Online publication; available at: https://jats-truffles.org/truffology/

Truffology 5 (2): 61–67 (2022)

Original peer-reviewed article （原著論文 ； 査読有）

Two new Elaphomyces species (Elaphomycetaceae, Eurotiales,
Ascomycota) from eastern USA
アメリカ合衆国東部において採集されたツチダンゴ属 （子嚢菌門
ユーロチウム目ツチダンゴキン科） の 2 新種
Michael A. Castellano
US Department of Agriculture, Forest Service, Northern Research Station, 3200 Jeﬀerson Way, Corvallis, OR 97331, USA
E-mail: mcastell33@hotmail.com

Abstract
The hypogeous, sequestrate Elaphomyces has a long taxonomic history in Europe and was formally describedby Nees von Esenbeck in 1820 from Europe. Until recently, names of European taxa were assigned to most
Elaphomyces specimens in North America due to lack of specialists working on this group and the diﬃculty ofusing pre-modern species descriptions. The genus Elaphomyces from North America has received increased
attention for the last decade and this has led to numerous new Elaphomyces species being described from eastern
North America. Here are described two new species of Elaphomyces discovered in eastern USA, E. gouldhoytii thathas similarities to E. septatus and E. maculatus, and E. ramulospinus with similarities to E. muricatus.

要旨
ツチダンゴ属は地中生のシクエストレート型の子実体を形成する子嚢菌類で、 1820 年にヨーロッパから Nees vonEsenbeck によって正式記載され、 ヨーロッパにおいて長い分類学史を有する。 近年まで北米のツチダンゴ属の標本の多くに対しヨーロッパ産種の種名が適用されてきたが、 これは北米に本属の専門家が不在で、 かつ、 古典的な記載方法に基づいた種の扱いが困難であったことに起因している。 北米のツチダンゴ属菌は過去 10 年間ににわかに注目を集め、北米東部から多数の新種が記載された。 本研究では、 E. septatus と E. maculatus に類似する新種 E. gouldhoytii、 ならびに E. muricatus に類似する新種 E. ramulospinus をアメリカ東部から記載する。
Article Info: Submitted: 12 March 2022

Accepted: 8 September 2022

Published: 31 December 2022

Introduction

from North America: E. verrucosus C.W. Dodge (Dodge, 1929),

The hypogeous, sequestrate genus Elaphomyces Nees

E. appalachiensis Linder (Linder, 1939), E. viridiseptum Trappe &(Elaphomycetaceae, Eurotiales, Ascomycota) has received increased

Kimbr. (Trappe & Kimbrough, 1972), E. subviscidus (Zeller) Trappe

attention over the last ten years with numerous new species added& Guzmán (Trappe & Guzmán, 1971), and E. spinoreticulatus Zhang

and a number of previously described species revised with modern& Minter (Zhang & Minter, 1989). A number of other Elaphomyces

descriptions (Castellano et al., 2011, 2012a, 2012b, 2012c, 2016,

species with European names had been variously reported across2018, 2021; Castellano & Stephens, 2017; Molia et al., 2020; Paz etNorth America. Since 2012, 24 new Elaphomyces species haveal., 2012, 2017; Shirakawa & Tanaka, 2020). Elaphomyces ascomata

been presented from North America by Castellano and colleagues

usually have a thick, complex peridium often encased in mycelium,(Castellano et al., 2012b, 2018, 2021; Castellano & Stephens, 2017),

roots, soil and debris, and a powdery gleba.

primarily from eastern USA.

Prior to 2012, there were five Elaphomyces species described© The Japanese Association for Truffle Science (JATS), 2022

The rich diversity of Elaphomyces species in the eastern portion of61Truffology - Volume 5, Issue 2, 2022

Castellano: Two new Elaphomyces species from eastern USA
North America, east of the Mississippi River, is attributed to the highlove for forestry and the natural world.

diversity of Betulaceae, Pinaceae, and particularly Fagaceae species in

Type: USA, New York, Thompkins Co., Danby, Michigan Hollow,

this region. In this paper, we extend our knowledge of Elaphomyces

1 Oct. 1980, J.M. Trappe & R. Korf (holotype OSC 40649; isotypes

species from eastern USA and report two new Elaphomyces species.

