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Endophytic Streptomyces spp. as Biocontrol Agents of Rice Bacterial Leaf Blight Pathogen (Xanthomonas oryzae pv. oryzae) HASTUTI, RATIH DEWI; LESTARI, YULIN; SUWANTO, ANTONIUS; SARASWATI, RASTI
HAYATI Journal of Biosciences Vol 19, No 4 (2012): December 2012
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (402.746 KB) | DOI: 10.4308/hjb.19.4.155

Abstract

Xanthomonas oryzae pv. oryzae (Xoo), a causal agent of bacterial leaf blight (BLB), is one of the most important pathogens of rice. The effectiveness of ten Streptomyces spp. isolates in suppressing Xoo disease was assessed in planta and in vitro. In planta experiments were carried out in a greenhouse and arranged in a randomized completely block design (RCBD) with three replications. Twenty treatments were tested which included plants inoculated with both Streptomyces spp. and Xoo, and plants inoculated with only Streptomyces spp. Plants inoculated with Xoo and sprayed with a chemical bactericide, and plants inoculated with only Xoo served as positive controls, whereas plants not inoculated with either Streptomyces spp. or Xoo were used as negative controls. The results showed that the effect of endophytic Streptomyces spp. on BLB disease expressed as area under disease progress curve (AUDPC) was not significantly different to that on control plants (P > 0.05). However, plants inoculated with endophytic Streptomyces spp. were significantly taller and produced higher tiller number than control plants (P < 0.05). Streptomyces spp. isolate AB131-1 gave the highest plant height. In vitro studies on biocontrol mechanisms of selected Streptomyces spp. isolates showed that isolate LBR02 gave the highest inhibition activity on Xoo growth, followed by AB131-1 and AB131-2. Two isolates (AB131-1 and LBR02) were able to produce chitinase, phosphatase, and siderophore which included biocontrol characteristics. Morphological and colonization studies under SEM and light microscopy confirmed that the three isolates were endophytic Streptomyces spp. from different species. These studies found that the paddy plant which was inoculated with endophytic Streptomyces spp. AB131-1 and infected by Xoo could increase the height of plant and number of tillers.
Potential Pseudomonas Isolated from Soybean Rhizosphere as Biocontrol against Soilborne Phytopathogenic Fungi SUSILOWATI, ARI; WAHYUDI, ARIS TRI; LESTARI, YULIN; SUWANTO, ANTONIUS; WIYONO, SURYO
HAYATI Journal of Biosciences Vol 18, No 2 (2011): June 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (105.133 KB) | DOI: 10.4308/hjb.18.2.51

Abstract

Plants are liable to be attacked by soilborne fungal pathogens which are responsible to reduce plant growth and losses in yield. In Indonesia, indigenous soybeans’ rhizobacteria such as antifungal producing Pseudomonas sp. have not many been reported yet. Therefore, the potential of the Pseudomonas sp. as biocontrol agent should be deeply explored. The aim of this study was to screen the indigenous soybeans’ rhizobacteria Pseudomonas sp. that possessing biocontrol characters against soilborne mainly i.e. Sclerotium rolfsii, Fusarium oxysporum, and Rhizoctonia solani, in vitro and in planta. Eleven isolates identified Pseudomonas sp. CRB numbered by CRB-3, CRB-16, CRB-17, CRB-31, CRB-44, CRB-75, CRB-80, CRB-86, CRB-102, CRB-109, and CRB-112 were affirmed to be candidates of biocontrol agents toward the soilborne fungal pathogens. Pseudomonas sp. CRB inhibited growth of the pathogenic fungi approximately 11.1-60.0% in vitro. Among of them, 7 isolates were also produced siderophore, 2 isolates produced chitinase, and 4 isolates produced hydrogen cyanide. Seed coating with the Pseudomonas sp. CRB accomplished disease suppression in planta about 14.3-100% in sterile soil condition and 5.2-52.6% in non sterile soil condition. Consistency in high performance more than 30% of disease suppression in non sterile soil condition suggested that 5 isolates i.e. CRB-16, CRB-44, CRB-86, CRB-102, and CRB-109 isolates have great promising to be developed as biocontrol agents of soilborne pathogenic fungi.
Immobilization of Extracellular Xylanase from Streptomyces sp. 45 I-3 for Hydrolysis of Corncob Xylan Sunarti, Titi Candra; Mutia, Ferry; Gusmawati, Niken Financia; Lestari, Yulin; Meryandini, Anja
Jurnal Teknologi Dan Industri Pangan Vol 20, No 1 (2009): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (290.721 KB) | DOI: 10.6066/313

