butterflies – North Africa is the source of ancestral populations of all Pararg

12 Pages·2006·0.15 MB·English

SystematicEntomology(2006),31,621–632 DOI:10.1111/j.1365-3113.2006.00333.x
Speciation in Pararge (Satyrinae: Nymphalidae)
butterflies – North Africa is the source of ancestral
populations of all Pararge species
ELISABET WEINGARTNER, NIKLAS WAHLBERG and SO¨ REN NYLIN
DepartmentofZoology,StockholmUniversity,Stockholm,Sweden
Abstract. The genus Pararge comprises three species: P. aegeria, distributed in
Europe and North Africa; P. xiphia, endemic to Madeira; and P. xiphioides,
endemic to the Canary Islands. Two subspecies are recognized in P. aegeria,
P. a. tircis and P. a. aegeria, distributed in northern and southern Europe,
respectively. In the 1970s, P. aegeria appeared on Madeira. However, despite
the status of P. aegeria as a model species in ecological studies, the evolutionary
history of Pararge remains unknown. We studied the phylogenetic relationships
of the three Pararge species, using the mitochondrial gene cytochrome oxidase
subunit I and the nuclear gene wingless to infer modes and times of speciation. On
the basis of our analyses, Pararge forms a strongly supported monophyletic
group, with the DNA haplotypes of the three species also forming well-supported
monophyletic groups. We found that P. xiphia diverged first from the common
ancestor a maximum of five million years ago, with P. xiphioides and P. aegeria
being sister species that diverged a maximum of three million years ago. The two
subspecies, P. a. tircis and P. a. aegeria, were not distinguishable on the basis of
DNA haplotypes; instead, our data clearly distinguished between European speci-
mens and those from North Africa. Madeiran P. aegeria has North African
haplotypes and thus originated from there rather than from Europe. We hypothe-
size that the Mediterranean Sea forms a strong barrier to dispersal for Pararge
butterflies,andhasdonesoforapproximatelythepastonemillionyears.
Introduction (Nylin et al., 1993, 1995; Gotthard et al., 1994, 2000) and
thermoregulatory ability in different colour morphs (van
Thespeckledwoodbutterfly,Parargeaegeria(Linnaeus,1758), Dyck&Matthysen,1998).However,theevolutionaryhistory
isoneofthemostfrequentlyusedbutterflyspeciesinecological of the genus remains unstudied. This paper focuses on the
studies.Ithasbeenamodelspeciesinstudiesofbehaviourand speciation processes in Pararge Hu¨bner, 1819 to understand
territorialdefence(Davies,1978;Wickman&Wiklund,1983; better the differences found between species and populations
Shreeve,1984,1987),lifehistorytraits(Gotthardetal.,1994, and to provide a phylogenetic framework for comparative
2000; Sibly et al., 1997; van Dyck et al., 1997; van Dyck & studies.
Wiklund, 2002; Stevens, 2004) and the impact of climate The genus Pararge is considered presently to comprise
changeonthespeciesdistributions(Hilletal.,1999).Thetwo three species: P. aegeria, P. xiphia (Fabricius, 1775) and
subspecies,P.a.tircis(Godart,1821)andP.a.aegeria,have P. xiphioides Staudinger, 1871. Previously, these were
beenusedcommonlyincomparativestudiesoflifehistorytraits considered to form a single polytypic species with four
clearly distinguished subspecies, three of which differed in
their genitalia (Higgins, 1975). Today, P. xiphia and
P. xiphioides have separate species status, but P.a. aegeria
Correspondence: Niklas Wahlberg, Department of Zoology, andP.a.tircisaretreatedstillastwoconspecificsubspecies.
StockholmUniversity,10691Stockholm,Sweden.E-mail:niklas. ParargeaegeriaisdistributedthroughoutEurope(eastto
[email protected] the Urals), Asia Minor and the Middle East (Bozano,
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety 621
622 E.Weingartneretal.
1999), and from the southern slopes of the High Atlas in Lasiommatamaera(Linnaeus,1758),L.megera(Linnaeus,
northern Africa to Scandinavia. Pararge aegeria tircis and 1767), L. petropolitana (Fabricius, 1787) and Lopinga
P. a. aegeria are distributed north and south of the Alps, achine (Scopoli, 1763). The origin and voucher identifica-
respectively, with a zone of intermediates in France. The tions of the specimens used in this study are detailed in
two subspecies are distinguishable clearly on morphology: Appendix 1. Specimens are presently stored at (cid:1)20(cid:2)C at
P. a.tircishasayellowgroundcolourandP. a.aegeriahas the Department of Zoology, Stockholm University,
anorangegroundcolour.Since1976,P. a.aegeriahasbeen Sweden, and will be deposited at the Swedish Museum of
recorded also from Madeira, where it is now well estab- NaturalHistory,Stockholm,Sweden.
lished (Owen et al., 1986). Thus, the colonization of
Madeira by P. aegeria offers a unique opportunity to
study the interactions between the endemic P. xiphia and Molecularmethods
the newcomer (Owen et al., 1986; Jones, 1992; Shreeve &
Smith,1992;Jonesetal.,1998). We extracted DNA from two legs of the frozen or
The other two species, P. xiphia and P. xiphioides, dried butterflies using a QIAGEN (Hilden, Germany)
are endemic to Madeira and the Canary Islands, respec- DNeasy Extraction Kit following the manufacturer’s
tively. Pararge xiphioides has been recorded from the instructions.
CanaryIslandsofGranCanaria,Tenerife,LaGomeraand Wesequencedtwogeneregions:onemitochondrialgene,
La Palma. With such species distributions, the speciation cytochrome oxidase subunit I (COI), of 1450bp, and one
processinParargeislikelytohavetakenplaceallopatrically. nuclear gene, wingless, of 407bp. COI was sequenced for
As species of the genus Pararge frequently are used as all seventy-one individuals sampled. From these seventy-
models in ecological studies, it is of great importance to oneindividuals,wechosetwenty-threeindividualsshowing
understandtheirevolutionaryhistory.Ithasbeensuggested the most divergent COI sequences for which we then also
that Madeira was colonized first by individuals carried by sequenced wingless. Polymerase chain reactions (PCRs)
strong air currents or, in the case of P. aegeria, perhaps were performed in a 20ml reaction volume. The protocol
deliberately or accidentally introduced by humans (Owen used was: 1 ml DNA, 11.5 ml H O, 2 ml 10(cid:3) buffer, 2 ml
2
etal.,1986).Wehypothesizethatthecommon ancestorof MgCl2(25 mM),1 mlofeachprimer,0.4mldNTP(10 mM)
Pararge species colonized both Madeira and the Canary and 0.1ml Taq Gold. The cycling profile was 95(cid:2)C for
Islands, followed by speciation due to isolation. However, 5 min, 35 cycles of 94(cid:2)C for 30s, 47(cid:2)C for 30s, 72(cid:2)C
wereMadeiraandtheCanaryIslandscolonizedbyindivid- for 1min 30s and 72(cid:2)C for 10min. We used the primers
ualsfromNorthAfricaorfromtheIberianPeninsula?The ofBrower&DeSalle(1998)forwinglessandofWahlberg&
colonization of Madeira by P. aegeria might offer a suffi- Zimmermann(2000)forCOI.
