Detection of Unculturable Fungi in Soil Regions of Aziz Awa in Sulaimani province

Abstract

Unculturable and non-sporulting fungi represent a great challenge instudying biotrophic, endotrophic and mycorrhizal fungal groups. In thisstudy collection of soil sample from region Aziz Awa in sulaimaniprovince and using culture-dependent and culture in-dependenttechniques for cultivation of unculturable fungi for the purpose ofexpanding studies on fungal biodiversity in soil. Sabouraud DextroseAgar with supplement as pyridoxine by replicating master plate fromhigher dilution of soil three tinny colonies (less than 1mm in diameter)had grown on medium after incubation for 5-7 weeks at 280C but had notshown growth when replicated on traditional mycological media (PDA,Czapack Dox Agar and Sabouraud Dextrose Agar) were selected,purified, studied culturally and microscopically and identified bymolecular methods. Four specific primer sets (NSIF/ITS4r, NSIF/ LRIF,EF4F/ITS4r and EF60F/ITS4r) were used to amplify partial sequences offungal r RNA gene included ITS sequences. The partial sequences ofthree clones were aligned through the BLASTN phylogenetic analysis inNCBI were available at GenBank database and revealed higher scoresand identities with Fusarium sp, Penicillium sp and Uncultured soilfungus clone. These results led us to consider the clones as viable butnon culturable (VBNC) soil fungi like the common phenomenon in somebacterial species.