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

Abstract

Unculturable and non-sporulting fungi represent a great challenge in
studying biotrophic, endotrophic and mycorrhizal fungal groups. In this
study collection of soil sample from region Aziz Awa in sulaimani
province and using culture-dependent and culture in-dependent
techniques for cultivation of unculturable fungi for the purpose of
expanding studies on fungal biodiversity in soil. Sabouraud Dextrose
Agar with supplement as pyridoxine by replicating master plate from
higher 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 not
shown growth when replicated on traditional mycological media (PDA,
Czapack Dox Agar and Sabouraud Dextrose Agar) were selected,
purified, studied culturally and microscopically and identified by
molecular methods. Four specific primer sets (NSIF/ITS4r, NSIF/ LRIF,
EF4F/ITS4r and EF60F/ITS4r) were used to amplify partial sequences of
fungal r RNA gene included ITS sequences. The partial sequences of
three clones were aligned through the BLASTN phylogenetic analysis in
NCBI were available at GenBank database and revealed higher scores
and identities with Fusarium sp, Penicillium sp and Uncultured soil
fungus clone. These results led us to consider the clones as viable but
non culturable (VBNC) soil fungi like the common phenomenon in some
bacterial species.