Zoological Journal of the Linnean Society, 2009, 156, 785–800. With 9 figures
The Iberian Peninsula: ancient history of a hot spot of
mite harvestmen (Arachnida: Opiliones: Cyphophthalmi:
Sironidae) diversity
zoj_512
785..800
JÉRÔME MURIENNE1,2 and GONZALO GIRIBET1*
1
Department of Organismic and Evolutionary Biology and Museum of Comparative Zoology, Harvard
University, 26 Oxford Street, Cambridge, MA 02138, USA
2
UMR 5202 CNRS, Département Systématique et Evolution, case 50, Muséum national d’Histoire
naturelle, 45 rue Buffon, 75005 Paris, France
Received 15 April 2008; accepted for publication 9 July 2008
The Iberian Peninsula represents a hot spot of cyphophthalmid (mite harvestman) disparity, with four of the eight
genera currently recognized in the family Sironidae represented in the region – a generic diversity and morphological disparity not found in any other region of the World so far. From these, two genera (Iberosiro and
Odontosiro) are monotypic, and are restricted to the western side of the peninsula. Parasiro is restricted to the
north-east region, from the Catalonian Coastal Ranges and both sides of the Eastern Pyrenees, in areas where the
annual rainfall surpasses 1000 mm, and mostly restricted to areas with Paleozoic and Variscan rocks, with other
species of the genus extending to Corsica, Sardinia, and the Italian Peninsula. A second species of the genus
Paramiopsalis, Paramiopsalis eduardoi sp. nov. from Fragas do Eume, is described here along with a
re-diagnosis of the genus. Paramiopsalis species, together with Odontosiro, inhabit the north-west corner of the
Iberian Peninsula, an area with some of the highest recorded annual rainfall, and with Paleozoic rocks from the
Iberian Massif or Variscan granitoid rocks. A phylogenetic analysis of the members of the family Sironidae using
four molecular markers, despite not including all of the Iberian genera, clearly shows the non-monophyly of the
Iberian Cyphophthalmi, indicating that the Iberian Peninsula is home to multiple ancient lineages of mite
harvestmen. The two Paramiopsalis species form a sister clade to the Balkan genus Cyphophthalmus, whereas
Parasiro constitutes the first lineage of the sironids represented.
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800.
doi: 10.1111/j.1096-3642.2008.00512.x
ADDITIONAL
KEYWORDS: new species – Paramiopsalis – Portugal – Southern Europe – Spain
.
INTRODUCTION
Cyphophthalmi are present in virtually all continental land masses or on islands of continental origin.
Within these land masses, peaks of species diversity
have been described or observed in such places as
New Zealand, Sumatra, Borneo, or the Balkans.
These radiations often account for single lineages,
although in some cases up to three lineages of independent origin have been observed. Such is the case
for New Zealand, which has perhaps the beststudied cyphophthalmid fauna, from a taxonomic
*Corresponding author. E-mail: [email protected]
and evolutionary point of view (Forster, 1948, 1952;
Boyer & Giribet, 2003, 2007, 2009). Only two lineages occur in the much larger land mass of Australia (Boyer & Giribet, 2007), whereas places such
as Sumatra and Borneo may host a single or at
most two lineages each, despite the large number of
species that inhabit those large islands (Clouse &
Giribet, 2007). In this article we focus on a land
mass that has been isolated from other terranes
during a long period of history: the Iberian Peninsula (Fig. 1). This territory is home to just a few
species of Cyphophthalmi, which nonetheless belong
to four genera, exhibiting greater morphological disparity than any other land mass of equivalent size.
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
785
786
J. MURIENNE and G. GIRIBET
Paramiopsalis eduardoi*
Paramiopsalis sp.
O. lusitanicus
Paramiopsalis sp.
Paramiopsalis ramulosus*
O. lusitanicus
Paramiopsalis sp.
Iberosiro sp.
Odontosiro sp.
Paramiopsalis sp.
O. lusitanicus
Parasiro coiffaiti
O. lusitanicus
Paramiopsalis
ramulosus
Parasiro coiffaiti*
Parasiro coiffaiti
?
Parasiro
coiffaiti
Parasiro coiffaiti*
Parasiro coiffaiti
Odontosiro lusitanicus
Paramiopsalis ramulosus
Iberosiro distylos
Figure 1. Satellite view centered on the Iberian Peninsula showing the known localities for the Iberian species of
Cyphophthalmi. Type localities are identified with the bold font. An asterisk denotes the localities of the specimens used
in the molecular study. Localities identified as ‘Paramiopsalis sp.’ are based on specimens listed by Rambla & Fontarnau
(1984) as Paramiopsalis ramulosus, but have not been examined by the authors. The type locality for Parasiro coiffaiti
is indicated with a question mark because the locality listed for the type material is not from the Girona Province (see
Results).
Furthermore, a new species of Cyphophthalmi from
Galicia is described and illustrated.
HISTORY
OF CYPHOPHTHALMID RESEARCH IN
THE
IBERIAN PENINSULA
A recent catalogue of the Iberian Opiliones (Prieto,
2004)1 lists four species of Cyphophthalmi for this
biogeographical area, namely Parasiro coiffaiti Juberthie, 1956, Odontosiro lusitanicus Juberthie, 1961,
Paramiopsalis ramulosus Juberthie, 1962, and Iberosiro distylos de Bivort & Giribet, 2004. Detailed
accounts for these species have been provided else1
Although the publication date of this issue of the Revista
Ibérica de Aracnología indicates 31.xii.2003, it includes
species published in 2004 such as Iberosiro distylos de Bivort
& Giribet (2004), and therefore could have not appeared until
2004.
where (Rambla & Fontarnau, 1984, 1985, 1986; de
Bivort & Giribet, 2004). Older accounts also report
the presence of Cyphophthalmus duricorius Joseph,
1868 (as Siro duricorius) as a result of an error
perpetuated in the literature (Kraus, 1961; Rambla,
1974), which was corrected later on by Rambla &
Fontarnau (1984: 146–147; see also Prieto, 2004; de
Bivort & Giribet, 2004).
Our knowledge on the Iberian Cyphophthalmi is
mostly derived from the work of two of the most
prominent European opilionologists: the French
Christian Juberthie and the Catalan Maria Rambla.
The first three known species from the Iberian Peninsula were all described by C. Juberthie in three
different genera, two of which were monotypic (Juberthie, 1956, 1961, 1962). The fourth species also
belongs to a monotypic genus (de Bivort & Giribet,
2004). In this article we revise the Iberian cyphoph-
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
thalmid fauna, adding a second species for the former
monotypic genus Paramiopsalis Juberthie, 1962, and
provide a phylogenetic analysis of selected species of
the family Sironidae, using four molecular markers in
order to test for monophyly of the Iberian lineages.
MATERIAL AND METHODS
MORPHO-ANATOMY
The male holotype and a female paratype of the new
species, and the male lectotype of Parasiro coiffaiti,
were photographed in dorsal, ventral, and lateral
positions using a JVC KY-F70B digital camera
mounted on a Leica MZ 12.5 stereomicroscope. A
series of images was taken at different focal planes
and assembled with the dedicated software package
Auto-Montage Pro v5.00.0271 (Syncroscopy). A male
paratype was examined using an FEI Quanta 200
Scanning Electron Microscope (SEM), after sputter
coating with gold/palladium.