Abstract

PENGGUNAAN  XILANASE  Streptomyces sp. 45 I-3 AMOBIL UNTUK HIDROLISIS XILAN TONGKOL JAGUNG [Immobilization of Extracellular Xylanase from Streptomyces sp. 45 I-3 for Hydrolysis of Corncob Xylan ] Anja Meryandini1),2), Titi Candra Sunarti3), Ferry Mutia3), Niken Financia Gusmawati4), dan Yulin Lestari2) 1)Pusat Penelitian Sumberdaya Hayati dan Bioteknologi IPB,Gedung PAU, Kampus IPB Darmaga 16680 2)Departemen Biologi, FMIPA-IPB, Gedung Fapet Lt 5 Wing 1, Kampus IPB Darmaga 16680 3)Departemen Teknologi Industri Pertanian, FATETA-IPB, Kampus IPB Darmaga 16680 4)PS Bioteknologi-Sekolah Pasca Sarjana IPB,  Gedung PAU, Kampus IPB Darmaga 16680 Autor korespondensi: ameryandini@yahoo.com Diterima 15 Desember 2008/Disetujui 13 Juni 2009 ABSTRACT   Xylan extraction from corncob is done by using alkaline as solvent. Xylan extraction from corncob could give the yields as 10.9%. One percent of corncob xylan is used as substrate to produce the xylanase, compared to oatspelt xylan. Immobilization of xylanase was performed using 1% EudragitTM S100 solution (w/v), with 5:1 volume ratio of xylanase and 1 % EudragitTM S100 (w/v). Activity of the immobilized xylanase was decreased to 23.97% compared with free xylanase. Immobilized xylanase have optimum pH and temperature at 6.0 and 40°C  respectively, have also thermal stability at 30–40°C for an hour. Immobilized xylanase could be reused, but its activity decreased to 52.38% after 3 times application.   Key words : xilanase, Streptomyces, amobile
Genetic Diversity of Antifungi-Producing Rhizobacteria of Pseudomonas sp. Isolated from Rhizosphere of Soybean Plant SUSILOWATI1, SUSILOWATI1; WAHYUDI, ARIS TRI; LESTARI, YULIN; WIYONO, SURYO; SUWANTO, ANTONIUS
Microbiology Indonesia Vol 4, No 1 (2010): April 2010
Publisher : Indonesian Society for microbiology