CUP, ITCV).
Description: Ascomata subglobose to irregular, up to 15 mm tall,Materials and methods

32 mm broad, completely enveloped in a pale yellow-brown to dark

Descriptions of macromorphological characters are based onbrown mycelium; mycelium not staining. Peridium 2–3 mm thick,fresh material. Colors are described in general terms based on theouter surface 200–250 µm thick, dark brown, minutely verrucose

observations of the authors and collectors. Preserved specimens

to naked eye but actually of tall conical warts (200–250 µm tall,were rehydrated and examined in 3% KOH, Melzer's reagent, or

150–275 µm broad) embedded in the mycelium, subcutis 2–2.8 mm

cotton blue. Microscopic descriptions were based on 3% KOH

thick, uniform, white, leathery, overlain by a thin covering of brown-

mounts unless specified. Spore dimensions (mean) are from 20

black to black hyphae to give the appearance of a black peridium andascospores measured from the holotype collection, dimensions

obscuring the details of the warts. Gleba spore mass powdery, dark

include ornamentation. For scanning electron microscopy (SEM),

brown, with white web-like hyphae. Odor fungoid. Taste not recorded.

dried ascospores were mounted on aluminum pegs with double-sided

Peridium two-layered: epicutis of septate, disorganized, compact, red-tape, coated with gold, and then observed with an AmRay 3300 FE

brown to yellow-brown hyphae, 3–4 µm broad, walls ± 1 µm thick,ﬁeld emission scanning electron microscope (AmRay, Bedford, MA,

these hyphae grade into the subcutis by the lose of pigment; subcutis

USA). Specimens are deposited in the following herbaria (Indexof hyaline, septate, compact, elongate, interwoven, cells, up to 7 µm

Herbariorum 2022 [http://sweetgum.nybg.org/science/ih/], accessedbroad, walls 1–2 µm thick; outer hyphal layer is periclinal and of

on 25 Jan. 2022): Cornell University (CUP), University of Florida

parallel, short-segmented, septate, more or less uniform pale yellow-(FLAS), Instituto Technológico de Ciudad Victoria (ITCV), Oregonbrown hyphae, 2–4 µm broad. Gleba of spores and hyaline, septate,State University (OSC), and University of Wyoming (RMS).

sinuous to curly smooth hyphae, 2–3 µm broad, walls <0.5 µm thick.
Asci globose with a tapered attachment point, 35–45 µm broad,

Results

hyaline, 6-spored, walls ± 3 µm thick when young, thinning as spores

During the course of examining thousands of Elaphomyces

mature, arising from knots of short, irregularly curved or contorted

collections during the last 15 years, I have come across numerous

clustered hyphae, up to 4 µm broad. Spores ornamented, globose, 33–

undescribed Elaphomyces species based on significant quantitative

37 µm (mean = 34.9 µm), immature spores abundant and 25–26 µm

morphological differences. In addition, I have attempted to receive on

broad, also numerous dark pigmented small spores 20–25 µm broad;loan and study as many different previously described Elaphomyces

walls ± 1 µm thick, in KOH singly and in mass hyaline at ﬁrst thenspecies as possible. This background work has enabled me todark red-brown when mature, ornamentation a coarse labyrinth ofcompile a detailed record of macro- and micro-characters of nearlyanastomosed rods with holes at the tops of the ridges (only apparent

all described Elaphomyces species from either type, authentic or typewith SEM), 2–3 µm tall.

locality material. The two new Elaphomyces species presented below

Distribution, habit, habitat and season: Known fromare a recent product of this work.

Massachusetts, New York, North Carolina, and West Virginia;
hypogeous; under Picea rubra, Pinus strobus, and Tsuga canadensis;

Taxonomy

August to November.

Elaphomyces gouldhoytii Castellano, sp. nov.

Other specimens examined: USA, Massachusetts, Middlesex Co.,

[MycoBank ID: MB 843351]

3 miles east of Concord, 16 Nov. 1984, J. Trappe (OSC 149097).
Fig. 1.

New York, Essex Co., Newcomb, College of Environmental Science

Diagnosis: Elaphomyces gouldhoytii is distinguished from all

Forest, Huntington Wildlife Forest, Sept. 1989, K.F. LoBuglio 102

other species in North America by its black carbonaceous peridium(OSC 149095, OSC 149096). North Carolina, Mitchells Co., Pisgahcomposed of tall warts and the unusually large spores.

