Efficient genetic analysis of fungal samples

We present a method for rapid genetic analysis of small amounts of fungal m aterial. Sterile glass slides, sufficiently small to fit in a standard PCR tube, were placed on agar inside a Petri dish. After a few days, fungal cul tures start to overgrow the glass slides. Glass slides with attached myceli um were harvested, analysed microscopically, and placed into a standard PCR tube. Conserved primers flanking the ITS regions of rDNA repeat were used in a direct PCR with the fungal material. Sequence data were generated to b e included in phylogenetic analyses to investigate the relationships of the studied mycorrhizal fungi from orchids. The mycelium attached to glass sli des was also used for an in situ hybridization experiment using fluorescent labelled oligonucleotides as probes. Fluorescent signal was found througho ut the cytoplasm when a probe specific to a site in the nuclear small subun it rRNA is used.