ArticlePDF Available

Elaphomyces muricatus and Fischerula macrospora, two interesting hypogeous fungi from Greece

Elaphomyces muricatus and Fischerula macrospora,
two interesting hypogeous fungi from Greece
Aristidou 17 Sokratous 40
ΤΚ 51100, Grevena – Greece TK 19016, Artemis Attiki – Greece, 4 (5) : 95-98.
Octobre 2012
Mise en ligne le 22/10/2012
Summary: This paper discusses two hypogeous ascomycetes recently recorded in Greece, Ela-
phomyces muricatus and Fischerula macrospora. Descriptions based on the samples that were
found are provided, accompanied by macroscopic and microscopic images.
Keywords: Ascomycota, Eurotiales, Pezizales, taxonomy.
The arrival of the first truffle hounds from Italy in 2007 has
prompted an intensive research on hypogeous fungi in
Greece. This has resulted in the publication of several pa-
AGNELLO & KAOUNAS, 2010 and 2011), while for some species,
the Greek collections have helped clarifying important taxo-
nomical issues (ALVARADO et al., 2011; KAOUNAS et al., 2011).
Elaphomyces muricatus Fr. is widely recorded in Europe and
appears to be the most common representative of the genus
in Greece. By contrast, Fischerula macrospora Mattir. is
scarcely reported and appears to be rare.
Materials and methods
Macroscopic characters are described from fresh speci-
mens. For the microscopic examination a trinocular micro-
scope was used, Nikon Eclipse E100, with plan achromatic
objectives 4×, 10×, 40× and 100× in oil immersion. Micro-
scopic characters were determined from sections mounted
in water for pigments and measurements: Melzer’s reagent
and Cotton blue were used to highlight the ornamentation;
Congo red was used to highlight the walls of the elements.
Spore dimensions are based on at least 30 randomly selec-
ted spores, excluding ornamentation. The Elaphomyces spe-
cimens were discovered without the aid of hounds, while the
Fischerula specimens were discovered with the aid of
hounds. The primary sources used for identification are HAW-
KER (1953), PEGLER et al. (1993), ASTIER (1998), MONTECCHI &
SARASINI (2000), GORI (2005) and KONSTANTINIDIS (2009).
Elaphomyces muricatus Fr., Syst. mycol., 3 (1): 59 (1829)
Ceraunium muricatum (Fr.) Wallr., Fl. Crypt. Ger., 2: 407
=Elaphomyces variegatus Vittad., Monogr. Tuberac.:68
(1831); Lycoperdastrum variegatum (Vittad.) O. Kuntze,
Revis. gen. pl., 2: 858 (1891).
=Elaphomyces hirtus Tul. & C. Tul., Ann. Sci. Nat., Bot., sér.
2, 16: 23 (1841).
=Elaphomyces scaber J. Schröt., in Cohn, Krypt.-Fl. Schle-
sien, 3.2(1–2): 223 (1893) [1908].
Original description
Ε. muricatus, globosus, aculeis subtetragonis stipatis muricatus,
sporidiis nigricantibus.
Colore et statura priori similis, sed, differt forma magis aequali,
globosa (minore saepius); duritie demum fere lignosa ob peridium
junius saltim crassius; superficie tota obtecta aculeis (non pertu-
sis) stipatis acutis! tetra-pentagonis, unde muricata, sporidiis
minus intense atris et praecipue cellulis punctiformibus, Dothi-
dearum referentibus in peridio, quo evidentior fit analogia cum Tu-
bere; addito odore non ingrato. Color fulvus, in exoleto fuscus. Non
igitur miremur Linnaeum ex externa facie hanc speciem potius
cum Tubere, quam cum praecedente conjunxisse. Verum Tuber in
Suecia hactenus non lectum est. Cum priori tam in fagetis, pinetis,
at apud nos vulgatior. (v. v.) Obs. 1. Memorabiles sunt illae cellulae
Dothideaceae in hujus peridio et granula prioris Sphaeriaemorpha
superficialia, ob quae eum Haller Sphaeriam dixit. Adhuc magis
singulare videtur duas Sphaeriae species ex his tantum oriri, quare
pro illarum volvis habiti fuerunt. Hactenus equidem Sphaeriam ca-
pitatam ad El. granulosum, Spl. οphioglossoidem in El. muricato
legi; quod tamen οmnino constans esse, nondum affirmare audeo.
