Phylogenetic analysis and abundance of culturable Fungi from Equatorial; Lewis Glacier Mt. Kenya

Abstract

Equatorial and tropical glaciers of Africa are mainly found on mountains summit. Due to climatic change, the receding glaciers exposes cryoconites which are ideal spots for microbial growth. Industrially important fungi are among the psychrophilic microorganism inhabiting cryoconites. In this study, the characterization and determination of the abundant fungal taxa in the tropical glaciers; Lewis glaciers Mount Kenya was done. Randomly selected cryoconite holes were mapped out across the study area by grid method, fifty samples were collected in different reagent bottles. Each sample was then serially diluted and cultured in Sabouraud Dextrose Agar (SDA) mixed with a broad-spectrum antibiotic to inhibit bacterial growth. The culturable pure strains that successfully grew were twenty-three. The pure isolates were first morphologically characterized Genomic DNA was extracted using CTAB method followed by DNA quality and yield assessments using agarose gel electrophoresis and nanodrop. For the molecular identification of the isolated fungi at the species level, the extracted fungal DNA was amplified by PCR using specific internal transcribed spacer primer (ITS1/ITS4). The amplified products were sequenced, and the sequence data were trimmed using BioEdit. The trimmed sequences were then characterized and there was a total of hundred fungi species; most of which had a range of 80-100% similarity with other fungi when subjected to BLASTn. For evolutionary relationship studies, fungi species that had a percentage similarity ranging between 95-100% were downloaded from NCBI and saved in MEGA X for further diversity analysis. Nine filamentous fungal species were identified as Bjerkandera adusta, Coprinellus micaceus, Penicillium chrysogenum, Polyporales sp., Schizophyllum commune, Trametes hirsuta, Trametes polyzona, Trametes versicolor and uncultured fungi clone. The results showed that T. polyzona was the most abundant fungus revealed from all the locations. It was also noticed that our isolates were of two phyla identified as: Basidiomycota and Ascomycota. Most of the fungi belonged to Basidiomycota.