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Abstrak :
Universitas Indonesia Culture Collection (UICC) memiliki koleksi strain-strain Rhizopus oryzae Went dan Prinsen Geerligs yang diisolasi dari tempe dan telah diidentifikasi berdasarkan karakter fenotipik (morfologi dan fisiologi). Tujuan penelitian adalah melakukan re-identifikasi lima strain R. oryzae (UICC 10, UICC 85, UICC 119, UICC 120, dan UICC 135) berdasarkan data sequence daerah internal transcribed spacers ribosomal DNA dan analisis filogenetik untuk memperoleh identitas spesies yang akurat. Amplifikasi dan sequencing daerah ITS rDNA dilakukan menggunakan primer forward ITS 5 dan primer reverse ITS 4. Data sequence dikirim ke database DNA GenBank untuk pencarian homologi dengan program BLAST. Sequence alignment dilakukan menggunakan program Clustal X. Konstruksi pohon filogenetik dilakukan menggunakan metode Neighbor Joining, model dua parameter Kimura, dan nilai bootstrap 1000 kali pengulangan. Karakterisasi morfologi dan pengujian kemampuan tumbuh pada variasi suhu dilakukan untuk mendukung hasil re-identifikasi dan melengkapi deskripsi strain masing-masing. Hasil amplifikasi daerah ITS rDNA menunjukkan bahwa kelima strain R. oryzae UICC memiliki panjang fragmen dengan ukuran 600--700 pb. Hasil pencarian homologi BLAST menunjukkan bahwa kelima strain memiliki homologi 99,8% terhadap type strain R. oryzae CBS 112.07T. Hasil analisis filogenetik menunjukkan bahwa kelima strain berada dalam satu grup yang monofiletik dengan strain tipe R. oryzae CBS 112.07T dan strain-strain R. oryzae dari database DNA GenBank, dengan dukungan nilai bootstrap yang tinggi (84%). Re-identifikasi lima strain R. oryzae UICC secara molekuler menghasilkan identitas spesies yang sama, yaitu sebagai R. oryzae, dan didukung oleh karakter morfologi dan fisiologi. ...... Universitas Indonesia Culture Collection (UICC) has collection of Rhizopus oryzae Went and Prinsen Geerligs strains, which were isolated from tempeh. These strains were identified based on phenotypic characters (morphology and physiology). The aim of this study was to re-identify five strains of R. oryzae (UICC 10, UICC 85, UICC 119, UICC 120, and UICC 135), based on internal transcribed spacers (ITS) region of ribosomal DNA sequence data to obtain accurate identification at species level. Amplification and sequencing of ITS region of rDNA were performed using primer set, forward primer ITS 5 and reverse primer ITS 4. The sequence data was sent to GenBank DNA database for homology search using BLAST. Sequence alignment was carried out using Clustal X. Construction of phylogenetic tree was performed using Neighbor Joining method, Kimura's two parameter model and bootstrap values of 1000 iterations. Characterization of the five strains based on morphology and growth ability at temperature variations were carried out to support the description of each strain. Amplification result showed that the strains have fragment length of ITS region of rDNA about 600--700 bp. Results of BLAST homology search of the strains showed a very high similarity (99.8%) to the type strain R. oryzae CBS 112.07T. Phylogenetic tree showed that the five strains were clustered together in a monophyletic group with the type strain R. oryzae CBS 112.07T and other R. oryzae strains from GenBank DNA database, and supported by high bootstrap value (84%). Re-identification of five strains of R. oryzae UICC based on molecular method showed that they were identified as R. oryzae, similar to previous identification, and supported by morphological and physiological characterization.