ciently recent event, in which it is still possible to find the The same primers were used in the sequencing reaction.
samehaplotypeinpopulationsonthemainland.Thisisone Inaddition,forCOI,theinternalprimer‘Patty’(Wahlberg
oftheaimsofthispaper.Inaddition,wewanttodetermine & Nylin, 2003) was used. The sequencing reactions were
whether P. aegeria is divided into two genetically differen- performed in a total volume of 20ml: 13.5 ml H O, 1.5 ml
2
tiated subpopulations corresponding to the two currently 10(cid:3)buffer,2mlCEQ2000Mix,2mlprimerand1 mlDNA.
recognized subspecies, that is, are P. a. aegeria and Sequencing was undertaken using a Beckman Coulter
P. a. tircis separate entities, with different mitochondrial (Bromma, Sweden) CEQ8000 capillary sequencer. All
haplotypes,aswecanexpectfrommorphology? sequence chromatograms were checked in the program
BIOEDIT(Hall,1999)andalignedbyeye.
Materialsandmethods
Phylogeneticanalyses
Sampling
In total, three datasets were analysed: COI and
Pararge aegeria was sampled as widely as possible to winglessaloneandbothgenescombined.Forthecombined
maximize the number of represented haplotypes analysis,weusedonlythoseindividuals(twenty-threespeci-
(Appendix 1). Both subspecies are represented by several mens) for which both genes had been sequenced (see
populations: P. a. aegeria from populations in southern Appendix1).
Portugal, central Spain, southern France and southern The most parsimonious cladograms were searched for
Greece; P. a. tircis from populations in Sweden, the UK from theequally weightedandunordereddatamatrix using
andBelgium.Parargexiphioideswassampledfromthreeof a heuristic search algorithm in the program NONA 2.0
the four islands where it is present, but unfortunately we (Goloboff,1998)viatheprogramWINCLADA1.00.08(Nixon,
could not obtain specimens from the island of Gran 2002). The heuristic searches were conducted with 200
Canaria. Pararge xiphia was sampled from various local- randomadditionreplicatesusingtreebisection-reconnection
ities on Madeira. As outgroup taxa, we chose representa- (TBR)branchswapping,withtentreesheldduringeachstep
tives from other genera and species belonging to the same and a final swapping to completion. The robustness of the
tribe, Parargini: Kirinia roxelana (Cramer, 1777), phylogenywasevaluatedusingbootstrap(Felsenstein,1985)
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
PhylogenyofPararge 623
and Bremer supportvalues (Bremer, 1988, 1994). Bootstrap Dating of the phylogenies was made with the program
valueswereobtainedusing1000replicates.Whencalculating R8S (Sanderson, 2004). We used three algorithms to esti-
Bremer supports, TREEROT version 2 (Sorensen, 1999) was mate the ages of divergence: the Langley–Fitch method
used in conjunction with PAUP 4.0b10 (Swofford, 2001) to (which assumes a global molecular clock), nonparametric
defineconstrainttreesforeachnode.Themostparsimonious ratesmoothing(Sanderson,1997)andpenalizedlikelihood
trees without a particular node were found in PAUP using a (Sanderson,2002).Forthepenalizedlikelihoodmethod,we
heuristic search with random additions of twenty replicates estimated the value of the smoothing parameter using a
andonetreeheldduringeachstep.Maxtreeswassetto500 cross-validationprocedure.Inallcases,topreventthealgo-
andbranchswappingwasperformedusingTBR.Toevaluate rithms from converging on a local optimum, the searches
the contribution of each dataset to the combined result, we werebegunatfivedifferentinitialtimeestimates.Thelocal
used partitioned Bremer support values (Baker & DeSalle, stability of the solutions for each estimate was checked by
1997;Gatesyetal.,1999).Pairwisesequencedivergencesand perturbing them and restarting the search five times. We
base frequencies were calculated using MEGA version 3 compared our age estimates with those obtained from the
(Kumaretal.,2004). now standard constant mutation rate of 2.3% pairwise
sequencedivergencepermillionyears(Brower,1994).
Errors in age estimates resulting from the stochastic
natureofsubstitutionprocesseswereassessedusingaboot-
Timingofdivergences strap resampling procedure, with the help of Perl scripts
madeavailablebyEriksson(2005).Onehundredbootstrap
Fordatingpurposes,themostparsimonioustreefromthe replicates of the combined dataset were constructed using
combinedanalysiswasprunedtocontainonerepresentative the SEQBOOT program (Felsenstein, 1993), and branch
of each major Pararge lineage (i.e. one P. xiphia, one lengthswerecalculatedusingmaximumlikelihoodandthe
P. xiphioides from each island, one Moroccan P. aegeria GTR þG þ I model for each replicate and then input to
and one P. aegeria from Europe), together with the out- the R8S program. The divergence ages were estimated for
groups. Branch lengths for this tree were estimated from the bootstrapped datasets using nonparametric rate
the combined dataset using maximum likelihood and a smoothing and fixing the age of the split between the La
GTRþ Gþ Imodel.Branchlengthsandparametervalues Palma and La Gomera lineages of P. xiphioides at two
were estimated in PAUP 4.0b10 (Swofford, 2001) with and millionyears.Bootstrapestimatesofthestandarddeviation
without a molecular clock enforced. The evolution of the for eachnodewerecalculatedforthe agedistribution esti-
sequences in a clocklike manner was tested with the like- matesobtained.