Observation and description of Opiliones genitalia
are classically conducted using light microscopy. In
this paper, we used a novel method of spermatopositor observation using confocal laser scanning microscopy (CLSM; Klaus & Schawaroch, 2006). This study
presents the first use of CLSM to describe the structure of a new species. The method relies on the
natural fluorescence property of the arthropod cuticle
to obtain a stack of 2D sections that can be combined
into a 3D object. Regarding the extremely small size
and fragility of the structure, observation with light
microscopy and preparation for electron microscopy
was impossible. In addition, to be relatively fast and
787
easy (with the whole process taking approximately
1 h), this technique is also non-destructive, which is a
great advantage when working with collection specimens. After dissection, the spermatopositor was
mounted in glycerin between a glass slide and a cover
slip, as for observation with light microscopy. Details
of experimental protocols can be found in Klaus,
Kulasekera & Schawaroch (2003) and Schawaroch,
Grimaldi & Klaus (2005). The structure was imaged
on a Zeiss LSM 510 meta confocal microscope. Threedimensional visualization was carried out using
Maximum Intensity Projection in the Zeiss LSM
Image Browser software.
TAXON
SELECTION
In this study, we use 14 species represented by 16
specimens in the molecular data set, including all the
available representatives of the Iberian Peninsula, as
well as other selected Cyphophthalmi (see Table 1).
All previous analyses of cyphophthalmid phylogeny
show that Metasiro, once classified within Sironidae,
belongs to the family Neogoveidae, and therefore it is
used to root the trees. Furthermore, many of our
analyses indicate that the genera Parasiro and
Suzukielus do not form a clade with the other
sironids, or that they are early offshoots of the sironid
lineage (Boyer, Karaman & Giribet, 2005; Boyer et al.,
2007b), thereby justifying our choice of outgroups.
In addition to the phylogenetic analyses, we used
three additional specimens of Paramiopsalis ramulosus collected in May 2008 for calculating within- and
among-species molecular distances.
Table 1. Museum of Comparative Zoology (MCZ) accession numbers and species for all represented taxa used in the
molecular analyses, with GenBank accession numbers
Family
Species
Neogoveidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
Sironidae
DNA101532
DNA100487
DNA100499
DNA100494
DNA100493
DNA101878
DNA102080
DNA100459
DNA101383
DNA101877
DNA100488
DNA100489
DNA101611
DNA100457
DNA100461
DNA101543
Metasiro americanus
Cyphophthalmus duricorius
Cyphophthalmus ere
Cyphophthalmus martensi
Cyphophthalmus minutus
Paramiopsalis eduardoi sp. nov.
Paramiopsalis eduardoi sp. nov.
Paramiopsalis ramulosus
Parasiro coiffaiti
Parasiro coiffaiti
Siro acaroides
Siro exilis
Siro kamiakensis
Siro rubens
Siro valleorum
Suzukielus sauteri
18S rRNA
28S rRNA
16S rRNA
COI
DQ825542
AY639461
AY639462
AY639470
AY639473
EU638284
EU638285
AY639489
AY918872
EU638283
AY639490
AY639491
DQ513147
U36998
AY639492
DQ513138
DQ825595
DQ513120
AY639499
AY639505
DQ825591
EU638287
–
DQ513121
DQ513122
EU638286
DQ513128
AY639520
DQ513134
DQ825584
DQ513123
DQ513116
DQ825616
AY639526
AY639527
AY639535
AY639537
EU638281
EU638282
AY639550
AY918877
–
AY639551
–
–
–
AY639552
DQ518086
DQ825645
AY639556
AY639557
AY639562
AY639565
EU638288
EU638289
DQ825641
DQ825642
–
DQ825643
AY639579
DQ513115
DQ513111
AY639580
DQ513108
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
788
J. MURIENNE and G. GIRIBET
MOLECULAR
DATA
Molecular markers included two nuclear ribosomal
genes (complete 18S rRNA, 18S hereafter, and a 1-kb
fragment of 28S rRNA, 28S hereafter), one mitochondrial protein-encoding gene, cytochrome c oxidase
subunit I, and mitochondrial 16S rRNA (16S hereafter). These markers have proven to be informative in
many studies on Opiliones systematics (e.g. Boyer
et al., 2005, 2007b).
The DNEasy® tissue kit was used for tissue lysis
and DNA purification, following the manufacturer’s
protocol (Qiagen). Total DNA was extracted by incubating the entire animal or appendage in lysis
buffer overnight, as described in Boyer et al. (2005).
The intact cuticle of the animal was then removed
after the lysis step and kept in ethanol. Purified
genomic DNA was used as a template for PCR
amplification. The complete 18S (c. 1.8 kb) was
amplified in three overlapping fragments of c.
900 bp each, using primers pairs 1F-5R, 3F-18Sbi,
and 18Sa2.0-9R (Giribet et al., 1996; Whiting et al.,
1997). The first c. 1000 bp of the 28S rRNA was
amplified using the primer set 28SD1F/28SrD1a28Sb (Whiting et al., 1997; Park & Ó Foighil, 2000;
Edgecombe & Giribet, 2006), or alternatively with
the more internal forward primer 28Sa (Whiting
et al., 1997). COI was amplified using the primer
pair LCO1490/HCOoutout (Folmer et al., 1994; Schwendinger & Giribet, 2005), or alternatively with
the more internal reverse primer HCO2198 (Folmer
et al., 1994). 16S was amplified with the primer pair
16Sar-16Sb (Xiong & Kocher, 1991; Edgecombe,
Giribet & Wheeler, 2002). Polymerase chain reactions (PCR; 50 mL) included 2 mL of template DNA,
1 mM of each primer, 200 mM of dinucleotidetriphosphates (Invitrogen), 1¥ PCR buffer, containing 1.5 mM MgCl2 (Applied Biosystems), and
1.25 units of AmpliTaq DNA polymerase (Applied
Biosystems). The PCR reactions were carried out
using a GeneAmp PCR System 9700 thermal cycler
(Applied Biosystems), and involved an initial denaturation step (5 min at 95 °C), followed by 35 cycles
including denaturation at 95 °C for 30 s, annealing
(ranging from 44 to 49 °C) for 30 s, and an extension at 72 °C for 1 min, with a final extension step
at 72 °C for 10 min. The double-stranded PCR products were verified by agarose gel electrophoresis (1%
agarose), and were then purified with a Perfectprep
PCR Cleanup 96 system (Eppendorf). The purified
PCR products were sequenced directly with the
same primer pairs used for amplification. Each
sequence reaction contained a total volume of 10 mL,
including 2 mL of PCR product, 1 mM of one of the
PCR primer pairs, 2 mL ABI BigDye 5x sequencing
buffer, and 2 mL ABI Big Dye Terminator v3.0
(Applied Biosystems). The sequencing reactions
involved an initial denaturation step for 3 min at
95 °C, and 25 cycles (95 °C for 10 s, 50 °C for 5 s,
and 60 °C for 4 min). The BigDye-labelled PCR
products were then cleaned using Performa DTR
Plates (Edge Biosystems). The sequence reaction
products were then analysed using an ABI Prism
3730xl Genetic Analyzer. Chromatograms were
edited, and overlapping sequence fragments for each
individual were assembled using the software
SEQUENCHER 4.7 (Gene Codes Corporation
1991–2007), after BLAST searches (Altschul et al.,
1997), as implemented by the NCBI website
(http://ncbi.nlm.nih.gov/), were conducted to check
for putative contamination. The software package
MACGDE: Genetic Data Environment for MacOSX
(Linton, 2005) was used to determine fragments
based on internal primers and secondary structure
features (Giribet & Wheeler, 2001; Giribet, 2002).
