The importance of Anatolian mountains as the cradle of global diversity in Arabis alpina, a key ...

12 Pages·2011·0.82 MB·English

Annalsof Botany108:241–252, 2011
doi:10.1093/aob/mcr134,available onlineatwww.aob.oxfordjournals.org
The importance of Anatolian mountains as the cradle of global diversity
in Arabis alpina, a key arctic–alpine species
Stephen W. Ansell1,*, Hans K. Stenøien3, Michael Grundmann1, Stephen J. Russell2, Marcus A. Koch4,
Harald Schneider1 and Johannes C. Vogel1
1Department of Botany, The Natural History Museum, London SW7 5BD, UK, 2Department of Zoology, The Natural History
Museum, London SW7 5BD, UK, 3Systematics and Evolution Group, Section of Natural History, Museum of Natural History
andArchaeology,NorwegianUniversityofScienceandTechnology,Trondheim,N-7491,Norwayand4DivisionofBiodiversity D
o
and Plant Systematics, Institute of Plant Sciences, University of Heidelberg, Im Neuenheimer Feld 345, Heidelberg, w
n
D-69120, Germany lo
a
*For correspondence. E-mail [email protected] de
d
fro
Received:19December2010 Returnedforrevision:1March2011 Accepted:4April2011 Publishedelectronically:28June2011 m
h
ttp
†deBsacrcikbgerdouansdeiathnedrAaimcesnAtrenaotfoloiarigisinaabnidolloognigc-atlelyrmdiPvleerisseto,cbeunteprheyfluoggieuomg,raoprhaicsaallyceunntrdeefro-erxgpelonreetdicraemgiaolng.aImtais- s://ac
a
tion,fedfromdistinctneighbouringrefugia.Thesecontrastinghypothesesaretestedthroughaglobalphylogeo- d
e
graphic analysisofthe arctic–alpine herb,Arabisalpina. m
†Methods Herbarium and field collections were used to sample comprehensively the entire global range, with ic.o
specialfocusonAnatoliaandLevant.SequencevariationinthechloroplastDNAtrnL-trnFregionwasexamined u
p
in483accessions.Ahaplotypegenealogywasconstructedandphylogeographicmethods,demographicanalysis .c
o
anddivergencetimeestimationswereusedtoidentifythecentresofdiversityandtoinfercolonizationhistory. m
†KeyResultsFifty-sevenhaplotypeswererecovered,belongingtothreehaplogroupswithnon-overlappingdis- /ao
b
tributionsin(1)NorthAmerica/Europe/northernAfrica,(2)theCaucuses/IranianPlateau/ArabianPeninsulaand /a
(3)Ethiopia–easternAfrica.AllhaplogroupsoccurwithinAnatolia,andallintermediatehaplotypeslinkingthe rtic
three haplogroups are endemic to central Anatolia and Levant, where haplotypic and nucleotide diversities le
-a
exceeded all other regions. The local pattern of haplotype distribution strongly resembles the global pattern, b
andthe haplotypesbegan to divergeapprox.2.7Mya,coinciding withthe climate cooling ofthe early Middle stra
Pleistocene. c
†Conclusions The phylogeographic structure of Arabis alpina is consistent with Anatolia being the cradle of t/10
8
origin for global genetic diversification. The highly structured landscape in combination with the Pleistocene /2
climatefluctuationshascreatedanetworkofmountainrefugiaandtheaccumulationofspatiallyarrangedgeno- /2
4
types. This local Pleistocene population history has subsequently left a genetic imprint at the global scale, 1
/1
through four range expansions from the Anatolian diversity centre into Europe, the Near East, Arabia and 5
2
Africa. Hence this study also illustrates the importance of sampling and scaling effects when translating 1
1
globalfromlocal diversitypatterns duringphylogeographicanalyses. 8 b
y
g
Keywords: Anatolia, centre of origin, Pleistocene glaciations, chloroplast trnL-F, divergence times, alpine u
e
plants,Arabis alpina. s
t o
n
0
4
A
p
INTRODUCTION Anatolia is a hotspot for biological diversity in the ril 2
Mediterranean basin and is a biogeographically interesting, 01
9
Geneticdiversityisoftenstructuredwithinspeciesdistribution but under-explored region. It is situated between the
ranges (Taberlet, 1998; Hewitt, 2000, 2004; Me´dail and European and Asian continents and is a meeting point for
Diadema, 2009). Local hotspots of diversity may arise the European and Turko-Iranian floras, which broadly
through several means: the long-term survival of populations overlap in central Anatolia (Davis, 1965–85). During the
during past climatic fluctuations (glacial refugia; Hewitt, Pleistocene (approx. 2.5 Mya) glacier development within
2004; Schmitt, 2007), or through the recent amalgamation of Anatolia was limited to the higher mountain peaks (Atalay,
divergent lineages (Ferris et al., 1998; Petit et al., 2003; 1996), while the lowlands remained open, developing steppe
Walter and Epperson, 2005), or local ecological niche gradi- communities (Michaux et al., 2004; Magyari et al., 2008).
ents may promote the accumulation of lineages (Ricklefs, These provided suitable habitats for temperate species to
2007). Discriminating among the first two scenarios still survive the last glacial maximum (Rokas et al., 2003; Dubey
remains a challenge when interpreting phylogeographic et al., 2006; Fritz et al., 2009) and perhaps in earlier cycles.
history, particularly outside the much studied European and Late Pleistocene sea levels around the Mediterranean and
North American continents. Caspian seas were regularly lower than current levels
#The Author2011.Publishedby OxfordUniversity Presson behalfofthe AnnalsofBotanyCompany.Allrights reserved.
