Deteksi dan Identifikasi Patogen Karat Jingga pada Tanaman Tebu Menggunakan Metode DNA Barcoding

Detection and Identification of Orange Rust Pathogen on Sugarcane Plants Using DNA Barcoding Method

Abstract

Detection and Identification of Orange Rust Pathogen on Sugarcane Plants Using DNA Barcoding Method

Identifying the pathogenic fungi that cause leaf rust down to the species level is often inaccurate when relying solely on morphological characteristics. This is due to the highly variable and overlapping nature of their morphological traits, which are influenced by environmental factors and interactions with host plants. Therefore, further analysis such as molecular characterization is necessary to ensure a more valid identification. This study aims to identify leaf rust fungi from sugarcane using the DNA barcoding method. Fungal samples were collected from sugarcane plantations in Bondowoso and Blitar, East Java. Morphological identification of leaf rust fungi is difficult because, during observation using scanning electron microscopy, often only a single life stage is detected. Molecular identification was performed by amplifying the internal transcribed spacer (ITS) region and analyzing the homology of the resulting sequences against a database to identify the fungi down to the species level. The ITS region was selected because it is a molecular marker commonly used for identification and phylogenetic analysis in fungi, given its high level of variation between species but relative conservation within the same species. The amplification results yielded a 1350 bp DNA band at both sites. Sequencing results showed that samples from Bondowoso and Blitar were closely related to Puccinia kuehnii sequences from the National Center for Biotechnology Information. The phylogenetic tree revealed that samples from both locations belonged to the same group as P. kuehnii sequences from other countries.