18S, 28S, and 16S rRNAs were divided into 2, 9,
and 5 fragments, respectively, according to internal
primers and secondary structure features. The COI
was divided in six fragments. All new sequences
have been deposited in GenBank under accession
codes EU638281–EU638289 (see Table 1).
PHYLOGENETIC
ANALYSES
Phylogenetic analyses were conducted under Direct
Optimization (Wheeler, 1996; Wheeler et al., 2006)
with POY version 4.0 RC build 2615 (Varón, Vinh &
Wheeler, 2008). Tree searches were conducted under
parsimony with 20 random-addition sequences, followed by subtree pruning and regrafting (SPR) and
tree bissection and reconnection (TBR), and one
round of ratcheting (Nixon, 1999). A detailed description of the commands used to implement this search
strategy with the new version of POY can be found in
Murienne, Harvey & Giribet (2008). In order to assess
nodal stability (Wheeler, 1995; Giribet, 2003a), different parameter sets were used for a range of indel-totranversion and transversion-to-transition ratios. We
used ‘111’, ‘121’, ‘211’, and ‘3221’ transformation
matrices. It has been argued that the parameter set
‘111’ (often called ‘equal weighting’ or ‘unweighted
parsimony’) maximizes explanatory power (Frost
et al., 2001), but it requires all indel events to be
counted independently. Parameter set ‘3221’ was
implemented
using
the
commands
transform(tcm:(2,1), gap_opening:3). Nodal support was estimated by jackknifing (Farris et al., 1996;
Farris, 1997) with 100 replicates using the command
transform (auto_sequence_partition:true),
which increases the number of fragments – and thus
the number of characters – used for jackknifing. The
data for all genes were analysed simultaneously.
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
Molecular evolutionary analyses for estimating
p-distances and the standard error associated for COI
sequence data were conducted using MEGA v3.1
(Kumar, Tamura & Nei, 2004). For these analyses we
used three additional specimens of Paramiopsalis
ramulosus from northern Portugal.
PARASIRO
RESULTS
COIFFAITI JUBERTHIE, 1956 (FIG. 2)
Parasiro coiffaiti was described based on an unspecified series of specimens collected by M. H. Coiffait
A
B
C
in the Pyrénées-Orientales (France) and Girona
Province (Spain) (Juberthie, 1956). The species is
endemic to the north-east Iberian Peninsula (provinces of Girona and Barcelona) and south-east of
France (Pyrénées-Orientales) (Juberthie, 1957, 1958;
Rambla, 1974, 1986), with a relatively broad distribution. Parasiro coiffaiti is found on the slopes of
the Eastern Pyrenees, Montnegre, Montseny (a
mountain range west of the coastal hills north of
Barcelona), and Collserola (the mountain that
delimits the city of Barcelona in the west) (Rambla,
1974, 1986; Rambla & Fontarnau, 1985). It preferentially inhabits argillaceous soils or slate, and has
also been found in two caves in Tavertet (Osona)
and Parc Natural de Sant Llorenç del Munt i Serra
de l’Obac (Rambla, 1986). The type material (five
males and two females), deposited at the Muséum
national d’Histoire naturelle, Paris (MNHN), is supposedly from the Girona Province (Catalonia, Spain),
but we were unable to locate the labelled locality
‘Porto de Santigoso’ in Girona, and rather than
Catalan, the locality name looks Galician or Portuguese. Santigoso is indeed a rural town in the Galician Province of Ourense, in north-western Spain.
Interestingly, most of the remaining Iberian diversity of Cyphophthalmi is in Galicia, and we suspect
that the type locality of ‘Porto de Santigoso’ may be
a labelling error. Juberthie could not have mistaken
Parasiro coiffaiti for any of the Galician or Portuguese species, and therefore we suspect that the
labelling error did not affect the specimens contained in the vial. A male from this vial has been
designated as the lectotype here, and is illustrated
in Figure 2; the remaining four males and two
females are designated as paralectotypes. Some of
the historical collections that were examined
by the authors include the following localities: Olot,
Riells, Terrassa, Tavertet (Cova Vora Fosca),
and Sant Feliu de Pallerols, all of which are in
Catalonia.
New collections: Font del Vidre (Berga, Berguedà,
Barcelona, Catalonia, Spain), Vall de Fuirosos (Montnegre Massif, Girona, Catalonia, Spain).
Parasiro coiffaiti is related to Parasiro corsicus
(Simon, 1872) and Parasiro minor Juberthie, 1958,
from Corsica, Sardinia, and the Italian Peninsula
(Simon, 1872; Juberthie, 1958; Brignoli, 1968; de
Bivort & Giribet, 2004).
ODONTOSIRO
Figure 2. Parasiro coiffaiti Juberthie, 1956, lectotype
male. A, dorsal view. B, ventral view. C, lateral view. Scale
bars: 0.5 mm.
789
LUSITANICUS
JUBERTHIE, 1961
Odontosiro lusitanicus Juberthie, 1961 was originally
described from four specimens (one male, two
females, and one juvenille) collected by M. H. Coiffait
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
790
J. MURIENNE and G. GIRIBET
in Guimarães,2 Portugal (Juberthie, 1961), and was
given its own generic status because of the unique
combination of type-1 ozophores (as in Parasiro) and
the presence of a complete corona analis (as in Siro
and Cyphophthalmus).
Rambla & Fontarnau (1984) reported two new
localities for O. lusitanicus, this time in Spain (in the
provinces of Pontevedra and León), and more details
on these plus three additional localities in Ourense,
Lugo and Asturias were provided by Rambla & Fontarnau (1986). The species is therefore known from
its type locality in northern Portugal, and from
the notrthwestern Spanish autonomous regions of
Galicia, Asturias and León. We have been unable to
locate these specimens in the collection of Maria
Rambla or at the MNHN. During G. Giribet’s recent
collecting trips to Guimarães (Braga), little suitable
habitat for Cyphophthalmi was identified, as most
forested areas were Eucalyptus plantations. Nearby
localities in the Costa Verde of Portugal and Galicia
did not yield any specimens of this genus. Odontosiro
remains a monotypic genus to date. This is also the
only known genus of Sironidae for which no specimen
has been examined by the authors. O. lusitanicus has
often been collected with P. ramulosus, although the
latter tends to be much more abundant (Rambla &
Fontarnau, 1986).
IBEROSIRO
DISTYLOS DE
BIVORT & GIRIBET, 2004
A single male specimen from Algarve (cave) da Terra
da Rolha, Rocha Forte, Serra de Montejunto
(Estremadura, Portugal), collected by A. de Barros
Machado in April 1941, and deposited at the MNHN
(Paris), was described as yet another monotypic
Iberian genus (de Bivort & Giribet, 2004). This
species constitutes the southernmost distribution of
Iberian Cyphophthalmi.
With a length of c. 1.4 mm, similar to that of O.
lusitanicus, it combines character states observed in
Paramiopsalis and Odontosiro. As in Paramiopsalis,
I. distylos has the coxa of the second pair of legs fused
to the third coxa, a unique trait within the Sironidae.
However, it shares with Odontosiro the ozophore position (which is also similar to that of Parasiro), the
type of leg ornamentation, and the anal plate that is
devoid of modifications.
