Table Of Content©EntomologicaFennica. 22August2014
Restricted geographic distribution and low genetic
distinctiveness of steppic Crioceris quinquepunctata
(Coleoptera: Chrysomelidae) populations
in Central East Europe
MiloszA.Mazur,DanielKubisz&LukaszKajtoch
Mazur,M.A.,Kubisz,D.&Kajtoch,L.2014:Restrictedgeographicdistribution
andlow genetic distinctiveness ofsteppic Crioceris quinquepunctata (Coleop-
tera: Chrysomelidae)populations inCentralEastEurope. —Entomol. Fennica
25: 102—110.
CriocerisquinquepunctataisaEuropeanleafbeetle,rareandstrictlyassociated
withsteppe-likehabitatsinCentralandEasternEurope.Wesampledsuitablelo-
calitiesinCentralEastEuropetodeterminethecurrentdistributionandtoverify
whether populations isolated by the Carpathian Mountains (within Pannonian
andPonticarea)showgeneticdifferentiation.SequencesfromthebeetleC01and
ITS1 were amplified and compared. Furthermore, ftsZ and hcpA genes ofthe
endosymbiont Wolbaclzia were analysed as additional genetic markers. We
foundonlytwopopulationsofC.quinquepunctata(inMoraviaandPodolianUp-
land). Unusually low genetic differences between these populations were re-
vealed,whichis incontrasttoprevious studies onothersteppebeetles. Therea-
sonsforsuchlowdiversityarespeculativeandprobablyrelatedtorecentnatural
expansionorman-madetranslocationofC. quinquepunctata.
M. A. Mazur, Centerfor Biodiversity Studies, Department ofBiosystematics,
Opole University, Oleska 22 St, 45—052 Opole, Poland; E—mail.‘ milosz@uni.
opole.pl
D.Kubisz&L.Kajtoch,InstituteofSystematicsandEvolutionofAnimals,Polish
AcademyofSciences, 31—016, Cracow, Poland
Received13August2013, accepted13December2013
1. Introduction In the past, Crioceris quinquepunctata
(Scopoli, 1763)wasdistributedinEuropeinAus-
The genus Crioceris Geoffroy, 1762 (Coleop- tria, BosniaandHerzegovina, Bulgaria, Croatia,
tera:Chrysomelidae)consistsof62speciesfound central and southern European parts ofRussia,
all over the world (except for Neotropical and theCzechRepublic,France,Germany,Hungary,
Australian regions) (Monros 1960) and 22 spe- Italy, Moldova, Poland, Romania, Slovakia,
ciesfoundinthePalaearctic,includingeightspe- Ukraine, Serbia and Montenegro (Lobl &
cies known in Europe (Lobl & Smetana 2005). Smetana 2005: 361). Most records are old and
All ofthem are oligophagous or monophagous, werenotrepeatedso theymostlyhavehistorical
feeding on members of the genus Asparagus valueandhencethereisaneedfornewdata. For
(Aspargaceae) (Clarketa1. 2004). example,onlytwolocalitiesareknownfromGer-
ENTOMOL. FENNICAV01. 25 ° Criocerisquinquepunctatain CentralEastEurope 103
Table 1. Listofinvestigatedxerothermiclocalitiesbycountries,withcodesofUTMsquares,yearsand numbers
ofcollectedspecimens(frommostlocalitiesnonewasfound). Speciescodes: CA, Cn'ocerisasparagi;C5, C.
quinquepunctata; C12, C. duodecimpunctata;C14, C. quatuordecimpunctata.
