From the' Dept. of Mycology and Parasitology, Isfahan University of Medical Sciences, Isfahan
Abstract: (3352 Views)
In this study techniques for laboratory identification of dermatophyte fungi
through protoplast hybridization were established. Firstly, auxotrophic mutants of
different species of microsporum and trichophyton were induced and identified.
Secondly, protoplasts from these mutants were isolated by digestion of their
mycelium with Novozyme 234 using CaCI, (O.4M) as an osmotic stabilizer and
glycine + HCI (pH 4.5) as the buffer system. Thirdly, isolated protoplasts from
different species were fused using a solution containing 35% polyethylene glycol,
I M KCI, 0.05 M CaCI, and 0.05 M glycine with pH 6.1. Afterwards, the fusion
products were plated onto minimal and complete media. It was found that
protoplasts from different auxotrophic mutants from the same species hybridized
and complemented each other and grew on both minimal and complete media.
whereas mutants from different species did not have the ability to complement
each other and therefore grew only on complete media. Information obtained in
this study may prove useful for definite identification of suspected species of
dennatophytes other than morphological criteria in laboratories.