So far the holotype remains the only known specimens for this species, despite a recent visit (May
2008) to the alleged type locality by G. Giribet.
However, a male and a female of a congener have
2
Juberthie only mentioned that the specimens were collected
in Guimaraes by Coiffat in the North of Portugal. There are
two localities in the North of Portugal named Guimarães, one
in the Province of Braga, and another in the Province of
Viseu.
been recently collected by pitfall trapping in Illano
(Asturias), and the specimens were forwarded to the
authors by C. Prieto after the first set of proofs of this
article. These specimens will be described and discussed in a forthcoming paper.
PARAMIOPSALIS JUBERTHIE, 1962
Emended diagnosis: Small sironids with free coxae I;
coxae II fused to coxae III and IV. Eyes absent.
Ozophores of type II. Sternum absent. Opisthosomal
dorsal longitudinal sulcus absent. Posterior border of
opisthosoma truncated in males and rounded in
females. Opisthosomal tergite IX free, and sternites 8
and 9 medially fused, for most of their length, not
forming a corona analis. A single anal gland pore
opening on tergite VIII. Basal cheliceral article
without dorsal crest. Tarsus and metatarsus of legs I
and II smooth; metatarsus of legs III and IV ornamented, but tarsus smooth. Claws of walking legs
smooth, without any sort of ornamentation. Tarsus IV
of males entire. Adenostyle large, lamelliform, or
plumose. Spermatopositor short, with bifurcating
dorsal microtrichia with thick bases, and shorter
ventral microtrichia with thinner bases. Pars apicalis
with hooked mobile digits. Ovipositor long, with a
multibranched, sensitive process on each lobe.
This diagnosis corresponds to the one provided for
the genus by Juberthie (1962: 267), with the exception of the adenostyle type, which is plumose only in
the type species of the genus.
Included species: Paramiopsalis ramulosus and Paramiopsalis eduardoi sp. nov.
Phylogenetic position: In recent molecular analyses
(Boyer et al., 2007b; this analysis) the genus Paramiopsalis is currently seen as sister to the genus
Cyphophthalmus, but its position has fluctuated
within Sironidae in other molecular analyses (Boyer
et al., 2005). These analyses did not include the
genera Iberosiro or Odontosiro. Morphologically,
Paramiopsalis is the closest relative of Iberosiro (de
Bivort & Giribet, 2004), although with low support.
Unfortunately, Iberosiro specimens are not available
for molecular study.
PARAMIOPSALIS
RAMULOSUS
JUBERTHIE, 1962
The third species described for the Iberian Peninsula
was also described originally from Portugal, from
Pessegueiro, in the Aveiro Province, based on specimens collected by M. H. Coiffait in April 1960. Later,
its range was extended to the north-west Iberian
Peninsula, with several localities in the provinces of A
Coruña, Pontevedra, and León, in Spain (Rambla &
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
Fontarnau, 1984). We have not been able to locate the
type specimens of the species, and all illustrations
appearing in the literature (Rambla & Fontarnau,
1984; de Bivort & Giribet, 2004) are from specimens
collected in Moscoso. New collections (see below) from
the Costa Verde, in northern Portugal, also belong to
this species.
With c. 1.8 mm in carapace length, this is the
largest Iberian cyphophthalmid species. It was originally described as belonging to the subfamily Stylocellinae (sensu Hansen & Sørensen, 1904), for having
the coxa of leg II fused to that of leg III, as do the
members of the currently recognized families Stylocellidae, Ogoveidae, and Neogoveidae (Hansen &
Sørensen, 1904; Shear, 1980). Its systematic position
was later on emended by Shear (1980), who placed it
within its current family, Sironidae. The unique
plumose adenostyle (that gives origin to the specific
epithet; see Rambla & Juberthie, 1994: plate I, figs
4–6; de Bivort & Giribet, 2004: fig. 32i), the typical
Siro ozophores, anal plate, and anal gland, or the
typical Parasiro–Odontosiro anal region (with sternites 8 and 9 fused, but not fused with tergite IX; see de
Bivort & Giribet, 2004: fig. 38i), make an original
combination of characters that legitimately deserve
generic status. Although Paramiopsalis and Odontosiro overlap in their areas of distribution, they are
clearly different morphologically.
New collections: Ponte San Miguel, Parque Nacional
da Peneda-Gerês, Serra do Gerês (Braga Province,
Região Norte, Portugal), Ponte do Rio Omen, Parque
Nacional da Peneda-Gerês, Serra do Gerês (Braga
Province, Região Norte, Portugal), Bouza de Mo,
Parque Nacional da Peneda-Gerês, Serra do Gerês
(Braga Province, Região Norte, Portugal), near
Celeiro (Braga Province, Região Norte, Portugal),
Garfe, Póvoa de Lanhoso (Braga Province, Região
Norte, Portugal).
New Paramiopsalis species: Recent collections from
Fragas do Eume, A Coruña (Galicia), by E. Mateos in
April and November 2006 yielded seven male, five
female, and three juvenile specimens of a new species
of Paramiopsalis that we describe here, in his honour.
Fragas do Eume is a unique ecosystem in Europe.3
Bordering the river of the same name (84 km long),
the trees (black poplars, alders, and oaks) are the
main elements of an ecological system (Atlantic
Forest) that is very characteristic, thanks to the proximity of the sea and the deep riverbanks, which led to
the creation of a nature reserve in 1997.
3
Information about Fragas do Eume comes from a website to
promote tourism in the Park consulted in November 2006:
http://www.eumeturismo.org/eng/portada.html
791
According to legend, when God created the three
rivers that rise in the Xistral mountain range (Eume,
Landro, and Masma), he promised a man every year
to the river that reached the sea first. Betrayed by the
other two, which left it sleeping, the Eume had to
traverse valleys and mountains in order to win. That
is why it ended up being rough and wild, and why it
takes the life of a man every year, or used to, before
the reservoirs were built.
There are more than 9000 hectares of different
kinds of woodland along the banks of five municipalities. Nearest to the river grow riverbank forests of
black poplars and alders, with ferns and mosses,
characteristic of the park, and also two rare species of
daffodils. Oak forests appear further up the mountainous slopes. Although the flora is the best appreciated biota of Fragas, it also hosts some endemic and
endangered animal species, including small amphibians, reptiles, numerous birds of prey, and large
mammal species, such as the wolf.
PARAMIOPSALIS
EDUARDOI SP. NOV.
(FIGS 3–8)
Type material: Male holotype (Museum of Comparative Zoology, Harvard University, MCZ, ex MCZ
DNA101878; Fig. 3) from Fragas do Eume, A Coruña
Province (Galicia, Spain), collected 12 April 2006 by
E. Mateos. One male paratype (MCZ DNA101878; two
legs used for DNA extraction), same collecting data as
the holotype. One male, in SEM stubs, and one female
(Fig. 4) paratypes (MCZ, ex MCZ DNA101878), same
collecting data as the holotype. Four male and four
female paratypes (MCZ DNA102080) from Fragas do
Eume, A Coruña Province (Galicia, Spain), collected
10 November 2006 by E. Mateos.
Additional
material:
Three
juveniles
(MCZ
DNA102080; one specimen used for DNA extraction)
from Fragas do Eume, A Coruña Province (Galicia,
Spain), collected 10 November 2006 by E. Mateos.
