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Abstrak :
Penelitian ini bertujuan untuk memperoleh isolat Actinobacteria termofilik potensial dari tanah di sekitar geiser Cisolok yang dapat mendegradasi xylan dan mengetahui hubungan kekerabatannya dengan taksa terdekat dari Actinobacteria penghasil xylanase. Tujuh belas isolat Actinobacteria termofilik diisolasi dari tanah di sekitar geiser Cisolok, Jawa Barat. Penapisan kemampuan 17 isolat Actinobacteria dan type strain Actinomadura keratinilytica NBRC 105837T mendegradasi xylan dilakukan menggunakan medium Minimal (Mm) padat dengan penambahan substrat xylan 0,5, inkubasi selama 7 hari. Pewarnaan dengan Congo red 0,2 (b/v) menunjukkan terbentuknya zona bening di sekitar koloni isolat Actinobacteria yang dapat mendegradasi xylan 0,5 pada suhu 45 C (15 isolat), 50 C (14 isolat), 55 C (4 isolat), dan 60 C (3 isolat). Type strain NBRC 105837T dapat mendegradasi xylan 0,5 pada suhu 45 C hingga 60 C. Tiga isolat (SL1- 2-R-2, SL1-2-R-3, dan SL1-2-R-4) yang mendegradasi xylan 0,5 hingga suhu 60 C dipilih sebagai isolat potensial. Tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi substrat Remazol Brilliant Blue R-xylan (RBB-xylan) 0,1 pada medium Mm padat setelah 3 hari inkubasi pada suhu 45 hingga 60 C. Tiga isolat potensial telah diidentifikasi pada penelitian sebelumnya sebagai Actinomadura keratinilytica berdasarkan karakter genotip dan fenotip. Crude enzyme dari tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi xylan 0,5 dan RBB-xylan 0,1 pada medium Mm padat setelah 24 jam inkubasi pada suhu 45 hingga 60 C. Berdasarkan analisis filogenetik sequence gen 16S rRNA menggunakan metode neighbor-joining, minimum evolution, dan maximum likelihood, 3 isolat potensial membentuk clade yang monofiletik dengan dua spesies Actinomadura termofilik yang dapat mendegradasi xylan (A. keratinilytica dan A. miaoliensis). Tiga isolat potensial membentuk clade yang monofiletik dengan empat spesies Actinomadura termofilik (A. keratinilytica, A. miaoliensis, A. rubrobrunea, dan A. viridilutea). Tiga isolat potensial menghasilkan miselium substrat yang bercabang dan tidak berfragmen, serta miselium aerial yang menghasilkan spora pada medium modified Bennetts padat setelah 14 hari inkubasi pada suhu 45 C. Penelitian ini memberikan informasi tambahan mengenai kemampuan typestrain A. keratinilytica NBRC 105837 mendegradasi xylan.

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The aims of this study were to obtain the potential xylan-degrading thermophilic Actinobacteria isolates from soil of Cisolok geysers and to understand their relationship with the closely related taxa of xylanase-producing Actinobacteria. Seventeen thermophilic Actinobacteria isolates were isolated from soil collected around Cisolok geysers, West Java. Xylan-degrading ability of 17 Actinobacteria isolates and type strain Actinomadura keratinilytica NBRC 105837T were screened by using Minimal (Mm) agar medium with the addition of 0.5 xylan substrate, incubated for 7 days. Clear zone was formed around the colony of Actinobacteria isolates which showed xylan-degrading ability at 45 C (15 isolates), 50 C (14 isolates), 55 C (4 isolates), and 60 C (3 isolates) after staining by 0.2 (w/v) Congo red. Type strain NBRC 105837T was able to degrade 0,5 xylan at 45 to 60 C. Three isolates (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4) that showed xylan-degrading ability at 45 to 60 C were choosen as potential isolates. Three potential isolates and type strain NBRC 105837T were able to degrade 0,1 Remazol Brilliant Blue R-xylan (RBB-xylan) substrate on Mm agar after 3 days incubation at 45 to 60 C. In the previous study, these potential isolates were identified as Actinomadura keratinilytica based on genotypic and phenotypic characters. Crude enzyme of 3 potential isolates and type strain NBRC 105837T were able to degrade both 0.5 xylan and 0.1 RBB-xylan on Mm agar after 24 hours at 45 to 60 C. Phylogenetic analyses based on 16S rRNA gene using neighbor-joining, minimum evolution, and maximum likelihood methods showed the 3 potential isolates formed monophyletic clade with two thermophilic xylan-degrading Actinobacteria species (A. keratinilytica and A. miaoliensis). Three potential isolates formed monophyletic clade with four thermophilic Actinobacteria species (A. keratinilytica, A. miaoliensis, A. rubrobrunea, and A. viridilutea). These isolates produced non-fragmented branched substrate mycelia and spores produced from aerial mycelia after 14 days incubation at 45 C. This study reports a new information regarding the xylan-degrading ability of A. keratinilytica NBRC 105837.