Ascomata: subglobose to elongated, 0.5–2 (4) cm in dia-
meter, hard, dermatoid, becoming fragile at maturity, yellow
at first, then luminous tawny or orange, finally brownish-
orange, ochre-brownish or mustard-brown, with small pyra-
midal spikes on the exoperidium, usually wrapped in a tight
network of rhizomorphs arising from the mycelium. Peridium:
1–2.5 cm thick, variable in colour, yellowish or orange-yel-
low, sometimes with pink or brown areas, heterogeneous,
endoperidium darker, purplish-brown, with prominent whi-
tish veins. Gleba: whitish-pink or greyish-pink at first, with
pinkish tramal plates, finally purplish-black and powdery.
Smell insignificant or unpleasant in maturity (“of gas” ac-
cording to literature).
Spores: initially hyaline, later purplish-brown, (15) 15.7–21.7
(28) × 14.5–20.6 (26) µm, with spikes up to (1.5) 1.7–2.3
(2.5) μm, having the form of a walking stick. Asci: subglo-
bose to broadly ellipsoid, with thin walls, 30–40 μm in dia-
meter, transitory.
Habitat: widespread in the northern hemisphere, fruiting gre-
gariously throughout the year, hypogeously or semi-hypo-
geously under broadleaved trees (mainly Fagus sp. but also
Quercus sp., Castanea sativa and Betula sp.) and conifers
(mainly Pinus sp.), on acidic soils. Sometimes parasitized by
Elaphocordyceps capitata (Holmsk.) G.H. Sung, J.M. Sung &
Spatafora (2007).
Studied collections: 14.04.2007, Zagora, under Fagus sp.,
ca 1100 m, GK 2226; 28.04.2007, Elatia Drama, under
Fagus sp., GK 2199; 29.10.2007, Zagora, under Fagus sp.,
ca 1100 m, VK 48; 29.06.2008, Zagora, under Fagus sp., ca
1100 m, VK 432; 30.05.2009, Trikala, under Quercus sp.,
GK 3880; 04.07.2010, Zagora, under Fagus sp., ca 1100 m,
VK 1505; 01.03.2011, Platania Kozani, under Quercus sp.,
GK 5600; 12.12.2011, Kastoria, under Fagus sp., ca
1000 m, VK 2385. All above collections were determined by
G. Konstantinidis and V. Kaounas and are conserved in GK
and VK personal herbarium.
Some authors like PEGLER et al. (1993) consider Elapho-
myces reticulatus Vittad. synonymous with E. muricatus,
while others like LAWRYNOWICZ (2006) consider it as a form of
the latter. The most obvious and constant features differen-
tiating E. muricatus from E. granulatus and E. asperulus, are
the fleshy layer of marbling in the peridium and the different
size and ornamentation of its spores. The more similar E. de-
cipiens Vittad. is distinguished by the paler color of its peri-
dium consisting of flat warts on a white background and the
almost unseparable mycelial mass usually surrounding the
fruitbody. The Greek specimens by their morphological fea-
tures match well the descriptions in HAWKER (1953), MON-
TECCHI & SARASINI (2000) and GORI (2005).
Fig. 1 — Elaphomyces muricatus
Left: Collection 2010 from Zagora (Greece). Picture: V. Kaounas.
Right: Detail of peridium and gleba, featuring the mature gleba and thick peridium with white veins. Picture: G. Konstantinidis.
Fig. 2 — Elaphomyces muricatus
A. Peridium. B. Detail of the peridium, appear the dense
orange pyramidoidal ledges that are overage from rhizo-
morphs. C. Ascospores ×400. D. Ascospores ×1000.
Pictures: A, C, D = G. Konstantinidis, B = V. Kaounas.
Fischerula macrospora Mattir., Nuovo G. bot. ital., 34: 1348
Original description
Exposita Generis Diagnosis unica tantum cum hac hucusque co-
gnita specie convenit.1
Fischerula Mattirolo — Nov. Genus.
A Tuberaceis adhuc notis eximium hoc novum Genus, ascorum,
sporarumque habitu differt, dum facies universa cum aliis fungis
hypogeais (praesertim e Genere « Tuber ») plerumque convenit.
Peridio irregulari, difformi, circumvoluto, contextu pseudoparen-
chymatico; colore umbrino-luteo notato.