For Permissions,please email:[email protected]
242 Ansell et al. — Anatolian mountain plant phylogeography
(approx.130m,Kereyetal.,2004)andlandbridgesroutinely (Koch et al., 2006; Assefa et al., 2007; Ehrich et al., 2007).
formed across the Sea of Marmara and the Bosporus Strait TheimportanceofAnatoliaatthecentreoftheglobaldiversity
(Magyari et al., 2008). This allowed for frequent inter- range for A. alpina was recognized by a ‘three-times out of
continental population exchange, especially for temperate Asia-Minor’ hypothesis (Koch et al., 2006), but sampling
organisms (Kucˇera et al., 2006; Dubey et al., 2007; Gu¨ndu¨z was modest (n¼3). An alternative hypothesis of Anatolia as
et al., 2007). In the east, the high ‘Anatolia diagonal’ a centre for the amalgamation of European and Near East
(Davis, 1971) mountains (.2000m) have limited the mixing lineages is considered for other Brassicaceae (Kucˇera et al.,
of European and Near-Eastern biota (Ekim and Gu¨ner, 1986; 2006). These two phylogeographic hypotheses depend on the
Bilginetal.,2006),buttheirtopographymayalsohavefacili- availability of suitable ecological and geographical connec-
tated local survival and speciation, as suggested by the high tions between Anatolia and adjoining territories during the
level of endemism (Davis, 1971; C¸iplak, 2003). Together Pleistocene. This is especially true for alpine species like A.
this combination of geography and climatic history has pro- alpina, as colonizations between Anatolian and adjoining
D
motedboththelong-termlocalsurvivalofbiota,andrepeated mountain systems are limited to glacial periods when alpine ow
uni- or bi-directional colonization(s) from nearby areas, habitats are more widespread at lower elevations. As such nlo
makingAnatoliaacomplexbutimportantareaforunderstand- we recognize that two key geographic features may have hin- ad
e
ing the diversification of Eurasian biodiversity. dered or aided Pleistocene colonization of Anatolia: the d
Increasingly the biogeographic history of Anatolia and Thyracian plain and the ‘Anatolian diagonal’. Consequently fro
m
adjoining areas is being explored using molecular tools. we put forward two additional hypotheses: (1) that the low h
While data are still limited, bats, rodent, turtles and wasp laying areas around the Sea of Marmara and adjacent ttp
s
groups are now represented (e.g. Michaux et al., 2004; Thyracian plain (,1000m) with its Mediterranean climate ://a
Bilgin et al., 2006; Dubey et al., 2007; Gu¨ndu¨z et al., 2007; have presented an ecological barrier to European/Asian ca
d
Flanders et al., 2009; Furman et al., 2009; Fritz et al., migrations;(2)thatthehighmountainsofthe‘Anatoliandiag- e
m
2009). Plants remain relatively poorly represented, despite onal’haveallowedpopulationexchangebetweenAnatoliaand ic
.o
several studies focusing on trees (Bartish et al., 2006; adjacent Caucasus and Near East mountain systems during u
p
Kucˇera et al., 2006; Go¨mo¨ry et al., 2007; Naydenov et al., ecologically favourable glacial periods. .c
o
2007) or herbs (Jakob et al., 2007; Font et al., 2009; Parolly To explore these two alternative hypotheses the following m
/a
et al., 2010). Unfortunately low sampling densities have specific questions are raised. (1) Is Anatolia a centre of o
b
limited the ability to characterize the location of refugia, global genetic diversity for A. alpina ? (2) Is the local /a
hybrid zones and colonization routes (Gu¨ndu¨z et al., 2007). Anatolian diversity comprised of related haplotypes, as rtic
le
Nevertheless several phylogeographic trends emerge from expected under a scenario of in-situ survival and local -a
b
these studies, but with no clear consensus across all groups: lineage accumulation? (3) Is genetic diversity geographically s
(1)NearEastandEuropeanlineagesaregeneticallydivergent; partitioned around the Sea of Marmara? (4) Is there evidence tra
c
(2)AnatoliawasthesourceareaforEuropeancolonization;(3) of range expansion associated with the ‘Anatolian diagonal’. t/1
0
the Thracian Plain/Sea of Marmara is a barrier to European (5) In what way has the Anatolian diversity contributed to 8
/2
colonization; and (4) the high Anatolian diagonal mountains the diversity of Europe, the Near East and eastern Africa? /2
4
are a barrier to colonization from the Near East (Fig. 1). These questions are addressed by a global chloroplast 1
/1
Phylogeographic information from alpine species is currently DNAphylogeographicanalysisofA.alpina,usingtheherbar- 5
2
1
missing and would complement the existing information ium collections from the Flora of Turkey project (Davis, 1
8
from temperate species, especially in a context of the 1965–1985) as a sampling resource to place special focus b
y
Anatolian mountains as a refugium. on Anatolia and associated Levant. g
u
The arctic–alpine herb Arabis alpina was selected to e
s
explorethephylogeographichistoryoftheAnatoliamountains t o
MATERIALS AND METHODS n
and relationships with adjacent mountainous areas. Arabis 0
4
alpina is a short-lived perennial belonging to the mustard Leafmaterialfrom 182individualsof Arabisalpina L.,repre- A
p
fmamodielyl oBrgraasnsiiscmaceAaerab(iHdoegpis,is1t9h8a6l)ia,naand(Kaochreleattivael.,o2f00th0e, sfreonmtinhger1b3a5riupmrevspioeucsimlyenusnpduebploissihteeddilnocthaelitfioelsl,owwienrgehsearmbaprliead; ril 20
1
2001, 2007; Beilstein et al., 2008; Couvreur et al., 2010). Berlin-Dahlem (B), Copenhagen (C), the Natural History 9
Arabis alpina itself is currently being developed as a model Museum London (BM) and the Royal Botanical Gardens
for studying alpine and life-history adaptation (Wang et al., Edinburgh (E). This sampling targeted mainly Iran, Iraq,
2009; Poncet et al., 2010), although complicated by loss of Lebanon, Saudi Arabia, Syria, Turkey and Yemen. To a
self-incompatibility in parts of the Europe (Tedder et al., lesser extent, material was sampled from Armenia,
2011). The herb is distributed throughout the alpine habitats Azerbaijan, Crete, Crimea (Ukraine), Georgia, Greece,
in Europe, in the arctic zone of Greenland and North Ethiopia, Kenya, Tanzania, Uganda and former Yugoslavian
America, in the high mountains of northern and eastern states (Supplementary Data Table S1, available online). The
Africa and Anatolia; and in the ranges extending into the GLOBAL GAZETTER version 2.1 (http://www.fallingrain.
