Vermiculite as a culture substrate greatly improves the viability of frozen cultures of ectomycorrhizal basidiomycetes.

We assessed a new cryopreservation protocol that uses vermiculite as a culture substrate, called the vermiculite protocol (VP), by assessing the viability, recovery time of hyphae after revival, and colony diameter of cryosensitive ectomycorrhizal basidiomycete strains after storage for 2 weeks or 1 year in a vapour-phase liquid nitrogen tank. Twelve difficult-to-preserve strains of nine species (Amanita citrina, A. pantherina, A. rubescens, A. spissa, Kobayasia nipponica, Lactarius akahatsu, L. hatsudake, Sarcodon aspratus, and Tricholoma flavovirens) that did not achieve good revival after cryopreservation with our previous Homolka's perlite protocol and modified perlite protocol (MPP) experiments were used to assess the new methodology. Vermiculite and liquid medium were put into a cryotube and inoculated with an agar plug containing mycelia. The cryotube was cultured for various incubation times. After adequate mycelial growth, a mixture of cryoprotectants (5% dimethyl sulfoxide and 10% trehalose [5D10T] or 5% glycerol and 10% trehalose [5G10T]) was placed into the cryotube. The cryotube was frozen in a freezing container in a -80 °C freezer and then stored in vapour-phase liquid nitrogen. In the recovery test, 10 of 12 strains showed 100% revival after 2 weeks of storage in the 5G10T cryoprotectant, and all 12 strains showed 100% revival after 2 weeks of storage in the 5D10T cryoprotectant. Furthermore, all strains were viable after 1 year of storage in a vapour-phase liquid nitrogen tank. Thus, the VP is applicable to a wide range of ectomycorrhizal basidiomycete cultures, including highly cryosensitive strains.

[Amanita pantherina poisoning or brain stroke?]. / Zatrucie muchomorem plamistym czy udar mózgu?

A case of a 72-year-old female who ingested panther cap (Amanita pantherina) was presented. During the ingestion of mushrooms she developed short-time diarrhea and severe transient neurological disorders; short-term hallucination followed by deep coma, skeletal muscle flaccidity with hyporeflexia. Also significant facial asymmetry was noted. A head CT showed a small ischaemic focus in capsula interna. After a neurological consult it was concluded that this small malformation was not the cause of severe patient's condition. The sporological test revealed numerous spores of panther cap in patient's vomit. After the next 6 hours complete resolution of neurological symptoms was observed. Finally it was assumed that patient's symptoms and signs were most probably a result of panther cap poisoning complicated with transient ischaemia of the brain.

Amanita phalloides en bosques de pinus radiata de la IX región de Chile: taxonomia, toxinas, métodos de detección, intoxicación faloidiana / Amanita phalloides in pinus radiata forests from IX region of Chile: taxonomy, toxins, detection methods, phalloidian intoxication

Se cita por primera vez en Chile Amanita phalloides (Vaill. ex Fr) Secr., junto a su descripción macro y microscópica, y algunos aspectos de su ecología. Se aporta información sobre sus toxinas, métodos de detección y cuadro clínico de la intoxicación faloidiana