MOLECULAR CHARACTERIZATION OF SOIL BORNE FUNGI ASSOCIATED WITH THE RHIZOSPHERE OF TELFAIRIA OCCIDENTALS (HOOK F.) (UGU)

This study was carried out to identify the fungal species associated with the rhizosphere of Telfairia occidentalis H. using molecular techniques. Fungi were isolated from Telfairia occidentalis rhizosphere using standard Blotter and Potato Dextrose Agar (PDA) methods. Deoxyribonucleic acid (DNA) was extracted from pure cultures of fungal isolates using Quick-DNA Fungal/Bacteria Mini prep kit. DNA was successfully extracted from isolate with concentration of 107 ng/µl and 81.77ng/µl for isolate 1 and 2, respectively. Polymerase chain reaction (PCR) amplification and sequencing of internal transcribed space (ITS) region of the fungi isolates was carried out using fungal universal primer pairs ITS and ITS. Sanger sequencing was performed and deposited in the Genbank the sequences obtained were aligned and compared with sequences on the National Centre of Biotechnology information (NCBI) database. The result of the nucleotide sequence analysis revealed that sample 1 is Aspergillus aculeatus with 590 base pairs 100% similarity while sample 2 is Fusarium oxysporum with 610 base pairs und 100% similarity. The phylogenetic tree was constructed using best BLAST hits. The evolutionary tree was inferred using the Neighbor-Joining method. Molecular characterization through polymerase chain reaction amplification and sequencing of ITS regions was effective in the identification of the fungal species. For adequate classification and identification of subsequent microorganisms associated with different crops, the use of molecular tools is therefore recommended.