 123,67. Produksi IAA tertinggi ditunjukkan oleh isolat HR92 yaitu sebanyak 97,02 mg.mL. Penghasilan siderofor yang tinggi diperlihatkan oleh isolate HR154 dan HR14, masing­masing sebanyak 26,6 dan 28,1 % siderofor unit. Namun demikian. keenam isolat tidal bersifat antagonis terhadap fungi patogen Rhizoctonia solani, Fusarium oysporum, Cercospora sp., dan Aspergillus sp.. Hasil penelitian mengindikasikan bahwa keenam isolat Atospirillum spp asal lahan pasir besi memiliki potensi sebagai kandidt RPPT

Aktivitas Pelarutan Fosfat Isolat Azospirillum spp. asal Lahan Pasir Besi

Iron sand lands have a low organic matter content as it contains of Iron (Fe) found nearly all over the coastal or beach in Indonesia. Phosphorous is one of macronutrient with considered role for plant promotion. One group of bacteria that are in an environment of rooting plants on land sand iron was Azospirillum spp. The purpose of this study was is to know the phosphate solubilizing activities of Azospirillum spp. isolates from land sand iron. The results showed that some isolates of Azospirillum spp. origin of iron sand land capable of solubilizing phosphate in qualitative and quantitative

Effect of Organic Fertilizer Products on the Growth and Health of Acacia Crassicarpa Seedlings

Acacia crassicarpa seedlings face challenges with growth and health. Xanthomonas leaf blight, Fusarium wilt, and leaf spots caused by Pestalotiopsis or Phaeotrichoconis are all major nursery diseases of A. crassicarpa. This study was carried out to consider solutions for improving seedling conditions. The growth and health of A. crassicarpa seedlings were assessed using two commercial products: an organic biofertilizer (LOB-BS) and an organic multipurpose fertilizer (OMF). The LOB-BS + basal fertilizer (BF) treatment led to a higher germination rate than the other treatments, according to our findings. Similarly, at 12 weeks after sowing, the same treatment produced the tallest A. crassicarpa seedlings. In comparison to other treatments, the incidence of Xanthomonas leaf blight, Fusarium wilt, and leaf spot diseases was lower in the plots of OMF and OMF + BF treatments. Based on these findings, the LOB-BS product could be used to improve the growth of A. crassicarpa seedlings, while the MOF product could be used to reduce the incidence of major A. crassicarpa seedling diseases in the nursery. Keywords: Fusarium, leaf spot, Xanthomonas, organic biofertilizer (LOB-BS), organic multipurpose fertilizer (OMF)

Indole Acetic Acid-Producing and Phosphate-Solubilizing Bacteria From the Rhizosphere of Clove (Syzygium Aromaticum L.) in Bali, Indonesia

Clove plants are routinely fertilized with synthetic fertilizer to increase yield. The use of synthetic fertilizer reduces soil productivity. Biofertilizer can be used as an alternative for increasing soil fertility. The goal of this study was to determine the potency of bacteria isolates capable of producing indole acetic acid (IAA) hormone and solubilizing phosphate, and to identify bacteria species from the rhizosphere of clove plants. Soil samples were collected from the clove plants’ rhizosphere, environmental parameters were measured, the potency of IAA-producing and phosphate-solubilizing bacteria was analyzed, and bacteria were molecularly identified. After 48 hours of incubation, isolate TCKI 5 from Karangasem produced the highest IAA hormone levels (19.64 ppm), and isolate TCBP 6 from Buleleng had the highest index of solubilizing phosphate (1.91). A compatibility test between the three best isolates of IAA hormone-producing and phosphate-solubilizing bacteria revealed that TCKI 5 was able to associate with TCBP 6. Isolate TCKI 5 was identified as Leclercia adecarboxylata C107 with a 99.92% similarity, and isolate TCBP 6 as Burkholderia cepacia GJ8 with a 99.61% similarity. Keywords: Bacteria, clove, Indole Acetic Acid, phosphate, rhizospher

Penapisan dan Identifikasi Bakteri Agens Biokontrol Penyakit Layu Fusarium Hasil Isolasi dari Rizosfer Pisang

Banana wilt disease caused by Fusarium oxysporum f.sp.cubense is considered as one of the most destructive diseases on banana plants in the tropical region. Biological control agents (BCA’s) have become a promising solution to overcome this disease. The objective of this study was to find potential BCA’s for wilt disease of banana plants. Bacteria and actinomycetes were isolated from banana’s rhizosphere in Lampung and Cianjur. As much as 64 actinomycetes and 142 bacteria isolates were obtained. Antagonistic test against F. oxysporum f. sp. cubense of those isolates showed that 21 bacteria and 10 actinomycete isolates have abilities to inhibit F. oxysporum f. sp. cubense. Actinomycetes showed relatively higher inhibition against F. oxysporum f. sp. cubense compared to bacteria. Isolates which have positive antagonistic activities against F. oxysporum f. sp. cubense were then tested for their protease, chitinase, and selulase activities qualitatively on specific medium. All actinomycetes which were tested had chitinase enzyme activities, while only 5 bacterial isolates had chitinase activities. On the other hand, 13 bacterial isolates showed protease activities and only 1 actinomycete showed protease activity. Two bacterial isolates (L.II.4.ND and L.A.I-5.DW) and 3 actinomycetes (L.A.I.DW, L.3.1.DW and Ci.I.A5.DW) which showed high inhibition against F. oxysporum f. sp. cubense and lyses enzymes activities were identified based on 16S rRNA genes. Analysis based on GenBank data, those isolates have 99% homology to Klebsiella pneumonia (L.II.4.ND), Burkholderia sp. (L.A.I-5.DW), Streptomyces sp. (L.A.I.DW), Streptomyces sp. (L.3.1.DW) and Streptomyces sp. (Ci.I.A5.DW)