Caucasus and the Near East (Meusel et al., 1965). The com/world) was used to georeference herbarium voucher
global phylogeographic structure of A. alpina was outlined localities with the derived latitude/longitude co-ordinates
by an earlier chloroplast DNA studies, and three haplogroups reduced to two decimal places.
were recovered from the core areas adjacent to Anatolia: TotalgenomicDNAwasextractedfromtheleaftissueusing
Arctic/Europe/north Africa, the Near East and east Africa a combination of the Retsch Tissue Lyser and Biosprint 96
Ansell et al. — Anatolian mountain plant phylogeography 243
Crimea
Thyracian plain
Marmara Sea TPAaounnrttuii-cs moAunnattaoliinasn Diagonal D
Taurus o
w
n
lo
a
d
e
d
Western fro
Taurus m
Mt. Lebanon h
range ttp
s
400 km ://a
c
a
d
e
FIG. 1. SomeimportantAnatoliangeographicfeatures.DiagonaladaptedfromDavis(1971). mic
.o
u
p
BioRobot (Qiagen) workstation and the Biosprint 96 plant respectively after examination of the original electrophero- .c
o
DNA extraction Kit (Qiagen). The trnL-trnF (trnL-F) region grams. In total, 57 unique trnL-F haplotypes are recognized m
/a
including the trnL(UAA) gene and the trnL-trnF(GAA) inter- for A. alpina. The relationship among haplotypes was recon- o
b
genic spacer (IGS) was amplified using PCR and the ‘C’ and structed using statistical parsimony with a 95% connection /a
‘F’primers(Taberletetal.,1991),accordingtotheconditions threshold (Templeton, 1992) and the TCS program version rtic
specified in Ansell et al. (2007). Both DNA strands were 1.21 (Clement et al., 2000). All haplotypes were re-classified le-a
sequenced using BigDye Terminator Kit version 1.3 to obtain a unifying scheme (Supplementary Data Table S2) bs
(Applied Biosystems, ABI), and an ABI 3730 capillary DNA to overcome the previous adoption of both Arabic numeric tra
c
analyser. The trnL-F IGS sequences were assembled using and Roman numeral haplotype naming schemes (Koch et al., t/1
0
SEQMAN version 6.00 (Lasergene, DNAstar). 2006; Assefa et al., 2007). 8
/2
Sequence data from published studies, especially for Sampleswereorganizedintotengroupingsbasedonmoun- /2
4
European and eastern African specimens were also tain range and observed haplotype distributions (Fig. 2A and 1
/1
utilized. The following chloroplast trnL-F IGS sequences of SupplementaryDataTableS1).Haplotyperichness,haplotype 5
2
Arabis alpina were recovered from GenBank: DQ060112– diversity and nucleotide diversity was estimated for each 11
8
DQ060145 (Koch et al., 2006; 142 accessions world-wide, region. b
y
including 122 from Europe, Greenland and Canada), Haplotype richness (R) with correction for the smallest g
u
EF449508-EF449514 (Assefa et al., 2007; 66 accessions regional sample size (n¼16) through rarefraction (El e
s
from 11 populations in the eastern African mountains) and Mousadik and Petit, 1996) was calculated using CONTRIB t o
n
EU403083–EU403084 (Ansell et al., 2008; 105 accessions (Petit et al., 1998). Nucleotide diversity (p; Nei, 1987) and 0
4
from 34 populations in the Italian mountains and European genetic differentiation by analysis of molecular variance pro- A
p
Afrolpms).29T6helotcoatatilodnasta(Sseutpcpolemmpernisteadry4D83atatrnTLa-bFleIGS1S).sequences c3e.1d0ur(eE(xAcoMffiOeVrAet)awl.e,re20c0a5lc).ulDatieffderuesnitnigatiAonRLwEaQsUinIiNtiavlleyrsciaoln- ril 20
1
culated from haplotype frequencies (F ), subsequently 9
ST
incorporating haplotype pairwise genetic distances (F ), as
Data analysis ST
the number of character changes. The significance of differ-
Sequencesfromthenewandpublishedstudieswerealigned entiation was assessed by 1000 non-parametric permutation
using Clustal X algorithm (Thompson et al., 1994) and sub- tests (Excoffier et al., 1992), as performed by ARLEQUIN.
sequently manually corrected using MEGALIGN version G and N are analogous to F and F (Pons and
ST ST ST ST
6.00 (Lasergene, DNAstar). New haplotypes types were sub- Petit, 1996), and when the genealogical relationship of haplo-
mitted to GenBank (accession numbers JF705219–705252). types is significantly correlated to geographic distribution of
Alignment positions 419–435 comprised a highly variable haplotypes, estimates of N (F ) are predicted to be sig-
ST ST
microsatellite C T (Supplementary Data Table S2), nificantly greater than G (F ) (Pons and Petit, 1996).
(1–5) (8–12) ST ST
which was excluded during haplotype discrimination. Two Estimates of G and N were calculated using PERMUT
ST ST
haplotypes previously reported as being private to single version 2.0 (Pons and Petit, 1996), and the significance of
accessions from Croatia and Iraq (types 16 and 31; Koch difference between the two was assessed by 1000
et al., 2006) where reclassified as haplotypes 13 and 04 permutations.
244 Ansell et al. — Anatolian mountain plant phylogeography
A
1
2 7 C
4
3 5 6
8 D
o
w
9 n
lo
a
D de
B Common haplotype (n > 25) B11 10 1000 km d fro
Rare haplotype (n ≤ 12) m
B8 G4 D 400 km h
ttp
R3 G10 s
R13R2 BB34B5B6B7 B9B10 B12B13B14BB1B51167G3 G7 G8 G9G11 8 ://academic
R12 B2 B1 G1 G2 G6 G12 .ou
R10 R11R9R8R7 R6 RB310B29B28B27B2652B B23B22B2B12B0B1198 G1G413 9 p.com/aob/a
R4 rtic
B24 G5 le
-a
b
C R5 strac
t/1
7 0
8
/2
/2
4
n = 6 per n = 1 per 1
4 population population /15
2
6 1
1
8
5 b
y
g
u
e
s
t o
n
0
4
A
p
ril 2
0
1
9
n = 1 per population 400 km
FIG.2. (A)GlobaldistributionrangeofArabisalpina,andthethreemajorchloroplasttrnL-trnFIGShaplotypegroups.DistributionadaptedfromMeuseletal.
