Zoological Journal of the Linnean Society, 2010, 159, 1–21. With 7 figures
Biogeography and phylogenetic relations within
the Dinaric subgenus Monolistra (Microlistra)
(Crustacea: Isopoda: Sphaeromatidae), with a
description of two new species
SIMONA PREVORČNIK*, RUDI VEROVNIK, MAJA ZAGMAJSTER and BORIS SKET
University of Ljubljana, Biotechnical Faculty, Department of Biology, Večna pot 111, 1000 Ljubljana,
Slovenia
Received 2 December 2008; accepted for publication 31 March 2009
A phylogenetic review of Monolistra (Microlistra), a freshwater cavernicolous subgenus of isopod crustaceans,
distributed in the north-western part of the Dinaric karst, is presented. The distribution data and an identification
key are provided for known taxa. Seven species are reviewed and two new species are described: Monolistra
(Microlistra) fongi sp. nov. and Monolistra (Microlistra) jalzici sp. nov. Monolistra (Microlistra) pretneri
spinulosa Sket is synonymysed with the nominate subspecies because of the morphological variability in the type
subspecies and the genetic uniformity of the species. Two major, geographically vicariant and morphologically
different clades have been identified by molecular analysis. Low genetic differentiation within the subgenus, as well
as conspicuous dorsal sculpturing of animals, indicate their apparently recent colonization of the hypogean realm.
These indications are confirmed by the distribution of Microlistra species within the current river systems, rather
than palaeo-hydrographically defined basins, as is the case of other subterranean aquatic groups of crustaceans,
including other members of the genus Monolistra.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21.
doi: 10.1111/j.1096-3642.2009.00593.x
ADDITIONAL KEYWORDS: Dinarides – phylogeny – subterranean.
INTRODUCTION
In 1929, Racovitza described Microlistra spinosa, an
extraordinarily shaped subterranean sphaeromatid,
from the collections of ‘Društvo za raziskovanje jam’
(Cave Research Society in Ljubljana). Soon afterwards Microlistra spinosissima was described, a
species with an even more bizarre shape (Racovitza,
1929a, b). Although Racovitza recognized their affinities with the genus Monolistra Gerstaecker, 1856, he
established a new genus Microlistra for both new
species. He distinguished the new genus from Monolistra by the absence of grasping ‘pincers’ (subchelae)
*Corresponding author.
E-mail: simona.prevorcnik@bf.uni-lj.si
in male pereopods II, but ignored their very showy,
long tergal processes, anticipating their occurrence as
taxonomically irrelevant. In 1930 Stammer described
Monolistra (Typhlosphaeroma) schottlaenderi with
numerous, but very short dorsal processes, but moved
it to Microlistra in 1932, as the third species of the
genus. Some additional species of Microlistra have
been described since then, one even with an entirely
smooth dorsum (Sket, 1960, 1964, 1965, 1982).
Karaman (1954), Stoch (1984), Deeleman-Reinhold
(1971), and Sket (1982) contributed some new taxonomic and/or distribution data, but the latest taxonomic and distribution reviews of the entire genus
Monolistra and the related genus Caecosphaeroma
Dollfus, 1896, were provided by Sket (1967,
1986a).
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
1
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S. PREVORČNIK ET AL.
In his review of the ‘Monolistrini’, Sket (1964)
showed that the adequate position of Racovitza’s
genus Microlistra would be as a subgenus within the
genus Monolistra: namely, considering morphology,
Microlistra relates to M. (Monolistra) s.str., the same
way as do the subgenera Typhlosphaeroma Racovitza,
1910, Pseudomonolistra Hubault, 1937, and the
newly established Monolistrella Sket, 1964. To avoid
the identification problems in samples with no males,
he proposed considering all these taxa as subgenera
of Monolistra.
In our study, we investigate the affinities between
species within the subgenus Microlistra, which we
prove to be a monophyletic clade nested within the
genus Monolistra. We list all known taxa within
the subgenus, and investigate their phylogenetic and
biogeographic relations within the ‘north-western
merodinaric biogeographic area’ (sensu Sket, 1994). We
use morphological characteristics to provide an easyto-use identification key. Two new species are
described. We consider this subgenus worth presenting
because: it is a clearly established monophylum, with
an apparent morphological diversity and high molecular homogeneity; it is biogeographically well defined as
a typical north-western Dinaric element; it differs from
some other cave-dwelling groups by its species being
distributed within recent drainages (rather than by
palaeo-drainages). We succeeded to sample nearly all
taxa for DNA analysis. We do not, however, discuss the
phylogenetic values of other nominal subgenera here;
they will be discussed when the complete phylogenetic
tree for the genus Monolistra is provided.
MATERIAL AND METHODS
MATERIAL
We collected all known taxa and some undescribed
forms of Microlistra (and of other Monolistra groups).
All localities were georeferenced (see the Appendix)
and distribution maps have been produced for biogeographic studies. Most of the localities were sampled
repeatedly.
With the exception of samples of Monolistra
(Microlistra) schottlaenderi and undescribed Monolistra (Microlistra) sp. (cf. sketi) from the Rupećica
spring [Croatia; referred to as Monolistra (Microlistra) sp. – Rupećica hereafter in the text], samples
from all listed localities are stored in the invertebrate
collection of Oddelek za biologijo, Biotehniška
fakulteta, Univerza v Ljubljani (OB BF UL). The
Monolistra (Microlistra) sp. – Rupećica sample is kept
in the collection of Hrvatsko biospeleološko društvo
(Croatian Biospeleological Society), Zagreb; a description of this possibly new species is in the domain of
colleagues from Croatia. Monolistra (Microlistra)
schottlaenderi localities are listed in the checklist of
Italian fauna (Ruffo & Stoch, 2000).
We included a total of 24 Monolistra specimens
belonging to 12 species (Table 1) in our DNA analyses.
Two species of the related genus Caecosphaeroma
were used as out-groups. The in-group taxa were all
the available Microlistra spp. and the type species of
all other subgenera. To ensure the taxonomic identity,
we mainly used specimens from topotype populations:
this was not possible for Monolistra (Microlistra) spinosissima, and no material was available for DNA
analysis for Monolistra (Microlistra) calopyge.
DNA
EXTRACTION, AMPLIFICATION, AND
SEQUENCE ANALYSIS
Genomic DNA was extracted from specimens preserved in 96% ethanol using a modified Mouse Tail
protocol of the Nucleospin Tissue kit (MachereyNagel, Düren, Germany), as described in Verovnik
et al. (2003). For each of the 26 specimens (including
the out-group taxa) an approximately 550-bp fragment of the mitochondrial 12S rDNA gene was
amplified using the primers 5′-CCTACTTTGTTAC
GACTTAT-3′ and 5′-GCCAGCAGCCGCGGTTA-3′,
designed by comparing the available invertebrate 12S
rDNA sequences. Approximately 500 bp of the mitochondrial 16S rDNA were amplified using the universal primers 16Sar and 16Sbr (Simon, 1991).
Amplification of an approximately 1280–1420-bp-long
fragment of the nuclear 28S rRNA gene was performed using the primers 5′-AGGGAAACTTCGG
AGGGAACC-3′ and 5′-CAAGTACCGGTGAGGGAA
AGTT-3′ that were designed by comparing available
invertebrate 28S rDNA sequences. Purified PCR
products were sequenced on an Applied Biosystems
3730xl sequencer by Macrogen (Seoul, Korea).
Sequences were aligned using MUSCLE (Edgar,
2004).
PHYLOGENETIC
ANALYSIS
The 12S, 16S, and 28S sequence alignments were
analyzed separately using the neighbour-joining algorithm as implemented in MEGA4 (Kumar et al.,
2007). As all three data sets produced almost identical
trees (results not shown, see below), sequences were
concatenated for the final analysis.
The alignment of concatenated sequences is
2452-bp long. Sequence data were analyzed using
Bayesian inference. The program MrBayes 3.1 (Ronquist & Huelsenbeck, 2003) was used. Hierarchical
likelihood tests (Posada & Crandall, 1998) were
employed in order to test alternative models of
evolution, using MrModeltest 2.2 (Nylander, 2004).
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
3
Table 1. List of sequenced samples of Monolistra and Caecosphaeroma, with corresponding information about nucleotide
sequence accession numbers (Acc. N.)
Taxon name
Monolistra (Microlistra)
spinosa
Monolistra (Microlistra)
spinosa
Monolistra (Microlistra)
spinosa
Monolistra (Microlistra)
schottlaenderi
Monolistra (Microlistra)
spinosissima
Monolistra (Microlistra)
spinosissima
Monolistra (Microlistra)
sp.
Monolistra (Microlistra)
sketi
Monolistra
(Microlistra) fongi
sp. nov.
Monolistra (Microlistra)
bolei bolei
Monolistra (Microlistra)
bolei brevispinosa
Monolistra (Microlistra)
pretneri cf. spinulosa
Monolistra (Microlistra)
pretneri pretneri
Monolistra (Microlistra)
pretneri cf. spinulosa
Monolistra (Microlistra)
pretneri spinulosa
Monolistra
(Microlistra) jalzici
sp. nov.