lihoodratiotest.Ifthelikelihoodsofthemodelsaresignifi-
cantlydifferent,themolecularclockcanberejected.Owing
to practical problems with estimating branch lengths at the Results
root node (see accompanying instructions of Sanderson,
2004), after the branch lengths had been estimated, two The COI dataset (n¼ 71 individuals) consisted of 1450
outgroup taxa (Kirinia Moore, 1893 and Lopinga Moore, base pairs, of which 235 were parsimony informative. The
1893) were pruned from the tree, leaving Pararge and its base frequencies were: T, 40.9%; C, 15.4%; A, 30.1%; G,
putativesistergenusLasiommataWestwood,1841. 13.6%.Intheheuristicsearch,110treesoflength604steps
To obtain reliable age estimates for clades, the ages of were found (CI¼0.71, RI ¼0.92). The strict consensus
particular nodes need to be fixed or constrained using treewithbootstrapvaluesisshowninFig.1.
external information from fossils or geological events. In Thewinglessdataset(n ¼ 23)consistedof407basepairs,
thepresentcase,nofossilsofeitherParargeoritsputative ofwhichthirty-threewereparsimonyinformative.Thebase
sister genus Lasiommata are known. There are two geo- frequencies were: T, 19.2%; C, 28.4%; A, 22.6%; G,
logical events that potentially can be used to calibrate our 29.7%. The analysis found a single most parsimonious
age estimates. The age of Madeira, at five million years tree of length seventy steps (CI ¼0.91, RI¼ 0.94). The
(Geldmacher & Hoernle, 2000), can be used to place the tree,withbranchlengths,isshowninFig. 2.
maximum age of the divergence between the ancestor of Analysisofthecombineddataset(n ¼23)generatedone
P.xiphiaandtheothertwoParargespecies.However,this mostparsimonioustree(Fig.3)with647steps(CI ¼0.74,
estimateiscomplicatedbythefactthattheislandofPorto RI ¼ 0.83). Of the 1857 characters, 260 were parsimony
Santoismucholder(fourteenmillionyears;Geldmacher& informative.
Hoernle, 2000) and, although it does not support popula- ThegenusParargeissupportedstronglyasa monophy-
tionsofP.xiphianow,itmayhavedonesointhepast.The letic group by both the COI and wingless datasets (Figs 1
second calibration age is a more certain maximum age, and 2). We find that P. aegeria is the sister species to
namely the age of the island of La Palma at two million P. xiphioides and that P. xiphia is sister to these two. In
years(Ancocheaetal.,1994).Currentmethodsrequirethat theCOIanalyses,theP.xiphioidespopulationsondifferent
atleastonenode agebefixed.Wetherefore fixed thesplit islands are monophyletic with high bootstrap support
between the La Gomera lineage and the La Palma lineage values. The population from Tenerife is sister to those
ofP.xiphioidesattwomillionyears. from La Gomera and La Palma. However, using wingless,
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
624 E.Weingartneretal.
Kirinia roxelana
Lopinga achine
Lasiommata maera
Lasiommata megera
Lasiommata petropolitana
Pararge xiphia 24-32
100 Pararge xiphia 5-1
Pararge xiphia 24-29
Pararge xiphia 3-12
Pararge xiphia 5-2
Pararge xiphia 24-28
Pararge xiphia 24-31
Pararge xiphia 24-30
Pararge xiphia 24-27
100 Pararge xiphioides 3-10 Te
Pararge xiphioides 28-16 Te
70 100 Pararge xiphioides 28-17 Te
Pararge xiphioides 27-1 Lg
98 75 PParaarragreg ex ixpihpihoiiodiedse s2 72-76- 2L pLg
100 Pararge xiphioides 28-19 Lp
Pararge xiphioides 28-18 Lp
Pararge xiphioides 27-4 Lp
Pararge xiphioides 27-5 Lp
Pararge xiphioides 27-7 Lp
Pararge a. aegeria 6-4 Ma
Pararge a. aegeria 27-12 MC
Pararge a. aegeria 27-8 MC
Pararge a. aegeria 25-7 Ma
Pararge a. aegeria 6-5 Ma
99 Pararge a. aegeria 6-6 Ma
Pararge a. aegeria 27-13 MC
Pararge a. aegeria 27-15 MC
100 Pararge a. aegeria 27-11 MC
Pararge a. aegeria 27-9 MC
Pararge a. aegeria 6-7 Ma
Pararge a. aegeria 26-4 Ma
Pararge a. aegeria 26-5 Ma
Pararge a. aegeria 27-14 MC
Pararge a. aegeria 25-9 Ma
Pararge a. aegeria 25-10 Ma
Pararge a. aegeria 29-19 Ma
Pararge a. aegeria 29-20 Ma
Pararge a. aegeria 29-21 Ma
Pararge a. aegeria 25-8 Ma
100 Pararge a. aegeria 1-3 FR
Pararge a. tircis 10-8 BE
Pararge a. tircis 16-3 UK
Pararge a. tircis 17-13 SE
Pararge a. aegeria 25-27 GR
Pararge a. aegeria 1-1 FR
Pararge a. aegeria 1-6 FR
Pararge a. aegeria 1-4 FR Fig.1. Strict consensus tree of 110 most
Pararge a. tircis 16-1 UK parsimonious trees of equal length, 604
Pararge a. tircis 18-1 SE
Pararge a. aegeria 24-18 FR steps,basedonthecytochromeoxidasesub-
100 PPaararargrgee a a. .t iarceisg e1r8ia-3 1 S7-E8 FR unitI(COI)dataset(n¼71taxa).Bootstrap
Pararge a. aegeria 1-8 FR support is shown above the branches.
Pararge a. aegeria 17-9 FR
Pararge a. tircis 17-14 SE Branchlengthsareproportionaltothenum-
Pararge a. tircis 18-2SE
Pararge a. aegeria 25-28 GR ber of changes inferred from parsimony.
Pararge a. tircis 16-2 UK Geographic abbreviations: BE, Belgium;
Pararge a. aegeria 1-2 FR
Pararge a. aegeria 31-5 ES ES, Spain; FR, France; GR, Greece; Lg,
Pararge a. aegeria 30-7 GR
1 step Pararge a. aegeria 31-6 ES La Gomera; Lp, La Palma; Ma, Madeira;
Pararge a. aegeria 33-22 PT MC, Morocco; SE, Sweden; Te, Tenerife;
Pararge a. aegeria 33-23 PT
Pararge a. aegeria 33-21 PT UK,UnitedKingdom.
only the population from La Palma is resolved as mono- The phylogeny of the combined dataset unsurprisingly
phyletic and does not share haplotypes with the other correspondstothetopologies ofCOI andwingless(Fig.3).