(1965)andJalasandSuominen(1994),withdottedlinesdelimitingthetenregionsusedforgeneticanalysis.(B)Thegenealogicalrelationshipamongthe57
uniquechloroplasthaplotypes,withinferredhaplotypesequencesshowninblack.(C,D)RegionaldistributionsofhaplotypesinAsiaMinor(C)andtheArabian
PeninsulaandEthiopia(D).Locationswithmultiplesamplesareindicatedbyblackdotsandconnectinglinesdefineregions:(1)ArcticandNorthAmerica,(2)
westernandcentralEurope,(3)northAfricaandMacronesia,(4)south-eastEurope,(5)westernAnatolia,(6)centralAnatoliaandLevant,(7)Caucasusand
IranianPlateau,(8)ArabianPeninsula,(9)Ethiopia,(10)eastAfrica.n,Samplesize.
Demographichistorywasexploredbymismatchdistribution by ARLEQUIN. Prior to estimating among haplogroup diver-
analysesofobservedhaplotypepairwisedifferences,following gence time, each haplogroup was tested for deviations from
a sudden population expansion model (Rogers and neutral expectations by Tajima’s D test (Tajima, 1989), as
Harpending, 1992), employing 1000 bootstraps, as performed implemented in DnaSP version 4.50.3 (Rozas et al., 2003),
Ansell et al. — Anatolian mountain plant phylogeography 245
and intraspecific recombination was tested using the SITES Anetworkconsistingofthreemajorhaplogroupswasobtained
software (Hey and Wakeley, 1997). Since no deviations from fortheuniquesequencesandthesearecolouredblue(B),green
neutrality and no signs of intraspecific recombination were (G)andred(R)inFig.2.
found, time since divergence between haplogroups was esti- Thesampleswerethenorganizedintotengroupings,reflect-
mated by the IM program (Nielsen and Wakeley, 2001; Hey ing mountain systems and observed haplotype distributions
and Nielsen, 2004). IM implements a Bayesian analysis (Fig. 2A and Supplementary Data Table S1). Samples from
approach based on coalescent theory and using Markov Levant (Mt Lebanon complex) and central Anatolia were
ChainMonteCarlomethodologytosamplefromagivenprob- pooled (region 6) due to a sharing of rare related green and
ability distribution. It has been shown that sampling of single red haplotypes (Supplementary Data Table S1). The global
haplotypesfromeachofmanydemesshouldyieldgenealogies distribution of haplogroups is given in Fig. 2A, and detailed
with similar properties as belonging to random mating popu- distributions for Anatolia and eastern Africa are given in
lations (Wakeley, 1999, 2001; Wakeley and Aliacar, Fig. 2C, D. The B haplogroup occurs throughout the western
D
2001).Three major haplogroups were recognized: blue (B), Anatolian–European–Arctic parts of the species range, the ow
green (G) and red (R) (see below). Genetically non-admixed G haplogroup occurs in the eastern Anatolian–Caucasus– nlo
regions representing each haplogroup were selected to Iranian Plateau–Arabian Peninsula part, but also enters the ad
e
estimate the divergence times between the genetic clusters. northern Ethiopian mountains (Fig. 2D). The R haplogroup d
The B–G-haplogroup separation was estimated by comparing mainly occurs in the eastern African part of the species fro
m
region 3 with regions 7 and 8 (Fig. 2), the B–R separation by range (red lineage; Fig. 2B, D), but is narrowly distributed h
comparing regions 3 and 10, and the R–G-haplogroup separ- in southern parts of central Anatolia and Levant (pink and ttp
s
ation by comparing regions 7 and 8 with region 10. The R– light orange lineages; Fig. 2C). central Anatolia/Levant is ://a
G analysis should be treated cautiously, as the B haplogroup the only part of the species range were all three haplogroups ca
d
is more similar to both R and G, than R and G are to each occur, including the three important haplotypes B1, G1 and e
m
other, violating the IM assumption that no populations R1, which link the three haplogroups together (Fig. 2B, C). ic
.o
should be more genetically similar to the populations u
p
implemented in the model than to each another. .c
Phylogeographyand population genetics o
Three separateruns wereperformed foreach pairwise com- m
parison,oneshort(runtime≥1h)andtwolong(runtimelong The dataset was structured into ten geographic groups /ao
b
enough to achieve at least 30 million steps in chains after a (Table 1). Only four haplotypes, G6, R6, R9 and B1, were /a
burn-in of 106 steps), with minimum effective sample size widespread. Thirty-two of the 57 haplotypes were private to rtic
le
(ESS) recorded being 138, and most ESS values being single accessions (Appendix I), with 50 private to single -a
b
.1000. The average B–G estimate is based on four IM regions. Central Anatolia was the most genetically diverse s
runs, the B–R estimate is based on two IM runs, while the region, having the greatest haplotype diversity in absolute tra
c
R–G estimate is based on four IM runs with similar prior terms,with13of57haplotypes,thehighesthaplotyperichness t/1
0
values but different random number seeds. Average substi- atR¼6.378andthehighestnucleotidediversityatp¼0.423 8/2
tution rates was set to 1.3×1029 per nucleotide % (Table 2), exceeding the pooled global population value at /2
4
(Richardsonetal.,2001),averagegenerationtimewasarbitra- p¼0.319%. Haplotype B1 dominates throughout the 1
/1
rily set to 5 years (which does not influence divergence time B-haplogrouprange,andwesternAnatoliahadthehighesthap- 5
2
1
estimates), and historical migration rates was set to zero to lotype and nucleotide diversity within the B haplogroup 1
8
reduce the number of parameters in the model. Since no (Table 2). Both the haplotypic and nucleotide diversity b
y
migration is assumed, the estimated divergence times reduced progressively from south-east Europe towards the g
u
betweenhaplogroupsshouldbeviewedasminimumestimates. Arctic (regions 4–1 in Fig. 2A). Haplotype G6 dominates e
s
Intheinitial,shortIMruns,upperpriorvaluesofthemutation throughout the G-haplogroup range, with regions 7 and 8 t o
n
anddivergencetimeparametersweresetto100.Inthetwofol- (Fig. 2A) having similar low values of haplotype richness 0
4
lowing long runs, prior values were adjusted according to and nucleotide diversity (Table 2). Haplotypes R6 and R9 A
p
iwneitriealirnecsluuldtse,d.enPsuriroinrgvtahluatestheforcotmheplevteariloikueslihaonoadlyspersofialeres aarreeascoomfmthoen Rin-hEapthloiogproiaupanrdanegaes.teTrnheAEfrthiciao,pitahne mtwoounmtaainins ril 20
1
shown in Supplementary Data Table S3. had substantially higher haplotypic and nucleotide diversity 9
compared with the rest of eastern Africa (Table 2), reflecting
the local mixing for G and R haplotypes (region 9; Fig 2D).