Monolistra
(Typhlosphaeroma)
racovitzai racovitzai
Monolistra (Monolistra)
caeca caeca
Monolistra
(Pseudomonolistra)
h. hercegoviniensis
Monolistra
(Monolistrella)
velkovrhi
Caecosphaeroma virei
Dollfus
Caecosphaeroma
b. burgundum Dollfus
Designation in
phylogenetic tree
Location (in short)
Acc. N. 12S
Acc. N. 16S
Acc. N. 28S
FJ842006
FJ842007
FJ842008
spinosa Kočevje
Tominčev studenec,
SLO
Slovenska vas, SLO
FJ842059
FJ842060
FJ842061
spinosa Ribnica
Mobišaht, SLO
FJ842029
FJ842030
FJ842031
Schottlanderi
Doberdo/Doberdob, ITA
FJ842062
FJ842063
–
spinosissima Vrhnika
Retovje, SLO
FJ842035
FJ842036
FJ842037
spinosissima Vrhnika
& Logatec
sp. Rupečica
Gašpinova jama, SLO
FJ842026
FJ842027
FJ842028
Rupećica, CRO
FJ842048
FJ842049
FJ842050
sketi T
Pećina špilja, CRO
FJ842045
FJ842046
FJ842047
fongi T
Kuruzovića pećina,
CRO
FJ842032
FJ842033
FJ842034
b bolei T
Stobe, SLO
FJ842012
FJ842013
FJ842014
b brevispinosa T
Vinica, SLO
FJ851101
FJ851102
–
p cf spinulosa Obrovac
Kusa, CRO
FJ842043
FJ842044
–
p pretneri T
Pećina kod Vrane, CRO
FJ842015
FJ842016
FJ842017
p cf spinulosa Žegar
Milića špilja, CRO
FJ842051
FJ842052
–
p spinulosa T
FJ842041
FJ842042
–
jalzici T
Špilja kod mlina na
Miljacki, CRO
Čepić tunnel, CRO
FJ842009
FJ842010
FJ842011
r racovitzai Postojna
Postojnska jama, SLO
FJ842023
FJ842024
FJ842025
c caeca T
Podpeška jama, SLO
FJ842053
FJ842054
FJ842055
h hercegoviniensis T
Vjetrenica, BiH
FJ842038
FJ842039
FJ842040
velkovrhi T
Stobe, SLO
FJ842018
FJ842019
FJ842020
C. virei FRA-Doubs
Moullin des Iles, FRA
FJ842021
FJ842022
–
C. burgundum
FRA-Moselle
Gorze, FRA
FJ842056
FJ842057
FJ842058
spinosa T
Sequence names are identical to the ones depicted on the phylogenetic tree: ‘T’ with taxon designation, denotes topotypic
population; BiH, Bosnia and Herzegovina; CRO, Croatia; FRA, France; ITA, Italy; SLO, Slovenia; for the additional data
and for a list of all localities see the Appendix.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
4
S. PREVORČNIK ET AL.
A general time reversible (GTR) model of nucleotide
substitution, with gamma-distributed rate heterogeneity and a significant proportion of invariable sites,
was selected. Uniform or fixed default prior settings
were used. A Markov chain Monte Carlo search was
run with four chains for 4 ¥ 106 generations, taking
samples every 100 generations. The approximate
number of generations needed to obtain stationarity
of the likelihood values (‘burn-in’) of the sampled
trees was estimated graphically, and was set to 5000.
From the resulting trees, posterior probabilities were
assessed for individual clades based on their observed
frequencies.
MORPHOLOGICAL
EXAMINATION AND DESCRIPTION
OF NEW SPECIES
Specimens preserved in 70% ethanol were transferred
to glycerol, partly dissected, and the appendages
were mounted in glycerol on slides, for examination
and drawing. Measurements were made under an
Olympus SZX12 stereoscope equipped with a Sony®
DXC-390P colour video camera, by means of the
Windows-supported program analySIS®. Original
drawings were made with a camera lucida. The
appendages were transferred from the slides and
deposited in 70% ethanol, together with the specimens. For SEM, the whole specimen was dehydrated
in a graded ethanol series, followed by acetone and
1,1,1,3,3,3-hexamethyldisilazane (HMDS), coated
with gold, and then observed under a scanning electron microscope (JEOL® JSM-840A).
The terminology of setae (non-cuticular structures,
articulating), spines (cuticular structures, articulating), setules (cuticular in origin, being derived from
cuticular scales), and setulose fringe (a dense mass of
setules on the posterior margin of the pereopod
articles), on the appendages, follows that of Bruce
(1994). Particular terms have been coined to describe
the dorsal sculpturing of the species. The prominent
large structures on tergites and pleotelson are
referred to as processes, which may be ‘conical’,
‘sickle-shaped’, ‘spine-like’, and other. Small dorsal
embossments on dorsum are termed nodules, which
may be ‘rounded’, ‘conical’, or of another shape.
USE
OF TAXONOMIC CATEGORIES
We tried to implement the ‘biological species concept’
and ‘polytypic species concept’ of Mayr & Ashlock
(1991) and Mayden (1997), taking into consideration
the cogent arguments of the authors. As all studied
taxa at the rank of species are allopatric, there is no
direct evidence about their interfertility/intersterility.
Their generally low molecular differences, however,
do not indicate clear reproductive isolation. On the
other hand, the evident differences in dorsal sculpturing can certainly represent effective pre-mating
barriers. As we suppose that populations with low
morphological differences (like Monolistra (Microlistra) bolei bolei and Monolistra (Microlistra) bolei
brevispinosa) might potentially be interfertile, we
consider such populations as morphologically distinct
subspecies.
Out of convenience we also use the subgenus category. From the phylogenetic point of view, there is no
‘natural’ criterion for the distinction of categories.
Also, the ICZN (1999) provides no criteria. It is,
however, very useful for the user of the classification
(an ecologist, faunist, etc.) to be able to identify the
lowest possible taxonomic rank for collected biota. So,
even though in samples containing only females,
specimens could not be identified to the species level,
they could at least be identified to the genus level (as
Monolistra sp., containing ~40 species). On the other
hand, splitting of the genus Monolistra into five new
genera (the present subgenera) would result in the
identification of females as ‘Sphaeromatidae gen. sp.’,
i.e. as one of the innumerable and ecologically diverse
species.
RESULTS
REVIEW
OF SPECIES AND SUBSPECIES
Within the subgenus Microlistra seven species with
two additional subspecies have already been
described. Two new species are described herein,
whereas some further taxa have not yet been studied
in detail.
Genus Monolistra Gerstaecker, 1856
Subgenus Microlistra Racovitza, 1929
Diagnosis: A Monolistra group of species with body
smaller in males than in females. Male pereopod II
without subchela, its distal articles only slightly
shortened and swollen; mature male pleopod II with
appendix masculina as long as endopodite or longer,
strongly curved in its distal part; pleopod IV with
large area respirans (‘aire respiratoire’ of Racovitza,
respiratory area hereafter in the text); uropod
reduced to tiny tubercle on a shallow elevation
(‘socle’) of pleotelson surface; pleotelson caudal bulge
projecting beyond the ventrocaudal border.
Remarks: Within the genus Monolistra, most of the
characters described above are diagnostic (Sket,
1965). In the secondary characters of sexual dimorphism (size relationships, development of male pereopods II), specimens of the subgenus Microlistra
resemble, to some degree, specimens of the subgenera
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
Pseudomonolistra and Monolistrella. In both later
subgenera, however, males are only slightly smaller
or are of equal size as females, and in Monolistrella
male pereopods are more differentiated. Apomorphic
for Microlistra, and probably unique within the
family, are the specifically positioned and, to differing
extents, reduced uropods.
Type species, by original designation: Microlistra
spinosa Racovitza, 1929.
Other taxa: Monolistra (Microlistra) bolei bolei Sket,
1960; Monolistra (Microlistra) bolei brevispinosa
Sket, 1982; Monolistra (Microlistra) fongi sp. nov.;
Monolistra (Microlistra) jalzici sp. nov.; Monolistra
(Microlistra) pretneri pretneri Sket, 1964; Monolistra
(Microlistra) pretneri spinulosa Sket, 1965; Monolistra (Microlistra) schottlaenderi schottlaenderi
(Stammer, 1930); Monolistra (Microlistra) schottlaenderi ssp. (Trieste/Trst) Stoch, 2000; Monolistra
(Microlistra) sketi (Deeleman, 1971); Monolistra
(Microlistra) sp. (cf. sketi) (Rupećica) Bedek; Monolistra (Microlistra) spinosissima (Racovitza, 1929).
Besides the characters mentioned above, which are
not present in females, the sculpturing of tergites is
characteristic for the subgenus, and distinguishes
between species and subspecies. As a rule, the head
and pereonal tergites are armed with transversal
rows of up to nine nodules or spine-like processes,
and, in the majority of species and subspecies, such
rugosities are also present on the pleotelson. In some
species, some of the nodules are absent (probably
reduced), whereas in others they are elongated into
processes, the length of which may reach the trunk
width. The position of processes differs among
species. The most stable is the position of both processes placed just in front of the vestigial uropods.
The only exception from these trends is the dwarfish (~6-mm long) Monolistra (Microlistra) calopyge,
which has a smooth tergum, a dorsally smooth pleotelson, and uropods that do not jut out. On the other
hand, its caudal bulge on the pleotelson is particularly apparent (projecting). As molecular data are not
available, the position of this species remains ambiguous, but its classification to the subgenus was evidently erroneous (see below). Some specimens of the
morphologically variable Monolistra (Microlistra)
pretneri are also nearly smooth, but with uropods of
the Microlistra-type.
5
Croatia, Bosnia and Herzegovina, and Montenegro,
i.e. between Como (Italy) and Podgorica (Montenegro). The troglobiotic members of the genus Caecosphaeroma, with two species and a subspecies, are
West European (Sket, 1986a). Another sphaeromatid
genus with troglobiotic species is the ‘antipodean’
Bilistra Sket & Bruce, 2004, from New Zealand.