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Abstract

Antifungi-producing rhizobacteria have been recognized playing an important role in plant disease suppression. In our laboratory, 13 indigenous soybeans&acute; rhizobacteria Pseudomonas sp. that showed strong growth inhibition of root pathogenic fungi, Rhizoctonia solani, Fusarium oxysporum and Sclerotium rolfsii, have been isolated from rhizosphere of soybean plant. For further understanding, the genetic diversity of the antifungi-producing Pseudomonas sp. was investigated using Amplified 16S rDNA Restriction Analysis (ARDRA) and 16S rRNA gene sequences analysis. 16S rDNA were amplified by PCR technique and digested with restriction endonuclease HaeIII, RsaI and AluI. Sequences of 16S rRNA gene were analyzed using the BLAST program for similarity searches on sequence databases. ARDRA based dendrogram analysis was carried out by neighbor-joining of TREECON 1.3b software package. ARDRA indicated the variability of Pseudomonas sp. based on the digestion sites. Dendrogram clustering analysis based on the restriction enzymes profile of the amplified rDNA distinguished Pseudomonas sp. into 7 ribotype groups. The sequences of 16S rRNA gene confirmed that the isolates belonging to Pseudomonas sp. and the phylogenetic tree formed 4 clusters. There was a quite overlap among ARDRA groups and 16S rRNA sequence clusters. This finding suggested that antifungal producing Pseudomonas sp. were present in the rhizosphere of soybean plant and the level of genetic diversity exist within these species. Sequence analysis of the 16S rRNA gene of the Pseudomonas sp. with an identical ARDRA pattern confirmed that members of an ARDRA group were closely related to each other.
Characterization of Xylanase Streptomyces spp. SKK1-8 MERYANDINI, ANJA; HENDARWIN, TRIO; SAPRUDIN, DEDEN; LESTARI, YULIN
HAYATI Journal of Biosciences Vol 13, No 4 (2006): December 2006
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (134.007 KB) | DOI: 10.4308/hjb.13.4.151

Abstract

Streptomyces spp. SKK1-8 producing xylanase was isolated from soil sample from Sukabumi West Java. The xylanase have an optimum condition at pH 6 and 50 0C. Addition of 5 mM Cu2+ decreased the xylanase activity up to about 77%, whereas not by other cations. The xylanase was stable at 3 0C for 48 hours, and the enzyme half lifetime was 1 hour 45 minute at 50 0C. This xylanase showed the highest activity on oatspelt xylan, and their molecular masses were estimated approximately 16.80, 15.21, and 13.86 kDa. HPLC analysis showed that xylosa and arabinosa were the main hydrolytic product of birchwood xylan. Key words: xilanase, Streptomyces spp., characterization, zymogram and SDS-PAGE, stability
Population and Diversity of Endophytic Bacteria Associated with Medicinal Plant Curcuma zedoaria SULISTIYANI, TRI RATNA; LISDIYANTI, PUSPITA; LESTARI, YULIN
Microbiology Indonesia Vol 8, No 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5454/mi.8.2.4

Abstract

Traditionally Curcuma zedoaria (white turmeric) known as herbal medicine which possessing many biological activities. Many endophytic bacteria live in association with their host and may play an important biological roles. The main interest of this study was to investigate the endophytic bacterial diversity associated with white turmeric. White turmerics were collected from three locations in Bogor, West Java, Indonesia. The isolation of endophytic bacteria was carried out using 4 kinds media (Nutrient Agar (NA), NA contained white turmeric extract (NAT), Water Yeast Extract Agar (WYEA), WYEA contained white turmeric extract (WYEAT)), and 2 methods of spread plate and plant piece methods. The identification of selected isolates was conducted by molecular analysis based on 16S DNA. The suitable media and method of isolation endophytic bacteria were NA and spread plate method. A total of 207 bacterial colonies were isolated from rhizomes, stems, and leaves and 73 endophytic bacteria were selected based on morphological characteristics. From them, 32% isolates from Bojong Gede, 22% isolates from Cibinong and 46% isolates from Dramaga were obtained. Endophytic bacteria were predominated 38% in the rhizomes, 32% of stems, and 30% of leaves. Based on 16S rDNA sequence analysis, the isolates were belonging to the cluster Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria, with twenty three different genera includes Stenothropomonas, Pseudomonas, Enterobacter, Providencia, Klebsiella, Dickeya, Pantoea, Bacillus, Acinetobacter, Citrobacter, Mycobacterium, Cellulomonas, Microbacterium, Methylobacterium, Penylobacterium, Roseomonas, Agrobacterium, Bosea, Xanthobacter, Rhizobium, Burkholderia, Ralstonia, and Alcaligenes. The plant location, age, part of plant, media and method of isolation seem to influence the endophytic bacterial communities.
STUDI MIKROBIOLOGI DAN SIFAT KIMIA MIKROORGANISME LOKAL (MOL) YANG DIGUNAKAN PADA BUDIDAYA PADI METODE SRI (System of Rice Intensification) Suhastyo, Arum Asiyanti; Anas, Iswandi; Santosa, Dwi Andreas; Lestari, Yulin
SAINTEKS Vol 10, No 2 (2013): Jurnal Sainteks Volume X No 2 Oktober 2013
Publisher : LPPM UNIVERSITAS MUHAMMADIYAH PURWOKERTO