Abstrak :
Penelitian ini berfokus pada keanekaragaman taksonomi Class Ktedonobacteria, kemampuannya sebagai penghasil enzim, deskripsi takson baru, dan analisis whole genome. Tujuh belas isolat diperoleh dari sampel tanah di hutan dekat geyser, kawasan geotermal Cisolok, Jawa Barat. Sequence gen 16S rRNA dari semua isolat dibandingkan dengan spesies terdekat pada database EzBioCloud. Seluruh isolat memiliki nilai homologi yang rendah terhadap Dictyobacter aurantiacus S-27T (97,82-98,18%). Penapisan kemampuan enzimatik menunjukkan bahwa sebagian besar isolat (88,23%) menghasilkan amilase dan selulase. Komposisi bakteri dari enam sampel dianalisis dengan metode metabarcoding gen 16S rRNA pada daerah V1-V3 menggunakan Illumina Mi-Seq Next Generation Sequencing. Ktedonobacteria merupakan kelas yang paling mendominasi dalam Phylum Chloroflexota (17,74-89,49%) pada lima sampel; Namun, kelas tersebut tidak terdeteksi pada satu sampel (tanah di bawah batu besar). Empat puluh tujuh amplikon gen 16S rRNA dari taksa terdekat Ktedonobacteria berhasil diperoleh dari enam sampel yang mewakili garis keturunan baru pada tingkat takson yang tinggi. Strain S3.2.2.5 diisolasi dari tanah di dalam serasah batang bambu. Karakter fenotipik, genotipik, dan filogenetik menunjukkan bahwa strain tersebut mewakili spesies berbeda dari Genus Dictyobacter; sehingga diusulkan spesies baru Dictyobacter halimunensis sp. nov. ......This study focuses on the taxonomic diversity of the class Ktedonobacteria, their ability as enzyme producers, description of novel taxon, and whole genome analysis. Seventeen isolates were obtained from soil samples in the forest near geyser, Cisolok geothermal area, West Java. The 16S rRNA gene sequence of all isolates was compared with all related species in the EzBioCloud database. All isolates had low similarity values to Dictyobacter aurantiacus S-27T (97.82-98.18%). Primary screening of enzymatic abilities showed that most isolates (88.23%) were amylase- and cellulase-producing Ktedonobacteria. Bacterial composition analyses from six samples were performed based on the V1-V3 of 16S rRNA gene metabarcoding using Illumina Mi-Seq Next Generation Sequencing. Ktedonobacteria was the most dominating class within the phylum Chloroflexota (17.74-89.49%) in five samples; however, it was not detected in one sample (the soil under a big rock). Forty-seven 16S rRNA gene amplicons of Ktedonobacteria-related taxa were generated from six samples and represented the putative new lineages in high taxonomic rank. A strain S3.2.2.5 was isolated from soil inside a decayed bamboo stem. The phenotypic, genotypic, and phylogenetic data suggest this strain represents a distinct species of the Dictyobacter genera; hence, a new species, Dictyobacter halimunensis sp. nov. is proposed.