RESULTS At a global scale a significant proportion of genetic
variation was partitioned among populations, and F was
Haplotype diversity and distribution ST
0. 438 (P¼0.0001) when based solelyon haplotype frequen-
Thetotal chloroplasttrnL-F dataset comprised483 accessions cies, and F was 0.631 (P¼0.0001) when pairwise haplo-
ST
of A. alpina sampled from 296 locations, and 71 trnL-F type genetic distances were also considered. G and N are
ST ST
types were recognized, 34 of which are newly discovered analogous measures to F and F and were estimated to
ST ST
(types38–71).Thealignmentpositions419–438compriseda be 0.439 and 0.557, respectively, and N was significantly
ST
complex variable microsatellite C T (Supplementary larger(P¼0.05),indicatingrelatedhaplotypesaregeographi-
(1–5) (8–12)
DataTableS2),thesizeofwhichvariedindependentlyamong cally arranged. Again, N (0.521) was significantly higher
ST
unrelated sequences. After excluding this region, 57 unique than G (0.357) for the central Anatolian diversity hotspot
ST
haplotypes were recognized (Supplementary Data Table S2). and the neighbouring regions (regions 5–7), indicating local
246 Ansell et al. — Anatolian mountain plant phylogeography
R13 .. . .... .11 haplotype diversity is geographically arranged in this critical
R12 .. . .... ..1 area (P¼0.01).
R10R11 .... .. ........ ..5..1 0A.n0Ga0te0on1lei)a,tiacdnidvdieftrfhseiirtseywnhtaioasttsimpoonatinawmlyaosdncugoenrtesogiditohenreasdboilfeuftFesrieSdnTec¼oesf0at.hm7e7o3cne(gnPttrh¼ael
R8R9 .... .. ........ ..216.12 main B/G/R-haplogroup ranges FST¼0.745 (P¼0.0002),
R7 .. . .... ..3 rather than due to differences within B/G/R-haplogroup
s R6 .. . .... .721 ranges. This was supported by individual AMOVA tests on
alysi R4R5 .... .. ....21.. ..... t0h.0e41c,omPb¼ine0d.00r0eg1i)o,ntsheofG-thhaeplBog-hroapulpogrraonugpe r(aFnge¼(F0.S0T0¼6,
an R3 .. . ..1. ... P¼0.3450) and the R-haplogroup range (F ¼S0T.183, P¼
forgenetic G13G14R1R2 ........ .... ..........11.... 11.......... l40tah.t0eai0onS0nd1sea)5f.r,ooTFmfheSMsTroea¼urimtsh0a-.le0riaat9ts5lte(,FEigPuger.¼noe2pt0Aei.c0,a0ndC5di))f.,fweCwreesohntneitcivrahnetriAosaSenrnTelyaa,tsomeplpiooaanprg(uartleeapgdtoiioopbnnuyss- Download
ed G12 .. . ...1 ... from central Anatolia and the eastern Anatolia–Iranian ed
us G11 .. . ...1 ... Plateaux had higher genetic differentiation (regions 6 and 7; fro
viduals(n) G7G8G9G10 ........ .... ............2.11 .....2...... FtivSiTty¼v0ia.3t3h3e,APn¼ato0l.i0a0n0d),iadgeosnpaitlemgoreuanttearingesyosgtreamphs.ic connec- m https://ac
mbersofindi G2G3G4G5G6 .......... ..... ..........511318....52 ....14....11..... itDnheceSmlueospdugaidrnradagpteehnthicmeehixrsiepsmptaoranretsycsiehoannnatdantmaidvlioyevdseeAersglneoan(tPncoel¼tihae0d.B7iv1ea0rn,sidtPyG,¼dhi0da.p6nl6oo0gt)r,roeujaepncsdt, ademic.oup.c
nu G1 .. . ..2. ... both followed a uni-modal mismatch distribution (Fig. 3A, om
egionsand B27B28B29B30 ........ ...1 11....1......... ............ rsBRee)-jp.heacartpaeTltdoehglewyroh(uPespnu¼dtdhd0ieev.n0eAr0sf1iret;iycxFapin(gaPn.Rs3¼i-Dohn0a)..p0Ml5om0itsy;ompdFeaeitldgci.hvwe3arnCasasi)lt,yysarwinesdajoesncwatetnahdaselydaslfaesotodar /aob/article-a
r b
phic B25B26 .... .. ..15.... ...... dsieornivmedodfreolmfowrietihtihnerAtnhaetoGliahadpidlongoroturpeje(Pct¼the1.s0u0d0d),enorexthpeanB- strac
gra B24 .. 1 11... ... haplogroup (P¼0.270). Insufficient data (n¼6) prevented a t/10
ngeo B22B23 .... .. 1....... ...... meTahneinIgMfulanteasltysfoesr rtheseuAltendationliacoRnv-heargpelongcreouopf pdaivraemrseittye.resti- 8/2/24
thete B20B21 ...2 .. 2....... ...... mallatpeasi,rswoitfhlohniggh-ruEnSsS,avnadlupeoss,tearlmioorsetstiidmenatteicsanlodtisnteraibrutthieonesdgoef 1/1521
among B18B19 ...1 .. ..1..... ...... otlrofigburuontuiiofponsrsm(wFpeirrgei.oer4svt)i:amlutaheteesd. BTfoh–reGadllivhpeaarpigrlwoengiscroeeuctpiomsmebhiapndraotbitohanbesilihotifyghhdaeipss--t 18 by g
es B17 .. 1 .... ... probability for t¼0.66 Mya (averaged over four runs, 95% ue
plotyp B15B16 ...1 1. ........ ...... tChIebheitgwheeestnp0r.o2b5abainlidty3f.8o6r tM¼ya0).,73thMe yBa–(Ravhearapgloegdroouvpesr thwado st on 0
ha B14 .. . 2... ... runs 95% CI between 0.23 and 2.48 Mya), and the highest 4 A
onof B12B13 .12. .. ........ ...... Mproybaab(ailvietyragfoerdthoeveRr –foGu-rharpulnosgr9o5up%seCpIarabteiotwnewenas0t.8¼02a.n6d9 pril 20
buti B11 .. . 6... ... 6.43Mya).Thelatterincludedthehaplo-groupswiththegreat- 19
stri B10 .. . 1... ... est nucleotide distances, and therefore represents an estimate
Di B9 .. . 2... ... for the overall age of extant haplotype diversification.