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Abstract

Penggunaan larutan mikroorganisme lokal (MOL) dalam metode SRI budidaya padi yang dikembangkan di Indonesia dimulai awal sejak persiapan bibit fase vegetatif, pembentukan malai dan pengisian bulir padi. MOL adalah cairan yang dapat dibuat dari bahan yang tersedia di sekitar kita seperti sisa sayuran, rebung, keong mas, buah maja, daun gamal, bonggol pisang, nasi, urine kelinci, dan lain-lain. Cairan umumnya diberikan 10, 20, 30, 40 dan 60 hari setelah tanam (HST ) atau sesuai kebutuhan. Penelitian ini menggunakan larutan MOL yang terbuat dari bonggol pisang, keong mas dan urin kelinci. Tujuan penelitian adalah untuk menentukan mikrobaa, identifikasi mikrobaa dan sifat kimia dalam MOL bonggol pisang, keong mas dan urin kelinci. Penelitian ini dilakukan di Laboratorium Bioteknologi Tanah IPB. Penelitian ini menggunakan rancangan acak lengkap dengan satu faktor (waktu) dan tiga kali pengulangan. Hasil penelitian menunjukkan MOL bonggol pisang memiliki rata-rata nilai pH terendah (4,2-4,5) dan nilai EC tertinggi (10,44-12,82 mikrodetik/cm) selama proses fermentasi. MOL keong emas memiliki rata-rata pH tertinggi (4,5-6,55) dan dan yang paling oksidatif - reduktif nilai Eh [ 269-(-381) mV ] selama proses fermentasi. MOL urin kelinci memiliki nilai rata-rata terendah dari EC (2,18-2,23 mikrodetik/cm) dan mengandung lebih banyak unsur K, Ca, Mg, Cu, Zn, Fe dan Mg dari kedua jenis MOL lainnya. Selanjutnya, Bacillus sp., Aeromonas sp. dan Aspergillus niger diidentifikasi dalam MOL dari bonggol pisang. MOL dari keong mas mengandung Staphylococcus sp. dan Aspergillus niger, sedangkan MOL urin kelinci memiliki Bacillus sp. , Rhizobium sp. , Pseudomonas sp. , Aspergillus niger dan Verticillium sp.Kata kunci : SRI, MOL, sifat kimia MOL, identifikasi mikrobaa
Metagenomic of Actinomycetes Based on 16S rRNA and nifH Genes in Soil and Roots of Four Indonesian Rice Cultivars Using PCR-DGGE Mahyarudin, .; Rusmana, Iman; Lestari, Yulin
HAYATI Journal of Biosciences Vol 22, No 3 (2015): July 2015
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1377.267 KB) | DOI: 10.4308/hjb.22.3.113