Other Sphaeromatidae are epigean, and are entirely
or predominantly marine, with the exception of the
thermophilic genus Thermosphaeroma Cole & Bane,
1978, from the southern North America.
The distribution of the members of the subgenus
Microlistra (see the exact localities in the Appendix)
is limited to the so-called north-western Dinaric area
(Sket, 1994). Species are known from approximately
40 localities, spread from the extreme east of Italy,
through southern (Dinaric) Slovenia, to southwestern Croatia. The most south-eastern localities
are along the Dalmatian river Krka north of Šibenik,
the most northern localities are along the Slovenian
river Krka, and in the western distribution area they
are found in the Italian part of the region Kras/Carso.
The largest extension of the known distribution area
is approximately 300 km. All species inhabit fresh
karst-groundwater, and may sometimes be sampled
in springs or resurgences.
1. Monolistra (Microlistra) bolei Sket, 1960
Distribution: Localities of both subspecies within the
river Kolpa/Kupa drainage.
2. Monolistra (Microlistra) bolei bolei Sket, 1960
Distribution: South-eastern Slovenia, Bela Krajina,
Črnomelj–Otovec.
3. Monolistra (Microlistra) bolei brevispinosa Sket,
1982
Distribution: South-eastern Slovenia, Bela Krajina;
Dragatuš–Vinica, south of the type subspecies.
4. Monolistra (Microlistra) fongi sp. nov.
Distribution: Croatia, Kordun. Probably the river
Korana drainage.
Remarks: See below for the description of the species.
5. Monolistra (Microlistra) jalzici sp. nov.
DISTRIBUTION
The members of the genus Monolistra are distributed
in the European continental karst groundwaters:
along the Southern Calcareous Alps and Dinarides in
northern Italy, southern Switzerland, Slovenia,
Distribution: Croatia, south-eastern Istra (Istria).
Drainage of the brook Boljunščica, close to the
Adriatic.
6. Monolistra (Microlistra) pretneri Sket, 1964
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
6
S. PREVORČNIK ET AL.
IDENTIFICATION
KEY
Species and subspecies of Microlistra are easily distinguished by their dorsal ornamentation.
1.
Caudal pleotelson bulge longer than 45% total ventral pleotelson length or 75% pleotelson height; tergites
smooth; pleopods without respiratory areas.............................................Monolistra (?Microlistra) calopyge
1.* Length of caudal pleotelson bulge (spine-like processes not included) shorter than 30% of total ventral pleotelson
length, and never surpassing half pleotelson height..............................................................................2
2.
Pleotelson without a pair of terminal caudal processes; its proximal lateral processes, if developed, short and
thick, blunt ................................................................................................................................... 3
2.* Pleotelson with pairs of narrowly pointed lateral and caudal processes....................................................5
3.
Along the pereon, a median, and a pair of lateral rows of short and thick, setulose processes, nodules between
them much smaller ...................................................................... Monolistra (Microlistra) jalzici sp. nov.
3.* On pereonites, only ± equal nodules in transversal lines, the lateral ones at the most twice as long as the others
(exceptionally with smooth body).......................................................................................................4
4.
Pleotelson dorsocaudal bulge tapering in caudal direction, with a narrow apex; male pleopod II endopodite with
~20 marginal plumose setae ........................................................... Monolistra (Microlistra) schottlaenderi
4.* Pleotelson dorsocaudal bulge broadly rounded apically; male pleopod II endopodite with ~15 marginal plumose
setae .................................................................................................. Monolistra (Microlistra) pretneri
5.
Anterior pereonites without sculptures; pleotelson without a median process ............................................ 6
5.* Some anterior pereonites with spine-like processes...............................................................................7
6.
Pereonite VI with a lateral pair of short, sickle-shaped processes; two pairs of similar processes on pleotelson a
little longer than wide at base .................................................................... Monolistra (Microlistra) sketi
6.* Pereon without any processes, pleonal processes longer......................Monolistra (Microlistra) sp. – Rupećica
7.
Pleotelson without a median, only with paired sickle-shaped processes; lateral sickle-shaped processes on
pereonites I–III and V–VI, Monolistra (Microlistra) bolei.......................................................................8
7.* Pleotelson with two pairs of processes plus a median sickle-shaped process; lateral sickle-shaped processes on
pereonites I, II, IV, and VI...............................................................................................................9
8.
Processes longer, tips of lateral pleonal ones reaching the ventrocaudal border...........................................
......................................................................................................Monolistra (Microlistra) bolei bolei
8.* Processes shorter, tips of lateral pleonal ones (in adults) not reaching ventrocaudal border...........................
............................................................................................Monolistra (Microlistra) bolei brevispinosa
9.
Pereonal lateral processes shorter than distances between their bases....Monolistra (Microlistra) fongi sp. nov.
9.* Pereonal lateral processes longer than distances between their bases.....................................................10
10. Pleotelson with only five major processes; processes strongly curved .............. Monolistra (Microlistra) spinosa
10.* Besides five major processes, also some small processes on pleotelson; processes less curved and longer .........
...................................................................................................Monolistra (Microlistra) spinosissima
Syn. Monolistra (Microlistra) pretneri spinulosa Sket,
1965, new synonymy
Distribution: Croatia, northern Dalmacija–Kvarner
(Quarnero). Drainages of the (Dalmatian) rivers
Krka, Zrmanja, and the lake Vransko jezero; southern
tip of Cres Island.
Remarks: Specimens of the type population (cave
Pećina, Vrana, Zadar) are dorsally either completely
smooth or of the spinulosa type. All other populations
are of the spinulosa type, and the one with the most
prominent, cone-shaped spines (cave Kusa, Obrovac)
is molecularly closest to the type population. Bearing
in mind the morphological variability, we do not consider the named subspecies to be valid.
7. Monolistra (Microlistra) schottlaenderi (Stammer,
1930)
Monolistra (Microlistra) schottlaenderi schottlaenderi
(Stammer, 1930)
Syn. Monolistra (Typhlosphaeroma) schottlaenderi
Stammer, 1930
Microlistra schottlaenderi Stammer, 1932
Distribution: North-eastern Italy, Carso/Kras. Springs
and caves north-east and south-east of Monfalcone.
All localities are parts of a coastal karst aquifer at the
Adriatic.
8. Monolistra (Microlistra)
(Trieste/Trst) Stoch, 2000
schottlaenderi
ssp.
Distribution: North-eastern Italy, Kras/Carso. Spring
Fonte Oppia/Klinčica, Bagnoli/Boljunc, Trieste/Trst.
Probably a hydrographically separated part of the
coastal aquifer.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
7
Remarks: Only one specimen found (F. Stoch, pers.
comm.), not yet studied.
Table 2. List of species co-occuring with Monolistra
(Microlistra)
9. Monolistra (Microlistra) sketi (Deeleman-Reinhold,
1971)
Monolistra (Microlistra)
species
Co-occuring Monolistra
species
M. (Microlistra) bolei
brevispinosa
M. (Microlistra) bolei bolei
M. (Monolistra) caeca
Gerstaecker
M. (Monolistrella)
velkovrhi Sket
M. (Monolistrella) sp.
and M. (Monolistra)
caeca Gerstaecker
M. (Typhlosphaeroma)
racovitzai Stammer
M. (Monolistra) caeca
Gerstaecker
M. (Monolistra) caeca
Gerstaecker
M. (Typhlosphaeroma)
racovitzai Stammer
M. (Typhlosphaeroma)
karamani Sket
None
None
None
None
Syn. Microlistra sketi Deeleman-Reinhold, 1971
Distribution: Croatia, Lika. Drainage of the river
Gacka.
10. Monolistra (Microlistra) sp. (cf. sketi) – Rupećica
Distribution: Croatia,
Ivanac, Ogulin.
Kordun.
Rupećica
M. (Microlistra) fongi sp.
nov.
spring,
Remarks: Molecularly almost identical to M.
(Microlistra) sketi; morphologically similar to it, but
nevertheless distinct, without any projections on
pereon, and with longer anterior pleonal processes.
Collected by B. Jalžić. Headwaters of the river Dobra.
M. (Microlistra)
schottlaenderi
M. (Microlistra) spinosa
M. (Microlistra) spinosa ssp.
(Kočevje–Ribnica)
M. (Microlistra) spinosissima
11. Monolistra (Microlistra) spinosa (Racovitza, 1929)
M. (?Microlistra) calopyge
Syn. Microlistra spinosa Racovitza, 1929
M. (Microlistra) sketi
M. (Microlistra) sp.-Rupečica
M. (Microlistra) pretneri
M. (Microlistra) jalzici sp.
nov.
Distribution: South-eastern Slovenia. Springs and
caves in a belt along the river Krka (Dolenjska Krka).
12. Monolistra
(Microlistra)
Kočevje-Ribnica
spinosa
ssp.
–
Distribution: Southern Slovenia. Two localities within
a syncline, parallel with the river Krka (Dolenjska
Krka), but 15 km from it: both probably draining
separately towards the river.
assignment to the subgenus is doubtful. No specimen
could be obtained recently for DNA analysis.
CO-OCCURRENCE
Remarks: Morphologically very similar, and molecularly nearly identical with M. spinosa, but with
slightly longer and less curved spines.
13. Monolistra (Microlistra) spinosissima (Racovitza,
1929)
Syn. Microlistra spinosissima Racovitza, 1929
Monolistra spinosa spinosissima S. Karaman, 1954
Distribution: South-western Slovenia. An aquifer
including the subterranean parts of the river Ljubljanica between Postojna and Vrhnika.