National Forest, Craver’s Gap, 30 Aug. 1995, S. Loeb & F. Tainter

Etymology: Named in honor of the late Professor Gould Jennings(OSC 150046); Transylvania Co., Pisgah National Forest, Devil’s

Hoyt Jr. (1922–2012) of Paul Smith’s College, New York, forester

Courthouse, 15 Aug. 1996, S. Loeb & F. Tainter 130 (OSC 150045).

and outdoorsman extraordinaire that single-handedly instilled in me aWest Virginia, Tucker Co., Blackwater State Park, 12 Oct. 2007,C.

62© The Japanese Association for Truffle Science (JATS), 2022

Castellano: Two new Elaphomyces species from eastern USA

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Fig. 1. Elaphomyces gouldhoytii. A: Ascoma showing peridium in section. B: Ascoma surface showing conical warts embedded in dark brown
mycelium. C: Cross-sectional view of the peridium showing the patches of mycelium filling the spaces between warts. D: Immature ascospores with in an
ascus. E: Ascospores in cross-sectional view showing the ornamentation of rods and ridges. F: Ascospores in surface view showing the density of rods andridges. G: SEM micrograph of ascospores showing the rods and ridges. H: SEM micrograph of an ascospore showing the fused rods that compose the ridges. A–
H: OSC 40649 (holotype). Scale bars: A = 2.5 mm, B = 3 mm, C = 90 µm, D = 10 µm, E, F = 18 µm, G = 6 µm, H = 9 µm.

© The Japanese Association for Truffle Science (JATS), 2022

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Truffology - Volume 5, Issue 2, 2022

Castellano: Two new Elaphomyces species from eastern USA
Stihler (OSC 149093).

Diagnosis: Elaphomyces ramulospinus is distinguished from all other

Elaphomyces septatus collection studied: FRANCE, Larrau, St.

species in North America by its tall orange-brown peridial warts andJoseph Forest, 18 May 1985, M. Gérnard & F. Lescourrent (OSC

relatively small spores (20–25 µm, mean = 25.0 µm) that are clearly

149128).

smaller than other Elaphomyces in subsection Muricati that occur

Elaphomyces maculatus collection studied: FH in the Patouillard

in North America (E. americanus Castellano, E. holstii Castellano

herbarium (ex. herbarium O. Mattirolo) marked “Portion autotici C.

& R.A. Healy, E. verrucosus C.W. Dodge, and E. wazhazhensis

Vittadini”.

Crabtree & Castellano). The spore ornamentation of tall, forked spines

Note: Elaphomyces gouldhoytii is placed in Elaphomyces section

anastomosed to form a ﬁne punctuation to the spore surface is unique

Ceratogaster, subsection Sclerodermei because of the lack of yellow,

in this subsection and clearly differentiate it from other Elaphomyces

greenish or purplish mycelial patches on the peridial surface (Paz etspecies in Europe.

al., 2017).

Etymology: “ramulospinus” ramul – forked, spinus – spines, in

The black carbonaceous peridium composed of tall warts andreference to the forked spines of the spores.

the unusually large spores of E. gouldhoytii clearly separate it from

Type: USA, Florida, Hillsborough Co., Tampa, University of

all other North American Elaphomyces species. The European

Southern Florida campus, 31 Aug. 1977, J.M. Trappe (holotype OSC

E. maculatus Vittad. (spores 37–41 µm, mean = 39.1 µm) and E.

38881).

septatus Vittad. (spores 30–35 µm, mean = 33.0 µm) have similarly-

Description: Ascomata irregularly subglobose, up to 9 mm tall × 30

sized large spores but both of the European species have a smooth,mm broad. Peridium ± 2 mm thick, outer surface of orange-brown,

black carbonaceous peridium versus the black tall warts of E.

pyramidal (4–6 sided) warts, up to 400 µm tall, up to 600 µm wide atgouldhoytii.

base, darker near the tips than at base, contiguous with each other at

Castellano studied E. septatus collections from France andbase, warts not uniform over entire surface, warts are smaller on sides