1. B7B8 .... .. 11...... ......
E
L B6 .. . 1... ... DISCUSSION
B
A B5 .1 . .... ...
T ThisstudyconfirmstheimportanceofAnatoliaandassociated
B4 .. . 1... ...
B3 .6 . .... ... Levant in understanding the global diversification history of
B2 .. 2 ..1. ... the arctic–alpine Arabis alpina. From the dense sampling in
B1 2152 20 118127. ... the present study, extensive genetic diversity was recovered,
n 2265 26 149214458 164439 including substantial new chloroplast sequence variation not
Region ArcticWesternandcentralEuropeNorthAfricaandMacronesiaSouth-east.EuropeWesternAnatoliaCentralAnatoliaEasternAnatoliaandIranianPlataeuxArabianpenisulaEthiopiaEasternAfrica reseetcnaotliv.n,egr2e0md0u7bc;yhAopnfrseevtlhlioeeutwsaolsr.t,lud2d-0wie0is8d)e(,Krdaoencsghpeit.eetInthaetlos.,eta2slt0ut0hd6ei;esnAurmesspberfeear-
Ansell et al. — Anatolian mountain plant phylogeography 247
TABLE 2. Distributionofhaplotypediversitybysamplingregion
Haplogroupfrequencies R
nhaps
nblue ngreen nred Total(n nhapsprivate private %abundencenhaps
Region n ¼ ¼ ¼ haps) individual regional regionallyprivate 16 74 p
(1)ArcticandNorth 22 22 0 0 2 1 1 4.545 0.727 – 0.013
America
(2)Western-central 65 65 0 0 7 3 6 20.000 2.438 – 0.060
European
(3)NorthAfricaand 26 26 0 0 6 4 3 11.538 3.323 – 0.077
Macronesia D
(4)South-eastern 149 149 0 0 14 8 12 13.422 2.698 – 0.083 o
w
European n
(5)WesternAnatolia 21 21 0 0 6 4 5 42.857 4.048 – 0.110 loa
(6)CentralAnatoliaand 44 8 30 6 13 6 10 40.909 6.378 – 0.423 de
(L7e)vCanatucasusandIranian 58 0 58 0 6 4 5 10.344 1.583 – 0.034 d fro
m
Plateau h
(8)ArabianPeninsula 16 0 16 0 3 2 2 12.500 2.000 – 0.036 ttp
(9)Ethiopia 44 0 13 31 7 3 3 20.000 4.440 – 0.430 s
(10)EasternAfrica 39 0 0 39 6 2 3 13.158 3.037 – 0.113 ://a
Blueregions1–5 283 283 0 0 31 20 28 20.922 – 11.314 0.077 ca
Greenregions7–8 74 0 74 0 8 6 8 9.459 – 7.000 0.035 de
Redregions9–10 83 0 13 70 10 2 9 86.747 – 8.750 0.313 mic
Redregions9–10(no 70 0 0 70 8 1 8 100.000 – – 0.130 .o
u
greenhaplotypes) p
World-wide(regions 484 291 117 76 57 32 – – – – 0.3187 .co
1–10) m
/a
o
b
Samplesize(n),frequenciesofhaplotypesbyhaplotypeclades,numbersofhaplotypes,privatehaplotypes,haplotypediversitybyrarefraction(R)for /a
correctedforsmallestsamplesizeatregional(n¼16)andhaplogrouplevels(n¼74),nucleotidediversity(p). rtic
Regions1–10areshowninFig.2. le
-a
b
s
of haplotypes increased from 31 to 57 (Supplementary Data for A. alpina, with a long history of lineage accumulation, tra
c
Table S2) and, remarkably, on average one new haplotype functioning as a major Pleistocene refugium. t/1
was discovered for every 8.5 samples at the global scale, 08
increasing to every 3.6 samples in Anatolia. All three pre- /2/2
Species origin 4
viously described haplogroups and their global distributions 1
/1
were recovered (Koch et al., 2006; Assefa et al., 2007), and The evidence presented is consistent with previous hypoth- 5
2
their distributions were confirmed to extend into the interior eses of (1) Anatolia being the cradle for Brassicaceae diversi- 11
8
of Anatolia. Furthermore, it was shown that the genotypic fication (Koch and Kiefer, 2006; Franzke et al., 2009) and b
y
and nucleotide diversity of Anatolia, especially central more specifically, (2) that the genus Arabis originated in the g
u
Anatolia (including Levant), exceeded that of all other areas. Mediterranean or Turko-Iranian areas (Meusel et al., 1965; e
s
Centresofgeneticdiversityhaveoftenbeenassociatedwith Hedge, 1976). A molecular phylogeny of the genus Arabis t o
n
range expansion and population amalgamation (Petit et al., (R. Karl et al., University of Heidelberg, Germany, unpubl. 0
4
2003; Walter and Epperson, 2005). The present results, res.)supportstheseconclusionsandconfirmstheBhaplogroup A
p
hsiotyw.evFeirrs,t,strtohnergelywiansdincaotentahrarotwAlnyatdoelilaimisitathteedcrraadnlgeeooffdihvaepr-- iwsesatnecrnes-AtranlatfooliraAh.adaltphienah.igWheistthignentehteicBd-ihvaeprsloitgyro(uTpabrlean2g)e, ril 20
1
logroup frequency shift, as typified from hybrid zones of suggesting that Arabis alpina may have originated in this 9
trees(Ferrisetal.,1998;Bartishetal.,2006),orbygrasshop- region.Analternative‘outofAfrica’scenarioisnotsupported,
pers, shrews and various bats from central Anatolia, which as the ancestral R-haplotype (R1) and descendents (R2–R5)
have divergent European and Near East lineages (Cooper are private to the areas covering central Anatolia and Levant.
