MOLECULAR IDENTIFICATION OF ENDOPHYTIC FUNGI ISOLATED FROM Garcinia porrecta AND Garcinia forbesii

Radji, Maksum ( Laboratory of Microbiology and Biotechnology, Department of Pharmacy Faculty of Mathematics and Sciences, University of Indonesia. Depok, 16424, Indonesia)
Nugraheni, Femi Arifah ( Laboratory of Microbiology and Biotechnology, Department of Pharmacy Faculty of Mathematics and Sciences, University of Indonesia. Depok, 16424, Indonesia)
Sumiati, Atiek ( Laboratory of Microbiology and Biotechnology, Department of Pharmacy Faculty of Mathematics and Sciences, University of Indonesia. Depok, 16424, Indonesia)

JFIOnline
Vol 4, No 4 (2009)

Penelitian ini dilakukan untuk mengidentifikasi isolat-isolat kapang endofit yang telah diisolasi dari tanaman Garcinia porrecta dan Garcinia forbesii yang dikoleksi di Laboratorium Mikrobiologi dan Bioteknologi Departemen Farmasi Universitas Indonesia. Lima galur isolat yang telah terbukti menunjukkan bioaktivitasnya sebagai antimikroba dan antioksidan diidentifikasi secara molekuler menggunakan profil sekuen DNA yang berasal dari gen penyandi ribosomal RNA. Identifikasi isolat kapang endofit dilakukan dengan metode Polymerase Chain Reaction (PCR) menggunakan primer NS1 dan NS4. Hasil amplifikasi dengan PCR selanjutnya dianalisis dengan sekuensing DNA yang kemudian dibandingkan dengan data pada GenBank dengan program BLAST (Basic Local Alignment Search Tools). Berdasarkan hasil analisis sekuens DNA, 5 isolat berhasil diidentifikasi. Empat isolat kapang endofit teridentifikasi sampai tingkat spesies yaitu: Penicillium purpurogenum, Zasmidium cellare, Colletothricum gloeosporioides, dan Nectria curta. Sedangkan satu isolat lain teridentifikasi sampai tingkat genus yaitu Pestalotiopsis sp. ABSTRACT The objective of this study was to identify the endophytic fungi isolated from Garcinia porrecta and Garcinia forbesii deposited in Laboratory of Microbiology and Biotechnology, Department of Pharmacy, University of Indonesia by using molecular tools. Five isolates of endophytic strains which were displayed the strongest antibacterial and antioxidant activities were identified based on ribosomal RNA (rRNA) gene sequence analysis. The Polymerase Chain Reaction (PCR) was used to identify these endophytic fungi by using primer NS1 and NS4. Amplicon or PCR product was analyzed by sequencing method to determine the DNA sequence. The result of sequencing DNA then compared with GenBank’s database using Basic Local Alignment Search Tool (BLAST). The results revealed that five isolates of endophytic strains had been successfully identified by this method. Four of them were identified up to the level of species as Penicillium purpurogenum, Zasmidium cellare, Colletothricum gloeosporioides, and Nectria curta, while the other one was identified up to the level of genus namely Pestaloptiosis sp.