Carne fuliginea, venis luteis filiformibus dispersis.
Ascis amplis, meiribranis spissis, formam praebentibus ovatam
(130 usque ad 179 micra et ultra long. secundum sporarum in-
clusarum numerum) basi leviter attenuatis.
Sporis raro unicis; plerumque 2-3-4 in singulis ascis, magnis (50-
70 long. et ultra; 40-50 lat. et ultra); non aculeatis, nec alveolatis
(uti in Genere « Tuber »); sed dense verrucosis; verrucis difformi-
bus, irregularibus, latis; colore saturate castaneo-badio, illum Hyd-
nothriae quodammodo referente.
Genus hoc clarissimo Eduardo FISCHER in Bernensi Athenaeo Bo-
tanices Professori, de Tuberibus optime merito, libenter D. et D.,
dum posterius accuratam illustrationem in lucem edere censeo.
Ascomata: irregularly globose or lobate, 1–2 cm in diame-
ter, brownish, yellow-brownish or ochre-brownish, darkening
when touched or exposed to the air. Peridium: pseudopa-
renchymatic, covered in a reddish, tomentose layer, irregu-
larly cracking in maturity. Gleba: compact, light greyish
initially, finally dark brownish-grey or black with paler veins.
It has a sweetish flavor and a pleasant smell, reminiscent of
Tuber species.
Spores: ellipsoid, reddish brown, (38.7) 47.2–62.3 (77.5) ×
(25.8) 31.1–42.1 (55.5) µm, with the rough, dense, irregu-
lar, dark brown spikes in maturity. The dimensions of spores
are similar to those of GORI (2005). Asci: large, oblong, thick
Fig. 3 — Fischerula macrospora
Left: Collection 2011 from Mainalon (Greece). Picture: G. Proutzopoulos.
Right: Collection from Gavros Kastoria (Greece). Picture: G. Konstantinidis.
1This diagnosis was based on the only species of the new genus Fischerula Matt. we report fully below.
Fig. 4 — Fischerula macrospora
A. Gleba. B. Ascospores ×100. C. Ascospores ×400.
D. Ascospores ×1000.
Pictures: G. Konstantinidis.
walled, (120–180 × 60–120 μm), containing (1) 2–4 (6) as-
Habitat: fruiting hypogeously in fall under broadleaved trees.
Studied collections: 01.11.2007, Gavros Kastoria, under
Fagus sp., ca 850 m, GK 2679; 07.11.2007, Gavros Kasto-
ria, under Fagus sp., ca 850 m, GK 2748; 15.10.2011, Mai-
nalon, under Abies cephalonica, ca 1400 m, VK 2275 and
GK 5818. All collections were determined by G. Konstanti-
nidis and V. Kaounas and are conserved in the GK and VK
personal herbarium.
Fischerula macrospora was described by MATTIROLO (1928)
from central Italy. A second species, F. subcaulis, was des-
cribed by TRAPPE (1975); molecular data was obtained by
O’DONNELL et al. (1997) and TRAPPE et al. (2007). This spe-
cies belongs to the lineage of the Morchellaceae-Discina-
ceae (LÆSSØE & HANSEN, 2007; PERRY et al., 2007) families,
although its position has not yet been fully resolved. So far
it is reported only from Italy, Spain, Sweden and Greece. The
Greek specimens by their morphological features match well
the descriptions in TRAPPE (1979), VIDAL (1997), ASTIER (1998),
MONTECCHI & SARASINI (2000) and GORI (2005).
The authors are indebted to G. Setkos (Kastoria), D. Kleisiari
(Grevena), G. Prountzopoulos (Athens), A. Papatsanis (Thes-
saloniki), D. Papadopoulou (Thessaloniki) and D. Tabouras
(Trikala) for the collections of Elaphomyces and Fischerula
presented here and to M. Loizides (Lemesos), secretary of
the Cyprus Mycological Association for the editing of this
paper and Carlo Agnello (Italy) for his decisive directives and
AGNELLO C. & KAOUNAS V. 2010. — Ruhlandiella berolinensis, Gena-
bea cerebriformis and Helvella astieri: tre rarissime specie rac-
colte in Grecia. Micologia e Vegetazione Mediterranea, 25:
DIS G., BARSEGHYAM G. & VENTURELLA G. 2011. — First molecular data
on Delastria rosea, Fischerula macrospora and Hydnocystis pili-
gera. Boletín de la Sociedad Micológica de Madrid, 35: 75-81.
