Identifikasi Cendawan Endofit Menggunakan Teknik Polymerase Chain Reaction

Abstract

ABSTRAK Penyakit keriting kuning yang disebabkan oleh infeksi Begomovirus (Geminiviridae) merupakan salah satu penyakit penting pada tanaman cabai di Indonesia. Eksplorasi cendawan endofit dilakukan sebagai langkah awal untuk mencari agens biokontrol yang potensial menekan penyakit keriting kuning cabai. Cendawan endofit diisolasi dari pertanaman cabai di daerah Sleman, Yogyakarta dan isolat selanjutnya diidentifikasi menggunakan teknik polymerase chain reaction (PCR) dan perunutan DNA. Pita DNA berukuran ± 500 pb berhasil diamplifikasi dari 10 isolat cendawan endofit menggunakan pasangan primer ITS1/ITS4, tetapi urutan DNA hanya diperoleh dari 8 isolat. Berdasarkan pada analisis BLASTN diketahui kemiripan masing-masing isolat dengan spesies cendawan pada GenBank, yaitu isolat H1 dengan Pleosporaceae sp. (98%), isolat H5 dengan Cercospora nicotianae (100%), isolat H11 dengan Cercospora piaropi (98%), isolat H16 dengan Guignardia mangiferae (99%), isolat H17 dengan Geomyces pannorum (95%), isolat H18 dengan Diaporthe phaseoloru (99%), isolat K3 dengan Dothideomycete sp. (100%), dan isolat K10 dengan Alternaria longissima (99%). ABSTRACT Yellow leaf curl disease, caused by a member of Begomovirus (Geminiviridae), is one of important diseases of chilli pepper in Indonesia. Exploration of endophytic fungi was initiated in order to find biological control agents for an alternative control strategies of this disease. Isolates of endophytic fungi were collected from chilli pepper growing area in Sleman, Yogyakarta and further identification using molecular technique involving polymerase chain reaction (PCR) and DNA sequencing was performed. DNA fragments of ±500 bp were successfully amplified from 10 fungal isolates by PCR using primer pair ITS1/ITS4, but only 8 DNA sequences was obtained for further genetic analysis. Based on BLASTN analysis the endophytic fungi were identified as having the highest similarity with Pleosporaceae sp. (98%) for H1 isolate, Cercospora nicotianae (100%) for H5 isolate, Cercospora piaropi (98%) for H11 isolate, Guignardia mangiferae (99%) for H16 isolate, Geomyces pannorum (95%) for H17 isolate, Diaporthe phaseoloru (99%) for H18 isolate, Dothideomycete sp. (100%) for K3 isolate, and Alternaria longissima (99%) for K10 isolate.