Corresponding author: Gonzalo Guevara-Guerrero (
Academic editor: Marc Stadler
Knowledge of sequestrate
Peña-Ramírez R, Ge Z-W, Gaitán-Hernández R, Martínez-González CR, Guevara-Guerrero G (2019) A novel sequestrate species from Mexico:
The collections were discovered with a cultivator, digging around trees up to a depth of 15 cm. All encountered fruiting bodies were photographed fresh and then dried at 50 °C. The chosen material was cut by hand and rehydrated with 5% KOH for morphological studies. Thirty spores were measured. Peridial slices were made and observed under optical microscopy (
Genomic DNA was obtained with CTAB (Martínez-González et al. 2017)) or using Fungal DNA extraction Kit (Bio Teke Corporation, China) from 2–3 mg of dry tissue. DNA quantification was performed with Nanodrop (Thermo, USA). Each sample was diluted to 20 ng/uL for PCR amplification. LR0R and LR5 primers were used to amplify the LSU gene (
Sequencing was carried out in a genetic analyzer (Model 3130XL, Applied BioSystems, USA) at the Biology Institute of the National Autonomous University of Mexico (
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ITS and LSU sequences of 12 samples
Maximum probability phylogram of clades obtained with Bayesian inference. The posterior probabilities for each clade are shown on the branches. The accession numbers in the sequence labels indicate the GenBank accession numbers.
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MEXICO. State of Guanajuato, municipality of Guanajuato, Cuenca de la Esperanza Protected Natural Area, 7 November. 2016Peña-Ramírez 108 (Holotype:
Electronic photomicrograph of basidiospores
"
Basidiome 4.4–17×3.9–13.5×3.2–11.2 mm, globose or subglobose to irregular, sometimes compressed when growing together. Peridial surface white, pale brown, fibrillose or tomentose, often with cotton-like patches of white hyphae encompassing some debris soil, stones, leaves, and roots. Peridium Separable and fragile, exposing portions of the gleba. < 0.5 mm thick, mostly hyaline, outer portion pale brown, with a dark ring next to the gleba. Gleba brown, trama gelatinized, locules irregularly shaped, columella dendroid, translucent gray. Odor fungoid; taste not recorded. Basidiomata hard wen dried.
Peridium three layered, 165–240 µm thick. Epicutis 7.5–22.5 µm thick of hyaline to reddish brown, thin-walled, interwoven to repent or erect hyphae, 2–9 µm wide, forming scattered caespitose groups of erect, branched, setal hyphae up to 170 µm long, with abundant crystalline structures adherent on hyphal walls, clamp connections present. Mesocutis 55–105 µm thick, abundant hyaline, isodiametric, globose to subglobose, angular pseudoparenchyma like cells. 4–35×3–24 µm, also with some irregularly shaped, interwoven hyphae, 3–11 µm wide. walls 1–2 µm µm wide, clamp connection absent. Subcutis 22.5–95 µm thick, of interwoven prostrate hyphae, 3–4 µm broad, with scattered large pseudoparenchyma like cells up to 37.5×30 µm, clamp connection absent.
Trama of hyaline, interwoven hyphae 4 µm wide, embedded in a gelatinized matrix, clamp connections present.
Basidia fusoid to clavate, hyaline, 14–48 × 9–12 µm, mean = 32.5 × 10.3 µm, wall 1 µm thick. Basidiospores ellipsoid to broadly ellipsoid, symmetrical, hyaline to pale brown, slightly reddish in KOH, pale brown in mass, excluding utricle 9–13 × 6–7 µm long, mean = 11 × 6.1 µm, Q range = 1.5–2.17, Q mean = 1.8; with utricle 12–17 × 7–10 µm long, mean = 14.47 × 8.27 µm, Q = 1.5–2.13, mean = 1.76. Ornamentation of irregular crest contained within an inflated utricle, hilar appendage in cross-section appears rectangular, 1–3 × 4–6 µm, mean = 1.97 × 5.2 µm. Apex obtuse. Utricle inflated up to 3 µm from spore wall, mean = 1.43 µm, occasionally the utricle is asymmetrically inflated.
MEXICO, state of Guanajuato. Cuenca de la Esperanza Protected Natural Area. Hypogeous, under
Mexico, state of Guanajuato, Cuenca de la Esperanza Protected Natural Area:
In the Bayesian inference analysis, Various
The hilar appendage is larger in
The collections were discovered in the Cuenca de la Esperanza Protected Natural Area in Guanajuato, Mexico, located north of Michoacán and east of Jalisco. The presence of unidentified species in this region highlights the importance of this protected natural area and as an area to search for additional new fungal taxa.
Peña-Ramírez acknowledges TecNM (Tecnológico Nacional de México) for fellowship PRODEP 1264-2015-2017. Guevara-Guerrero thanks CONACyT and TecNM for research support. Martínez-González acknowledges Laura Márquez y Nelly López, LaNaBio, of Instituto de Biología, Universidad Nacional Autónoma de México and to María Eugenia Muñiz Díaz de León, for giving us access to the Laboratory of Molecular Biology. Zai-Wei was supported by the National Natural Science Foundation of China (Nos. 31670024 and 31872619).