14. Monolistra (?Microlistra) calopyge Sket, 1982
Distribution: South-eastern Slovenia. Boreholes in
Družinska vas, near Kronovo, Novo mesto. One locality in the drainage of the river Krka (Dolenjska
Krka); hypothermic water (16–18 °C).
Remarks: Because of the complete absence of sculpturing, the reduction of pleopod respiratory areas,
and very short article 6 of male pereopod II, the
All Microlistra species within Slovenia and Italy were
found to share the same locality with (at least) one
Monolistra species from another phyletic group
(Table 2). This was surprisingly not the case in most
(with one exception) Croatian species or localities. On
the other hand, all Microlistra species are strictly
allopatric. Also, the co-occurrence of species from
other subgenera is extremely rare (Sket, 1965).
PHYLOGENETIC
RELATIONSHIPS
In addition to the tree presented here (Fig. 1), we
calculated phylogenies including 75 Monolistra s.l.
samples, both Caecosphaeroma spp., and some
marine Sphaeromatidae. The result corroborated the
position of Caecosphaeroma as a sister clade to Monolistra s.l. To investigate the phylogeny of just the
species within the Microlistra clade, we used a
smaller set of taxa. The phylogeny of Monolistra s.l. is
not yet ready for publication, as the samples of some
important species are still missing.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
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S. PREVORČNIK ET AL.
Of 24 examined specimens of the genus Monolistra,
we found 21 with differences in the concatenated
alignment. Of 768 (31.2%) variable characters, 318
(12.9%) were parsimony informative. The obtained
Bayesian tree (Fig. 1) is highly resolved, with high
posterior probability support values. The monophyly
of the genus Monolistra received high support, but
the relationships among species belonging to different
subgenera (sensu Sket, 1965) remains unresolved,
possibly because of insufficient sampling in other sub-
genera. The subgenus Microlistra is monophyletic
and distinctly separated into two main clades: one
including M. Microlistra pretneri and Monolistra
(Microlistra) jalzici sp. nov., and the other including
the type species Monolistra (Microlistra) spinosa with
all other species.
Only two differences exist between the single gene
trees (data not shown) and the Bayesian tree (Fig. 1),
both with low bootstrap support: Monolistra
(Microlistra) jalzici forms a separate basal Microlistra
Figure 1. Phylogenetic relationships of Monolistra (Microlistra) species derived from 35 000 Bayesian trees in comparison to their tergal sculpturing. Values on major branches are Bayesian posterior probabilities. The concatenated
sequences of 12S, 16S and 28S rDNA are named as in Table 1: b = bolei; c = caeca; h = hercegoviniensis; p = preterni;
r = racovitzai; M. = Monolistra. ‘T’ after the taxon name denotes that specimen was collected at type locality.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
Figure 2. Geographical distribution of species of Monolistra (Microlistra). Black circles denote localities of
sequenced samples, the others are open. Solid lines
encircle the southern and the northern clade, respectively;
smaller ovals encircle single uniform species (solid lines)
or pairs of closely related species or subspecies (dashed
lines). Numbers are close to the type localities (see text)
and denote: M. – Monolistra (Microlistra), 1 – M. schottlaenderi, 2 – M. spinosissima, 3 – M. spinosa, 4 – M.
spinosa spp., 5 – M. bolei bolei, 6 – M. bolei brevispinosa,
7 – M. jalzici, 8 – M. sp.-Rupećica, 9 – M. sketi, 10 – M.
fongi, 11 – M. pretneri, 12 – M. pretneri ‘spinulosa’, 13 – M.
calopyge; BiH, CRO, ITA, SLO, – Bosnia-Herzegovina,
Croatia, Italy, Slovenia.
clade in the 12S tree, and Monolistra (Microlistra)
fongi sp. nov. is the sister taxon to the Monolistra
(Microlistra) sp. – Rupećica clade in the 16S tree.
The distinction of subspecies in Monolistra
(Microlistra) pretneri is not supported, as the
sequences of both morphotypes are almost identical.
The assignment of the insular (Cres Island) population to the mentioned species could not be verified
because of the lack of an appropriate sample. Monolistra (Microlistra) jalzici sp. nov. is positioned as a
sister taxon to Monolistra (Microlistra) pretneri in our
molecular analysis. This is in accordance with the
(southern) distribution of both species along the
Dinaric syncline, and across the gulf of Kvarner
(Quarnero), which was submerged in the Pleistocene
(Marković-Marjanović, 1971; Sket, 1988).
9
In the northern clade (Fig. 2), the expected sister
relationship between the two species with longest
processes (Monolistra (Microlistra) spinosa and Monolistra (Microlistra) spinosissima) is confirmed. The
unnamed variety of Monolistra (Microlistra) spinosa
from the cave Mobi šaht near Ribnica (morphologically between both species mentioned above) is nearly
identical to Monolistra (Microlistra) spinosa from the
type locality according to our molecular analysis: they
both occur in two parallel valleys ~15 km apart. The
Monolistra (Microlistra) sp. – Rupećica species group
and the newly decribed M. (Microlistra) fongi sp. nov.
are positioned as sister groups to the type-species
group, but their geographic distribution does not correspond entirely with the phylogenetic relationships.
Namely, they geographically encircle the Monolistra
(Microlistra) bolei species group positioned basally
within the northern clade. Despite geographic relations and morphological similarities with the southern clade, Monolistra (Microlistra) schottlaenderi
clearly belongs to the northern spinosa–spinosissima
clade in our molecular phylogeny.
Both newly described species are genetically distinct from all the rest, and their separation is confirmed by high posterior probabilities. Monolistra
(Microlistra) pretneri, the sister species of Monolistra
(Microlistra) jalzici sp. nov., was separated by an
average Kimura’s two-parameter (K2P) genetic distance of 3.4%. The sister relationship of Monolistra
(Microlistra) fongi sp. nov. is less clear, however, as its
position as a sister clade to the clade including
Monolistra (Microlistra) sketi, Monolistra (Microlistra) spinosa, and Monolistra (Microlistra) spinosissima, is poorly resolved. The average K2P genetic
distance between these species and Monolistra
(Microlistra) fongi sp. nov. was 1%.
Based on the two available molecular clock calibrations for 16S rDNA in Malacostraca (Crustacea)
(Sturmbauer, Levinton & Christy, 1996; Schubart,
Diesel & Hedges, 1998), the estimated time of the
split between the subgenus Microlistra and the rest of
the genus was 1.61 ± 0.35 or 2.23 ± 0.48 Mya, and the
split between the northern and the southern clades of
Microlistra dates to 1.11 ± 0.17 or 1.54 ± 0.23 Mya.
The timing of the final speciation events could be
represented by the split between the morphologically
very different, and geographically and hydrographically unrelated, species Monolistra (Microlistra)
schottlaenderi and its sister clade Monolistra
(Microlistra) spinosa – Monolistra (Microlistra)
spinosissima, which dates to 0.55 ± 0.25 or
0.76 ± 0.35 Mya.
DESCRIPTIONS
OF THE NEW SPECIES
Genus Monolistra Gerstaecker, 1856
Subgenus Microlistra Racovitza, 1929
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
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S. PREVORČNIK ET AL.
Figure 3. Monolistra (Microlistra) jalzici sp. nov.
tunnel Čepić, Istra, Croatia, female 11.6 mm, obliquelateral view (SEM JEOL JSM-840A by K. Drašlar). * – *,
median row of dorsal spines.
MONOLISTRA (MICROLISTRA)
(FIGS 3–5)
JALZICI SP. NOV.
Holotype: Adult male, 7.9 mm, Croatia, Istra Peninsula, springs in the drainage tunnel Čepić, near
Kršan, Labin; leg. B. Sket and S. Legović. Inv. No.
‘Malacostraca 2240 ht’, partly dissected, preserved in
ethanol, deposited in the collection of the OB BF UL.
Paratypes: Same data as holotype. Three adult
females (one ovigerous, two non-ovigerous, 11.6–
6.4 mm). Collection of OB BF UL Inv. No. Malacostraca 2240 pt.
One paratype deposited in the Hrvatski prirodoslovni muzej (Croatian Natural History Museum),
Zagreb, Croatia.
Etymology: The species is named after our good colleague Branko Jalzić, a deserving Croatian caver and
cave-fauna researcher.
Diagnosis: Microlistra species with short, but stout,
densely setulose massive cones on its dorsal surface.
Pleotelson irregularly nodular, with large lateral protuberances and uropod rudiments elevated on small
elevations (‘socles’), projecting over pleotelson outline
(Fig. 4). Anterior coxae truncated; posterior ones
blunt and slightly turned up distally.
Description of holotype: Male, 7.9-mm long. Body
width 44% of body length. Head (cephalothorax) dorsally with transverse row of four conical processes.
Pereonites I–VI each with one median and two lateral
conical processes. Pereonite I with three, and pereonites II–VI each with two conical paramedian nodules
on each side; protuberances on epimeral apex larger,
gradually increasing in size from epimeron II–VI;
protuberances on epimeron V–VI similar to median
and lateral conical processes. Pereonite VII without
lateral and epimeral processes. Pleonite I without
lateral and epimeral nodules. Pleotelson width 150% of
pleotelson length, highly vaulted, and its surface
densely and irregularly covered with conical and
rounded nodules. Only paired lateral conical processes
(resembling those on epimeron VI) protruding remarkably over pleotelson outline, and followed by raised
socles carrying uropod rudiments. Pleotelson ventrocaudal border without a groove, with a comparatively
narrow dorsocaudal bulge reaching far beyond it. All
large dorsal protuberances covered with thick mat of
very long setules, except at the very base and apex.
Antenna I length 32% of body length; peduncle
article 1 bent perpendicularly; 6 flagellar articles,
articles 3–6 each with single, long aestethasc.
