Identification of Post Harvest Fungi Associated Telfairia occidentalis Hook F. Leaves Using Basic Molecular Techniques

Background and Objective: The leaf of Telfairia occidentalis, a highly nutritious and economically important vegetable in Nigeria, is threatened by fungal diseases that affect its quality and yield. This study was conducted to isolate and identify the fungal species associated with the leaves of Telfaira occidentalis using both conventional microbiological techniques and molecular methods. Materials and Methods: Fungal species were isolated from Telfairia occidentalis obtained from the Botanic Garden of the University of Port Harcourt, Choba Port Harcourt, Rivers State, Nigeria. Potato dextrose agar (PDA) was used as a growth medium for the fungi. Deoxyribonucleic Acid (DNA) was extracted from pure cultures of fungal isolates using Quick-DNA Fungal/Bacteria Mini Prep Kit. Polymerase Chain Reaction (PCR) amplification of the internal transcribed spacer (ITS) region of the fungal isolates was carried out using fungal universal primer pairs ITS4 and ITS5. Sanger sequencing was performed and the sequences obtained were aligned and compared with sequences on the National Centre of Biotechnology Information (NCBI) database. Results: The result of the nucleotide sequence analysis revealed that isolate 1 is Trichorderma reesei with 590 base pairs and 99.66% similarity while isolate 2 is Aspergillus aculeatus with 610 base pairs and 99.62% similarity. A phylogenetic tree was constructed using the best BLAST hits. The evolutionary tree was inferred using the neighbor-joining method. Conclusion: The study identifies Trichoderma reesei and the newly discovered Aspergillus aculeatus on post-harvest Telfairia occidentalis leaves, raising concerns about potential health risks from consuming contaminated leaves due to harmful metabolites.