ABSTRAK
Universitas Indonesia Culture collection (UICC) memiliki koleksi strain-strainRhizopus arrhizus Fischer yang diisolaasi dari tempe dan telah diidentifikasiberdasarkan karakter morfologi dan fisiologi. Penelitian bertujuan memperolehidentitas yang akurat pada tingkat spesies dari lima strain R. arrhizus (UICC 26,UICC 36, UICC 39, UICC 55, dan UICC 121) berdasarkan data sequence daerahinternal transcribed spacers ribosomal DNA dan analisis filogenetik. Amplifikasidan sequencing daerah ITS rDNA dilakukan menggunakan primer forward ITS5dan primer reverse ITS4. Pencarian homologi sequence dilakukan denganprogram BLAST. Sequence alignment dilakukan menggunakan program ClustalX. Konstruksi pohon filogenetik dilakukan menggunakan metode neighborjoining dengan model dua parameter Kimura dan nilai bootstrap 1000pengulangan. Karakterisasi morfologi dan fisiologi (pengujian pertumbuhan padavariasi suhu) dilakukan untuk melengkapi deskripsi strain. Panjang fragmendaerah ITS rDNA kelima strain R. arrhizus sekitar 600--700 pb. Hasil BLASTmenunjukkan kelima strain UICC memiliki homologi dengan type strain R.oryzae CBS 112.07T pada kisaran 98,9--99,8%. Pohon filogenetik menunjukkanstrain UICC 36 dan strain UICC 55 berada dalam satu grup yang monofiletikdengan type strain R. oryzae CBS 112.07T dan neo type R. arrhizus NRRL1469NT. Saat ini R. arrhizus merupakan sinonim dari R. oryzae, sehingga keduastrain tersebut diidentifikasi sebagai R. oryzae. Strain UICC 26, UICC 39, danUICC 121 berada dalam satu grup yang monofiletik dengan type strain R.delemar CBS 120.12T, sehingga ketiga strain diidentifikasi sebagai R. delemar.Rhizopus oryzae dan R. delemar merupakan dua spesies yang berkerabat sangatdekat, sehingga secara morfologi tidak dapat dibedakan. Re-identifikasi limastrain R. arrhizus UICC secara molekuler menghasilkan identitas spesies yangberbeda menjadi R. oryzae dan R. delemar, namun karakter morfologi danfisiologi lima strain tersebut menunjukkan karakter sebagai R. oryzae.
ABSTRAK
Universitas Indonesia Culture Collection (UICC) has collection of Rhizopusarrhizus Fischer strains, which were isolated from tempeh and were identifiedbased on morphological and physiological characters. The aim of this study wasto obtain accurate identification at species level of five strains of R. arrhizus(UICC 26, UICC 36, UICC 39, UICC 55, and UICC 121) based on internaltranscribed spacers (ITS) region of ribosomal DNA (rDNA) sequence data andphylogenetic analysis. Amplification and sequencing of ITS region of rDNA wereperformed using forward primer ITS5 and reverse primer ITS4. Sequencehomology search was performed using BLAST. Sequence alignment was carriedout using Clustal X. Construction of phylogenetic tree was performed usingneighbor joining method, Kimura?s two parameter model and bootstrap values of1000 iterations. Characterization of morphological features and growth atvariation of temperature were carried out to support the description of the strains.The fragment length of ITS region rDNA from five strains of R. arrhizus wasabout 600--700 bp. Results of BLAST homology search of the strains showed98.9--99.8% similarities to the type strain R. oryzae CBS 112.07T. Phylogenetictree showed that UICC 36 and UICC 55 were clustered together in amonophyletic group with the type strain R. oryzae CBS 112.07T and neo typestrain R. arrhizus NRRL 1469NT. Currently, R. arrhizus is a synonym of R.oryzae, therefore both strains were identified as R. oryzae. Strains UICC 26,UICC 39, and UICC 121 were clustered together in a monophyletic group withthe type strain R. delemar CBS 120.12T, therefore they were identified as R.delemar. Rhizopus oryzae and R. delemar are very closely related species andmorphologically similar. Re-identification of five strains R. arrhizus UICC basedon molecular has difference of species identity as R. oryzae and R. delemar, butthe five strains show similar morphological and physiological characters as R.oryzae. |
S64952-Yura Vebliza.pdf :: Unduh