Country& Locality UTM Years CA C5 C12 C14
Poland
Nida vic. D862 2005—2013 >10 — 10 6
Gogolin vic. 8A89 2003—2012 >10 — 5 —
GOraZdZe 8880 2008—2012 7 — 8 —
Prudnik XR87 2008—2012 5 — 2 —
Kamien Slaski 8890 2003—2013 7 — — —
Checiny D862 2011 — — 7 —
Duszniki Zdréj WR98 2008, 2012 — — — —
Dziewcza GOra F877 2006, 2008, 2011 — — — —
Golebia GOra WR95 2008 — — — —
GOra Bukowa F830 2012 — — — —
Grédekad Hrubieszéw G802 2012 — — — —
Hebdéw DA55 2012 — — — —
JagienieckieLaki XS70 2008—2011 — — — —
Kajaséwka DAO4 201O, 2013 — — — —
Kalwaria Paclawska FV29 2011 — — — —
Katy ll F841 2006, 2013 — — — —
Kietrz 8A85 2011—2013 — — — —
Kikéw DA98 2010, 2012 — — — —
Klucze CA97 2012 — — — —
KrakéwSkotniki DA13 2006—2013 — — — —
Ligota Dolna 8A99 2004—2013 — — — —
Morsko DA75 2008—2013 — — — —
Niewiatrowice DA58 2010 — — — —
Rybna XS83 2007—2012 — — — —
Sandomierz E851 2006—2012 — — — —
Stobrawa XS83 2008—2012 — — — —
Szymiszéw C809 2004—2013 — — — —
Walyad Miechéw DA48 2012 — — — —
Wieprzecka GOra F851 2006, 2012 — — — —
Winna GOra YSO4 2007—2012 — — — —
Zubrzyce XR95 2005—2008 — — — —
Czech Rep.
Pouzdrany XQ12 2012 — 7 7 7
Sedlec VQ69 2012 — — 3 7
Popice XQ22 2012 — — — 6
Mohelno WQ83 2012 — — — —
Slovakia
Cenkovska Step CT28 2012 — — >10 6
8ratyslawa XP54 2011 — — — 7
Devinska Kobyla XP43 2011 — — — 6
Dlha n. Orava CV85 2011 — — — —
Dunajske Luhy XP8O 2011 — — — —
Plast’ovce CU43 2012 — — — —
Skalica-Veternik XQ61 2012 — — — —
VelkeVrchy CU19 2011 — — —
Ukraine
Miodobory MQ46 1993—1998 >10 — >10 8
Lomachyntsi NQ22 2010 — 6 — 6
Ustia n. Dniestrem MP77 2010 — — 5 8
Subiczn. Dniestrem MP88 2010 — — 3 9
8ilcheZolote MQ10 2010 — — 5 —
Kryve MQ27 2010 — — >10
104 Mazuretal. ' ENTOMOL. FENNICAVol. 25
Table 1, continued
Country& Locality UTM Years CA C5 C12 C14
Novodnistrovsk NP33 2010 — — 8 —
Zvenihorod LQ83 2010 — — >10 —
Biala Gora ad Zolochiv LR31 2007 — — — —
Czartowa Gdra ad Rohatyn LQ27 2010, 2011 — — — —
Dzwinogréd CB6? 2010 — — — —
Halicz LQ34 2011 — — — —
Huciska ad Lviv LR45 2011 — — — —
Kasova Gdra LQ25 2011 — — — —
ad Bursztyn
Suraz FD36 2012 — — — —
Zlata Gora ad Brody LR53 2007 — — — —
Austria
EdelstahI-Spitzenberg VQOO 2011 — — — —
Hainburg XP43 2012 — — — —
Leitha Gebirge XP10 2012 — — — —
Mannersdorf XP21 2012 — — — —
Hungary
Vackisujfalu CT78 2012 — — 5 7
Nagyvisnyo DU53 2012 — — 7 —
Verpelét DT49 2012 — — 5 —
Latorpuszta DU71 2012 — — — —
Lénérddardc DU53 2012 — — — —
Noszvaj DU6O 2012 — — — —
Pénzesgyor YN13 2010 — — — —
Topolca XM88 2010 — — — —
Velence CT23 2010 — — — —
manyandatleastoneofthemreferstoartificially abundantlyhostplants(Asparagusspp.),thespe-
settled populations on Asparagus plantations cieshasonlybeencollectedfromspecificdryand
(Schmitt & Ronn2011). In Poland, C. quinque— warmlocalities, mainlythe steppes, exceptfora
punctatawas recordedinthe 19th century andat single locality in Germany (Schmitt & Ronn
thebeginningofthe20thcentury(Borowiecetal. 2011). These habitats probably originatedinthe
2011), but it has not been found since then. It Pleistocenesteppe-tundra—anenvironmentcom-
raises the possibility that its populations, espe- prised ofdry grasslands andwetbogs (Nehring
ciallyinthenorthoftheCarpathiansandtheAlps 1890)
are presently extinct. Schmitt and Ronn (2011) Currently,steppesaredistributedintheconti-
reportedabout20localitiesforthis speciesinthe nentalzones ofEasternEurope(Pontic steppes),
PannonianBasin.However,thesedataarefroma the Pannonian Basin and the Iberian Peninsula;
broadtime-span(spanning the 19th and20th cen- however,theyhavebeenhighlyfragmented. The
tury), so the present distribution ofC. quinque— steppes are presently restrictedto “warm-stage”
punctata in the south-west ofthe Carpathians is refugia(Stewartetal. 2010). Thesethreenatural
poorlyknown.InUkrainethisspecieswasfound steppe areas in Europe are highly isolated from
only around the Black Sea (e.g. in Crimea), eachotherbymountains,forestsandanthropoge-
whereasincentralandwesternpartofthecountry nicbarriers.
