The first record of Urnula mediterranea (Pezizales) in

Continental Europe

Nenad MILOSAVLJEVIĆ
Ascomycete.org, 9 (5) : 156-163.
Septembre 2017
Mise en ligne le 27/09/2017
Abstract: All previous collections of the rare fungus Urnula mediterranea were recorded on the Mediterra-
nean coast or Atlantic coast of Europe. Our discovery in Serbia represents the first ever recorded in conti-
nental part of Europe. We present herein photographs made on this new locality, along with microscopic
and macroscopic descriptions.
Keywords: Ascomycota, Sarcosomataceae, Serbia, taxonomy.
Izvod: Svi dosadašnji nalazi retke gljive Urnula mediterranea su zabeleženi na obalama Mediterana. Ovaj
nalaz je prvi zabeleženi iz konntinentalnog dela Evrope. Prilažemo fotografije snimljene na mestu nalaza
kao i njen makroskopski i mikroskopski opis sa odgovarajućim ilustracijama.
Ključne reči: Ascomycota, Sarcosomataceae, Srbija, taksonomija.

Introduction One of our Serbian collections is genetically confirmed and it has

Urnula mediterranea is considered as a rare species, only known
from Europe. According to KAOUNAS et al. (2015), until now it was
recorded in three countries (Italy, France and Greece) for a total of
15 locations, all on the Mediterranean and Atlantic coasts, even if
three (two from Corsica and one from Tuscany) were not microscop-
ically confirmed. Its presence in Tuscany (Italy) has been then con-
firmed by MARZIANI et al. (2016). Another collection has been recently
recorded from a new place in Ligury, Italy (M. Carbone, pers. comm.).
Considering that it has only recently been described (CARBONE et al.,
2009) and considered very rare, further researches are needed, es-
pecially to know the ecological conditions required for its growth.
For these reasons it is also necessary to conduct a monitoring of this
species in known localities in order to assess its level of vulnerabil-
ity.
not shown any difference with the sequenced ones both published
and unpublished (M. Carbone, pers. comm.).
Materials and methods
The macrophotographs were taken in situ using a Cannon A630
camera.The material was studied on fresh and dried samples using
the Biolam C11 optical microscope with a 100 to 1350 magnifica-
tion. Microscope samples were cut manually using a scalpel, and the
observations of the ornamentation were made in water, Cotton blue
and Congo red. The microphotographs were taken through the mi-
croscope eyepiece using a Cannon A530 camera. The amyloid reac-
tion of the ascus tip was tested using Lugol’s solution. All
measurements were obtained in water medium using Piximetre