Antenna II length about 40% of body length; 11 flagellar articles. Length ratio of peduncular articles and
flagellum in antenna I 100 : 73 : 80 : 135, in antenna
II, 68 : 100 : 123 : 191 : 653.
Left mandible stout, incisor and lacinia mobilis
both unicuspidate, bluntly rounded (spatulate); apical
spine row of 13 spiniform processes on a long stalk;
molar process with prominent serrations around
smooth mesial surface. Right mandible without
lacinia mobilis. Other mouth appendages and maxilliped as in type species, Monolistra (Monolistra) caeca
Garstaecker (Racovitza, 1910).
Pereopod I propodus (article 6) width 36% of propodus length; merus (article 4) with two serrate
spines at anterodistal angle; carpus (article 5) with
two biserrate spines at posterodistal angle; propodus
with three biserrate spines on posterior margin and
two at posterodistal angle; setulose fringe near continuous on posterior margin of basis (article 2) to
beginning of unguis (distal part of article 7, claw);
setules lengths on ischium (article 3) about 50% of
article width, diminishing towards unguis; setulose
fringe also present on anterior margin of basis to
merus, length of setules increasing towards merus;
secondary unguis finely serrate. Pereopod II basis
with three long plumose setae on anterior margin;
merus with two serrate spines at anterodistal angle;
carpus with one biserrate spine at posterodistal
angle, and one on distal margin; propodus with two
short serrate spines on posterior margin, one serrate
spine at posterodistal angle, and one long plumose
seta at anterodistal angle; setulose fringe present on
posterior margin of distal third of ischium to beginning of unguis, nearly continuous on anterior margin
of basis to unguis; secondary unguis finely serrate.
Pereopod VII basis with one long plumose seta on
anterior margin; merus with two serrate spines
at posterodistal angle; carpus with one serrate
spine at posterodistal angle, one biserrate spine at
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
11
Figure 4. Monolistra (Microlistra) jalzici sp. nov., tunnel Čepić, Istra, Croatia, dissected ovigerous female: MdbL,
left mandible (apical part); MdbR, right mandible with palp (apical parts); Mx I, II, maxilla I and II (apical parts); Mxlp,
maxilliped; PMxlp, maxilliped palp. Holotype male, 7.9 mm: A I, II, antennae I and II; F, frons: with ep, epistome and la,
labrum; Plt, pleotelson, ventral view: with U, uropod rudiment on its socle.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
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S. PREVORČNIK ET AL.
Figure 5. Monolistra (Microlistra) jalzici sp. nov., tunnel Čepić, Istra, Croatia, holotype male, 7.9 mm: Plp I–V, pleopods
I–V; Pp I, II, VII, pereopods I, II, VII.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
anterodistal angle, and two on distal margin; propodus with one serrate spine on posterior margin, one at
posterodistal angle, and one long plumose seta at
anterodistal angle; setulose fringe as in pereopod II,
except setules shorter; secondary unguis finely
serrate. Relative lengths of pereopods I, II, and VII:
33, 41, 48% of body length; length ratio of articles
(coxa excluded) in pereopod I 100 : 49 : 31 : 19 : 54 :
40, in pereopod II 100 : 51 : 32 : 48 : 53 : 38 in pereopod VII 100 : 91 : 34 : 53 : 63 : 33.
Pleopod I protopodite with fine long setules along
internal and external margins, and two strong spines
at internal angle; exopodite elliptical, with 7 plumose
setae along terminal margin, and setules at proximoexternal angle; endopodite about half as wide and
nearly as long as exopodite, parallel sided, with two
plumose setae on rounded terminal margin and setules
at proximo-internal angle. Pleopod II similar to I, but
with more numerous plumose setae; endopodite as
wide as exopodite, slightly widened distally, with 17
setae; appendix masculina distally curved and terminally blunt, its length 120% of endopodite length;
exopodite with 9 plumose setae. Pleopod III exopodite
elongate, subovoid, without respiratory area, with
short transverse suture at the external margin, and
with a trace of the same suture at the internal margin.
Pleopod IV shape similar; respiratory area length 55%
of exopodite length, surface 35% of exopodite surface.
Pleopod V exopodite irregularly elliptical with thick
sclerotized ridge along proximal half of external
margin; three patches on intero-distal half densely
scaled; respiratory area on intero-proximal half, its
length 30% of exopodite length. Uropods vestigial, as
in the subgenus type species Monolistra (Microlistra)
spinosa Racovitza (1929).
Description of paratypes: Females of 11.6, 8.2, and
6.4 mm in length; larger than male if adult; body
width 67% of body length. Pleotelson dorsocaudal
vault surpassing ventrocaudal pleotelson border to
greater extent than in male.
Antenna I length 29–39% of body length; of
between six and nine flagellar articles, articles 4–6
and 8, or 3–5 and 7, each with single, long aestethasc.
Antenna II length 38–43% of body length, flagellum of
between 10 and 12 articles.
Pereopod I spines and setulose fringe as in male,
but propodus with between three and five biserrate
spines on posterior margin, and two at posterodistal
angle. Pereopod II spines and setulose fringe as in
male, but basis with one or two long plumose setae on
anterior margin; carpus with one or two biserrate
spines at anterodistal angle, and one or two biserrate
spines on distal margin; propodus with one or two
short serrate spines on posterior margin. Pereopod
VII spines and setulose fringe as in male, but basis
13
with between two and five long plumose setae on
anterior margin; merus with one or two biserrate
spines at anterodistal angle; carpus with between one
and three biserrate spines at anterodistal angle, one
on distal margin and two at posterodistal angle; propodus with one or two short serrate spines on posterior margin. Relative length of pereopods I, II, and
VII, 30–34, 36–40, and 43–50% of body length, respectively; length ratio of their articles (coxa excluded)
in pereopod I 100 : 47 : 28 : 13 : 48 : 35/100 : 54 : 37 :
16 : 57 : 49/100 : 54 : 39 : 19 : 58 : 48, in pereopod
II 100 : 49 : 31 : 45 : 49 : 34/100 : 55 : 34 : 53 : 61 : 35/
100 : 79 : 46 : 67 : 81 : 60, in pereopod VII 100 : 94 :
39 : 57 : 64 : 33/100 : 93 : 35 : 58 : 61 : 32/100 : 101 :
39 : 62 : 64 : 38.
Pleopods I and II as in male, but with different
number of plumose setae: pleopod I with one or two
and six or seven plumose setae; pleopod II with 13–18
and 8–11 plumose setae. Pleopods III and IV shapes
as in male. Pleopods IV and V respirtory area length
54–63 and 32–42% of exopodite length, pleopod V
respiratory area surface 26–41% of exopodite surface.
Distribution and ecology: Numerous specimens were
found in small freshwater springs, appearing in the
tunnel draining the karst polje (a large depression
within karst) Čepićko polje, north of Labin, eastern
Istra Peninsula, Croatia. They were accompanied by
the large cirolanid isopod Sphaeromides virei virei
(Brian), and by an atyid cave shrimp Troglocaris sp.
In the past, numerous cave salamanders, Proteus
anguinus Laurenti (Amphibia: Proteidae), used to be
washed up from the springs into the tunnel (Sket,
1997). Some specimens of the new Monolistra were
dorsally nearly black, presumably as a result of bacterial iron deposition.
Remarks: The pleopods of M. (Microlistra) jalzici sp.
nov. are nearly identical to those of M. (Microlistra)
pretneri Sket, 1964. In most populations of the latter
species, dorsal protuberances are present on pereonites; however, they are all of equal length, and are
conical and smooth. We believe that prominent differences in dorsal sculpturing might play the role of
a reproductive barrier, and therefore we consider
M. (Microlistra) jalzici sp. nov. to be a true species,
in accordance with the molecular tree and with
the biological species concept.
MONOLISTRA (MICROLISTRA)
(FIGS 6, 7)
FONGI SP. NOV.
Holotype: Adult male of 13.5 mm in length, Croatia,
Kordun, cave Kuruzovića pećina, Vaganac, near Rakovica; leg. B. Sket. Inv. Collection of OB BF UL, Inv.
No. ‘Malacostraca 2245 ht’, partly dissected, preserved in ethanol.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
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S. PREVORČNIK ET AL.
Figure 6. Monolistra (Microlistra) fongi sp. nov., cave Koruzovića pećina, Croatia, holotype male, 13.5 mm: A I, II,
antennae I and II; B, body oblique-lateral view; E, epimera; F, frons: with ep, epistome and la, labrum; Pe VII, pereomere
VII; Plp I–II, pleopods I–II; Plt(1), pleotelson, lateral view; Plt(v), pleotelson, ventral view: with U, uropod rudiment.
Paratypes: Same data as for holotype, two adult males
(11.2- and 10.9-mm long) and two adult females (nonovigerous, 13.3- and 11.9-mm long), Inv. No. ‘Malacostraca 2245 pt’. One paratype deposited in the
Hrvatskio prirodoslovni muzej (Croatian Natural
History Museum), Zagreb, Croatia.
Other material: Croatia, Kordun, spring of the river
Slunjčica, Slunj, one ex. leg. B. Jalžić, deposited in
collection of Hrvatski prirodoslovni muzej, Zagreb.
Etymology: The species is named after our colleague
Daniel W. Fong, a renowned American speleobiologist.
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BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
15
Figure 7. Monolistra (Microlistra) fongi sp. nov., cave Koruzovića pećina, Croatia, holotype male, 13.5 mm: Plp
III–V, pleopods III–V; Pp I, II, VII, pereopods I, II, VII.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
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S. PREVORČNIK ET AL.