islands. In contrast, the two subspecies of P. aegeria are The monophyly of all three species receives moderate to
notrecoveredasmonophyletic.Instead,accordingtoCOI, strong Bremer support values, although this is mainly due
the species is divided into two clades, with continental to the COI partition. The sister group relationship of
European specimens forming one group, and the African P. aegeria and P. xiphioides is supported strongly by COI
andMadeiranspecimensformingtheother.Theanalysisof datapartition,butweaklybythe winglesspartition.Within
winglesssupportstheEuropeanclade,buttheAfricanand P. xiphioides, the populations from different islands receive
Madeirangroupisunresolved. highsupportfromtheCOIpartition,butlowornosupport
Thegenusitselfreceiveshighbootstrapsupportforboth fromthewinglesspartition.Thehighlysupporteddivisionof
genes (Figs 1 and 2), as do P. aegeria and P. xiphia. P. aegeria into European and North Africanþ Madeiran
However, the value is less for P. xiphioides, 70% for COI cladesisduealmostentirelytotheCOIpartition.
and 87% for wingless. The sister group relationship of PairwisesequencedivergencesareshowninTable1.For
P. aegeria and P. xiphioides is strongly supported by COI COI,anaverageofabout1.9%differencebetweenthetwo
(bootstrapvalueof99%),butlesssobywingless(bootstrap major P. aegeria lineages is detected. The pairwise COI
valueof70%). sequencedivergencewithinthetwolineagesiscloseto0%
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
PhylogenyofPararge 625
Kirinia roxelana
Lopingaachine
Lasiommata maera
Lasiommata megera
Lasiommata petropolitana
89
Pararge xiphia 5-1
Pararge xiphia 24-29
Pararge xiphioides 3-10 Te
Pararge xiphioides 27-1 Lg
93 87
Pararge xiphioides 27-2 Lg
Pararge xiphioides 28-16 Te
Pararge xiphioides 28-17 Te
80 Pararge xiphioides 27-6 Lp
70
Pararge xiphioides 28-19 Lp
Pararge a. aegeria 6-4 Ma
Fig.2. Mostparsimonioustreefoundforthe Pararge a. aegeria 27-12 MC
winglessdataset(treelength,70steps;n¼23 98 Pararge a. aegeria 27-8 MC
taxa). Bootstrap support is shown above the
Pararge a. aegeria 25-7 Ma
branches. Branch lengths are proportional to
Pararge a. aegeria 1-3 FR
the number of changes inferred from parsi-
mony. Geographic abbreviations: BE, Pararge a.tircis 10-8 BE
62
Belgium; ES, Spain; FR, France; GR, Pararge a.tircis 16-3 UK
Greece; Lg, La Gomera; Lp, La Palma; Ma,
Madeira; MC, Morocco; SE, Sweden; Te, 1 step Pararge a.tircis 17-13 SE
Tenerife;UK,UnitedKingdom. Pararge a. aegeria 25-27 GR
(i.e. haplotypes tendto differby onlyonebase pair outof (1994)molecularclockestimateof2.3%pairwisesequence
1450 sequenced). The most common haplotype in Europe divergence per million years to Pararge gives similar ages
also is the most widespread, being found in individuals for the younger splits, including the assumed two million
collected in southern Portugal, central Spain, southern yearsforthesplitbetweenP.xiphioidesonLaGomeraand
France, the UK and Sweden. Most haplotypes in LaPalma(Table2).
Morocco and Madeira are identical to each other. The
highest intraspecific diversity in COI is found within
P.xiphioides,withamaximumof4.4%differencebetween
specimens.Haplotypediversitywithinthemajorlineagesis Discussion
verypoor(closeto0%),suggestingrepeatedbottlenecksor
selective sweeps purging the populations of mitochondrial Speciationinbutterfliesmaybepromotedbyanexpanded
haplotypediversity. host plant range (Janz et al., 2001; Weingartner et al.,
Clockwise evolution of the sequences in the combined 2006).However,speciesofthegenusParargearespecialists
datasetwasrejected(P <0.05),andsowebaseourdiscus- of grasses and show no evidence for an expanded host
sion on the nonclock results. However, the choice of algo- range.Nevertheless,hostplantusecouldstillbeanimport-
rithm did not change the age estimates very much antfactorinthediversificationprocess.Ofthetenendemic
(Table2). Calibrating the split between the La Palma and butterflyspeciesoftheCanaryIslands,fivearegrass-feeding
La Gomera lineages of P. xiphioides at two million years satyrines (Hipparchia wyssii Christoph, 1889, H. bacchus
provides us with a maximum estimate of the ages of the (Higgins,1967),H.gomera(Higgins,1967),H.tilosi (Manil
othersplitsinthetree(Fig.4).Theageofthesplitbetween 1984)andP.xiphioides).Oftheothers,oneisastrongflier
P.xiphiaandtherestofParargeisestimatedatamaximum (Vanessa vulcanica Godart, 1819; Nymphalidae), one is a
of 5.6 million years with a standard deviation of 0.9 hesperid (Thymelicus christi Rebel, 1894), which also feeds
(Table2). This age is consistent with the age of Madeira ongrass,twobelongtothefamilyPieridae(Pierischeiranthi
and suggests that the ancestor of P. xiphia may have (Hu¨bner, 1808), Gonepteryx cleobule (Hu¨bner 1825)) and
reached the island soon after it emerged from the ocean. one is a lycaenid (Cyclyrius webbianus (Brulle´, 1840)). The
TheageofthesplitbetweenP.xiphioidesandP.aegeriais dominance of grass feeders on the Canary Islands might
estimatedatamaximumof3.1millionyears((cid:4) 0.46),while reflect their good colonization ability. Grass is a resource
thesplitbetweenNorthAfricanpopulationsandEuropean availablealmosteverywhere,anditmightbeeasytocolonize
populations of P. aegeria may have occurred around 0.87 new habitatseven though the butterflies themselvesare not
millionyears((cid:4) 0.24)ago.Interestingly,applyingBrower’s verygooddispersers.
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
626 E.Weingartneretal.
Kirinia roxelana
Lopinga achine
Lasiommata maera
100 Lasiommata megera
100 Lasiommata petropolitana
Pararge xiphia
27,2
100 Pararge xiphia
58
Pararge xiphioides Te
24,0
100 Pararge xiphioides Te
Pararge xiphioides Te
10,3 3,2
99 86 Pararge xiphioides Lg
20,0
100 Pararge xiphioides Lg
4,0
Pararge xiphioides Lp
80 20,2
100
Pararge xiphioides Lp
11,1
Pararge a. aegeria Ma
99 Fig.3. Most parsimonious tree of the
Pararge a. aegeria Ma combineddataset[cytochromeoxidasesub-
13,0
unit I (COI)þwingless]. Tree length, 647
100 Pararge a. aegeria MC steps(n¼23).PartitionedBremersupport
valuesare shown abovethebranches.The
Pararge a.aegeria MC firstvaluereferstothesupportcontributed
17,3 byCOIandthesecondtothesupportvalue
100 Pararge a. aegeria FR of the wingless dataset. Bootstrap support
is shown below the branches. Branch
Pararge a. aegeria GR
lengths have been estimated using maxi-
11,1 mum likelihood (GTRþGþI model).