(e.g. in PodolianUpland) itwas foundonly in a Steppes contain plant communities rich in
singlelocalityintheDniestervalley (KubiszD., rare and endemic species ofplants and animals
unpublished). (mainlyinsects).Despitetheprotectionofsteppe-
Criocerz'squinqaepunctataisstronglyassoci- like habitats within the framework of Natura
atedwiththesteppeandsimilardrygrasslandsin 2000 network and many nature reserves, these
Central and Eastern Europe. Despite having habitatsarestillseriouslythreatened.Inmany10-
ENTOMOL. FENNICAVol. 25 ° Criocerisquinquepunctatain CentralEastEurope 105
calities, populations ofsteppe species ofplants areas, namely in Ukraine and Pannonia (Mora-
andanimalshavebeenlost,andmanyotherareon via). We used also endosymbiotic bacterium
the brink of extinction (Michalik & Zarzycki Wolbachia genotyping as an additional marker
1995) forestimationofpopulationdifferentiation.
The main threats include natural succession,
leading to sites being overgrown by common
plantspeciesnonspecifictosteppes,andchanges 2. Material and methods
inlanduse,withmanynaturalsteppeplantcom-
munities being replaced by farmlands, orchards 2.1. Sampling
orwoodplantations.Moreover,steppesareextra-
zonal elements in Central Europe; also, they are There is no detailed information on the present
highly susceptible to natural andman-made cli- distributionofC. quinquepunctata inUkraine, it
matic and environmental changes. All these is absentinPoland(Mazur&Kubisz2013), and
threats lead to fragmentation and isolation of there is only one map ofthe distribution ofthe
steppe areas. For these reasons, steppes require speciesinthePannonianBasin(Schmitt&Ronn
activeprotection, such as regular cutting andre- 2011).However,thismapisbasedondatafroma
moval ofplants (especially shrubs and trees) or broad time-span (the 19th and 20th centuries).
grazingby sheep orgoats. Consequently, currentdistributionofthisspecies
Despite having many suitable habitats C. ispoorlyknowninCentralandEasternEurope.
quinquepunctata populations in Central and Specimens of Crioceris were sought during
Eastern Europe are strongly isolated, with the severalfieldtripsinCentralEastEuropein 1993—
Carpathianrangeformingadistinctivebarrierbe- 2013 (Table 1). Inthecourseofthisresearch,we
tweenthenorth-eastemandsouth-westempopu- investigatedafewdozensteppeanddrygrassland
lations (Fig. 1). Previous genetic studies ofsimi- sites,wherethis specieswasdetectedinthepast,
larly distributed species, e.g. leaf-beetles Crio— as well as in many other steppe localities which
ceris quatuordecimpunctata (Scopoli), Cheilo— potentiallycouldsustaintheirpopulations(Table
tomamusciformis(Goeze),weevilCentricnemus 1). Wepaidspecialattentionto searchingbeetles
leucogrammus (Germar) (Curculionidae) and inAsparagusplants (checking all foundplants).
buprestid Coraebus elatus (Fabricius) (Bupre- Allfieldtripswereperformedduringthemiddle
stidae), revealcleardifferencesbetweenpopula- ofthe growing season (15 May—15 July) and in
tions fromthe southern andnorthern arch ofthe goodweatherconditions.Beetleswerecaughtus-
Carpathian mountains (Kajtoch & Lachowska- ing sweep-nets and by hand from their host
Cierlik 2009, Kajtoch 2011, Kubisz et al. plants.