ASTIER J. 1998. — Truffes blanche et noires (Tuberaceae et Terfe-
ziaceae). Louis–Jean, Gap, 127 p.
DIAMANDIS S. & PERLEROU C. 2008. — Recent records of hypogeous
fungi in Greece. Acta Mycologica, 43: 139-142.
FRIES E.M. 1829. — Systema mycologicum, sistens fungorum or-
dines, genera et species, huc usque cognitas. Vol. III. Gryphis-
waldiae, Ernesti Mauritii, 524 p.
HAWKER L.E. 1953. — British hypogeous fungi. Department of Bo-
tany, University of Bristol, 449p.
GORI L. 2005. — Funghi ipogei della Lucchesia, di altre province ita-
liane e dall’estero. Pacini Fazzi, 320 p.
KAOUNAS V., ASSYOV B. & ALVARADO P. 2001. — New data on hypogeous
fungi from Greece with special reference to Wakefieldia ma-
crospora (Hymenogastraceae, Agaricales) and Geopora clausa
(Pyronemataceae, Pezizales). Mycologia Balcanica, 8: 105-113.
KONSTANTINIDIS G. 2009. — Mushrooms, a photographic guide for col-
lectors. Athens, Published by the Author, 526 p. (In Greek).
LÆSSØE T. & HANSEN K. 2007. — Truffle trouble: What happened to
the Tuberales? Mycological Research, 111: 1075-1099.
LAWRINOWISZ M. 2006. — Hypogeous fungi collected in Estonia in
1989 and 1999. Folia Cryptogamica Estonica, 42: 67-71.
MATTIROLO O. 1927. — Secondo elenco dei “Fungi Hypogaei” raccolti
nelle Foreste di Vallombrosa (1900-1926). Nuovo Giornale Bo-
tanico Italiano, n.s, 34: 1343-1358.
MONTECCHI A. & SARASINI M. 2000. — Funghi ipogei d’Europa. AMB -
Fondazione CSM, Vicenza, 714 p.
genetic relationships among ascomycetous truffles and the true
and false morels inferred from 18S and 28S ribosomal DNA se-
quence analysis. Mycologia, 89: 48-65.
PEGLER D. N., SPOONER B.M. & YOUNG T.W.K. 1993. — British Truffles
– a revision of British Hypogeous Fungi. Royal Botanic Garden
Kew, 216 p. + 26 pl.
PERRY B.A., HANSEN K. & PFISTER D.H. 2007. — A phylogenetic overview
of the family Pyronemataceae (Ascomycota, Pezizales). Mycolo-
gical Research, 111 (3): 549-571.
TRAPPE M., EVANS F. & TRAPPE J.M. 2007. — Field Guide to North Ame-
rican Truffles. Ten Speed Press, Berkeley, 136 p.
TRAPPE J.M. 1975. — The genus Fischerula (Tuberales). Mycologia,
67 (5). 934-941.
TRAPPE J.M. 1979. — The orders, families and genera of hypogeous
Ascomycotina (truffles and their relatives). Mycotaxon, 9 (1): 297-
VIDAL J.M. 1997. — Algunus hongos hipogeos nuevos o poco cita-
dos de Cataluna (Zygomycotina, Ascomycotina y Basidiomyco-
tina). Revista Catalana de Micologia, 20: 25-62.
... Those species occur throughout Europe, and are also known from Asia and North America. The finding of Elaphomyces muricatus in Armenia along with recent findings of this and other Elaphomyces species from other countries (KONSTANTINIDIS & KAOUNAS 2012, LACHEVA 2012, WANG 2011, LAESSØE & al. 2009, KUTORGA & KATARŽYTĖ 2008 shows that geographical distribution and ecological adaptation of these fungi are wider than known so far. Recent publications show that Elaphomyces, particularly E. muricatus and E. granulatus, grow also in northernmost countries (e.g., Norway), can reach up to middle and high mountainous zones (e.g. ...
... Recent publications show that Elaphomyces, particularly E. muricatus and E. granulatus, grow also in northernmost countries (e.g., Norway), can reach up to middle and high mountainous zones (e.g. E. muricatus and E. granulatus in Armenia and E. granulatus in Taiwan) and that the preference of host plants and soil conditions are wider (for details see also PEGLER & al. 1993, LAWRYNOWICZ 1988, HAWKER 1954, KONSTANTINIDIS & KAOUNAS 2012, CAZARES 1992, WANG 2011. ...