et al., 1995; Bilgin et al., 2006, 2008; Furman et al., 2009). Foralpinespecies,theavailabilityofcoldhabitatsareinflu-
Secondly, the Anatolia haplotype diversity includes the most ential on species and population ranges and the accumulation
common European and Near East haplotypes (B1, G6), plus of genetic diversity (Comes and Kadereit, 2003). Using
all intermediate haplotypes linking the European, Near East coalescence analysis, it was estimated that divergence
and eastern African lineages (Fig. 2B, C). Of the remaining between the most distantly related haplogroups (R, G) com-
genealogically important haplotypes, only the red eastern menced around 2.69 Mya (95% CI 0.80–6.43 Mya). These
AfricandiversityismissingfromAnatolia,andtheseprobably results must be interpreted cautiously, reflecting the single
originated in situ due to geographic isolation from the main gene approach of the present study, and the violation of IM
species range (Koch et al., 2006; Assefa et al., 2007). Hence assumption, specifically that no other populations are more
the prsent results indicate thatAnatolia is the centre of origin closely related to the sampled populations than they are to
248 Ansell et al. — Anatolian mountain plant phylogeography
30000 for the separation of Euro-Anatolian/Near Eastern lineages
e) A Observed of yellow-necked house mouse, the white-toothed shrew and
olut25 000 Expected oak gallwasps (Michaux et al., 2004; Dubey et al., 2006;
s20000
b Challis et al., 2007), suggesting the widespread involvement
a
y (15 000 of early Middle Pleistocene climate fluctuation onthe diversi-
c
en10 000 fication of Anatolian biota.
u
eq 5000
Fr
0
1 2 3 4 5 6 7 Pleistocene glacial survival and the accumulation of diversity
Pairwise difference
Anatolia’s dissected mountainous landscape has been
5000 crucial for the long-term survival of A. alpina and its ability
B D
absolute) 3344050500000000 tPEolueriosatpcoeccauenmneAullapitcseeo-srgheSinecleadtnicdienqdauviivivaearlsaeinntydt.otnoTlyhettrhheoesehwigacshoevrnemorinogumnatatjhoiner ownload
uency ( 122505000000 p1e9a9k6s),apbroovveidi2n2g0o0pmporwtuenreitiegslafcoiartelodca(lEprionpc¸u,la1t9io7n8;suArvtaivlaayl., ed from
Freq 1500000 Fatuhrtihgehremroerlee,vtahtieonmso,uonfftaseinttisnygsttehmesdprireorvcidliemdamteotihstatcopnredviatiiolends http
0 1 2 3 4 5 6 7 8 9 during the glacial periods (Webb and Bartlein, 1992). Alpine s://a
species like A. alpina would have potentially had broad ca
Pairwise difference d
glacial distributions in Anatolia, albeit around the alpine belt e
m
absolute) 111802400000000 C aeoTssfhpeseaocuifsartiahtlulgeyamrtnlieioknAnetnleydawtioonnluieatlthdwe(oDhmrakavovireosef,mp1leo9orc7imas1tli;tctsEeoudkarsivmtialvolacaTlanaldciuzerGenudtus¨renMsae.lrot,iTut1unh9dtia8sini6nai)ssl. ic.oup.com
Frequency ( 2460000000 mwdmiiivitggehrrriaasnttiiitooyannsrmshetirsofuptdcionetnulgsr(eeIassbl.ptroainhTteihhmeibsPeellteti,saisltt.ho,aecn1rea9enlb9oey6gt)oemummsoposerttelroyaotufrtrteheetenaflinua‘ipcpntphgulaaielltaoidoncnxtaos’l /aob/article-ab
1 2 3 4 5 6 7 8 9 the survival of temperate biota in the southern European s
Pairwise difference mountain refugia (Hewitt, 2004; Schmitt, 2007). trac
For European alpinesthere are extensive palaeo-ecological, t/1
1400 0
ute) 1200 D ggeraoplohgicicascleannadrifoosss(iTlrdibatsachtoasnudppSocrhto¨thneswdeetvteerlo,p2e0d0p3h;yCloogmeoes- 8/2/2
bsol 1000 and Kadereit, 2003). Similar data are currently lacking for 41/1
cy (a 680000 APlneaisttoolciaennealgplianceiasl, daynndamwiecsmtousstupreployrtoonurgehnyeptoicthedsaitsa. Iatnids 5211
n 8
eque 240000 rbeeacsoomnaebslelimtoitaesdsufmoer tahlaptintehse advuariilnagbiltihtey owfasrumitaibnlteerh-galbaictaiatsl by g
Fr 0 phases in Anatolia. Regular range fragmentation during the ues
1 2 3 4 5 6 inter-glacials (geographic isolation) may potentially restrict t o
Pairwise difference genetic exchange and promote the accumulation of local gen- n 0
4
FIG. 3. Mismatch distribution analyses of pairwise haplotype differences, otypes among the surviving populations. This should result in Ap
o(Abs),erNveedarvaElausets,(gexrepeenc)teldinveaalgueess((Bas),inAdsiicaante–dA),frfiocranEu(rroepde)alnin(ebalguees)l(iCne)aagneds tgheenowtyidpeessprteoadbdeisstrpiabtuiatilolyn ocflulsotceareldg.enHoatypploetsypaensd fporrivraetleatetdo ril 20
Africanonly(red)lineages(D). 1
single localities were scattered throughout the Anatolian 9
rangeofA.alpina(Fig.2C),andgenealogicallyrelatedhaplo-
each other (R–G vs. R-B or G-B comparisons). Interestingly, types (G2–5, R2–4) were locally arranged in the southern
our estimate is similar to a previous estimate of 2.1 Mya esti- Anti-Taurus and Mount Lebanon complexes (Fig. 2C).
