Genomic and Molecular Identification and PCR-Based Validation of the Fungal Luciferase Gene from Armillaria mellea Reveals Bioluminescence Genetic Potential

Abstract

Fungal bioluminescence is a genetically regulated biochemical phenomenon driven by a conserved luciferin-luciferase system. This study aimed to genomically identify and molecularly validate the fungal luciferase (Luz) gene from Armillaria mellea to confirm its inherent bioluminescent genetic potential. The fungal strain was cultured under standardized conditions on Malt Extract Agar and in broth media to obtain sufficient mycelial biomass for genomic DNA extraction. High-quality genomic DNA was isolated and used for sequencing library preparation followed by Illumina-based genome sequencing. Gene-specific primers were designed to amplify a conserved fragment of the Luz gene. PCR amplification yielded a distinct 785 bp amplicon corresponding to the expected size of the luciferase gene fragment. Agarose gel electrophoresis confirmed specific amplification without nonspecific bands or primer-dimer formation. The amplified product was further validated by Sanger sequencing, and BLASTn analysis demonstrated high sequence homology with previously reported fungal luciferase genes associated with the caffeic acid-dependent bioluminescence pathway. Sequencing chromatograms displayed clear nucleotide peaks, confirming high-quality sequence data.