Pararge a. tircis SE
100 Geographic abbreviations: BE, Belgium;
Pararge a. tircis UK ES, Spain; FR, France; GR, Greece; Lg,
La Gomera; Lp, La Palma; Ma, Madeira;
Pararge a. tircis BE MC, Morocco; SE, Sweden; Te, Tenerife;
0.01
UK,UnitedKingdom.
In the case of Pararge, it thus appears that speciation Where these ancestors came from is thus of interest. The
happened through processes not directly related to host islandofMadeiraisabout850kmsouth-westofPortugal,
plants. Quite likely, P. xiphia and P. xiphioides speciated about540kmwestofMoroccoandabout450kmnorthof
after their ancestors had colonized their respective islands. theCanaryIslands.TheislandofGranCanaria(theclosest
Table1. AverageuncorrectedpairwisedistancesofcytochromeoxidasesubunitI(COI)haplotypesforeachofthemajorlineagesofPararge.
P.xiphioides P.xiphioides P.xiphioides P.aegeria P.aegeria
Taxon Lasiommata P.xiphia (Te) (Lg) (Lp) (Africa) (Europe)
Lasiommata 0.061 0.089 0.098 0.093 0.093 0.095 0.092
P.xiphia 0.001 0.060 0.057 0.056 0.057 0.064
P.xiphioides(Te) 0.002 0.043 0.043 0.047 0.049
P.xiphioides(Lg) 0.001 0.032 0.048 0.045
P.xiphioides(Lp) 0.000 0.047 0.048
P.aegeria(Africa) 0.000 0.019
P.aegeria(Europe) 0.001
Lg,LaGomera;Lp,LaPalma;Te,Tenerife.
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
PhylogenyofPararge 627
Table2. EstimatedagesforsplitswithinPararge.
Node Constant Clock NPRS PL BootSD Bootrange
Root 4.1 9.07 9.96 9.88 1.59 4.26–12.50
Pararge 2.6 5.16 5.63 5.64 0.90 3.02–8.13
P.aegeriaþP.xiphioides 2.1 3.09 3.06 3.08 0.46 2.14–4.56
P.aegeria 0.8 0.94 0.86 0.87 0.24 0.42–1.58
P.xiphioides 1.9 2.45 2.43 2.44 0.26 2.00–3.09
LaPalmaþLaGomeraa 1.4 2.00 2.00 2.00 0.00 2.00
Constant,assumingaconstantmutationrateof1.2%permillionyears(Brower,1994);NPRS,nonparametricratesmoothing;PL,penalized
likelihoodwithasmoothingrateof0.0032;BootSD,standarddeviationofbootstrappeddatasets;Bootrange,minimumandmaximumages
estimatedfrombootstrappeddatasets.
aNodefixedattwomillionyears.
islandtotheAfricanmainlandwhichhasParargeonit)is islands of the archipelago, most probably due to the lack
about190kmwestoftheAfricancontinent.Thesourceof ofsuitablehabitatsontheseislands.
the ancestors of P. xiphia is not clear and may have been One explanation for the short basal branch lengths for
eitherEuropeorNorthAfrica. the P. xiphioides haplotype clade (see Fig. 3) could be a
Most likely, the Canary Islands were colonized from rapid diversification process on these islands. This would
NorthAfrica,fromwhichthenearestislandisonly95km. produce the observed basal branches with few character
Although we have yet to sample P. xiphioides from Gran changes, but large genetic distances between islands, indi-
Canaria,thepatternofcolonizationseemstobefromolder cating genetic isolation between them (Table1). High
to younger islands, which corresponds to an east to west genetic divergences amongst the populations on different
direction, with the population on Tenerife sister to the islandsintheCanaryIslands,comparedwiththemainland
populationsonLaGomeraandLaPalma.Thisresultagrees populations, are found also in psyllids (Percy, 2003). The
with other studies on Canary Islands colonizations estimatedmaximumageofP.xiphioidesis3.1millionyears
(Carranza et al., 2000; Emerson et al., 2000a; Rees et al., ((cid:4)0.46)(Fig. 4).Thus,P. xiphioides ismuch younger than
2001). With regard to butterflies, there is one study of the theformationofTenerife(11.6millionyearsago)and,once
genusGonepteryxLeach,1815,inwhichtheauthorssuggest it colonized the Canary Islands, it was able to spread
that the colonization of the Canary Islands has occurred amongsttheislandsquickly.
fromAfrica(Brunton&Hurst,1998).However,itiscrucial Contrary to what might be expected from morphology,
toincludethepopulationonGranCanariainordertodraw we have not discovered any indication that P. a. aegeria
any conclusions about the colonization pattern of P. andP. a.tircisareseparateevolutionaryentitiesbasedona
xiphioides. The species has not been recorded on the two mitochondrial gene and a nuclear gene. Instead, we found
islands Lanzarote and Fuerteventura, the easternmost twoseparatecladeswithinP.aegeriacorrespondingtothe