Abstrak :
Universitas Indonesia Culture Collection UICC memiliki koleksi Rhizopus spp. yang diisolasi dari tempe dan telah diidentifikasi berdasarkan karakter morfologi dan fisiologi. Tujuan penelitian adalah re-identifikasi lima strain R. oligosporus koleksi UICC UICC 27, UICC 40, UICC 51, UICC 67, dan UICC 116 berdasarkan data sequence daerah internal transcribed spacers rDNA dan analisis filogenetik. Amplifikasi dan sequencing daerah ITS rDNA menggunakan pasangan primer forward ITS5 dan primer reverse ITS4. Data sequence daerah ITS rDNA Rhizopus koleksi UICC dikirim ke database GenBank untuk pencarian homologi BLAST terhadap spesies terdekat dengan pembatasan pencarian hanya pada type material. Pembuatan pohon filogenetik menggunakan metode neighbor joining dengan model evolusi dua parameter Kimura, serta bootstrapping dilakukan sebanyak 1000 kali pengulangan. Karakterisasi morfologi makroskopik dan mikroskopik dan fisiologi pertumbuhan pada variasi suhu dilakukan untuk menunjang hasil identifikasi molekuler. Hasil elektroforesis gel produk PCR daerah ITS rDNA menunjukkan kelima strain memiliki ukuran fragmen ITS rDNA pada kisaran 500--700 pb, dan hasil sequencing lengkap daerah ITS rDNA kelima strain menunjukkan panjang berkisar antara 655--658 pb. Hasil BLAST menunjukkan strain UICC 27, UICC 40, UICC 51, dan UICC 67 memiliki homologi 99,2 ; sedangkan strain UICC 116 memiliki homologi 99,8 terhadap type strain R. oryzae CBS 112.07T. Berdasarkan konsep spesies filogenetik, strain UICC 27, UICC 40, UICC 51, dan UICC 67 dapat diidentifikasi sebagai R. delemar karena pada pohon filogenetik berada dalam satu kelompok yang monofiletik dengan type strain R. delemar CBS 120.12T, dengan dukungan nilai bootstrap 90 ; sedangkan strain UICC 116 diidentifikasi sebagai R. oryzae karena pada pohon filogenetik berada dalam satu kelompok yang monofiletik dengan type strain R. oryzae CBS 112.07T , dengan dukungan nilai bootstrap 82 . Hasil penelitian menunjukkan bahwa kedua spesies tidak dapat dibedakan secara morfologi, kelima strain menunjukkan karakter morfologi yang sesuai dengan R. oryzae. Namun demikian, terdapat perbedaan karakter fisiologi antara R. oryzae dan R. delemar, yaitu pertumbuhan pada variasi suhu. ...... Universitas Indonesia Culture Collection UICC has strains collection of Rhizopus spp. which were isolated from tempeh and identified based on phenotypic characters morphology and physiology . The aim of this study was to re identify five strains of R. oligosporus UICC 10, UICC 85, UICC 119, UICC 120, and the UICC 135 , based on internal transcribed spacers region of ribosomal DNA sequence data and phylogenetic analysis. Amplification and sequencing of ITS region of rDNA were performed using primer set forward primer ITS 5 and reverse primer ITS 4. The sequence data was sent to GenBank for homology search using basic local alignment search tool BLAST program, restricted only to type materials. Construction of phylogenetic tree was performed using Neighbor Joining method with Kimura's two parameter evolution model and bootstrapping with 1,000 replicates. Characterization of morphological and physiological growth at variation of temperature data was carried out to support molecular identification results. Gel electrophoresis showed that five strains have fragment length of ITS region of rDNA, about 500 700 bp. Full sequence data of ITS rDNA of five strains showed the length of their ITS rDNA was ranged 655 658 bp. BLAST homology search results showed that UICC 27, UICC 40, UICC 51, and UICC 67 strain have similarity 99,2 to the type strain of R. oryzae CBS 112.07T, whilst UICC 116 strain showed a higher similarity 99,8 to the type strain of R. oryzae CBS 112.07T. Based on phylogenetic species concept, four strains UICC 27, UICC 40, UICC 51, and UICC 67 were identified as R. delemar. They were clustered with the type strain of R. delemar CBS 120.12T in a monophyletic group, and supported by 90 bootstrap value. Another one strain UICC 116 was identified as R. oryzae. It was clustered with the type strain of R. oryzae CBS 112.07T in a monophyletic group and supported by 82 bootstrap value. Morphological characterization results indicated that the two species are indistinguishable and they showed morphological characters that correspond to R. oryzae. However, the physiological characters i.e. growth at temperature variations differ between of R. delemar and R. oryzae.

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.