FLUORESCENCE MICROSCOPIC INVESTIGATION OF ASPERGILLUS-AWAMORI GROWINGON SYNTHETIC AND COMPLEX MEDIA AND PRODUCING XYLANASE

Several fluorescence techniques were used to detect cellular component s within the hyphae of Aspergillus awamori. Single-stranded RNA (ss RN A) and double-stranded nucleic acids (ds RNA and DNA) were localized i n mycelial hyphae and pellets of Aspergillus awamori by means of acrid ine orange (AO) staining. Pyronine Y/methyl green (PY/MG) staining all owed the localization of ds RNA and DNA, but no ss RNA in the cells. T he replicating DNA was localized by the detection of the incorporated thymidine analogue (BrdU) via immunofluorescence. The ss RNA marks the local protein production in the cells. It indicates that the main pro tein production occurs in subapical and branching zones of the hyphae. In pellets no protein synthesis was observed in the corpus, but only in the hyphae on the pellet periphery. The replicating DNA observed in subapical and branching zones and nucleus indicates the cell propagat ion. On account of the intensive yellow fluorescence of the wheat bran , it was not possible to identify the cells with: replicating DNA grow ing on a complex medium. By means of the vitality staining by fluoresc ein diacetate (FDA) and propidium iodide (PI) no internal hyphal struc tures were observed. The ethidium bromide (EB) is more suitable for st ructural investigations. The protein staining with fluorescein isothio cyanate (FITC) does not give useful information on the protein product ion, because of low fluorescence intensity, rapid fading and low selec tivity.