Lasiommata maera
Lasiommata megera
Lasiommata petropolitana
Pararge xiphia
Pararge aegeria North Africa
Pararge aegeria Europe
Pararge xiphioides Tenerife
Pararge xiphioides La Gomera
*
Pararge xiphioides La Palma
Fig.4. Chronogram for Pararge. The
9 8 7 6 5 4 3 2 1 0
asteriskmarksthenodewhichwasfixedat
twomillionyears. Millions of Years Before Present
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
628 E.Weingartneretal.
European and African þ Madeiran populations. This TheevolutionofParargebutterfliesisthusanexampleof
pattern is consistent in both genes. However, the morpho- aseeminglysimplecaseofspeciationthat,infact,showsa
logical differences indicate that other genes are differen- rather complicated pattern. In order to understand this
tiated between P. a. tircis and southern European pattern, we must rely on the phylogeny. Five observations
populations known as P. a. aegeria. The implication of canbemade:(1)thesistergrouptoParargeis(orappears
this finding will affect comparative studies of P. a. tircis tobe)thePalaearcticgenusLasiommata(infact,allpoten-
and P. a. aegeria in which Madeiran specimens have been tial sister groups to Pararge are Palaearctic; see Bozano,
used as representatives of the subspecies P. a. aegeria (e.g. 1999); (2) the two island species of Pararge are not sister
Nylin et al., 1994, 1995). It cannot be concluded that life species, suggesting that Madeira and the Canary Islands
historydifferencesbetweenthetwosubspeciesareaconse- werecolonizedindependently;(3)thedirectionofcoloniza-
quenceofrecentadaptationtothehabitatonMadeira(e.g. tionontheCanaryIslands(fromwesttoeast)accordswith
Nylinetal.,1995).Instead,theMadeiranpopulationsmay other studies and suggests that the source of the colonists
carry with them adaptations to habitats in North Africa, wasNorthAfrica;(4)therecentcolonizationofMadeiraby
whicharebeneficialalsoonMadeira.Itisthuspreliminary P.aegeriatookplacefromNorthAfrica;and(5)thehaplo-
to conclude that over 100 generations is enough time to types of P. aegeria form a monophyletic group to the
allow life history traits to evolve on Madeira (Nylin et al., exclusion of the two other species and, importantly, the
1995), without a comparative study of populations from haplotypes in North Africaþ Madeira and in Europe are
NorthAfricaandMadeira. reciprocallymonophyletic.
It is of great interest to determine when the two mito- The foregoing observations suggest strongly the follow-
chondrialcladesofP.aegeriadiverged,asthiscanhelpus ing scenario to us. The ancestor of the genus Pararge
tounderstandtheprocessthatmighthavepromoteddiver- colonized North Africa from the Palaearctic, perhaps dur-
sification. The results from dating should, however, be ingtheMessiniansalinitycrisis(assumingthatP.xiphiais
interpreted carefully. When using the age of islands to atmostfivemillionyearsold).Theancestralpopulationsin
calibrate nodes, we obtain the maximum age of a clade if Europesubsequentlywentextinct.Whenthisoccurred,we
we assume that divergence has taken place on the island. cannot say, but at least prior to the recolonization of
Most likely, the ancestral butterflies colonized the islands Europe by P. aegeria some four million years later (evi-
later, maybe much later. However, the much used 2.3% denced by no endemic European species or ancient haplo-
pairwise sequence divergence per million years (Brower, typesinEuropeanpopulationsofP.aegeria).Theancestral
1994) gives a very similar age (1.4 million years) for the species then colonized Madeira from North Africa soon
split between La Gomera and La Palma lineages as our after the island appeared above sea level, and speciated
assumed two million years (the age of the island of La subsequently and evolved into what we know currently as
Palma),lendingsomecredencetotheageswediscuss. P. xiphia. The ancestral populations in North Africa con-
Accordingtoourdating,thesplitbetweentheEuropean tinuedtoevolvethroughtimeuntilasecondcolonizationof
P. aegeria and the North African populations occurred at theislandsandspeciationeventtookplace,thistimeinthe
about0.87millionyears((cid:4)0.24)ago(Table2,Fig.4).This Canary Islands. The subsequent spread of P. xiphioides
fallswithintheQuaternary,whichinEuropewasdominated amongsttheislandsappearstohavebeenrapid.Thecolo-
byrepeatedintervalsofglaciationsthataffectedtheevolu- nizationoftheCanaryIslandsapparentlyalsowasasingle
tion of European species (Hewitt, 1996). During glacial event, evidenced by the monophyly of current haplotypes
periods,northernpopulationswereextinguishedorpushed ofP.xiphioides.
furthersouth.Whentheclimatebecamewarmer,thepopu- After the divergence of P. xiphioides from ancestral P.
lations expanded their ranges northwards. The aegeriapopulations,theNorthAfricanpopulationscontin-
Mediterranean has been a strong barrier to migration ued to develop in isolation until perhaps 0.8 million years
(Taberlet et al., 1998). During the Messinian salinity crisis ago, when Europe was colonized. This colonization also
(5.57–4.93millionyearsago),whentheMediterraneanwas appears to be a single event, evidenced by the deep diver-
desiccated,EuropeandAfricawerejoinedintoasingleland gence between, and reciprocal monophyly of, North
mass(DeJong,1998,andreferencescitedtherein),andthis AfricanandEuropeanpopulations.Subsequenttothecolo-
possiblyenabledmigrationbetweenthetwocontinents. nizationofEurope,thespecieseventuallyspreadtooccupy
The phylogenetic patterns we have uncovered present a its current range. It is quite likely that the European
difficult case for investigating the historical biogeography P.aegeriahasspreadandretracteditsrangeseveraltimes,
of the group. Three of the four major lineages show auta- astherehavebeenseveralglaciationperiodsduringthepast
pomorphic distributions (Madeira, Canary Islands and one million years. Recent observations in the U.K. have
NorthAfrica),whereastheonelineagesharingitsdistribu- shown that P. aegeria has the ability to expand its range
tion with the sister group (Europe) is in a highly derived rapidly in response to climate changes (Hill et al., 1999).
position, suggesting convergence. Conventional methods GiventherelativelydeepdivergencebetweentheEuropean
would suggest that Europe is the ancestral range of the andNorthAfricanlineages,itispossiblethattheEuropean
genus Pararge, and this is undoubtedly true. However, it P.aegeriasurvivedglacialmaximainrefugiaawayfromthe
doesnotshedlightonthesourcefromwhichtheislandsor Iberian peninsula, such as the Balkan peninsula. Further
NorthAfricawerecolonized. evidence for the scenario described above comes from the
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
PhylogenyofPararge 629
colorationofthewings.Fourtaxonomicentitiesretainthe entities within P. aegeria, one in North Africa (and
ancestral orange wing colour (which is also found in Madeira), one in Europe south of the Alps and one in
Lasiommata):P.xiphia,P.xiphioides,P. a.aegeria(North EuropenorthoftheAlps.