Diagnosis: As for the genus. Adenostyle lamelliform,
as opposed to the plumose type found in Paramiopsalis ramulosus. Anal plate with a longitudinal ridge,
thinner than that of Paramiopsalis ramulosus.
Description: Total length of male holotype (in mm),
1.40; largest body width in prosoma behind ozophores, 0.72; width across ozophores, 0.67; body
length/width ratio, 1.94. Body orange-brown when
preserved in 96% ethanol. Anterior margin of dorsal
scutum straight without lateral projections; prosomal
region trapezoidal (Figs 3A, 4A). Eyes absent
(Figs 3C, 4C). Ozophores conical (slightly round;
Figs 3A, 4A, 5B), of type II (facing laterally slightly
above edge of carapace; Figs 3C, 4C), with subtermi-
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
792
J. MURIENNE and G. GIRIBET
nal ozopore (‘plugged’, as in de Bivort & Giribet,
2004); ornamentation uniform and non-directional
(Fig. 5B). Transverse prosomal sulcus inconspicuous;
transverse opisthosomal sulci inconspicuous (Figs 3A,
4A). Dorsal scutum convex; maximum width in prosomal area, behind ozophores (Figs 3A, 4A). Opisthosomal part of dorsal shield wider than ventral side
(Figs 3B, 4B).
Ventral prosomal complex of males with coxae I, II,
and IV meeting in the midline, the latter for a distance longer than the gonostome length; coxae III not
meeting in the midline (Fig. 5A, C). Coxae I, II, and
IV with broad endites; coxal pore between coxae III
and IV not observed (Fig. 5A, C). Coxae II and III
with small processes running along their sutures;
coxae III and IV with conspicuous processes running
along their sutures (Fig. 5A, C). Small projections of
coxae IV endite present in the anterior portion of the
gonostome wall (Fig. 5C). Coxa I free, coxa II fused to
coxae III and IV. Sternum absent (Fig. 5C). Male
gonostome pentagonal, almost triangular, wider than
long (99 ¥ 69 mm), with an almost straight (slightly
concave) posterior margin, and delimited laterally
and anterolaterally by the elevated endites of coxa IV
(Fig. 5A, C).
Spiracles oval in shape, closed, with a maximum
diameter of 51 mm (Fig. 5E). Ventral opisthosomal
region without conspicuous modifications or gland
openings (Fig. 5A). Opisthosomal tergite IX free, and
sternites 8 and 9 medially fused for most of their
length, not forming a corona analis (Fig. 5A, D). Anal
plate slightly oval (180 ¥ 144 mm), with a thin medial
ridge; ridge with conspicuous setae (Fig. 5D). Anal
glands not observed with SEM because of the putative secretion accumulated on the dorsal part of
tergite VIII. Cuticle ornamented (with a tuberculatemicrogranular surface; Murphree, 1988) in all ventral
areas, except on the spiracle and coxal endites; anal
plate without tuberculate-microgranular ornamentation, but with a honeycomb pattern (Fig. 5A, C, D).
A
A
B
B
C
C
Figure 3. Paramiopsalis eduardoi sp. nov., holotype
male. A, dorsal view. B, ventral view. C, lateral view. Scale
bars: 0.5 mm.
Figure 4. Paramiopsalis eduardoi sp. nov., paratype
female. A, dorsal view. B, ventral view. C, lateral view.
Scale bars: 0.5 mm.
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CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
793
A
B
D
C
E
Figure 5. Paramiopsalis eduardoi sp. nov., scanning electron microgarphs of a male paratype. A, ventral view of
whole body. B, ozophore. C, prosomal ventral complex. D, anal region. E, spiracle.
Chelicerae (Fig. 6A, B) relatively short and robust;
basal article 362-mm long, 157-mm wide, without conspicuous ventral processes, and without a dorsal
crest; second article 379-mm long, 123-mm wide;
movable finger 182-mm long; widest part of cheliceral
distal article near articulation with mobile digit; all
articles with few setae; proximal article almost
entirely granulated. Second cheliceral segment only
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
794
J. MURIENNE and G. GIRIBET
Table 2. Leg measurements (in mm) given as length/width and total length
Leg
Trochanter
Femur
Patella
Tibia
Metatarsus
Tarsus
Total
I
II
III
IV
101/115
103/103
114/99
174/105
422/126
304/113
293/119
357/115
181/112
153/124
165/113
166/120
249/116
231/123
203/120
224/132
145/92
130/91
101/83
108/96
383/130
370/126
323/105
372/120
1481
1291
1199
1401
A C
B
D
Figure 6. Paramiopsalis eduardoi sp. nov., scanning electron microgarphs of a male paratype. A, right chelicera. B,
cheliceral fingers. C, left pedipalp. D, pedipalpal claw.
partly and slightly ornamented. Cheliceral denticles
of the uniform type (Fig. 6B).
Palp 1.026-mm long, smooth, slightly covered with
scale-like processes on all articles (Fig. 6C). Measurements of palpal articles, length/width (L/W ratio) in
mm: trochanter 123/65 (1.89), femur 284/70 (4.06),
patella 173/67 (2.58), tibia 225/63 (3.57), tarsus
221/57 (3.88); claw 30-mm long (Fig. 6D). Palp
trochanter with a conspicuous ventral process. Ornamentation present in trochanter; absent on second
palp article.
Legs relatively long and robust; see measurements
in Table 2. Leg I longer than leg II. Solea in tarsus I
absent (Fig. 7A). Tarsus and metatarsus smooth in
legs I (Fig. 7A) and II (Fig. 7C); metatarsus almost
completely ornamented in leg III (Fig. 7E), and
entirely ornamented in leg IV (Fig. 7G); all other
articles ornamented on legs I–IV. Tarsus IV of male
entire (Fig. 7G); with a broad lamelliform adenostyle
(102/93 mm), not plumose, subcylindrical at the base,
with a lateral pore; distal margin at 44% of the tarsal
length (not in the most basal region of the tarsus;
Fig. 7G, H). Claws hooked, smooth, and without dentition or lateral pegs (Fig. 7B, D, F, I).
Spermatopositor (Fig. 8) small (180-mm long), with
3/3 dorsal long bifurcating microtrichia with enlarged
bases, and shorter ventral microtrichia without
enlarged bases. Pars apicalis with hooked mobile
digits. Ventral part with four pairs of shorter microtrichia; ventral plate clearly surpasses the pars distalis, only visible from the dorsal side. Ovipositor not
studied.
Etymology: The species is named after our colleague
Eduardo Mateos from the Departament de Biologia
Animal, Universitat de Barcelona, who collected the
specimens of the new species.
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
795
B
A
C
D
E
G
F
H
I
Figure 7. Paramiopsalis eduardoi sp. nov., scanning electron microgarphs of the legs of a male paratype. A,
metatarsus and tarsus I. B, tarsal claw I. C, metatarsus and tarsus II. D, tarsal claw II. E, metatarsus and tarsus III.
F, tarsal claw III. G, metatarsus and tarsus IV. H, detail of the adenostyle. I, tarsal claw IV.
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
796
J. MURIENNE and G. GIRIBET
Figure 8. Paramiopsalis eduardoi sp. nov., confocal
laser scanning micrograph of the spermatopositor of a
male paratype, dorsal view. Total length is 180 mm.
