Ds ri

The morphological analyses were performed on specimens, including types, deposited in the Fungal Collection of the Federal University of Rio Grande do Norte (UFRN Herbarium), the collection of fungi of the Real Jardín Botánico of Madrid (MA-Fungi), the cryptogamy collection (PC) at the Herbarium of the Muséum national d'Histoire naturelle (MNHN—Paris), and the Fungarium of the Royal Botanic Gardens, Kew (K), (Table 1). Macromorphological studies were based on 26 exsiccates using a Nikon H600L stereomicroscope coupled with a Nikon DS-Ri camera for image capture. Colour descriptions followed Kornerup and Wanscher [25 ]. For micromorphological features, such as basidiospores, eucapillitium and exoperidial hyphae, a Nikon Eclipse Ni light microscope (LM) coupled with a Nikon DS-Ri camera was used. Basidiospore measurements were made at 1000× magnification following Sousa et al. [21 ] and include ornamentation. Scanning electron microscopy (SEM) was used to observe the patterns of ornamentation on basidiospores, eucapillitium and endoperidial surfaces.

Cells fixed in 4% paraformaldehyde at 4°C for 10 min were washed in 1× phosphate-buffered saline (PBS), and then permeabilized in PBS with 0.5% Triton X-100 for 5 min at room temperature (RT). Cells were then blocked with 3% goat serum in PBS containing the primary antibody TUJ1 (anti-rabbit, 1:500; Abcam) and incubated overnight at 4°C. Cells were then washed with PBS and incubated with Alexa Fluor^TM goat anti-rabbit 594 sary antibody for 1 h at RT. Individual cell nuclei were confirmed using Prolong Diamond antifade mountant with 4’,6-diamidino-2-phenylindole (DAPI; Life Technologies). Imaging was undertaken on a Nikon TE2000E inverted microscope (Nikon) equipped with a Nikon DS-Ri camera. Images were converted to greyscale in ImageJ, scaled and the cell soma size measured using the polygonal selection tool. Neurite number, neurite length and the number of branch points were determined using the NeuronJ plugin in Fiji. Morphological characterization was undertaken from 30 cells per donor age group.