Africa) and P. a. aegeria (Europe). Our phylogenetic There are names available for each of the three entities:
hypothesis suggests that the yellow colour of the northern meoneStoll,1780(typelocality:Algeria),aegeriaLinnaeus,
P. a. tircis has evolved later, perhaps as an adaptation to 1758 (type locality: South Europe and North Africa) and
colderclimates. tircis Godart,1821(typelocality: France).However, using
Analternativebiogeographicalhypothesis,andonemore thesenamesiscomplicatedbythefactthatthetypelocality
parsimoniousbasedonourphylogenetichypothesis,would ofaegeriaisgivenas‘SouthEuropeandNorthAfrica’and
be that Madeira was colonized first from Europe by the notypematerialexists(seeHoney&Scoble,2001).Fixing
ancestralPararge.TheCanaryIslandswerecolonizedsub- thenametopopulationsinsouthernEuropewouldrequire
sequently from Madeira by the ancestor of P. xiphioides a neotype, and such an act is beyond the scope of this
andP.aegeria,andthenNorthAfricawascolonizedfrom paper. On the other hand, the names aegeria and tircis
the Canary Islands (before the differentiation of have been used consistently for southern and northern
P.xiphioidesontheislands).Finally,Europewascolonized European populations, respectively, and taking the name
from North Africa, completing a circle that took perhaps meone for the North African and Madeiran populations
five million years. Both hypotheses require that the ances- would not cause much taxonomic instability. We thus
tral Pararge went extinct in Europe at some stage before raise the name meone Stoll, 1780 from synonymy to sub-
the present. It is impossible to differentiate between these speciesstatus(Parargeaegeriameonestat.rev.)andrestrict
two hypotheses based on the evidence we have on hand. its use to populations in North Africa and Madeira. The
However,webelievethatindirectevidencefavoursthefirst name Pararge aegeria aegeria refers to populations in
hypothesisoverthesecond.Studiesonotherinsectssuggest southern Europe with an orange background colour on
thattheislandgroupshavebeencolonizedfromcontinental theforewings,andthenameParargeaegeriatircistopopu-
sources (Brunton& Hurst, 1998; Emerson etal., 2000a,b; lationsinnorthernEuropewithalightyellowbackground
Reesetal.,2001;Hundsdoerferetal.,2005).Althoughthe colourontheforewings.
colonization of Madeira by P. aegeria from North Africa Different genetic lineages in Europe and Africa have
cannot be seenas direct evidence of the same routeby the been found in several studies (Burban et al., 1999; Percy,
ancestorofP.xiphia,intheabsenceofotherevidence,itis 2003; Hundsdoerfer et al., 2005). Quite clearly, the
suggestive. There is some evidence of inter-island group Mediterranean remains a barrier for genetic exchange,
colonizations, but so far the direction of colonization has even though the Iberian Peninsula and Morocco are sepa-
been inferred to be from the Canary Islands to Madeira ratedbyonly14kmatonepoint(Dobson&Wright,2000).
(e.g.Emersonetal.,2000b;Hundsdoerferetal.,2005). This does seem paradoxical, as Pararge clearly crossed
Our results have important taxonomic implications for large bodies of water to reach Madeira and the Canary
Pararge.Firstofall,P.aegeria,P.xiphiaandP.xiphioides Islands. However, a general pattern of intraspecific diver-
clearlyformthreegoodspeciesthatproducemonophyletic gencebetweenthetwocontinentsmayexist,butstudies,as
clades with both mitochondrial and nuclear genes. This yet,arefew,especiallyinflyinginsects.Asimilarpatternis
implies that the three species have had a relatively long foundinthesatyrineLasiommatamegera(E.Weingartner,
period of independent evolution, an implication which is unpublished),aswellasinthenymphalinesMelitaeacinxia
corroboratedbyouranalysisoftheagesoftheclades.The (Linnaeus, 1758) and Melitaea phoebe (Denis &
second taxonomic implication is within the species Schiffermu¨ller, 1775) (N. Wahlberg, unpublished).
P. aegeria, for which there is conflicting information However, in the strong flyer Polygonia c-album (Linnaeus,
regarding morphological and geneticdata. On the basis of 1758)(Nymphalidae),deepdivergencebetweenEuropeand
morphology, there are two clear subspecies found north Morocco could not be found (E. Weingartner, unpub-
andsouthoftheAlps,respectively.Basedonthesequences lished).Moretaxaneedtobestudiedinordertoseeifthe
oftwogenes,therearetwoclear lineageswhich arehighly patternisconsistent.Ifthispatternoccursinothertaxa,it
diverged from each other, one in North Africa and the would suggest that there has been an event that has facili-
other in Europe. DNA taxonomy is enjoying great popu- tated movement between North Africa and Europe at one
larityatthemoment(see,forexample,Hebertetal.,2003), point in the recent geological history of the region. What
although calls are being made not to discard traditional this event could have been remains to be discovered
morphological data (see, for example, Wahlberg et al., throughcomparativestudiesofotherspecieswithdistribu-
2005). In this case, there are two COI haplotype lineages tionsinNorthAfricaandEurope.
which have apparently not interbred for a long period of
time(perhapsaslongas0.8millionyears,althoughtheyare
able to produce fertile offspring in the laboratory; Nylin Acknowledgements
etal.,1994),andtwomorphologicallineagesthatdointer-
breed but form a narrow hybrid zone and retain their WearegratefultoEnriqueGarcı´o-Barros,KarlGotthard,
morphological distinctiveness outside the hybrid zone. Bengt Karlsson, Georg Nygren, Thomas Schmitt,
This would suggest that there are in fact three taxonomic Constantı´ Stefanescu, Michel Tarrier and Christer
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632
630 E.Weingartneretal.
Wiklund for providing specimens for this work. We thank Emerson, B.C., Oromi, P. & Hewitt, G.M. (2000a) Colonisation
AndrewBrower,IanKitchingandananonymousreviewer and diversification of the species Brachyderes rugatus
for constructive comments on a previous version of the (Coleoptera) on the Canary Islands: evidence from mitochon-
manuscript. drialDNACOIIgenesequences.Evolution,54,911–923.
Emerson, B.C., Oromi, P. & Hewitt, G.M. (2000b) Interpreting
colonization of the Calathus (Coleoptera: Carabidae) on the
Canary Islands and Madeira through the application of the
References
parametricbootstrap.Evolution,54,2081–2090.
Eriksson,T.(2005) BergianskaTra¨dga˚rdenSoftwareURLhttp://
Ancochea,E.,Hernan,F.,Cendrero,A.,Cantagrel,J.M.,Fuster,J.M., www.bergianska.se/index_kontaktaoss_torsten.html [accessed
Ibarrola, E. & Coello, J. (1994) Constructive and destructive on24May2005].
episodes in the building of a young oceanic island, La Palma, Felsenstein, J. (1985) Confidence limits on phylogenies: an
CanaryIsland,andgenesisoftheCalderadeTaburiente.Journal approachusingthebootstrap.Evolution,39,783–791.
ofVolcanologyandGeothermalResearch,60,243–262. Felsenstein, J. (1993) Phylip (Phylogeny Inference Package),
Baker, R.H. & DeSalle, R. (1997) Multiple sources of character Version 3.63. Available from author at URL http://evolution.
information and the phylogeny of Hawaiian drosophilids. gs.washington.edu/phylip.html[accessedon12May2005].