Diagnosis: Microlistra species with pairs of sickleshaped lateral processes on pereonites I, II, IV, and
VI, and median sickle-shaped process on pereonites
I–VII; lateral processes length shorter than distances
between their bases. Epimera apically narrowly
rounded to bluntly pointed. Pleotelson with pair of
long sickle-shaped lateral processes, a pair of shorter,
straight, narrowly pointed terminal caudal processes,
and a sickle-shaped median process. Uropod rudiments elevated on small bulges, projecting over pleotelson outline.
Description of holotype and paratype males (data in
parentheses): Holotype male 13.5-mm long (paratype
males 11.2- and 10.9-mm long). Body width 45% (48–
53%) of body length. Head (cephalothorax) dorsally
with two conical nodules. All pereonites with one
median sickle-shaped process; pereonites I, II, IV, and
VI each with a pair of longer sickle-shaped lateral
processes, gradually increasing in size in caudal
direction. Pleonite I without dorsal structures. Pleotelson width 140% (150–157%) of pleotelson length,
highly vaulted, with pair of sickle-shaped lateral processes and pair of shorter straight terminal processes;
lateral process length 152% (160–182%) of terminal
process length. Lateral processes followed by two
raised socles with uropod rudiments. Pleotelson ventrocaudal border without a groove, comparatively
narrow dorsocaudal bulge projecting far beyond it. All
processes remarkably shorter than distances between
them. Anterior epimera apically narrowly rounded,
posterior ones bluntly pointed.
Antenna I length 27% (26–28%) of body length;
peduncle article 1 bent perpendicularly; ten (9) flagellar articles, articles 5–7 (3–6 and 8) with single long
aestethasc each. Antenna II length about 30% (41%)
of body length, seven (13–15) flagellar articles.
Length ratio of peduncular articles and flagellum in
antenna I 100 : 92 : 114 : 155 (100 : 26 : 93 : 113 :
167), in antenna II 100 : 110 : 113 : 233 : 453
(100 : 114 : 140 : 211 : 703).
Mouth parts as in Monolistra (Microlistra) jalzici
sp. nov. Pereopod I propodus (article 6) width 33%
(39–42%) of propodus length; merus (article 4) with
two serrate spines at anterodistal angle; carpus
(article 5) with two biserrate spines on posterodistal
margin, and one at posterodistal angle; propodus with
seven biserrate spines on posterior margin and 2 (1)
at posterodistal angle; setulose fringe near continuous
on posterior margin of ischium (article 3) to beginning
of dactylus (article 7); setules length on ischium only
one third article width, diminishing towards dactylus;
setulose fringe also present on anterior margin of
basis to merus, setules longest on ischium; secondary
unguis finely serrate. Pereopod II basis with two
medium long plumose setae on anterior margin;
merus with two serrate spines at anterodistal angle;
carpus with one serrate spine at posterodistal angle,
and one on anterodistal angle; propodus with 4 (3)
short serrate spines on posterior margin; setulose
fringe present on posterior margin of ischium to
beginning of unguis (distal part of article 7, claw),
near continuous on anterior margin of ischium to
unguis; secondary unguis finely serrate. Pereopod VII
basis with one short plumose seta on anterior margin
and one at anterodistal angle; ischium with one short
plumose seta on anterior margin; merus with one
serrate and one biserrate spine at anterodistal, angle
and one short plumose seta on posterodistal angle;
carpus with two serrate spines on posterior margin,
one at posterodistal angle, one biserrate spine at
anterodistal angle, and two on distal margin; propodus with three serrate spines on posterior margin,
one at posterodistal angle and one short plumose seta
at anterodistal angle; setulose fringe present from
posterodistal angle of ischium to beginning of unguis,
anterior margins mainly without setules, except for
distalmost parts of ischium, merus, and carpus,
setules extremely short; secondary unguis hidden
within thick mat of unresolved structure. Relative
length of pereopods I, II, and VII: 29, 40, and 55% (30,
52, and 55%) of body length; length ratio of articles
(coxa excluded) in pereopod I 100 : 63 : 45 : 20 :
76 : 46 (100 : 61 : 41 : 23 : 64 : 35), in pereopod II
100 : 56 : 38 : 57 : 62 : 36 (100 : 62 : 40 : 56 : 67 : 37),
in pereopod VII 100 : 95 : 41 : 67 : 81 : 35 (100 :
93 : 43 : 64 : 78 : 32).
Pleopod I protopodite with many fine long setules
along external margin, fewer and shorter setules
along internal margin, and and two strong spines at
internal angle; exopodite elliptical, with six (7)
plumose setae along terminal margin, scarce slender
short spines on upper surface, and setules at
proximo-external angle; endopodite about half as
wide and nearly as long as exopodite, proximally
almost parallel sided, distally slightly tapering, with
three plumose setae on rounded terminal margin,
without setules at proximo-internal angle. Pleopod II
similar to I, but with more numerous plumose setae;
endopodite as wide as exopodite, slightly widened
distally, with 29 (28) plumose setae on terminal
margin; appendix masculina distally sicle-shaped,
apically pointed, its length 120% (116–119%) of
endopodite length; exopodite with 15 (13) plumose
setae on terminal margin, and slender short spines
on the upper distal surface. Pleopod III exopodite
elongate subovoid, without respiratory area, with
long transverse suture at its external margin and
short suture at the internal margin. Pleopod IV of
similar shape, transverse suture at the lateral
margin almost reaching respiratory area; area
length 71% (76%) of exopodite length, surface 50%
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
(54%) exopodite surface. Pleopod V exopodite irregularly elliptical without thick sclerotized ridge, but
with three slender spines along proximal half of
external margin; three sclerotized patches on mediodistal half differently shaped and less densely scaled
than in M. (Microlistra) jalzici sp. nov.; respiratory
area on interoproximal half, its length 39% (48%) of
exopodite length, surface 30% (36%) of exopodite
surface. Vestigial uropods as in M. (Microlistra)
jalzici sp. nov.
Description of paratype females: Females of 11.9 and
13.3 mm in length; larger than males; body width
43–44% of body length. Pleotelson width 126–128% of
pleotelson length, dorsocaudal vault surpassing pleotelson ventrocaudal border to greater extent than in
males.
Antenna I length 25% of body length; of ten flagellar articles, articles 3–6 and 8, each with single long
aestethasc. Antenna II length 34–38% of body length,
flagellum of 14 or 15 articles. Pereopod I spines and
setulose fringe as in males, but carpus with two
biserrate spines at posterodistal angle; propodus with
four biserrate spines on posterior margin and one or
two at posterodistal angle. Pereopod II spines and
setulose fringe as in males, but merus with one or two
serrate spine at anterodistal angle. Pereopod VII
spines and setulose fringe as in males, but merus
with one serrate spine at anterodistal angle; carpus
with one serrate spine at posterodistal angle and two
biserrate spines at anterodistal angle; propodus with
between one and three serrate spines on posterior
margin, none or one at posterodistal angle, and one
short plumose seta at anterodistal angle. Relative
length of pereopods I, II, and VII 27–29, 38–41, and
50–54% of body length, respectively; length ratio of
their articles (coxa excluded) in pereopod I
100 : 45 : 35 : 17 : 61 : 35/100 : 52 : 34 : 17 : 58 : 35, in
pereopod II 100 : 57 : 34 : 51 : 62 : 35/100 : 58 : 37 :
53 : 64 : 34 in pereopod VII 100 : 77 : 35 : 57 :
73 : 30/100 : 90 : 42 : 64 : 83 : 32.
Pleopod I and II as in males, but pleopod II with 24
(25) plumose setae along distal endopodite margin.
Pleopods III and IV shapes as in males. Pleopods IV
and V respiratory area length 71 and 28% of
exopodite length, respectively; pleopods IV and V
respiratory area surface 42 and 40% of exopodite
surface, respectively.
Distribution and ecology: Specimens were found
scarcely in the residual pools in deeper parts of the
cave Kuruzovića pećina (= Kukuruzovićeva pećina,
K. špilja), functioning periodically as a boiling
spring, near Vaganac, Kordun, Croatia. They were
accompanied by a few specimens of two additional
17
species, M. (Monolistra) caeca and M. (Monolistrella)
sp., some shrimps (Troglocaris sp., Decapoda),
slightly troglomorphic specimens of Synurella ambulans O.F. Müller (Amphipoda), more numerous
unidentified Cyclopoidea (Copepoda), and single
specimens of troglobiotic Proasellus sp. (Isopoda)
and Niphargus steueri Schellenberg (Amphipoda).
Besides crustaceans, numerous snails, Hydrobioidea
(Gastropoda), few Oligochaeta and Nematoda, and
only three specimens of certainly trogloxene Chironomidae larvae (Diptera) were present in the
pools. The other locality is the big karst spring of
Slunjčica, ~20 km in the north-western, i.e. ‘Dinaric’,
direction.
Remarks: The anterior and the posterior margins of
the pereopod articles are covered with an unidentified
layer of varying thickness, considerably hindering
the observation and illustration of the pereopods; the
lining may represent an extremely dense crust of the
interlaced short setulae, and/or mats of bacteria
attached to setae, or of something else.
Monolistra (Microlistra) fongi sp. nov. is most
similar to the related type species Monolistra
(Microlistra) spinosa by its appearance: its dorsal
processes are similarly arranged, but remarkably
shorter. Its epimera are apically blunt, whereas they
are sharply pointed in Monolistra spinosa and some
other species with long processes. The huge respiratory areas on exopodites of pleopods IV and V are
even larger than in Monolistra (Microlistra) spinosa,
Monolistra (Microlistra) spinosissima, and Monolistra
(Microlistra) sketi.