Fig. 1 – Urnula mediterranea, 06.05.2014, Žeželj. Photo: N. Milosavljević

156
 Fig. 2 – Different stages of development of U. mediterranea. Photos: N. Milosavljević
program, and the photographs were processed with Adobe Photo-
shop CC. The exsicates are housed in author’s private fungarium.
Taxonomy
Urnula mediterranea (M. Carbone, Agnello & Baglivo) M. Car-
bone, Agnello & P. Alvarado, Ascomycete.org, 5 (1): 6 (2013).
≡ Plectania mediterranea M. Carbone, Agnello & Baglivo, Riv. Micol.,
52 (3): 252 (2009).
Macroscopic features
Ascoma initially spherical, then deeply cup-shaped, with the rim
folded inwardly, 0.8–2 cm deep, 3.5–4.5 (9) cm wide and 2.5–5 cm
high, sometimes caespitose. The margin can be slightly indented
and cleft or it can be thick-edged. It becomes a bit deeper while rip-
ing, and opens up like a shallow plate at the end of the growth pe-
riod, but it is never fully open. Flesh 1.5–5.7 mm thick, with three
clearly distinct layers on a cross section. The upper layer is hymenial
and dark red in color, the middle one is gelatinous, ocher-gray col-
ored, and the lower layer is black. Flesh structure is compact, odor-
less and tasteless. Hymenium radially wavy from center towards the
edges, reddish-brown with tones of copper colour, darker to almost
black in the center; when riping it becomes brown, then dark brown,
and black in the end; it can be dull velvety to shiny, especially in wet
conditions. External surface subtly velvety, wrinkled, sometimes
with small cracks, creating irregular segments 1.5–5 mm wide that
are darker compared to the lighter background, dark red, grayish-
brown to dark brown and finally black. Stem 1.5–2.5 (5) cm high,
0.5–2.5 cm wide, the lower half is covered in the mycelium, plant re-
manants and soil. Spore print creamy-white.
Microscopic features
Asci cylindrical, 480–580 × 12.5–16 μm, with wall 0.8–1.5 μm
thick, 8-spored, inamyloid, without crozier, carminophilic, opercu-
late. On some ascomata, more than 50% of asci can be sterile. Para-
physes similar to or a bit longer than the asci, 2–3 μm wide,
light-gray colored, cylindrical, septate, sometimes moniliform, with
branching on the bottom and often on the top, with tip segments
that vary from flat to curved and branched, and often contain yel-
lowish granules of pigment. Hymenial hairs present, sparse, 3–
4 μm wide. Ascospores ellipsoid, smooth, hyaline, containing
multiple oil drops of different sizes. Initially roundish with a thick
wall, they later become ellipsoid with a thinner wall, and contain 3–
5 guttules, sometimes tapered at the poles, when fully mature, with
a thin wall and filled with plasma or numerous small oil drops. The
spore size is: (26.6) 27.9–32.3 (42) × (9.8) 10.9–14 (17) μm, Q = (1.7)
2.1–2.7 (2.9) ; N = 50, Me = 30.3 × 12.6 μm ; Qe = 2.4. Subhymenium
of textura intricata, 130–150 μm wide, brown-coloured, composed
of densely interwoven hyphae 5-7 μm wide. Medullary excipulum
of textura intricata, 600–1200 μm thick, gelatinous, light grayish
brown in colour, composed of sparse interwoven hyphae 4–7 μm
wide, especially in the middle part of the layer. Ectal excipulum of
textura angularis, 280–310 μm wide, composed of cells measuring
7–13 × 10–15μm, which are deep brown coloured. Chains (catelu-
nate) of elliptical (fusiform) cells 20–37 × 8.5–16.5 μm as well as elon-
gated bean-shaped cells (allantoid) sometimes covered by crystals
of the same dimensions were noticed. Two kinds of excipular hairs
were observed: the first one is made of transparent hyphae with thin
wall, 3 μm wide, coated with crystals outside, and the second one
composed of hyphae with thick wall, 5–8 μm wide, with a brownish
black pigment. Basal tomentum composed of hairs, 5.5–7.5 μm
thick, thin-walled, septate, brownish-black.
Ascomycete.org 157
 Fig. 3 – Urnula mediterranea
A: habitat; B: micellar hairs; C: cross section of the ascoma. Photos: N. Milosavljević
Ecology and habitat
The specimens studied in this work were observed in a locality
that was continually prospected from 2014 to 2017. It was first dis-
covered on 6th May 2014, on the southwest side of Žeželj hill near
Kragujevac city, at 320 m asl. All specimens were recorded on a sin-
gle location, in a young mixed forest composed of oak (Quercus ssp.),
hornbeam (Carpinus ssp.), hawthorn (Crataegus ssp.), beech (Fagus
ssp.) and ash (Fraxinus ssp.) trees on a limestone terrain. The exam-
ined samples were found on destructed tree stumps or on the
ground, that is, on tree remains covered with soil on the surface of
15 m radius, and they were grouped in eight nests (cespitose) with
3-5 specimens per group. In 2015, the first findings were recorded
almost a month later, on 30 May 2015 on the same place, in 4 nests
with 3-5 specimens. In 2016, the first findings were recorded on 23
May 2016 in 6 nests with 3-5 specimens. The final findings on this
location were recorded on 25 May 2017 in 8 nests with 3-5 speci-
mens per nest. The population of U. mediterranea is stable on this
location, but the most prolific year was 2014, which was one of the
hottest and rainiest years in this region.
Considering very slow maturation process, fruitbodies have en-
dured for more than 20 days, depending on the environmental con-
ditions. We can conclude that fruitbodies appear earlier, at the
beginning of May 06.05.2014 in conditions of higher temperature
and increased humidity, whilst it, in the normal conditions for May,
appears roughly 20 days later (between 30.05.2015 and 23.05.2016).
158 Ascomycete.org
Discussion
Urnula mediterranea was first described in 2009 by CARBONE et al.
(2009) as a member of the genus Plectania. Upon a detailed phylo-
genetic analysis, it was then transferred into the genus Urnula (CAR-
BONE et al., 2013).
Among the similar European species, Urnula craterium (Schwein.)
Fr. is deeply cup shaped, 2–5 cm wide and 3–6.5 cm high, with short
or long stipe; the hymenium is smooth, blackish brown, matte or a
little shiny; the external surface is blackish-brown; the ascospores
are ellipsoid and measure (21) 25–33 (36) × 11–14 μm (DISSING, 1981).
Urnula hiemalis Nannf. looks like a deep plate, with a short stem
or stemless, 1–5 cm wide, and 2–5 cm high; the hymenium is
smooth, brownish-black; the external surface is concolorous; the as-
cospores are ellipsoid or subcylindrical, and measure 24–31 × 12–
15 μm (CARBONE & AGNELLO, 2012).
Urnula groenlandica Dissing, is only known from Greenland, and
it is smaller than the former ones; the hymenium is brownish-black
with a lighter edge; the ascospores are ellipsoid and measure 21.5–
27.1 × 10.0–13.2 μm (DISSING, 1981).
In examined samples, spores ripen in asci for a very long time are
changing thereby their shape from spherical to elliptical with pointy
tips (fusoid-ventricose) and finally elliptical. The content of the as-
cospores also changes with ripening: at the beginning 3 to 10 oil
drops can be observed, then more oil droplets are present in the
plasma. “Abnormal giant” spores were also seen, measuring 45–
50 μm in length. Germinated spores were also present, both inside
and outside asci, within mature bodies.
Fig. 4 – U. mediterranea, 18.05.2014, Žeželj hill. Photo: N. Milosavljević