Hungary, no authentic or type material of this species could be

and below, in cross-section the warts are mottled yellow and yellow-

located. Castellano data (on file at Forestry Sciences Laboratory,

brown, more evenly yellow near base, grading from pale white toCorvallis, Oregon, USA) from OSC 149128 is as follows: Peridial

pale gray where inner marbled area begins; subcutis 1.4–1.6 mm

surface black, appearing smooth to the naked eye, covered by dark

thick, outer portion of subcutis irregularly marbled, matrix somewhat

brown hyphae, spores globose, 30–35 µm, mean = 33.0 µm including

globose-shaped, 330–400 µm broad, paler than inter-venation, veinsornamentation that appears finely granulate in patches under light

30–60 µm broad, slightly but distinctly darker than matrix, inner

microscopy, in section appearing as short spines imbedded in an

portion of subcutis distinctly darker overall but of similar structure,

uneven, lumpy matrix conveying a dark outline to the spore, SEM

darker inner portion 200–225 µm in thickness. Gleba spore mass

reveals the spore ornamentation as spines or rods overlain withpowdery, dark brown to nearly black, with numerous off-white toamorphous, rounded, irregular plates, plate surface roughened.

pale brown mycelial strands arising from an inner peridial wall, andCastellano studied an E. maculatus collection from Italy (FH in thetraversing the powdery spore mass. Odor not recorded. Taste not

Patouillard herbarium (ex. herbarium O. Mattirolo) marked “Portion

recorded. Peridium two-layered: epicutis, 400–600 µm thick, of

autotici C. Vittadini”. Castellano data (on file at Forestry Sciences

golden brown-yellow compact, septate, disorganized, interwoven

Laboratory, Corvallis, Oregon, USA) on the FH collection in part ishyphae, 4–5 µm broad, walls ± 2 µm thick, distinctly and abruptly

as follows: Peridial surface black, nearly smooth, with tiny, widelydifferent from interwoven hyphae, interwoven hyphae stacked

spaced, low papillae with ﬂoccose, green mycelium, spores globose,

perpendicular, hyaline, straight, septate, 3–4 µm broad; subcutis(35–)37–41(–42) µm, mean = 39.1 µm including ornamentation that

1.4–1.6 mm thick, of hyaline (somewhat brown near gleba), septate,appears in surface view as a ﬁnely wrinkled bumpy surface, in section

compact, interwoven hyphae, 3–5 µm broad, walls ± 2 µm thick.

appearing bumpy under light microscopy, SEM reveals the spore

Gleba of spores and hyaline, septate, smooth, curly and loosely

ornamentation as coarse spines, up to 2 µm tall (see Fig. 2g, h ininterwoven hyphae, 1–3 µm broad, walls ± 1 µm thick. Asci hyaline,

Castellano et al., 2021).

walls 2 µm thick, variably sized and shaped depending on spore
number within, 1–spored asci globose, 27–33 µm broad, 2–spored

Elaphomyces ramulospinus Castellano, sp. nov.

asci ovate, 28–31 µm broad, 3–spored asci irregular, 29–35 µm[MycoBank ID: MB 843352]

broad, 4–spored asci subglobose, 32–37 µm broad, all arising from
Fig. 2.

64

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Castellano: Two new Elaphomyces species from eastern USA

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Fig. 2. Elaphomyces ramulospinus. A: Ascomata showing peridial surface of warts and peridium in section showing the powdery spore mass of the gleba.
B: Ascoma surface showing the tall pyramidal warts. C: Cross-sectional view of the peridium showing the marbled tissue of the inner peridium. D: Asci showing
single, double and four-spored asci. E: Ascospores in cross-sectional view showing the ornamentation of tall forked spines forming a continuous surface. F:
Ascospores in surface view showing the fine punctation of the spore surface. G: SEM micrograph of ascospores showing the fine spines that anastomosed toform a fine punctate spore surface. H: SEM micrograph of an ascospore showing composition of the fused spines to form the punctate surface. A: T32037; B–H:
OSC 38881 (holotype). Scale bars: A = 15 mm, B, C = 500 µm, D–F = 20 µm, G = 6 µm, H = 7.5 µm.

