Bulk isolation of basidiospores from wild mushrooms by electrostatic attraction with low risk of microbial contaminations

Kiran Lakkireddy, Ursula Kües

The basidiospores of most Agaricomycetes are ballistospores. They are propelled off from their basidia at maturity when Buller's drop develops at high humidity at the hilar spore appendix and fuses with a liquid film formed on the adaxial side of the spore. Spores are catapulted into the free air space between hymenia and fall then out of the mushroom's cap by gravity. Here we show for 66 different species that ballistospores from mushrooms can be attracted against gravity to electrostatic charged plastic surfaces. Charges on basidiospores can influence this effect. We used this feature to selectively collect basidiospores in sterile plastic Petri-dish lids from mushrooms which were positioned upside-down onto wet paper tissues for spore release into the air. Bulks of 10(4) to >10(7) spores were obtained overnight in the plastic lids above the reversed fruiting bodies, between 10(4) and 10(6) spores already after 2-4 h incubation. In plating tests on agar medium, we rarely observed in the harvested spore solutions contaminations by other fungi (mostly none to up to in 10% of samples in different test series) and infrequently by bacteria (in between 0 and 22% of samples of test series) which could mostly be suppressed by bactericides. We thus show that it is possible to obtain clean basidiospore samples from wild mushrooms. The technique of spore collection through electrostatic attraction in plastic lids is applicable to fresh lamellate and poroid fruiting bodies from the wild, to short-lived deliquescent mushrooms, to older and dehydrating fleshy fruiting bodies, even to animal-infested mushrooms and also to dry specimens of long-lasting tough species such as Schizophyllum commune.