DISCUSSION
TAXONOMY, BIOGEOGRAPHY
Monolistra calopyge was included in the subgenus
Microlistra (Sket, 1982) because of its pleotelson
shape, the absence of subchelae on male pereopods,
reduced uropods, and presence of a long and apically
curved appendix masculina, but its assignment to
Microlistra is dubious. It differs from all other
Microlistra spp. because males are larger than
females, male pereopod II is more differentiated, pleopods IV and V have reduced respiratory areas, and
there is no sculpturing on tergites, including those
surrounding the uropods. Molecular data for this
species are not available.
Excluding Monolistra (? Microlistra) calopyge, the
subgenus Microlistra appears quite homogeneous in
less obvious morphological characters, i.e. pereopod
and pleopod morphology. This is in strong contrast
with the differentiation seen in M. (Monolistra) caeca.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
18
S. PREVORČNIK ET AL.
In its area of distribution, less than 150 km in linear
extension, races with moderately large to reduced or
absent respiratory areas on pleopods are encountered
(Sket, 1964). In contrast, pleopod respiratory areas
remain virtually unchanged within more than 300 km
of the distribution range of Microlistra species.
Microlistra species exhibit extreme diversity in their
overall appearance and dorsal sculpturing; the sculpturing may be absent, present in the form of small
tubercles (nodules), or present as large/long processes. Nodules are rare among nearly 30 other
species and subspecies of the genus Monolistra, occurring only in Monolistra (Monolistra) monstruosa Sket,
1970.
Both major clades revealed by our molecular
study are well separated geographically, and we can
roughly designate them as the southern and the
northern clades. The species belonging to the southern clade are distributed along the outermost
Dinaric mountain chains, draining towards the Adriatic. Monolistra (Microlistra) pretneri is found near
Šibenik, crossing to Cres Island, whereas Monolistra
(Microlistra) jalzici sp. nov. inhabits the Istra Peninsula. The distribution of Monolistra (Microlistra)
schottlaenderi could represent an extension of the
southern area, although slightly displaced. Nevertheless, in our molecular phylogeny, this species
belongs to the spinosa–spinosissima species group in
the northern clade. The latter clade is orographically less defined, covering a long, wide area from
Italy, through South Slovenia, to Croatia, between
its borders with Slovenia and Bosnia. Monolistra
(Microlistra) bolei is positioned centrally, partly surrounded by related taxa. The area inhabited by the
species belonging to the northern clade is partly
drained towards the Adriatic [M. (Microlistra) schottlaenderi and Monolistra (Microlistra) sketi], but
mainly towards the Black Sea. Each species of the
subgenus is related to the subterranean parts of
present river systems; this is characteristically not
the case in some other crustacean species, including
some other taxa within the genus Monolistra (Sket,
1986b, 2002; Trontelj et al., 2007).
Both main clades are also morphologically welldefined. The bulk of species of the northern clade
possess smooth processes, gradually tapering towards
their tips, and extended pereonal epimera, the elongation of their pointed tips progressing simultaneously with the progressive elongation of processes.
Members of the southern clade never have smooth
processes: ornamentation elements are either small
nodules or large warts, widened in the middle and not
narrowly pointed, and pereonal epimera are short and
wide, always obtuse. The exception is the ‘northern’
species Monolistra (Microlistra) schottlaenderi, which
is morphologically of the ‘southern’ type.
PHYLOGENY
Our study indicates that the previous calculations
(Trontelj et al., 2007) regarding the timing of splits
within the subgenus might have been inaccurate. The
Microlistra group had already separated from its sister
clades in the beginning of the Pleistocene (see above),
but persisted without further splitting for a comparatively long time afterwards. The first split within the
group most probably occurred within the period of
1.5–1.1 Mya, which means that it might even be
slightly younger than the split between the putative
subspecies of the related M. (Monolistra) caeca (2.0–
1.5 Mya; Trontelj et al., 2007). Such a late split may
perhaps explain the great similarity of pleopods within
the Microlistra clade, in contrast with the pleopods
within M. (Monolistra) caeca. On the other hand, the
habitus of the latter is very homogeneous because of
the lack of any prominent ornamentation.
The youngest speciations within the subgenus
Microlistra, resulting in a weak molecular divergence and little variability in pleopod morphology,
are corroborated further by the restriction of species
to recent river drainages. The persistence of both
the genetic and morphological characters in the
Microlistra clade, despite its occurrence in a geographically extremely heterogeneous area, can only
mean that its predecessors lived and survived in
surface waters even after the formation of recent
drainages. Namely, areas of most other troglobiotic
species and even subspecies are bound to past
(palaeo-) river drainages. Therefore, distributions of
Microlistra species are in strong contrast to the
palaeo-hydrographically defined distributions of M.
(Monolistra) caeca races. These races are not only
morphologically and/or molecularly distinct, but they
occur in a number of separated current drainages
(Sket, 2002).
Different dorsal sculpturing (see above) of both the
main Microlistra clades (the southern and the northern clades) could only have evolved after the first
split. An extreme case of different sculpturing is the
difference between the sister species Monolistra
(Microlistra) schottlaenderi and Monolistra (Microlistra) spinosa. Furthermore, Monolistra (Microlistra)
bolei has its processes positioned as no other species
within its sister lineage, which also indicates independent evolution of its dorsal structures. All this
shows that the development of the sculpturing is a
result of convergent evolution, which resulted in patterns differing in details. Less revealing are the
absence of sculpturing (in some Monolistra (Microlistra) pretneri) and a weak development of processes
[in Monolistra (Microlistra) sketi] in some terminal
branches, as these might be the results of secondary
reductions of an originally richer ornamentation. As
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
all species are allopatric, we do not consider the
development of dorsal structures to be enforced by the
need for reproductive isolation (i.e. by a selective
reinforcement). Nevertheless, as sphaeromatids are
copulating animals, such differences in body shapes
could certainly prevent interbreeding. This is the
reason why we consider the named taxa as ‘biological’
species, regardless of their molecular similarities.
It is a fact that dating with a molecular clock
results in quite broad potential time-spans for speciations. Nevertheless, with the possible exception of the
first intrageneric split, all divergence events clearly
fall within the geologically (particularly hydrographically) active and climatically highly fluctuating Pleistocene. Taking into consideration our previous
statements about the speciation in Monolistra (Sket,
1986b), these late speciation events could only mean
that some Monolistra populations had to survive glaciations in surface freshwaters, comparatively close to
the Alpine glacier.
ACKNOWLEDGEMENTS
We are very thankful to D.C. Culver for revising the
English language of the manuscript. France Velkovrh,
Branko Jalžić, Jana Bedek, Tonći RaËa, and Hrvatsko
Biospeleološko Društvo provided some samples. The
study was supported by the Research Agency of
Slovenia.
REFERENCES
Bruce NL. 1994. The Cassidininae Hansen, 1905 (Crustacea:
Isopoda: Sphaeromatidae) of Australia. Journal of Natural
History 28: 1077–1173.
Deeleman-Reinhold CL. 1971. Deux Monolistrini nouveaux
(Crustacea: Isopoda) des eaux souterraines de Croatie.
International Journal of Speleology 3: 205–213.
Edgar RC. 2004. MUSCLE: multiple sequence alignment
with high accuracy and high throughput. Nucleic Acids
Research 32: 1792–1797.
ICZN (International Commission on Zoological Nomenclature). 1999. International code of zoological nomenclature, 4th edn. London: The International Trust for Zoological
Nomenclature. On-line version. Available at: http://
www.iczn.org/iczn/index.jsp
Karaman SL. 1954. Über die jugoslavischen Arten des Genus
Monolistra. Acta Musei Macedonici Scientiarum Naturalium 2: 125–143.
Kumar S, Tamura K, Nei M. 2007. MEGA4: Molecular
Evolutionary Genetics Analysis (MEGA) Software Version
4.0. Molecular Biology and Evolution 24: 1596–1599.
Marković-Marjanović J. 1971. Coast lines, Pleistocene sediments and fauna of the eastern part of Adriatic in Yugoslavia. Quaternaria 15: 187–195.
Mayden RL. 1997. A hierarchy of species concepts: the
19
denouement in the saga of the species problem. In: Claridge
MF, Dawah HA, Wilson MR, eds. Species: the units of
biodiversity. London: Chapman & Hall, 381–424.
Mayr E, Ashlock PD. 1991. Principles of Systematic Zoology,
2nd edition. New York: McGraw-Hill.
Nylander JAA. 2004. MrModeltest v2.2. Program distributed
by the author. Uppsala University: Evolutionary Biology
Centre.
Posada D, Crandall KA. 1998. Modeltest: testing the model
of DNA substitution. Bioinformatics 14: 817–818.
Racovitza EG. 1910. Sphéromiens (premiere série) et révision des Monolistrini. Archives de Zoologie expérimentale et
générale, 5e Sér. 4: 625–758, Tables 18–31.
Racovitza EG. 1929a. Microlistra spinosa n. g., n. sp.,
isopode sphéromien cavernicole nouveau de Slovénie.
Buletinul Societăţii de ştiinţe din Cluj 4: 89–96.
Racovitza EG. 1929b. Microlistra spinosissima, n. sp.,
isopode sphéromien cavernicole nouveau de Slovénie.
Buletinul Societăţii de ştiinţe din Cluj 4: 107–115.
Ronquist F, Huelsenback JP. 2003. MRBAYES 3: Bayesian
phylogenetic inference under mixed models. Bioinformatics
19: 1572–1574.