Full-text available
Elaphomyces muricatus, new for Armenia, and E. granulatus have been found in the forests of Vanadzor and Dilijan in Armenia. Morphology, anatomy, as well as ecology and geographical distribution of E. muricatus are described. The ecological adaptation and geographical distribution of the genus Elaphomyces are wider than known before.
... Some of the items found seem to be fairly rare in our country, as demonstrated by the results of six years of research. This has led to the publication of several papers (DIAMANDIS & PERLEROU, 2008; KONSTANTINIDIS, 2009; AGNELLO & KAOUNAS, 2010 KONSTANTINIDIS & KAOUNAS, 2012) and, in some cases, the Greek collections helped to clarify certain important taxonomical issues ( KAOUNAS et al., 2011; ALVARADO et al., 2012). It seems that the species here presented are rare in our country because DIAMANDIS & PERLEROU (2008) have only reported one collection of Mattirolomyces terfezioides (Mattir.) ...
Full-text available
Article, 6 (1) : 1-4. Mars 2014 Mise en ligne le 15/03/2014 Summary: Two hypogeous Ascomycetes recently observed in Greece are presented: Hydnotrya tulasnei and Mattirolomyces terfezioides. Descriptions based on specimens found are given, accompanied by macrosco-pic and microscopic images. Σύνοψη: Παρουσιάζονται δύο υπόγειοι Ασκομύκητες που παρατηρήθηκαν πρόσφατα στην Ελλάδα: Hyd-notrya tulasnei και Mattirolomyces terfezioides. Δίνονται περιγραφές που βασίστηκαν σε ευρεθέντα δείγματα, συνοδευόμενες από απεικονίσεις μακροσκοπικών και μικροσκοπικών χαρακτηριστικών.
Full-text available
Technical Report
The report describes and suggests the development of a new/updated legislative regulation for the collection, certification, and trade of Wild Edible Mushroom (WEMs) in Greece after an initiative taken by the Ministry of Environment and Energy, and the Ministry of Rural Development and Food.
Full-text available
Some new or rare hypogeous fungi from Catalonia (Spain) (Zygomycotina, Ascomycotina and Basidiomycotina): 54 species and 3 varieties of hypogeous fungi are reported. 25 of them are new to Catalonia (1 zygomycete, 13 ascomycetes and 11 basidiomycetes), and 6 of them are new records in the Iberian Peninsule: Elaphomyces citrinus, E. granulatus var. asperulus, Genea vagans, Sphaerozone ostiolatum, Arcangeliella borziana and Gautieria morchelliformis var. globispora. Most of them have been described and illustrated by O.M. photographs, and new data about their ecology and phenology have been supplied. Some new synonyms are proposed.
Full-text available
Th is work provides new information about fi ve interesting and uncommon hypogeous fungi from Greece – Balsamia vulgaris, Geopora clausa, Hydnocystis piligera, Sclerogaster compactus and Wakefi eldia macrospora. Descriptions of the fi ve species are included based upon Greek collections, accompanied by colour macro-and microphotographs, and molecular data of four of them. On the basis of molecular results, the genus Wakefi eldia seems to be closely related to Hebeloma in the Hymenogastraceae, while Geopora clausa appears to be related to Geopora in the Pyronemataceae.
Full-text available
Phylogenetic relationships among ascomycetous truffles and the true and false morels were examined by using sequences from two nuclear encoded ribosomal DNA genes. The data consist of 18S rDNA and partial 28S rDNA sequences for 29 taxa. Individual and combined data sets were analyzed by maximum parsimony (MP), neighbor-joining (NJ) and maximum likelihood (ML) methods. Parsimony analysis of the combined data set, which contained 3 published 18S sequences and consisted of 2358 nucleotide characters, yielded a single most parsimonious tree of 1728 steps. The results indicate that the hypogeous ascomycetous truffle and trufflelike taxa studied represent at least 5 independent lineages within the Pezizales. Results also suggest that several epigeous and most hypogeous taxa have been misplaced taxonomically. Bootstrap analyses show strong support for a Tuberaceae-Helvellaceae clade which is a monophyletic sister group of a Morchellaceae-Discinaceae clade. Rhizina is a sister group to both of these clades in the MP and ML trees while in the NJ tree it is a sister of the Morchellaceae-Discinaceae clade. MP, ML, and NJ tree topologies indicate that Rhizina should be removed from the Helvellaceae and classified in a monotypic family, the Rhizinaceae. A phylogenetically-based classification for these fungi is proposed together with emendations of 4 families.