SystematicBiology,46,654–673. Gatesy,J.,O’Grady,P.&Baker,R.H.(1999)Corroborationamongdata
Bozano,G.C.(1999)SatyridaePartI,SubfamilyElymniinae,Tribe setsinsimultaneousanalysis:hiddensupportforphylogeneticrelation-
Lethini.OmnesArtes,Milan. shipsamonghigherlevelartiodactyltaxa.Cladistics,15,271–313.
Bremer, K. (1988) The limits of amino acid sequence data in Geldmacher, J. & Hoernle, K. (2000) The 72 Ma geochemical
angiospermphylogeneticreconstruction.Evolution,42,795–803. evolutionoftheMadeirahotspot(easternNorthAtlantic):recy-
Bremer,K.(1994)Branchsupportandtreestability.Cladistics,10, cling of Paleozoic ((cid:5)500 Ma) oceanic lithosphere. Earth and
295–304. PlanetaryScienceLetters,183,73–92.
Brower,A.V.Z.(1994)Rapidmorphologicalradiationandconver- Goloboff, P.A. (1998) NONA, Version 2.0. Published by author.
gence among races of the butterfly Heliconius erato inferred URLhttp://www.cladistics.com[accessed2001].
frompatternsofmitochondrialDNAevolution.Proceedingsof Gotthard,K.,Nylin,S.&Wiklund,C.(1994)Adaptivevariationin
theNationalAcademyofSciencesUSA,91,6491–6495. growthrate:lifehistory costsand consequences inthe speckled
Brower, A.V.Z. & DeSalle, R. (1998) Patterns of mitochondrial woodbutterfly,Parargeaegeria.Oecologia,99,281–289.
versus nuclear DNA sequence divergence among nymphalid Gotthard,K.,Nylin,S.&Wiklund,C.(2000)Matingopportunity
butterflies: the utility of wingless as a source of characters and the evolution of sex-specific mortality rates in a butterfly.
forphylogeneticinference.InsectMolecularBiology,7,73–82. Oecologia,122,36–43.
Brunton, C.F.A. & Hurst, G.D.D. (1998) Mitochondrial DNA Hall, T.A. (1999) BioEdit: a user-friendly biological sequence
phylogeny of brimstone butterflies (genus Gonepteryx) from alignmenteditorandanalysisprogramforWindows95/98/NT.
the Canary Islands and Madeira. Biological Journal of the NucleicAcidsSymposiumSeries,41,95–98.
LinneanSociety,63,69–79. Hebert, P.D.N., Cywinska, A., Ball, S.L. & deWaard, J.R. (2003)
Burban,C.,Petit,R.J.,Carcreff,E.&Jactel,H.(1999)Rangewide BiologicalidentificationsthroughDNAbarcodes.Proceedingsofthe
variation of the maritime pine bast scale Matsucoccus feytaudi RoyalSocietyofLondon,SeriesB,BiologicalSciences,270,313–322.
Duc. (Homoptera: Matsucoccidae) in relation to the genetic Hewitt, G.M. (1996) Some genetic consequences of ice ages, and
structureofitshost.MolecularEcology,8,1593–1602. theirroleindivergenceandspeciation.BiologicalJournalofthe
Carranza, S., Arnold, E.N., Mateo, J.A. & Lo´pez-Jurado, L.F. LinneanSociety,58,247–276.
(2000) Long-distance colonisation and radiation in gekkonid Higgins, L.G. (1975) The Classification of European Butterflies.
lizards,Tarentola(Reptilia:Gekkonidae),revealedbymitochon- WilliamCollinsSons&Co,London.
drial DNA sequences. Proceedings of the Royal Society of Hill, J.K., Thomas, C.D. & Blakeley, D.S. (1999) Evolution of
London,SeriesB,BiologicalSciences,267,637–649. flightmorphologyinabutterflythathasrecentlyexpandedits
Davies,N.B.(1978)Territorialdefenceinthespeckledwoodbut- geographicrange.Oecologia,121,165–170.
terfly (Pararge aegeria): the resident always wins. Animal Honey, M.R. & Scoble, M.J. (2001) Linnaeus’s butterflies
Behaviour,26,138–147. (Lepidoptera: Papilionoidea and Hesperoidea). Zoological
DeJong,H.(1998)Insearchofhistoricalbiogeographicpatterns JournaloftheLinneanSociety,132,277–399.
in the western Mediterranean terrestrial fauna. Biological Hundsdoerfer,A.K.,Kitching,I.J.&Wink,M.(2005)Thephylo-
JournaloftheLinneanSociety,65,99–164. genyoftheHyleseuphorbiaecomplex(Lepidoptera:Sphingidae):
Dobson,M.&Wright,A.(2000)Faunalrelationshipsandzoogeo- molecular evidence from sequence data and ISSR-PCR finger-
graphicalaffinitiesofmammalsinnorth-westAfrica.Journalof prints.OrganismsDiversityandEvolution,5,173–198.
Biogeography,27,417–424. Janz,N.,Nyblom,K.&Nylin,S.(2001)Evolutionarydynamicsof
van Dyck, H. & Matthysen, E. (1998) Thermoregulatory differ- host-plantspecialization:acasestudyofthetribeNymphalini.
ences between phenotypes in the speckled wood butterfly: hot Evolution,55,783–796.
perchersandcoldpatrollers?Oecologia,114,326–334. Jones, M.J. (1992) The speckled wood butterflies Pararge xiphia
van Dyck, H. & Wiklund, C. (2002) Seasonal butterfly design: and P. aegeria (Satyridae) on Madeira: distribution, territorial
morphological plasticity among three developmental pathways behaviour and possible competition. Biological Journal of the
relative to sex, flight and thermoregulation. Journal of LinneanSociety,46,77–89.
EvolutionaryBiology,15,216–225. Jones,M.J.,Lace,L.A.,Harrison,E.C.&Stevens-Wood,B.(1998)
vanDyck,H.,Matthysen,E.&Dhondt,A.A.(1997)Theeffectof Territorial behaviour in the speckled wood butterflies Pararge
wingcolouronmalebehaviouralstrategiesinthespeckledwood xiphiaandP.aegeriaofMadeira:amechanismforinterspecific
butterfly.AnimalBehaviour,53,39–51. competition.Ecography,21,297–305.
#2006TheAuthors
Journalcompilation#2006TheRoyalEntomologicalSociety,SystematicEntomology,31,621–632