Molecular Identification of Fungal Isolates from Textile Wastewater by ITS-RFLP Method

Abstract

Eight strains isolated from textile post-finishing wastewaters were subjected to molecular identification by ITS-RFLP (Internal Transcriber Spacer-Restriction Fragment Length Polymorphism) method, using ITS1 (5’-TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’) DNA amplification primers for ITS region of the nuclear small-subunit rRNA gene. Restriction fragments analysis was conducted post PCR amplification, and cleaved with HaeIII, HinfI, HhaI, MseI and SduI restriction enzymes. Differences in the lengths of the obtained amplicons were detected after gel electrophoresis, and restriction analyses of the PCR fragments led to identification of six out of eight isolates as belonging to fungi T. parceramosum/ T. reesei/ T. longi, Polyporus squamosus, Fusarium oxysporum and Trichoderma atroviride groups. The PCR ITS-RFLP molecular identification technique can be used successfully for routine screening of fungal isolates from various environments.