Ascomycete.org 159
 Fig. 5 – U. mediterranea
A: Ascospores; B-B1: Immature ascospores; C: Mature ascospores in the ascus; D: “Abnormal giant” ascospores; E-E1: Germination of the
ascospores; F: Ascospores from left to right: in water mounts, Cotton blue and Congo red; A,B1,C,I,D in water mounts ×400; B,E1 in Congo
red ×400. Scale bars = 20 μm. Photos: N. Milosavljević.

160 Ascomycete.org
 Fig. 6 – U. mediterranea
A, A1: Ascus; A2: Ascus tips; B: Germination of ascospores; C: Ascus bases; D: Moniliform paraphyses; D1, D2, D3, D4, D5, D6: Tips of para-
physes; E, E1: Setae and paraphyses; A, A1, A2, D3, D4 in Congo red 400×; D2 in Cotton blue 400×; B, C, D, D5, D6, E, E1 in water mounts
400×. Scale bars = 20 μm except A1 = 100 μm. Photos: N. Milosavljević.

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 Fig. 7 – U. mediterranea
A: Apothecium cross section; B: Subhymenium; C: Medullary excipulum; D: Ectal excipulum; E: Excipular hairs; A, B, C in Congo red 400×;
D, E in water mounts 400×. A Scale bar = 100 μm; B, C, D, E Scale bars = 20 μm. Photos: N. Milosavljević.

162 Ascomycete.org
 Fig. 8 – U. mediterranea
A: Hairs of the basal tomentum; B: Cell chains of ectal excipulum; C: Ectal excipulum with allantoid cells; A, C in water mounts ×400 ; B in
Cotton blue ×400. Scale bar = 20 μm. Photos: N. Milosavljević.

All previous known habitats of U. medierranea were located on References

the Mediterranean and Atlantic coasts. The occurrence in Serbia is
interesting because this locality does not present the Mediterranean
or Atlantic climate type. An explanation for this could be that global
climate change, which is expressive in the Balkan Peninsula, condi-
tioned the emergence of new species. Presently, many typically
Mediterranean plants and fungi can be found in the vicinity of the
city of Kragujevac, which confirms this theory and justifies the belief
that the finding of U. mediterranea was not a mere chance.
Conclusion
U. mediterranea is certainly a rare fungus and likely an European
endemic species. This study was proposed as an addition for a better
knowledge of this species, both in macroscopic and microscopic
features, as well as some ecological data. Further studies are still nec-
essary to assess its conservation status in its natural habitats.
Acknowledgements
The author owes special thanks to Carlo Agnello (Italy) for his crit-
ical review of the manuscript and Matteo Carbone (Italy) for impor-
tant and useful information and his comments.
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ef
Nenad Milosavljević
Ivana Milutinovića 23
34 000 Kragujevac
Serbia
nenadivesna@yahoo.com

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