2012a,b,Kajtochetal.2013a,b).Apartfrommo- We used the same number ofspecimens (5)
lecularanalysesbasedonDNAmarkers isolated from each ofthe two known populations of C.
frombeetlegenomes, ahelpfultoolinestimation quinquepunctata for further genetic analyses.
ofpopulationdifferentiationcouldalsobegeno- Therestofthe available specimens were leftfor
typing ofendosymbionts or parasites. Notably, museum collections, because DNA isolation
genotyping ofthe bacterium Wolbachia was re- damagedbodiesofthesebeetles. This isbecause
cently used as an additional marker for genetic we isolated DNA from the whole specimens as
population studies ofinsects, e.g. forsteppe spe- only this procedure allows one to sequence
cies Crioceris quatuordecimpunctata and Wolbachia genes (aswellas host-plantbarcodes
Polydrusus inustusGermar(Kajtochetal. 2012, frominsecthemolymph,whicharenotpresented
Kubiszetal. 2012a). inthisstudy).Thespecimenswerefirstpreserved
Ourstudyseekstodeterminecurrentdistribu- in99% ethanolandafterwards storedat—22°C.
tion ofC. quinquepunctata in Central East Eu- All specimens, also the slightly damagedby
rope,notablyintheCarpathianBasinanditssur- DNAisolation,weredepositedattheDepartment
roundings. Moreover, we aim to verify whether ofCollections, InstituteofSystematics andEvo-
genetic differences existbetweenpopulations of lutionofAnimals,PolishAcademyofSciencesin
C. quinquepunctata currently localized in two Cracow, Poland.
106 Mazureta]. - ENTOMOL. FENNICAVol. 25
0 Localitywith Crioceris quinquepunctata
1' Extinct population of C. quinquepunctata in Poland
El Localitywith Criocerisspp. otherthan C. quinquepunctata
D Localitywithout Criocerisspp.
(Wu 2 f)
Fig. 1. Crioceris
quinquepunctataand
otherCriocerisspp. in
Central EastEurope.
PictureofC. quinque-
punctataprovided
kindlybyProfessor
L. Borowiec(Icono-
graphiaColeopterorum
Poloniae: http://coleop-
tera.ksib.p|/kfp/search.p
hp?img=38958).
2.2.Laboratoryprocedure distanceswerecalculatedusingtheKimura2pa-
rametermodel(K2P).Haplotypenetworkprepa-
Two DNA markers were used for population ration and Wolbachia strain identification were
studies of C. quinquepunctata: Cytochrome carried out as in the study ofC. quatuordecim-
OxidaseI(COI)mitochondrialgeneandInternal punctata (Kubiszetal. 2012a) usingthe follow-
TranscribedSpacerofnuclearrDNA(ITS1).Ad- ing programs: DnaSP v.5 (Librado & Rozas
ditionally,twoDNAmarkerswereusedtodetect 2009), MEGA v.5 (Tamura et a]. 2011), TCS
thepresence ofWolbachia: cell divisionprotein 1.21 (Clement et a]. 2000) and BLASTn algo-
gene (ftsZ) and hybrid cluster protein gene rithm(Altschuletal. 1990).
(hcpA) givereference for these. Wolbachiawas Phylogenetic analyses were omitted in this
used here as an additional, unlinked genetic work as the phylogeny of Crioceris species
marker forpopulation genetic studies ofits host (among others, the relationship of C. quinque-
(like in Kajtoch et a]. 2012 and Kubisz et al. punctatawithotherEuropeancongeners)wasthe
2012a,b).Alllaboratoryprocedureswerecarried topic of a previous research by Kubisz et al.
out exactly like in the studies of Crioceris (2012b) andthephylogenyofWolbachiawithin
barcodingandC. quatuordecimpunctatapopula- infected Crioceris species was presented by
tion genetics (Kubisz etal. 2012a,b). All newly Kubiszetal. (2012a).
obtained sequences were deposited in GenBank
(Accessionnos. KC123330—KC123344).