© The Japanese Association for Truffle Science (JATS), 2022

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Truffology - Volume 5, Issue 2, 2022

Castellano: Two new Elaphomyces species from eastern USAµm thick. Spores ornamented, globose, highly variable in size (20–25

of Wyoming, D. Mitchell of West Virginia, C. Stihler of West(–27) µm, mean = 25.0 µm) or (16–18 µm, these spores appear to

Virginia, and J.M. Trappe of Corvallis, Oregon. I also appreciate thebe immature or aborted), color brown to dark brown in KOH singly

opportunity to study specimens from the following herbaria: CUP,

and in mass, ornamentation 2–3 µm tall, ornamentation of tall, forked

FLAS, ITCV, OSC, and RMS.

interconnected spines, spore surface appears finely punctate underlight microscopy with a distinct dark inner wall.

References

Distribution, habit, habitat and season: Known from Florida andCastellano M.A., Beever R.E., Trappe J.M. (2012a) Sequestrate

Virginia; hypogeous; under Betula alleghaniensis, B. lenta, Quercus

fungi of New Zealand: Elaphomyces (Ascomycota, Eurotiales,

hemisphaerica, Q. laurifolia, Q. prinus, Q. rubra, and Q. virginiana;

Elaphomycetaceae). New Zealand Journal of Botany 50: 423–433.

January, February, April, June through September, and November.

Castellano M.A., Crabtree C.D., Healy R.A., Mitchell D. (2021)

Other specimens examined: USA, Florida, Alachua Co., Newnans

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Lake, just south of Windsor, Owens-Illinois County Park, 17 Sept.

Ascomycota) from eastern North America. Fungal Systematics and1980, J. Trappe (OSC 46707); near Gainesville, 27 Aug. 1977, J.

Evolution 7: 113–132.

Trappe (OSC 38946); southwest of Gainesville, Sugarfoot Hammock,
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farmland near City Golf course, 20 Nov. 1982, S. Miller 598 (RMS).
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peridial surface and the marbled inner peridium (Paz et al., 2017).
The brown, warty peridial surface and marbled inner peridiumof E. ramulospinus resembles all the other species in Elaphomyces
section Elaphomyces subsection Muricati except E. decipiens Vittad.
from Europe. The small spores (20–25 µm, mean = 25.0 µm) of
E. ramulospinus are clearly smaller than other Elaphomyces insubsection Muricati that occur in North America (E. americanus, E.

Castellano M.A., Henkel T.W., Miller S.L., Smith M.E., Aime M.C.
(2012c) New Elaphomyces species (Elaphomycetaceae, Eurotiales,
Ascomycota) from Guyana. Mycologia 104: 1244–1249.
Castellano M.A., Stephens R.B. (2017) Elaphomyces species(Elaphomycetaceae, Eurotiales, Ascomycota) from Bartlett
Experimental Forest, New Hampshire, USA. IMA Fungus 8: 49–63.
Castellano M.A., Trappe J.M., Vernes K. (2011) Australian species of
Elaphomyces (Elaphomycetaceae, Eurotiales, Ascomycota). Australian
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145–184.

holstii, E. verrucosus, and E. wazhazhensis). The spore ornamentation

Linder D.H. (1939) A new species of Elaphomyces from the Great Smokyof tall, forked spines anastomosed to form a ﬁne punctuation to theMountains National Park. Journal of the Elisha Mitchell Scientificspore surface is unique in this subsection and clearly differentiate E.
ramulospinus from other Elaphomyces species in this subsection.
Some specimens were parasitized by Tolypocladium ophioglossoides(J.F. Gmel.) C.A. Quandt, Kepler & Spatafora.

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para la Península Ibérica y una clave de identiﬁcation de las especies

included in this study: the late J. Kimbrough of Gainesville, Florida,del género para Europa. Boletín Micológico de FAMCAL 7: 85–104.

K. LoBuglio of New York, S. Loeb of North Carolina, S.L. Miller

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66© The Japanese Association for Truffle Science (JATS), 2022

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Castellano: Two new Elaphomyces species from eastern USA
Larsson E., Ibarguren I.O., Jeppson M., Læssøe T., Sauve M., RichardF., Moreau P.-A. (2017) The genus Elaphomyces (Ascomycota,
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Zhang B.C., Minter D.W. (1989) Elaphomyces spinoreticulatus sp. nov.,

Elaphomyces marmoratus and Elaphomyces fuscus spp. nov., from awith notes on Canadian species of Elaphomyces. Canadian Journal of

secondary forest in Japan. Mycoscience 61: 315–322.

Botany 67: 909–914.

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