Phylogentics: Our molecular phylogenetic analysis
resulted in a well-resolved and stable phylogeny. We
show the single most parsimonious tree of 2914 steps
obtained under equal weighting (Fig. 9), and represent the nodal stability using sensitivity plots. Except
for Suzukielus sauteri (Roewer, 1916), which appears
as sister to the genus Siro under alternative parameter sets, all nodes appear highly stable. The two
specimens of Paramiopsalis eduardoi sp. nov. form a
clade (100% jackknife proportion), and group with
Paramiopsalis ramulosus (100% bootstrap). The
genus Paramiopsalis appears as sister to the genus
Cyphophthalmus with strong support (93% jackknife
proportion), and under all of the parameter sets
examined.
DISCUSSION
PHYLOGENETICS
The different Iberian species of Cyphophthalmi have
been included in several phylogenetic analyses, both
using morphological (Giribet & Boyer, 2002; Giribet,
2003b; de Bivort & Giribet, 2004) and molecular
(Giribet & Boyer, 2002; Boyer et al., 2005, 2007b)
data. The first morphological analysis included Parasiro coiffaiti and Paramiopsalis ramulosus, along with
representatives of most other cyphophthalmid genera
(Giribet & Boyer, 2002), and placed them in two
independent lineages. A subsequent morphological
analysis including sironid species from all genera
placed O. lusitanicus as sister to the genus Parasiro,
whereas Iberosiro was placed as sister to Paramiopsalis. These two clades were not sisters to each
other.
Molecular data are available for only two of the four
Iberian genera: Paramiopsalis and Parasiro. Whereas
most analyses place Paramiopsalis with other
sironids of the genera Siro and Cyphophthalmus
(Giribet & Boyer, 2002; Boyer et al., 2005, 2007b;
Schwendinger & Giribet, 2005), Parasiro tends to
branch more basally, or even outside Sironidae (Boyer
et al., 2007b). In our analysis, using only one nonsironid outgroup, Metasiro americanus (Davis, 1933),
Parasiro branches out before the core sironid clade
and its sister genus Suzukielus, which is also often
found not to form a clade with the other sironids
(Boyer et al., 2007b) (Fig. 9).
Although the results from those analyses have
increased the amounts of taxa and data, we are still
far from having a comprehensive picture of the phylogeny of the family Sironidae. The resolution from
morphological data is somewhat limited within
Cyphophthalmi because of the low degree of morphological variation of the group (e.g. Giribet & Boyer,
2002; Giribet, 2003b; de Bivort & Giribet, 2004; Boyer
& Giribet, 2007), and therefore molecular data have
proven essential for reconstructing the phylogeny of
the group (e.g. Boyer et al., 2005, 2007b; Schwendinger & Giribet, 2005; Boyer & Giribet, 2007; Clouse
& Giribet, 2007). It is therefore imperative to add
molecular data from Iberosiro and Odontosiro.
The new data strongly suggest that Paramiopsalis
eduardoi sp. nov. is sister to Paramiopsalis ramulosus, as expected by morphology, suggesting that the
plumose adenostyle of Paramiopsalis ramulosus is
apomorphic for the type species and not for the genus.
All other relevant characters for distinguishing the
genus from other sironid genera (e.g. ozophore position, fusion of coxae, leg ornamentation, and anal
region) are identical for both species. At the molecular
level, the two species differ in one nucleotide in the
complete 18S rRNA (out of 1800 bp), a number identical to that found between two cryptic species of
centipedes of the genus Craterostigmus in New
Zealand and Tasmania (Edgecombe & Giribet, 2008).
The average p-distances for COI within Paramiopsalis eduardoi sp. nov. (N = 2) and within Paramiopsalis
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
797
DNA101532 Metasiro americanus
100
DNA101877 Parasiro coiffaiti
DNA101383 Parasiro coiffaiti
DNA101543 Suzukielus sauteri
100
74
DNA100489 Siro exilis
56
89
DNA101611 Siro kamiakensis
DNA100488 Siro acaroides
63
95
DNA100461 Siro valleorum
DNA100457 Siro rubens
41
DNA100459 Paramiopsalis ramulosus
100
100
DNA102080 Paramiopsalis eduardoi sp. nov.
DNA101878 Paramiopsalis eduardoi sp. nov.
93
DNA100499 Cyphophthalmus ere
99
111
121
211 3221
DNA100487 Cyphophthalmus duricorius
90
55
DNA100494 Cyphophthalmus martensi
DNA100493 Cyphophthalmus minutus
Figure 9. Phylogeny of selected members of the family Sironidae, based on the combined analysis of 18S, 28S,
cytochrome c oxidase subunit I (COI), and 16S under direct optimization and equal weighting. The support values on
branches indicate the jackknife frequencies. Each weighting scheme is assigned a code corresponding to the ratio of
indel/transversion, transversion/transition, and transition values. Tree lengths for the different parameter sets are as
follows: 111, 2914; 121, 4585; 211, 3228; 3221, 6168. Black squares indicate monophyly; grey squares indicate that either
the group is paraphyletic or the internal relationships are different.
ramulosus (N = 4) are 0.000 (±0.000 SE) and 0.049
(±0.006 SE), respectively, whereas average p-distance
between the two species is 0.145 (±0.013 SE). For
pairs of Paramiopsalis ramulosus, the p-distances
range between 0.014 and 0.076. This level of withinspecies genetic divergence in COI is much lower than
the average genetic divergences for the pettalid
species Aoraki denticulata (Forster, 1948) reported by
Boyer, Baker & Giribet. (2007a).
Paramiopsalis ramulosus was originally described
from the Aveiro Province in Região Norte of Portugal,
and had been cited from Galicia and León, in northwestern Spain. Recent collections in several localities
across north-western Portugal, in the Braga Province,
by G. Giribet shows that the River Miño/Minho has
not acted as a barrier for this species. Although some
of the citations for Galicia and León could belong to
cryptic species, we have not been able to examine
these specimens (indicated as Paramiopsalis sp. in
Fig. 1). It is clear that the specimens from southern
Galicia and northern Portugal belong to the same
morphospecies, and form a clade (G. Giribet, unpubl.
data), although they exhibit p-distances of up to 0.076
for COI.
As suggested by previous phylogenetic analyses,
the genus Paramiopsalis is sister to Cyphophthalmus,
distributed in the eastern part of Southern Europe
and Turkey, instead of being sister to the Western
Europe/North American genus Siro. This relationship
is supported in 93% of the jackknife replicates, and is
recovered with all of the analytical parameters examined. As seen in previous analyses, our data strongly
suggest that the genus Parasiro represents yet
another lineage of sironids, not directly related to the
Siro clade or to the Cyphophthalmus/Paramiopsalis
group.
PHYSICAL
SETTING
Cyphophthalmi are found in most land masses that
have suitable habitat, generally in temperate to tropical rain forests. Temperate rain forests are most
common in the north-western and northern Iberian
Peninsula, where the annual rainfall is greater than
1000 mm (Ninyerola, Pons & Roure, 2005). However,
cyphophthalmid populations are not distributed
evenly along the northern edge of the Iberian Peninsula. Instead, there is a bimodal distribution: with
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
798
J. MURIENNE and G. GIRIBET
Parasiro found exclusively in the easternmost
Pyrenees and the Catalonian Coastal Ranges, and
with Paramiopsalis and Odontosiro concentrated on
the north-western Iberian Peninsula (northern Portugal, Galicia and León). However, another important
requirement of Cyphophthalmi is the age of the land
mass where they reside (or its immediate adjacency),
as they are extremely poor dispersers (Boyer et al.,
2007a). Not surprisingly, the two areas where
cyphophthalmids are found correspond to regions
with Variscan granitoid rocks and Paleozoic (and Precambrian) rocks of the Iberian Massif of Galicia/
northern Portugal, and in the Variscan granitoid
rocks and Paleozoic inliers of the Eastern Pyrenees
and the Catalonian Coastal Ranges (CCR) (Gibbons &
Moreno, 2002).
