Isolation and Amplification DNA on Endophytic Fungi of Local Aromatic Rice Enrekang

Abstract

Local rice is one of the biodiversity that is currently not explored in detail. The exploration of endophytic fungi of local rice in South Sulawesi was carried out to find out the types of endophytic fungi in the local rice plant. Plant samples were taken from local rice plantations in the Tator, Enrekang, and Luwu regencies. DNA isolation and PCR amplification were the initial stages that need to be carried out in molecular identification. DNA fungi extraction was performed by using DNesay DNA extraction KIT. The result of DNA fungi isolation was electrophoresed with 0.8% agarose gel in TAE buffer (Tris Acetate EDTA). DNA isolation was successfully carried out by the formation of DNA bands. DNA amplification of two endophytic fungi using the primary pairs of ITS1 and ITS4, with condition PCR of predenaturation at 95°C for 5 minute, denaturation at 94°C for 1 minute, annealing at 55,4°C for 1 minute, elongation at 72°C for 1 minute, final elongation at 72°C for 10 minute. The denaturation stage until the extension was repeated 35 times and produced DNA fragments with a molecular size of 700 bp and 600 bp.