Abstract

The research was conducted to study the metagenomic of actinomycetes based on 16S ribosomal RNA (rRNA) and bacterial nifH genes in soil and roots of four rice cultivars. The denaturing gradient gel electrophoresis profile based on 16S rRNA gene showed that the diversity of actinomycetes in roots was higher than soil samples. The profile also showed that the diversity of actinomycetes was similar in four varieties of rice plant and three types of agroecosystem. The profile was partially sequenced and compared to GenBank database indicating their identity with closely related microbes. The blast results showed that 17 bands were closely related ranging from 93% to 100% of maximum identity with five genera of actinomycetes, which is Geodermatophilus, Actinokineospora, Actinoplanes, Streptomyces and Kocuria. Our study found that Streptomyces species in soil and roots of rice plants were more varied than other genera, with a dominance of Streptomyces alboniger and Streptomyces acidiscabies in almost all the samples. Bacterial community analyses based on nifH gene denaturing gradient gel electrophoresis showed that diversity of bacteria in soils which have nifH gene was higher than that in rice plant roots. The profile also showed that the diversity of those bacteria was similar in four varieties of rice plant and three types of agroecosystem. Five bands were closely related with nifH gene from uncultured bacterium clone J50, uncultured bacterium clone clod-38, and uncultured bacterium clone BG2.37 with maximum identity 99%, 98%, and 92%, respectively. The diversity analysis based on 16S rRNA gene differed from nifH gene and may not correlate with each other. The findings indicated the diversity of actinomycetes and several bacterial genomes analyzed here have an ability to fix nitrogen in soil and roots of rice plant.
PENGGUNAAN XILANASE Streptomyces sp. 45 I-3 AMOBIL UNTUK HIDROLISIS XILAN TONGKOL JAGUNG [Immobilization of Extracellular Xylanase from Streptomyces sp. 45 I-3 for Hydrolysis of Corncob Xylan ] Sunarti, Titi Candra; Mutia, Ferry; Gusmawati, Niken Financia; Lestari, Yulin; Meryandini, Anja
Jurnal Teknologi Dan Industri Pangan Vol 20, No 1 (2009): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (290.721 KB)

Abstract

Xylan extraction from corncob is done by using alkaline as solvent. Xylan extraction from corncob could give the yields as 10.9%. One percent of corncob xylan is used as substrate to produce the xylanase, compared to oatspelt xylan. Immobilization of xylanase was performed using 1% EudragitTM S100 solution (w/v), with 5:1 volume ratio of xylanase and 1 % EudragitTM S100 (w/v). Activity of the immobilized xylanase was decreased to 23.97% compared with free xylanase. Immobilized xylanase have optimum pH and temperature at 6.0 and 40°C  respectively, have also thermal stability at 30–40°C for an hour. Immobilized xylanase could be reused, but its activity decreased to 52.38% after 3 times application.
Isolation of actinomycetes from maize rhizosphere from Kupang, East Nusa Tenggara Province, and evaluation of their antibacterial, antifungal, and extracellular enzyme activity Fatmawati, Umi; Lestari, Yulin; Meryandini, Anja; Nawangsih, Abdjad Asih; Wahyudi, Aris Tri
Indonesian Journal of Biotechnology Vol 23, No 1 (2018)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.33064

Abstract

Actinomycetes are the one of the components of the rhizospheric microbial population and useful for producing secondary metabolites such as lytic enzymes, antibiotics, and antifungal. The aim of the study was to isolate the actinomycetes from maize rhizosphere collected from Kupang, East Nusa Tenggara. The screening was focused on the actinomycetes that showed the ability to produce antibacterial, antifungal, and extracellular enzymes such as amylase, cellulase, and protease. The actinomycetes were isolated using Humic-Acid Vitamin B (HV) agar media. The antagonistic assay was tested against Escherichia coli, Staphylococcus aureus, Sclerotium rolfsii and Fusarium oxysporum. Isolate JKP-8 was an isolate that showed the highest activity in inhibiting the growth of E. coli and S. aureus bacteria. Isolate JKP-5 showed the highest activity in inhibiting the growth of F.oxysporum. There were no actinomycetes isolates that showed an ability to inhibit the growth of S. rolfsii fungus based on dual culture assay. JKP-3 and JKP-4 isolates exhibited the highest ability to hydrolyze amylum, while JKP-5 and JKP-8 isolates exhibited the highest ability to hydrolyze CMC. The results of the amplification of 16S rRNA gene in selected potential isolates JKP 5 and JKP 8 indicated that both isolates belong to the genus Streptomyces.