Ruffo S, Stoch F. (eds). 2000. Checklist of the species of the
Italian fauna. On-line version 2.0. Available at: http://
www.checklist.faunaitalia.it/checklist/index-html
Schubart CD, Diesel R, Hedges SB. 1998. Rapid evolution
to terrestrial life in Jamaican crabs. Nature 393: 363–365.
Simon C. 1991. Appendix 3. In: Hewitt GM, Johnston A,
Young JP, eds. Molecular techniques in taxonomy. Berlin:
Springer-Verlag, 345–355.
Sket B. 1960. Einige neue Formen der Malacostraca aus
Jugoslawien, III. Bulletin Scientifique 5: 73–75.
Sket B. 1964. Östliche Gruppe der Monolistrini. I. Systematischer Teil. International Journal of Speleology 1: 163–
189, Tab. 22–44.
Sket B. 1965. Östliche Gruppe der Monolistrini. II. Biologischer Teil; Nachtrag. International Journal of Speleology 1:
249–267, Tab. 60–66.
Sket B. 1967. Crustacea Isopoda (aquatica). Catalogus faunae
Jugoslaviae, Cons. Acad. Sc. RPSF Jugoslaviae 3: 1–21.
Sket B. 1970. Presenetljive novosti v jamski favni Bosanske
Krajine (Überraschende Novitäten der Höhlenfauna in der
Bosanska Krajina). Naše jame 11: 93–100.
Sket B. 1982. Some news about the subgenus Microlistra
(Isopoda, Sphaeromatidae) in the subterranean waters of
Yugoslavia. Biološki vestnik 30: 143–154.
Sket B. 1986a. Isopoda: Sphaeromatidae. In: Botosaneanu L,
ed. Stygofauna Mundi. Leiden: Brill & Backhuys, 423–
427.
Sket B. 1986b. Evaluation of some taxonomically, zoogeographically, or ecologically interesting finds in the hypogean
waters of Yugoslavia (in the last decades). Comunicacions,
9. Congreso Internacional de Espeleologia 1: 126–128.
Sket B. 1988. Zoogeografija sladkovodnih in somornih rakov
(Crustacea) v kvarnersko-velebitskem območju (Zoogeography of the freshwater and brackish Crustacea in the
Kvarner-Velebit islands (NW Adriatic, Yugoslavia). Biološki
vestnik 36: 63–76.
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
20
S. PREVORČNIK ET AL.
Sket B. 1994. Distribution patterns of some subterranean
Crustacea in the territory of the former Yugoslavia. Hydrobiologia 287: 65–75.
Sket B. 1997. Distribution of Proteus (Amphibia: Urodela:
Proteidae) and its possible explanation. Journal of Biogeography 24: 263–280.
Sket B. 2002. The evolution of the karst versus the distribution and diversity of the hypogean fauna. In: Gabrovšek F,
eds. Evolution of karst: from Prekarst to cessation.
Ljubljana-Postojna: Založba ZRC, 225–232.
Sket B, Bruce NL. 2004. Sphaeromatids (Isopoda, Sphaeromatidae) from New Zealand fresh and hypogean waters,
with description of Bilistra n. gen. and three new species.
Crustaceana 76: 1347–1370.
Stammer HJ. 1930. Eine neue Höhlensphäromide aus
dem Karst, Monolistra (Typhlosphaeroma) schottlaenderi,
und die Verbreitung des Genus Monolistra. Zoologischer
Anzeiger 88: 291–304.
Stammer HJ. 1932. Die Fauna des Timavo. Zoologische
Jahrbücher, Abteilung Systematik 63: 521–656.
Stoch F. 1984. Su una nuova ‘Monolistra (Crustacea,
Isopoda)’ delle acque sotterranee del Friuli e osservazioni
sulla distribuzione dei Monolistrini nell’Italia nordorientale.
Atti Museo civico Storia naturale Trieste 36: 61–67.
Sturmbauer C, Levinton JS, Christy J. 1996. Molecular
phylogeny analysis of fiddler crabs: test of the hypothesis of
increasing behavioral complexity in evolution. Proceedings
National Academy Sciences of the United States of America
93: 10855–10857.
Trontelj P, Gorički Š, Polak S, Verovnik R, Zakšek V,
Sket B. 2007. Age estimates for some subterranean taxa
and lineages in the Dinaric Karst. Acta carsologica 36:
183–189.
Verovnik R, Sket B, Prevorčnik S, Trontelj P. 2003.
Random amplified polymorphic DNA diversity among
surface and subterranean populations of Asellus aquaticus
(Crustacea: Isopoda). Genetica 119: 155–165.
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Name
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
M.
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
(Mic.)
fongi
jalzici
pretneri
pretneri (spinulosa)
pretneri
pretneri
pretneri
pretneri
pretneri
pretneri
pretneri
sketi
sketi
sp. (cf. sketi)
schottlaenderi
schottlaenderi
schottlaenderi
schottlaenderi
schottlaenderi ssp.
bolei bolei
bolei bolei
bolei brevispinosa
bolei brevispinosa
bolei brevispinosa
spinosa
spinosa
spinosa
spinosa
spinosa ssp.
spinosa ssp.
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
spinosissima
calopyge
DNA
Loc. typ.
Locality name
Nearby settlement
Country
WGS_Xdd
WGS_Ydd
D
D
D
D
D
T
T
T
(T)?
cave Kuruzovića pećina
spring in Tunnel Čepić
cave Pećina kod Vrane
cave Miljacka spilja 1
cave Milića špilja
spring 1 km east Žegar
spring below cave Miljacka spilja 1
spring at power plant Jaruga
cave Kusa nad Manastirskom lukom
cave Kusaća jama
well in Punta Križa
cave Pećina kod sela Pećina
cave Markovac jama
spring of Rupećica
springs Moščenice
cave Abisso Massimo
spring at Lago di Pietrarossa/Laško jezero
springs at Monfalcone
spring Fonte Oppia/Klinčica
spring Otovški zdenec
cave Stobe
cave Džud
cave Jama v kamnolomu – 2950
cave Lobešnica
spring Tominčev studenec
cave Bobnova jama
spring below Rivčja jama
spring Rupnica
water works in Slovenska vas
cave Mobišaht
cave Logarček
springs Bistra
cave Gašpinova jama
resurgence-cave Izvir pod orehom
resurgence-cave Malo Okence
cave near Mirke
cave Najdena jama
resurgence-cave Retovje
resurgence-cave Veliko okence
resurgence-cave in D9Verd
well in Kronovo
Pašina Luka, Drežnik Grad, Rakovica
Kožljak, Čepićko polje, Podpićan
Pećina, Vrana, Pakoštane
Miljacka mlin, Kistanje, Knin
Milići, Bogatnik, Kaštel Žegarski
Žegar, kaštel Žegarski, Obrovac
Miljacka mlin, Kistanje, Knin
Skradin, Šibenik
Manastir Krupa, Pirevište, Obrovac
Žegar, Kaštel Žegarski
Cres (island)
Pećina, Ličko Lešče
Kangrge, Podum, Otočac
Ivanci, Ogulin
San Giovanni/Štivan, Monfalcone
Sgonico/Zgonik, Prosecco/Prosek
Monfalcone/Tržič
San Giovanni/Štivan, Monfalcone
Bagnoli/Boljunec, Trieste/Trst
Otovec, Črnomelj
Petrova vas, Črnomelj
Belčji Vrh, Črnomelj
Vinica
Stara Lipa, G. Suhor pri Vinici, Črnomelj
Žužemberk
Stavča vas, Novo mesto
Male Rebrce, Šmihel pri Žužemberku
Rupe, Stična
Slovenska vas, Kočevje
Dolenja vas, Ribnica
Laze, Logatec
Vrhnika
Logatec
Verd
Verd, Vrhnika
Verd, Vrhnika
Laze, Planina
Verd, Vrhnika
Verd, Vrhnika
Vrhnika
Šmarješke toplice
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
CRO
ITA
ITA
ITA
ITA
ITA
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
SLO
15,7025507
14,15998546
15,55492234
16,01861835
15,80311235
15,84712646
16,01861835
15,96267831
15,88169165
15,84249204
14,49492522
15,33076608
15,30447971
15,22480539
13,58426023
13,73284909
13,56025
13,59154
13,87227759
15,16412441
15,17073562
15,20062456
15,24430696
15,17706432
14,96855801
14,94481174
14,87005342
14,79747913
14,83022311
14,78954216
14,26821292
14,33310623
14,23573699
14,29966281
14,29541835
14,29309948
14,24548199
14,2958184
14,2958284
14,31112931
15,25622146
44,91472811
45,17021515
43,95979217
44,00313313
44,14297815
44,15461241
44,0031331
43,80726946
44,1904967
44,13953176
44,6394318
44,79670227
44,86137889
45,18782454
45,80086511
45,71827308
45,81481
45,79123
45,62050931
45,59227634
45,60426056
45,50239495
45,45443663
45,49778204
45,79628292
45,81039298
45,85604346
45,95186554
45,6676524
45,70822136
45,86494137
45,94603575
45,91070877
45,95250727
45,95144627
45,95429292
45,87630166
45,94969846
45,94969446
45,95558924
45,85633622
D
D
D
D
D
T
T
D
D
D
T
T
D
T
D
D
T
D
D
D
D
T
BIOGEOGRAPHY AND PHYLOGENY OF MONOLISTRA
© 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 159, 1–21
APPENDIX 1
List of known localities of Monolistra (Microlistra) spp. with coordinates. Abbreviations: M., Monolistra; Mic., Microlistra; D, sequences for phylogenetic analysis;
T, type locality; CRO, Croatia; ITA, Italy; SLO, Slovenia.
21