As Iberosiro distylos has only been found in one
cave in Montejunto, near Lisbon, rainfall does not
seem to be determinant. Given the geology and rainfall, one could expect Cyphophthalmi in the Pyrenees,
directly north of Pamplona, or in the Internal Betics
(see Gibbons & Moreno, 2002), two areas for which no
species has been reported so far.
ACKNOWLEDGEMENTS
We are indebted to Eduardo Mateos for providing the
specimens for the description of the new species and
to Carlos Prieto for providing specimens of a new
Iberosiro from Asturias. Dave Smith assisted with the
CLSM, and the Center for Nanoscale Systems (CNS)
at Harvard provided SEM support. Ligia Benavides
kindly generated the Automontage photographs.
Arturo Muñoz-Cuevas (Paris) kindly lent the type
specimens of Parasiro coiffaiti and Iberosiro distylos.
M. A. Arnedo helped with the evolutionary analyses
in MEGA. M. A. Arnedo and an anonymous reviewer
provided comments that helped to improve this
manuscript. This material is based upon work supported by the National Science Foundation under
Grant no. 0236871. GG was supported in part by a
sabbatical grant to the Centre d’Estudis Avançats de
Blanes, CSIC, from the Ministerio de Educación y
Ciencia (Spain). JM was supported by a Lavoisier
fellowship from the French Ministry of Foreign
Affairs, and by a Marie Curie fellowship from the
European Commission.
REFERENCES
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z,
Miller W, Lipman DJ. 1997. Gapped BLAST and PSIBLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402.
de Bivort BL, Giribet G. 2004. A new genus of cyphophthalmid from the Iberian Peninsula with a phylogenetic
analysis of the Sironidae (Arachnida: Opiliones: Cyphophthalmi) and a SEM database of external morphology. Invertebrate Systematics 18: 7–52.
Boyer SL, Baker JM, Giribet G. 2007a. Deep genetic divergences in Aoraki denticulata (Arachnida, Opiliones,
Cyphophthalmi): a widespread ‘mite harvestman’ defies
DNA taxonomy. Molecular Ecology 16: 4999–5016.
Boyer SL, Clouse RM, Benavides LR, Sharma P, Schwendinger PJ, Kuranarathna I, Giribet G. 2007b. Biogeography of the world: a case study from cyphophthalmid
Opiliones, a globally distributed group of arachnids. Journal
of Biogeography 34: 2070–2085.
Boyer SL, Giribet G. 2003. A new Rakaia species (Opiliones,
Cyphophthalmi, Pettalidae) from Otago, New Zealand.
Zootaxa 133: 1–14.
Boyer SL, Giribet G. 2007. A new model Gondwanan taxon:
systematics and biogeography of the harvestman family
Pettalidae (Arachnida, Opiliones, Cyphophthalmi), with a
taxonomic revision of genera from Australia and New
Zealand. Cladistics 23: 337–361.
Boyer SL, Giribet G. 2009. Welcome back New Zealand:
Regional biogeography and Gondwanan origin of three
endemic genera of mite harvestmen (Arachnida, Opiliones,
Cyphophthalmi). Journal of Biogeography 36: 1084–1099.
Boyer SL, Karaman I, Giribet G. 2005. The genus Cyphophthalmus (Arachnida, Opiliones, Cyphophthalmi) in Europe: a
phylogenetic approach to Balkan Peninsula biogeography.
Molecular Phylogenetics and Evolution 36: 554–567.
Brignoli PM. 1968. Note su Sironidae, Phalangodidae e
Trogulidae Italiani, cavernicoli ed endogei (Opiliones). Fragmenta Entomologica 5: 259–293.
Clouse RM, Giribet G. 2007. Across Lydekker’s Line – first
report of mite harvestmen (Opiliones: Cyphophthalmi: Stylocellidae) from New Guinea. Invertebrate Systematics 21:
207–227.
De Laet JE. 2005. Parsimony and the problem of inapplicables in sequence data. In: Albert VA, ed. Parsimony,
phylogeny, and genomics. Oxford: Oxford University Press,
81–116.
Edgecombe GD, Giribet G. 2006. A century later – a total
evidence re-evaluation of the phylogeny of scutigeromorph
centipedes (Myriapoda: Chilopoda). Invertebrate Systematics
20: 503–525.
Edgecombe GD, Giribet G. 2008. A New Zealand species of
the trans-Tasman centipede order Craterostigmomorpha
(Arthropoda: Chilopoda) corroborated by molecular evidence. Invertebrate Systematics 22: 1–15.
Edgecombe GD, Giribet G, Wheeler WC. 2002. Phylogeny
of Henicopidae (Chilopoda: Lithobiomorpha): A combined
analysis of morphology and five molecular loci. Systematic
Entomology 27: 31–64.
Farris JS. 1997. The future of phylogeny reconstruction.
Zoologica Scripta 26: 303–311.
Farris JS, Albert VA, Källersjö M, Lipscomb D, Kluge
AG. 1996. Parsimony jackknifing outperforms neighborjoining. Cladistics 12: 99–124.
Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek RC.
1994. DNA primers for amplification of mitochondrial
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
CYPHOPHTHALMI IN THE IBERIAN HOT SPOT
cytochrome c oxidase subunit I from diverse metazoan
invertebrates. Molecular Marine Biology and Biotechnology
3: 294–299.
Forster RR. 1948. The sub-order Cyphophthalmi Simon in
New Zealand. Dominion Museum Records in Entomology 1:
79–119.
Forster RR. 1952. Supplement to the sub-order Cyphophthalmi. Dominion Museum Records in Entomology 1: 179–
211.
Frost DR, Rodrigues MT, Grant T, Titus TA. 2001. Phylogenetics of the lizard genus Tropidurus (Squamata: Tropiduridae: Tropidurinae): direct optimization, descriptive
efficiency, and sensitivity analysis of congruence between
molecular data and morphology. Molecular Phylogenetics
and Evolution 21: 352–371.
Gibbons W, Moreno T, eds. 2002. The geology of Spain.
London: The Geological Society.
Giribet G, Carranza S, Baguñà J, Riutort M, Ribera C.
1996. First molecular evidence for the existence of a
Tardigrada + Arthropoda clade. Molecular Biology and Evolution 13: 76–84.
Giribet G. 2002. Relationships among metazoan phyla as
inferred from 18S rRNA sequence data: a methodological
approach. In: DeSalle R, Giribet G, Wheeler WC, eds.
Molecular systematics and evolution: theory and practice.
Basel: Birkhäuser Verlag, 85–101.
Giribet G. 2003a. Stability in phylogenetic formulations and
its relationship to nodal support. Systematic Biology 52:
554–564.
Giribet G. 2003b. Karripurcellia, a new pettalid genus
(Arachnida: Opiliones: Cyphophthalmi) from Western Australia, with a cladistic analysis of the family Pettalidae.
Invertebrate Systematics 17: 387–406.