Full-text available
Uncertainty among Greek farmers who are in search of new and profitable crops has increased interest in truffle cultivation. Recent research has come up with 23 taxa of hypogeous fungi new for Greece including gastronomically valuable species. Natural ecosystems of Quercus pubescens, Q. frainetto, Q. ilex and Q. coccifera seem to be rich in hypogeous species. Ecosystems with Corylus avellana, Carpinus betulus, Mediterranean pines and even poplar plantations were found to also host hypogeous fungi. These records, supported by historical information about the existence of truffles in Greece, seem to be encouraging hints for systematic truffle cultivation.
Full-text available
Several samples of Delastria rosea, Fischerula macrospora and Hydnocystis piligera from the Mediterranean basin are studied, and the first molecular data of these species are reported. A colour plate showing the external morphology of their ascomata is also included.
Three very rare species: Ruhlandiella berolinensis, Genabea cerebriformis and Helvella astieri are descriibed. The collections were made under different plants in the Attica region of mainland Greece, in winter. The descriptions are accompanied by colour photographs of ascomata and photographs as well as line drawings of the relative microanatomical features
Partial sequences of nuLSU rDNA were obtained to investigate the phylogenetic relationships of Pyronemataceae, the largest and least studied family of Pezizales. The dataset includes sequences for 162 species from 51 genera of Pyronemataceae, and 39 species from an additional 13 families of Pezizales. Parsimony, ML, and Bayesian analyses suggest that Pyronemataceae is not monophyletic as it is currently circumscribed. Ascodesmidaceae is nested within Pyronemataceae, and several pyronemataceous taxa are resolved outside the family. Glaziellaceae forms the sister group to Pyronemataceae in ML analyses, but this relationship, as well as those of Pyronemataceae to the other members of the lineage, are not resolved with support. Fourteen clades of pyronemataceous taxa are well supported and/or present in all recovered trees. Several pyronemataceous genera are suggested to be non-monophyletic, including Anthracobia, Cheilymenia, Geopyxis, Humaria, Lasiobolidium, Neottiella, Octospora, Pulvinula, Stephensia, Tricharina, and Trichophaea. Cleistothecial and truffle or truffle-like ascomata forms appear to have evolved independently multiple times within Pyronemataceae. Results of these analyses do not support previous classifications of Pyronemataceae, and suggest that morphological characters traditionally used to segregate the family into subfamilial groups are not phylogenetically informative above the genus level.
An overview of truffles (now considered to belong in the Pezizales, but formerly treated in the Tuberales) is presented, including a discussion on morphological and biological traits characterizing this form group. Accepted genera are listed and discussed according to a system based on molecular results combined with morphological characters. Phylogenetic analyses of LSU rDNA sequences from 55 hypogeous and 139 epigeous taxa of Pezizales were performed to examine their relationships. Parsimony, ML, and Bayesian analyses of these sequences indicate that the truffles studied represent at least 15 independent lineages within the Pezizales. Sequences from hypogeous representatives referred to the following families and genera were analysed: Discinaceae-Morchellaceae (Fischerula, Hydnotrya, Leucangium), Helvellaceae (Balsamia and Barssia), Pezizaceae (Amylascus, Cazia, Eremiomyces, Hydnotryopsis, Kaliharituber, Mattirolomyces, Pachyphloeus, Peziza, Ruhlandiella, Stephensia, Terfezia, and Tirmania), Pyronemataceae (Genea, Geopora, Paurocotylis, and Stephensia) and Tuberaceae (Choiromyces, Dingleya, Labyrinthomyces, Reddellomyces, and Tuber). The different types of hypogeous ascomata were found within most major evolutionary lines often nesting close to apothecial species. Although the Pezizaceae traditionally have been defined mainly on the presence of amyloid reactions of the ascus wall several truffles appear to have lost this character. The value of the number of nuclei in mature ascospores as a delimiting family character is evaluated and found to be more variable than generally assumed.