Isolasi Gen Pi1 Penyandi Kelas Protein NBS-LRR pada Padi Siporang (Oryza sativa var. Siporang) dari Sumatera Utara sebagai Gen Ketahanan Patogen

Abstract

Siporang is a superior local rice variety from North Sumatra known to be resistant to blast disease. Previous studies have successfully identified several blast resistance (R) genes in Siporang. However, no research has yet reported information on the Pi1 gene from this variety. This study aimed to isolate and analyse the sequence of the Pi1 gene from Siporang. The methodological stages in this research included DNA isolation using CTAB method, qualitative analysis by 1% agarose gel electrophoresis at 70V for 30 minutes, quantitative analysis with a nanophotometer, PCR amplification with Pi1 gene-specific primers, gel extraction using the GenepHlow Gel/PCR kit from Geneaid, Sanger sequencing, and Pi1 gene sequence analysis using BLASTn and MEGA12. The BLASTn analysis results confirmed that the candidate gene successfully isolated from Siporang rice is the Pi1 gene, as it showed a 99.40% identity percentage with accession numbers JF312001.1, JF312002.1, and JF312003.1. Phylogenetic analysis using the maximum likelihood method generated a dendrogram with an evolutionary distance of 0.07 relative to five other Pi1 gene collections from GenBank. The sequence information obtained is expected to be beneficial for further research in the development of blast-resistant crop lines with high yield potential.