Giribet G, Boyer SL. 2002. A cladistic analysis of the
cyphophthalmid genera (Opiliones, Cyphophthalmi). The
Journal of Arachnology 30: 110–128.
Giribet G, Wheeler WC. 2001. Some unusual small-subunit
ribosomal RNA sequences of metazoans. American Museum
Novitates 3337: 1–14.
Hansen HJ, Sørensen W. 1904. On two orders of Arachnida:
opiliones, especially the suborder cyphophthalmi, and
Ricinulei, namely the family cryptostemmatoidae. Cambridge: Cambridge University Press.
Juberthie C. 1956. Une nouvelle espèce d’Opilions Sironidae
de France et d’Espagne: Parasiro coiffaiti n. sp. Bulletin du
Muséum, 2e série 28: 394–400.
Juberthie C. 1957. Notes sur le biotope et la répartition
géographique de quelques opilions français. Bulletin de la
Société zoologique de France 82: 331–336.
Juberthie C. 1958. Révision du genre Parasiro (Opilions,
Sironidae) et descriptions de Parasiro minor n. sp. Bulletin
du Muséum, 2e série 30: 159–166.
Juberthie C. 1961. Étude des Opilions Cyphophthalmes
(Arachnides) du Portugal: description d’Odontosiro lusitanicus g. n., sp. n. Bulletin du Muséum National d’Histoire
Naturelle 33: 512–519.
Juberthie C. 1962. Étude des opilions cyphophthalmes Stylocellinae du Portugal. Description de Paramiopsalis
799
ramulosus gen. n., sp. n. Bulletin du Muséum National
d’Histoire Naturelle 34: 267–275.
Klaus AV, Kulasekera VL, Schawaroch V. 2003. Threedimensional visualization of insect morphology using confocal laser scanning microscopy. Journal of Mircroscopy 212:
107–121.
Klaus AV, Schawaroch V. 2006. Novel methodology utilizing confocal laser scanning microscopy for systematic analysis in arthropods (Insecta). Integrative and Comparative
Biology 46: 207–214.
Kraus O. 1961. Die Weberknechte der Iberischen Halbinsel
(Arach., Opiliones). Senckenbergiana biologica 42: 331–
363.
Kumar S, Tamura K, Nei M. 2004. MEGA3: integrated
software for molecular evolutionary genetics analysis
and sequence alignment. Briefings in Bioinformatics 5: 150–
163.
Linton EW. 2005. MacGDE: genetic data environment for
MacOS X, 2.0 edn. Software. Available at http://
www.msu.edu/~lintone/macgde/
Murienne J, Harvey MS, Giribet G. 2008. First molecular
phylogeny of the major clades of Pseudoscorpiones (Arthropoda: Chelicerata). Molecular Phylogenetics and Evolution
49: 170–184.
Murphree CS. 1988. Morphology of the dorsal integument of
ten opilionid species (Arachnida, Opiliones). The Journal of
Arachnology 16: 237–252.
Ninyerola M, Pons X, Roure JM. 2005. Atlas climático
digital de la Península Ibérica. Metodología y aplicaciones
en bioclimatología y geobotánica. Cerdanyola del Vallès:
Universitat Autònoma de Barcelona.
Nixon KC. 1999. The Parsimony Ratchet, a new method for
rapid parsimony analysis. Cladistics 15: 407–414.
Park JK, Ó Foighil D. 2000. Sphaeriid and corbiculid clams
represent separate heterodont bivalve radiations into freshwater environments. Molecular Phylogenetics and Evolution
14: 75–88.
Prieto C. 2004. [for 2003]. Primera actualización de la
check-list de los Opiliones de la Península Ibérica e Islas
Baleares. Revista Ibérica de Aracnología 8: 125–141.
Rambla M. 1974. Consideraciones sobre la biogeografía de los
Opiliones de la Península Ibérica. Miscellanea Alcobé,
45–56.
Rambla M. 1986. Els opilions. In: Armengol J, Blas M, eds.
Artròpodes I. Barcelona: Fundació Enciclopèdia Catalana,
168–182.
Rambla M, Fontarnau R. 1984. Les Opilions Cyphophthalmes (Arachnida) de la faune iberique: I. Sur Paramiopsalis ramulosus Juberthie, 1962. Revue Arachnologique 5:
145–152.
Rambla M, Fontarnau R. 1985. Les Opilions Cyphophthalmes (Arachnida) de la faune ibérique: II. Sur Parasiro
coiffaiti Juberthie, 1956. Annales de Spéléologie, Paris.
Rambla M, Fontarnau R. 1986. Les Opilions Cyphophthalmes (Arachnida) de la faune ibérique: III. Sur Odontosiro lusitanicus Juberthie, 1961. Mémoires de la Societe
Rayale belge d’Entomologie 33: 171–178.
Rambla M, Juberthie C. 1994. Opiliones. In: Juberthie C
© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800
800
J. MURIENNE and G. GIRIBET
and Decu V, eds. Encyclopaedia Biospeologica. 1 ed. MoulisBoucarest: Société de Biospéologie. 215–230.
Schawaroch V, Grimaldi D, Klaus AV. 2005. Focusing on
morphology: applications and implications of confocal laser
scanning microscopy (Diptera: Campichoetidae, Camillidae
and Drosophilidae). Proceedings of the Entomological
Society of Washington 107: 323–335.
Schwendinger PJ, Giribet G. 2005. The systematics of the
south-east Asian genus Fangensis Rambla (Opiliones:
Cyphophthalmi: Stylocellidae). Invertebrate Systematics 19:
297–323.
Shear WA. 1980. A review of the Cyphophthalmi of the
United States and Mexico, with a proposed reclassification
of the suborder (Arachnida, Opiliones). American Museum
Novitates 2705: 1–34.
Simon E. 1872. Notice sur les Arachnides cavernicoles et
hypogés. Annales de la Société entomologique de France, Sér.
5 2: 215–244.
Varón A, Vinh LS, Wheeler WC. 2008. POY 4.0.0. New
York: American Museum of Natural History. Available at
http://research.amnh.org/scicomp/projects/poy.php
Wheeler WC. 1995. Sequence alignment, parameter sensitivity, and the phylogenetic analysis of molecular data.
Systematic Biology 44: 321–331.
Wheeler WC. 1996. Optimization alignment: the end of multiple sequence alignment in phylogenetics? Cladistics 12:
1–9.
Wheeler WC, Aagesen L, Arango CP, Faivovich J, Grant
T, D’Haese C, Janies D, Smith WL, Varón A, Giribet G.
2006. Dynamic homology and phylogenetic systematics: a
unified approach using POY. New York: American Museum
of Natural History.
Whiting MF, Carpenter JM, Wheeler QD, Wheeler WC.
1997. The Strepsiptera problem: phylogeny of the holometabolous insect orders inferred from 18S and 28S ribosomal
DNA sequences and morphology. Systematic Biology 46:
1–68.
Xiong B, Kocher TD. 1991. Comparison of mitochondrial
DNA sequences of seven morphospecies of black flies
(Diptera: Simuliidae). Genome 34: 306–311.
SUPPORTING INFORMATION
Additional Supporting Information may be found in the online version of this article:
Appendix S1. Voucher details for specimens used in molecular analyses. Repositories abbreviated as follows:
Museum of Comparative Zoology, Harvard University (MCZ).
Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials
supplied by the authors. Any queries (other than missing material) should be directed to the corresponding
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© 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156, 785–800