3. Results
2.3.Dataanalysis
3.1. Criocerisquinquepunctata distribution
Weused standardmethodwhich allow forcom-
parison of genetic diversity and differentiation WecaughtseveraldozenCriocerisbeetlesrepre-
between studied populations. Pairwise genetic senting four species, mostly C. duodecim-
ENTOMOL. FENNICAVol. 25 - Criocerisquinquepunctata in CentralEastEurope 107
ftsZ
PONTIC
O
PANNONIAN
Fig.2. HaplotypenetworksofC. quinquepunctatapopulations(Ponticand Panno-
nian)drawnforCOIand ITS1 beetlemarkersandforftsZand hcpAWolbachia
genes.Sizeofthecirclecorrespondstonumberofspecimenssharing particular
haplotype. Numberofcirclesinparticulargeneticmarkerscorrespondstonumberof
haplotypesfound instudiedpopulationsinthosemarkers. Small blackdotsindicate
haplotypesmissinginsampled individuals(theseareintermediatehaplotypesjoining
haplotypesfound insampled individuals).A, Band B’—namesofWolbachia
supergroupstowhichidentifiedgenevariantsbelong.
punctata(Linnaeus) andC. asparagi(Linnaeus), 3.2. Genetics ofCrioceris
sometimes also C. quatuordecimpunctata; all10- quinquepunctatapopulations
calities of particular species are presented in
Table1.Ontheotherhand,wemanagedtofindC. Noindelsweredetectedinthe788bpoftheCOI
quinquepunctata inonlytwo localities, onesitu- gene andinthe606bpofITS1 gene.
ated to the north-east ofthe Carpathianrange in ITS1 sequenceswereidenticalinalltheindi-
the Podolian Upland (south-western Ukraine— viduals from the both regions while COI se-
Pontic steppes; 48°35’10.14”N and 27°08’ quence fromthePannonianandPontic individu-
2.39”E, 6 specimens) in 2010, and the other alsdifferedby00—04%.COIsequencesforindi-
found to the south-west of these mountains in vidualswithinthePannonianpopulationdiffered
Moravia (the Czech Republic—Pannonian by00—04%andwithinthePonticpopulationby
Steppes; 48°47’36.80”N and 16°41’40.20”E, 7 00—01%. Three variable sites in the COI gene
specimens) in2012 (Fig. 1). Wedidnotfindthe fragmentwerefound,twoparsimonyinformative
species in other localities in western Ukraine, were identifiedwithinthe Pannonianpopulation
Slovakia, Austria and Hungary, although we (haplotypediversityh= 0.800 (i0.164 SD); nu-
foundotherCriocerisspeciesthere(Table1).We cleotide diversity TE 2 0.00183 (i 0.00129 SD)),
also tried to find C. quinquepunctata in south- whilethePonticpopulationhadonlyonevariable
westernandsouth-eastemPoland,wherethespe- and parsimony informative site (h = 0.600 (i
cies was collected over 100 years ago, and in 0.175 SD); 7: = 0.00076 (i 0.00061 SD)). As
otherareaswithsuitablehabitats.However,these ITS1 was monomorphic in both populations,
populations now appear to be extinct (SW P0- therewasonlyonehaplotype inthismarker.
land) or they have neverbeenpresent there (SE The most common haplotype of C01 was
Poland). foundintwoPannonianandthreePonticindivid-
108 Mazuretal. ' ENTOMOL. FENNICAVol. 25
uals. Two other haplotypes were found in the entlyextinct. InwesternUkraine, itis stillfound
PannonianpopulationandoneinPontic.AsITS1 in only one locality in the Dniestervalley. This
wasmonomorphicinbothpopulations,therewas information strongly suggests that this species
only onehaplotypeinthismarker(Fig. 2). shouldbetreatedasvulnerableorendangeredin
allCentralEuropeancountries, anditispossible
thatinsomeareasitisextinct.Moreover,thespe-
3.3. Wolbachiascreening ciesshouldbeincludedintheRedListofanimals
threatenedwithextinctiondevelopedinSlovakia,
No indels were observed in 472 bp ofthe ftsZ Austria andHungary— it is included in the Red
geneandin453 bpofthehcpAgene. List ofthe Czech Republic as endangered; it is
AllspecimensfrombothC. quinquepunctata criticallyendangeredinGermany;inPoland,itis
populationswere foundpositive forinfectionby labelled as probably extinct (Binot et al. 1998,
Wolbachia(Fig. 2). Two Wolbachia strainswere Pawlowski et al. 2002, Farkac et al. 2005). Fi-
found in both populations and most individuals nally, this species, similar to other steppe leaf-
had double infections (all specimens from the beetles (e.g. Cheilotoma muscz'formis (Goeze),
Pontic population and four out offive from the Kajtoch et al. 2013b), should be added to the
Pannonian population). Wolbachia strains from Habitat Directive of the European Union and
thePannonianandPonticpopulationshadsimilar Special Areas ofConservation shouldbe desig-
but not identical ftsZ sequences oftwo distinct natedforits conservation.
