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Pezizomycetes, Pezizaceae): a New Genus and Species Near the Pachyphlodes–Amylascus Lineage

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Pezizomycetes, Pezizaceae): a New Genus and Species Near the Pachyphlodes–Amylascus Lineage See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/297661410 Luteoamylascus aculeatus (Pezizomycetes, Pezizaceae): a new genus and species near the Pachyphlodes–Amylascus lineage Article in Mycological Progress · April 2016 DOI: 10.1007/s11557-016-1177-9 CITATIONS READS 3 481 4 authors: Julio Cabero Pablo Alvarado ALVALAB 8 PUBLICATIONS 86 CITATIONS 127 PUBLICATIONS 2,219 CITATIONS SEE PROFILE SEE PROFILE Rosanne Healy Gabriel Moreno University of Florida University of Alcalá 158 PUBLICATIONS 1,160 CITATIONS 331 PUBLICATIONS 3,057 CITATIONS SEE PROFILE SEE PROFILE Some of the authors of this publication are also working on these related projects: Fungal pathogens in "Weeds and Wilds" View project A comparison of ectomycorrhizal fungi on red oak roots in an arboretum and a forest View project All content following this page was uploaded by Pablo Alvarado on 15 March 2016. The user has requested enhancement of the downloaded file. Mycol Progress (2016) 15:33 DOI 10.1007/s11557-016-1177-9 ORIGINAL ARTICLE Luteoamylascus aculeatus (Pezizomycetes, Pezizaceae): a new genus and species near the Pachyphlodes–Amylascus lineage Julio Cabero1 & Pablo Alvarado2 & Rosanne Healy3 & Gabriel Moreno4 Received: 20 November 2015 /Revised: 17 February 2016 /Accepted: 19 February 2016 # German Mycological Society and Springer-Verlag Berlin Heidelberg 2016 Abstract The new taxon Luteoamylascus aculeatus de- several distinct genetic lineages with independent and some- scribed in this article is proposed to accommodate two collec- times convergent evolutive trends (Hansen et al. 2005;Læssøe tions of a hypogeous ascomycete from central Spain, charac- and Hansen 2007). The ascomata of some Pezizaceae lineages terized by a tomentose yellowish peridium, labyrinth-like have evolved sequestrate forms to varying degrees. Some of gleba filled with whitish hyphae, and intensely reacting amy- them have a folded hymenium within the ascoma, an interme- loid asci. ITS, 28S, and RPB2 data suggest that this new taxon diate status referred to as a ptychothecium, while others de- is an independent lineage proposed here as the new genus velop truffle-like ascomata with a solid gleba and no orga- Luteoamylascus. Until now, this lineage was only known from nized hymenium, referred to as a stereothecium (Weber et al. ectomycorrhizal root tips and mitotic spore mats. In phyloge- 1997;Morenoetal.2014). netic analyses, the Luteoamylascus lineage is placed close to Recently, a lineage of ectomycorrhizal Pezizaceae putative- the genera Amylascus, Pachyphlodes,andScabropezia. ly close to the Amylascus–Pachyphlodes lineage was inferred Morphological data suggest an affinity with Amylascus. through phylogenetic analysis of rDNA sequences from root tips and mitotic spore mats, but there were no ascomata to Keywords Hypogeous . Australia . Mediterranean . Truffle . inform on their fruiting form as apothecial or truffle-like Ectomycorrhizal . Spore mat (Healy et al. 2013). Recent collections of truffle-like ascomata from Spain provide the first glimpse into the type of fruiting bodies produced within this lineage, which we are calling the Introduction Luteoamylascus (gen nov.) lineage. The Pachyphlodes–Amylascus lineage as defined by The Pezizaceae includes both epigeous and hypogeous gen- Hansen et al. (2005) and Læssøe and Hansen (2007)isone era. The polyphyletic genus Peziza sensu lato applies to of the major lineages in the Pezizaceae. It is known to include epigeous apothecia (Scabropezia Dissing & Pfister), as well as hypogeous ptychothecia (Amylascus) and stereothecia Section Editor: Franz Oberwinkler (Pachyphlodes Zobel = Pachyphloeus Tul. & C. Tul.). Healy et al. (2013) produced abundant molecular data in the * Julio Cabero Pachyphlodes–Amylascus lineage from ascomata, mitotic [email protected] spore mats, and ectomycorrhizal root tips, suggesting that Amylascus represents a monophyletic sister lineage to Pachyphlodes. Unfortunately, the type species of Amylascus, 1 C/ El Sol 6, 49800 Toro, Spain A. herbertianus (Cribb) Trappe, has not yet been sequenced to 2 ALVALAB, La Rochela 47, 39012 Santander, Spain verify their actual relationships. Amylascus herbertianus was 3 Department of Plant Pathology, University of Florida, first proposed as Hydnobolites herbertianus Cribb, to accom- Gainesville, FL 32611, USA modate a rare hypogeous fungus from Australia (Cribb 1957) 4 Departamento de Ciencias de la Vida (Botánica), Facultad de with a verrucose peridium formed by a layer of large cells (up Biología, Universidad de Alcalá, 28805 Alcalá de Henares, Spain to 100 μm), covered with a loose tomentum and bearing a 33 