Abstrak :
ABSTRAK
Penelitian ini bertujuan untuk memperoleh identitas spesies kapang dari dua manuskrip Cina lama yang mengalami deteriorasi asal plot 5 Ruang Naskah Perpustakaan Pusat Universitas Indonesia PP-UI , Depok berdasarkan data sekuens daerah internal transcribed spacers ribosomal DNA ITS rDNA . Pengambilan sampel pada manuskrip menggunakan metode swab dengan cotton bud steril. Isolasi kapang menggunakan metode culture-dependent. Polymerase chain reaction PCR dan DNA sequencing menggunakan primer forward ITS5 dan primer reverse ITS4. Pencarian homologi sekuens daerah ITS rDNA menggunakan program basic local alignment search tool BLAST dan pembuatan pohon filogenetik menggunakan metode Neighbor Joining, model dua parameter Kimura, serta bootstrap sebanyak 1.000 kali replikasi. Penentuan spesies terdekat dan posisi taksonomi menggunakan analisis filogenetik dan didukung oleh data morfologi. Isolasi kapang menghasilkan enam isolat kapang terpilih berdasarkan tipe morfologi yang berbeda dengan kapang dari manuskrip Cina lama asal plot 1, 2, 4, dan 6 Ruang Naskah PP-UI, Depok. Berdasarkan elektroforesis gel, panjang fragmen daerah ITS rDNA dari enam isolat kapang bervariasi antara 600--700 pb. Hasil DNA sequencing lengkap menunjukkan panjang daerah ITS rDNA enam isolat berkisar 582--625 pb. Enam strain UICC merupakan anggota dari tiga kelas Dothideomycetes, Eurotiomycetes dan Sordariomycetes , tiga ordo Capnodiales, Eurotiales dan Hypocreales , dan empat famili Cladosporiaceae, Nectriaceae, Ophiocordycipitaceae, dan Pleosporaceae . Strain UICC 1107 memiliki homologi 99,32 dengan type strain Purpureocillium lilacinum sin. Paecilomyces lilacinus ATCC 10114T. Lima strain UICC tidak dapat ditentukan spesiesnya. Strain UICC 1106 adalah Cladosporium sp. dengan homologi 100 terhadap type strain Cladosporium oxysporum CBS 125991T dan Cla. tenuissimum CPC 14235T. Strain UICC 1105 adalah Curvularia sp.1 dengan homologi 93,80 dan strain UICC 1108 adalah Curvularia sp.2 dengan homologi 94,70 terhadap type strain Curvularia carica-papayae CBS 135941T. Strain UICC 1109 adalah Rectifusarium sp. dengan homologi 85,87 terhadap type strain Rectifusarium robinianum CBS 430.91T. Strain UICC 1104 adalah Sarocladium sp. dengan homologi 97,13 terhadap type strain Sarocladium bifurcatum UTHSC 05-3311T. Enam strain UICC merupakan fungi anamorf dan bersifat xerofilik.

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ABSTRACT The aim of this study was to determine the species identity of moulds from two deteriorated old Chinese manuscripts from plot 5 Ruang Naskah Central Library Universitas Indonesia, Depok based on internal transcribed spacers region of ribosomal DNA ITS rDNA . Samples from the manuscripts were collected by using swab method with sterile cotton swabs. Mould isolates were obtained by culture dependent method. Polymerase chain reaction PCR and DNA sequencing were performed using forward primer ITS5 and reverse primer ITS4. Homology search of ITS rDNA sequences was carried out using basic local alignment search tool BLAST program and phylogenetic tree construction was performed using Neighbor Joining method, Kimura rsquo s two parameter model, and bootstrap 1,000 replicates. The closest species and taxonomic position were obtained by phylogenetic analysis and supported by morphological data. Six mould isolates were selected based on morphological type differences compared to mould isolates from old Chinese manuscripts from plot 1, 2, 4, and 6 Ruang Naskah Central Library UI, Depok. Based on gel electrophoresis, the lengths of ITS rDNA fragments of six mould isolates varied between 600 700 bp. Full sequence data of ITS rDNA of six isolates showed that the lengths of their ITS rDNA varied between 582 625 bp. Six UICC strains belonged to three classes Dothideomycetes, Eurotiomycetes and Sordariomycetes , three orders Capnodiales, Eurotiales and Hypocreales , and four families Cladosporiaceae, Nectriaceae, Ophiocordycipitaceae, and Pleosporaceae . UICC 1107 strain showed 99.32 homology to the type strain, Purpureocillium lilacinum syn. Paecilomyces lilacinus ATCC 10114T. Five UICC strains were not able to be determined to the species level. UICC 1106 strain was identified as Cladosporium sp., with 100 homology to the type strains, Cladosporium oxysporum CBS 125991T and Cla. tenuissimum CPC 14235T. UICC 1105 strain was identified as Curvularia sp.1, with 93.80 homology and UICC 1108 strain was identified as Curvularia sp.2, with 94.70 homology to the type strain, Curvularia carica papayae CBS 135941T. Strain UICC 1109 was identified as Rectifusarium sp., with 85.87 homology to the type strain, Rectifusarium robinianum CBS 430.91T. Strain UICC 1104 was identified as Sarocladium sp., with 97.13 homology to the type strain, Sarocladium bifurcatum UTHSC 05 3311T. Six UICC strains were anamorphic and xerophilic fungi.