types, onedifferingbyonemutationandanother Thetwofoundpopulationsareapproximately
by two mutations (substitutions). Wolbachia 700kmdistantfromeachotherinastraightline,
strainshadidenticalonetypeofhcpA sequences approx. 800 km distant around the northern
inbothpopulationsandanotherhcpAtypediffer- Carpathiansandapprox. 1,300kmdistantaround
ing by one mutation (substitution) between the the southern Carpathians. They are separatedby
PannonianandPonticpopulations. Accordingto unfavorable mountainous environments in the
ftsZ, one ofthe Wolbachia strains belonged to Carpathians (high altitude and the lack ofboth
supergroup A and the other to supergroup B. suitablehabitats andhostplants). In spite ofthat
However, for the hcpA marker this was not so isolationandtherarityofthespecies,thereareno
clear, as one strain belonged to supergroup B considerable genetic differences between the
whiletheotherseemedtobeanintermediatebe- populationsfoundondifferentsidesoftheCarpa-
tween supergroups A andB, butrather closerto thians. In both populations, specimens with the
supergroupB (calledB’). samemtDNAhaplotypeswere found(fourfrom
thePonticareaandtwofromthePannonianarea),
andthereisonlylimitedevidenceforcorrelation
4. Discussion ofmtDNA variation with the geographic local-
ization ofthe samples: one haplotype from one
Presently Crioceris quinquepunctata is an ex- individualisuniquetothePonticpopulationand
tremelyrarespeciesknownonlyfromfewpopu- twohaplotypesfromthreeindividualsareunique
lationsinthePannonianandPonticrefugias(Fig. to the Pannonian population; however, these
1). Its distribution is probably even more re- haplotypesdifferbyonly 1—2mutationsfromthe
stricted than it can be deduced from the Fauna most common andwidespread haplotype. Vari-
Europaea web-site (http://www.faunaeur.org) ability within nuclear DNA was not detected,
andSchmittandRonn(2011).Inmanyhistorical whichmaybeduetoeitherlowerpolymorphism
localities, the species has not been found for of the ITS1 marker, which may not have had
many years, despite intensive search, and it is enough time to accumulate mutations, or its ho-
probably extinct there (Poland, Slovakia). In mogenization (ITSs are tandemly repeatedunits
other Pannonian countries, it is probably very belonging to the family ofrDNA genes andun-
rare species and in at least halfofthe localities dergoconcertedevolution,Nei&Rooney2005).
specified by Schmitt and Ronn (2011) in the Thesamepattern(thelackofdifferencesinITS1
CzechRepublic, AustriaandHungary itispres- despite COI variability) was found in C.
ENTOMOL. FENNICAVol. 25 ° Criocerisquinquepunctatain CentralEastEurope 109
quatuordecimpunctata (Kubisz et al. 2012a). therresearchonpopulationgeneticsandmolecu-
Furthermore, the lack of genetic isolation be- lar ecology ofother steppe insects and other in-
tweenthe Pontic andthe Pannonianpopulations vertebrates isrequiredinthefuture.