Page 2 of 7 Mycol Progress (2016) 15:33 basal tuft of hyphae. Trappe (1971) proposed the new genus was proposed by Dissing and Pfister (1981) to combine Amylascus for this species, because of its thick-walled asci Peziza scabrosa Cooke as S. scabrosa (Cooke) Dissing & that have a diffuse bluing reaction to iodine over the entire Pfister, an epigeous apothecial species provided with a verru- ascus wall (a feature common at the tips of Pezizaceae asci but cose outer surface. A second species, S. flavovirens (Fuckel) a rarity in truffles), spiny non-reticulated spore ornamentation, Dissing & Pfister, was combined from Plicaria flavovirens and verrucose reddish peridium formed by inflated cells. Fuckel, and differs from the type because of its larger spores Hydnobolites asci are not amyloid unless pretreated in KOH (>14.0 μm) ornamented with minute warts. Scabropezia (Kimbrough et al. 1991), and the amyloid reaction is weak. In echinophora (Donadini) Donadini was combined from addition, the Hydnobolites spores are alveolate–reticulate Peziza echinophora Donadini and is considered a synonym rather than spiny, and the peridium is smooth rather than wart- of S. scabrosa by some authors (Dougoud 2012)becauseof ed. Trappe also observed hypha-filled canals, lined with a the very subtle morphological differences between them. The reddish peridium-like layer, that opens at the surface of the spores of Scabropezia are highly similar to Pachyphlodes,and ascoma, and globose spores ornamented with straight to not Amylascus. The ornamentation is of uniform height and curved spines 3 μm high. The peridium is finely tomentose spacing (Dissing and Pfister 1981). in some areas, the glebal trama is solid when wet (Beaton and The aim of the present work was to use molecular phylo- Weste 1982) to chambered when dry (Trappe 1975), and the genetic analyses to place two rare fungal collections asci are inordinate. (Luteoamylascus gen nov.) made in central Spain, and to com- The only other species described, A. tasmanicus (Rodway) pare this new taxon morphologically and molecularly with its Trappe, was originally proposed by Rodway (1926)as putative closest relatives for which ascomata are available: Terfezia tasmanica Rodway, and combined by Trappe Amylascus, Pachyphlodes,andScabropezia. (1975). This species has a minutely verrucose chestnut brown tomentose peridium, veined gleba, and labyrinth-like cham- bers filled with hyphae. Spores are similar in size to those of Materials and methods A. herbertianus, but their ornamentation is half the length, 1(– 2) × 0.5–1.5 μm, and made up of rods and blunt cones that are Herbarium material and morphological observations covered by a perispore. Amylascus tasmanicus has an outer peridium layer of large cells (30–90 μm) similar to Samples of hypogeous fungi were located with the aid of a A. herbertianus. The small differences between these species trained Spanish Water Dog. Micrographs were taken with an were previously noted by Trappe (1975) and Beaton and Olympus BHS 100 W microscope (APO optics, multiplica- Weste (1982), who considered that additional collections were tion coefficient NFK 1.67) and an Olympus E-M1 image cap- needed to define them more satisfactorily. ture device. Mounts were prepared in Lugol’s, Melzer’s, or The excipulum of Amylascus resembles that of 5 % KOH. Spore measurements were taken in Mycometre Scabropezia and of those species of Pachyphlodes that have 2.01 software (Georges Fannechère). The holotype collection a brownish-reddish verrucose peridium. These include is deposited in the herbarium of the Universidad de Alcalá P. melanoxanthus (Tul. & C. Tul. ex Berk.) Doweld, which (AH). The isotype is kept at the personal herbarium of the first also resembles Amylascus in having diffusely amyloid asci. author (CJ 4102831NR 03). Two collections were kindly However, the amyloid reaction of Pachyphlodes asci is weak loaned by the National Herbarium of Victoria (MEL): (Lange 1956)comparedtoAmylascus. Amylascus cf. herbertianus (MEL2364119) and All species in Pachyphloeus were transferred by Doweld A. tasmanicus (MEL2339255). In addition, an authentic (2013)intoPachyphlodes, a name proposed by Zobel (in A. herbertianus specimen (OSC 34757) was studied and Corda 1854)toseparateP. ligericus Tul. & C. Tul. The name imaged. Pachyphloeus Tul. & C. Tul. was illegitimate for fungi be- cause the name Pachyphloeus Göppert was previously given DNA extraction, amplification, and sequencing for a taxon in the Pteridophyta. The spores of Pachyphlodes differ from Amylascus in the uniformity of height and Total DNAwas extracted from dry specimens and polymerase spacing of the spore spines, and in their tendency to chain reaction (PCR) was performed following the procedures remain rigidly straight (Healy 2013). The spore spines described in Alvarado et al. (2015). PCR amplification
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