of C. quinquepunctata is also corroborated by
smalldifferencesbetweenthe Wolbachiastrains. Acknowledgements. ThisworkwassupportedbygrantN
It is worth noting that Wolbachia strains com- N303 311137 from the Polish Ministry ofScience and
HigherEducation(toDanielKubisz,forpopulationgenet-
monlyvaryacrosspopulationsfoundindifferent ics),UMO—2013/11/D/NZ8/00583 fromtheNationalSci-
regions (e.g., in C. quatuordecimpunctata, Ku- ence Centre, Poland (to Lukasz Kajtoch, for Wolbachia
biszetal. 2012a). Howewer, theyareuniformin screening). WearegratefultoLechBorowiecforprovid—
recently expanded species (e.g. weevil Poly— ingapictureofaspecimen(Fig. 1)andtoMichaelSchmitt
drusus inustus (Curculionidae), Kajtoch et al. andanonymousreviewerfortheirhelpfulcomments and
supportinginformation.
2012). Highly similar strains in populations lo-
catedover700kmapartsuggestthatthebacteria
andthemitochondrialgenomeswereinheritedin
References
the course ofrecent sweeps by each strain, each
fromacommonsource.
Altschul,S.F.,Gish,W.,Miller,W.,Myers,E.W.&Lip—
Interestingly, the results differ considerably
man,D.J. 1990:Basiclocalalignmentsearchtool.7
from those reported in previous studies on a JournalofMolecularBiology215:4037410.
closelyrelatedspecies, C. quatuordecimpunctata Binot,M.,Bless,R.,Boye,P.,Gruttke,H.&Pretscher,P.
(Kubiszetal. 2012a,b).Thisspecieshasthesame (eds) 1998: Rote Liste gefahrdeter Tiere Deutsch—
habitat,foodpreferencesandrange(theyarealso lands. 7 Schriftenreihe fur Landschaftspflege und
Naturschutz55: 17434.
dividedby the Carpathians), but in spite ofthat
Borowiec, L., Scibior, R. & Kubisz, D. 2011: Critical
our study revealed distinctive differences in the check-list ofthe Polish Chrysomeloidea, excluding
mitochondrial C01 andITS1 markers as well as Cerambycidae (Coleoptera: Phytophaga). 7 Genus
inthe Wolbachiagenesinthedifferentpartsofits 22(4): 5797608.
range. All these populations have been strongly Clark, S. M., LeDoux, D. G., Seeno, T. N., Riley, E. G.,
Gilbert, A. J. & Sullivan, J. M. 2004: HostPlants of
isolated for a long time and it is likely that the
LeafBeetles Species Occurring inthe United States
Wolbachiasweepsoccurredlongtimeago.How- andCanada.7ColeopteristsSocietySpecialPublica—
ever, the situationofC. quinquepunctata is quite tionNo.2,Sacramento,California,UnitedStates:476
different despite similar habitats, the same host PP-
plants andthe general distribution, the latterbe- Clement,M.,Posada,D. &Crandall,K.A. 2000: TCS: a
ing more compact as itconsists ofonly two iso- computerprogram to estimate gene genealogies. 7
MolecularEcology9: 165771659.
latedareas. Farkac, J., Kral, D. & Skorpik, M. (eds) 2005: Cerveny
Generally,verylowgeneticvariabilityofthe seznamohroienychdruhfiCeske’republiky.Bezobrat—
beetleandits Wolbachiawithinandamongpopu- 1i.7AgenturaochranyprirodyakrajinyCR,Praha,
lationsisunusual.Otherstudiedspeciesofsteppe 760pp. [InCzech]
Kajtoch, L. 2011: Conservation genetics ofxerothermic
beetles (Crioceris quatuordecimpunctata, Chei—
beetles in Europe: the case ofCentricnemus [euca-
lotoma musczformis, Centricnemus leucogram—
grammus.7JournalofInsectConservation15:7877
mus, Coraebuselatus)havehighinterpopulation 797.
genetic diversity (Kajtoch 2011, Kubisz et al. Kajtoch, L., Korotyaev, B. & Lachowska-Cierlik, D.
2012a,Kajtochetal.,2013a,b).Forthesespecies, 2012: Genetic distinctness ofparthenogenetic forms
of European Polydrusus weevils of the subgenus
differentphylogeneticlineageswerefoundinthe
Scythodrusus.7InsectScience 19: 1837194.
north, east and south-west fromthe Carpathians
Kajtoch, L, Kubisz, D., Gutowski, J. M. & Babik, W.
andthispatternis commonto all species studied 2013a: EvolutionaryunitsofCoraebuselatus(Cole—
so far. optera: Buprestidae) incentral andeasternEurope7
The example of C. quinquepunctata shows implications for origin and conservation. 7 Insect
Conservation and Diversity. DOI: 10.1111/icad.
thatpopulation genetics ofsteppe leaf-beetles is
12031
highlydiversifiedandthatthehistoryandcurrent
Kajtoch,L.,Kubisz,D.,Lachowska-Cierlik,D.&Mazur,
populationstructureofstudiedspeciesmaydiffer M. A. 2013b: Conservation genetics ofendangered
fromwhatwehaveknownsofar.Thatiswhyfur- leaf—beetle Cheilotoma musczformis populations in
110 Mazuretal. ' ENTOMOL. FENNICAVol. 25
Poland.7JournalofInsectConservation 17(1): 677 Michalik, S. &Zarzycki, K. 1995: Managementofxero-
77. thermicgrasslandsinPoland:botanicalapproach. 7
Kajtoch,L. &Lachowska—Cierlik,D. 2009: Geneticcon— ColloquesPhytosociologiques24: 8817895.
stitutionofparthenogeneticformofPolydrususinus- Nehring,A. 1890: UeberTundrenundSteppenderJetzt—
tus(Coleoptera: Curculionidae)7hintsofhybridori— undVorzeit. 7F. Dummler, Berlin, Germany. 257
ginandrecombinations.7FoliaBiologica(Cracow) PP-
57: 1497156. Nei,M. &Rooney,A. P. 2005: Concertedandbirth—and
Kubisz,D.,Kajtoch,L.,Mazur,M.A.,Lis,A. &Holeco— Heathevolutionofmultigenefamilies.7AnnualRe-
va,M.2012a:Conservationgeneticsofhighlyisolated viewofGenetics39: 1217152.
populationsofthexerothermicbeetle Criocerisqua- Pawlowski,J Kubisz,D.&Mazur,M.2002:Coleoptera.
tuordecimpunctata (Chrysomelidae). 7Invertebrate 7In:Glo.,wacinski,Z.(ed.),Redlistofthreatenedani-
Biology 131(4): 3337344.
malsinPoland:887110.PolishAcademyofSciences,
Kubisz,D.,Kajtoch,L.,Mazur,M.A.&Rizun,V.2012b:
InstituteofNatureConservation,Krakow. 155pp.
Molecular barcoding for central-eastem European
Schmitt, M. & Ronn, T. 2011: Types of geographical
Crioceris leaf—beetles (Coleoptera: Chrysomelidae).
distributionofleafbeetles(Chrysomelidae)inCentral
7CentralEuropeanJournalofBiology7(1): 69776.
Librado, P. &Rozas, J. 2009: DnaSPv5: Asoftware for Europe.7Zookeys 157: 1317158.
comprehensiveanalysisofDNApolymorphismdata. Stewart,J.R.,Lister,A.M.,Barnes,1.,&Dalen,L.2010:
7Bioinformatics25: 145171452. Refugiarevisited:individualisticresponsesofspecies
Lobl,I.&Smetana,A.(eds.)2005:CatalogueofPalaearc— inspaceandtime.7ProceedingsoftheRoyalSociety
ticColeoptera.Volume6.Chrysomeloidea.7Apollo B: Biologicalsciences277: 6617671.
Books, Stenstrup. 924pp. Tamura, K., Peterson, D., Peterson,N., Stecher, G.,Nei,
Mazur,M.&Kubisz,D.2013:Distributionandmigration M.&Kumar, S.2011:MEGA5:MolecularEvolutio—
ofthexerothermicbeetles(Coleoptera)intheVistula naryGeneticsAnalysisusingMaximumLikelihood,
Rivervalley.7InstituteofSystematicsandEvolution Evolutionary Distance, and Maximum Parsimony
ofAnimalsPAS,Krakow. 254pp. Methods. 7 Molecular Biology and Evolution 28:
Monros,F.1960:LosgenerosdeChrysomelidae(Coleop— 273172739.
tera).7OperaLilloana, 3: 17336 [InSpanish.]