mated from synonymous mutation rates (Koch et al., 2006), Notably, haplotype B26 was distributed among the southern
and approximates to an origin at the Pliocene–Pleistocene TaurusMountainlocalities(Fig.2C),whicharealsoimportant
transition, when the climate rapidly cooled (Webb and for their endemic Arabis species (R. Karl et al., University of
Bartlein, 1992) and suitable alpine growing conditions would Heidelberg, Germany, unpubl. res.) and locally differentiated
have become more widespread. The subsequent haplogroup lineages of Lebanese cedar and ground squirrels (Gu¨ndu¨z
diversifications were estimated around 0.70 Mya and roughly et al., 2007; Fady et al., 2008). Furthermore, the southern
coincidedwiththeonsetoftheCromerianComplexofglacia- Anatolian systems were recently recognized as molecular
tion cycles (approx. 300–850 Kya), when the Anatolian diversity hotspots (Me´dail et al., 2009), and local ‘micro-
climate was 4–58C cooler than at present (Erinc¸, 1978; refugia’ for pond turtles (Fritz et al., 2009), again consistent
Furman et al., 2009). Similar divergence times were derived with a hypothesis of multiple survival centres.
Ansell et al. — Anatolian mountain plant phylogeography 249
4·5
Blue region 3–Green region
4·0
Blue region 3–Green region
3·5 Blue region 3–Red region
3) 3·0 Green region 7–Red region
–
0
1
× 2·5 Green region 8–Red region
d (
o
o 2·0
h
eli
k 1·5
Li
1·0
D
o
w
0·5 n
lo
a
0 d
0 1 2 3 4 5 6 7 8 9 ed
Time since divergence (t ) from
FIG. 4. ResultsinoFfigth.e2AIM,anrudnhs,apcloomtyppaeriinngfodrmivaetrigoenngceivetinmieneTsatibmleat1e.sTbiemtweeseinncehadpilvoegrrgoeunpcepiasirminegassuarsedinidnicmatielldi,onbsasoefdyoeanrsthseinscaempprelisnegntr.egions outlined https
://a
c
a
Out of Anatolia alpine habitats were presumably at lower elevations, creating d
e
greater ecological connectivity between the two continents. m
Most strikingly the internal haplogroup arrangement of ic
Furthermore, the progressive loss of nucleotide diversity .o
Anatolia closely resembles the wider global pattern for u
from western Anatolia to northern Europe (Table 2) suggests p
A. alpina (Koch et al., 2006) (Fig. 2C). Hence, we argue .c
that gene flow has predominantly occurred from Anatolia o
that the global haplogroup arrangement was achieved by m
into Europe. /a
expansionsfromalready internallystructuredAnatolian diver- o
The mountains of the ‘Anatolian diagonal’ in eastern b
sity. This ‘inside-out’ scenario is compatible with an existing /a
‘three-times out of Asia Minor’ hypothesis for A. alpina Anatolia are substantially different (.2000m), providing rtic
extensive areas for species preferring cool moist climates to le
(Koch et al., 2006), but we note that the G1-dominated growathigherelevations.Indeed,thediagonalischaracterized -ab
Arabian Peninsula and Caucasus/Iranian Plateau are separated s
byG2-toG5-dominatedinterveningAnti-Taurus/MtLebanon by numerous plant and insect endemics (Davis, 1971; Ekim tra
ranges (Fig. 2C). This suggests these regions were indepen- and Gu¨ner, 1986; C¸iplak, 2003), which may have persisted ct/1
locally or nearby during recent glaciations (Gu¨ndu¨z et al., 0
dently colonized, and thus a ‘four-times out of Anatolia’ 8
2007). Importantly the diagonal mountains have a north-east/ /2
scenario seems likely. /2
south-west orientation, whereas most Anatolian mountain 4
Two geographic features are widely acknowledged to have 1
systems have east–west orientations, and this provides a /1
influenced the expansions of plants and animals around 5
continuous mountain chain connection between the 21
Anatolia: the Sea of Marmara and associated Thyracian 1
Mediterranean and Caspian seas. This links the southern 8
Plain in European Turkey, and the high ‘Anatolian diagonal’ b
Taurus diversity hotspot (Me´dail and Diadema, 2009) to y
mountains in the east (Davis, 1971; Ekim and Gu¨ner, 1986; g
nearby Caucasus and Near East mountain systems. Thus, we u
Rokas, et al., 2003; Bilgin et al., 2009). The hills around the e
Sea of Marmara reach only 1100m, and currently have a postulate the diagonal has been a migratory corridor for st o
A. alpina range expansion out of Anatolia during favourable n
Mediterranean climate and flora (Davis, 1965–85), but dry 0
glacial periods. This is supported by populations near to the 4
steppe communities dominated this area during Pleistocene A
Mediterranean Sea harbouringhaplotypessisterto those com- p
rgelgacuilaatriloynsfo(rMmiecdhaaucxroesst athl.e, 2S0e0a4)o.fPMleaisrtmocaerane(Klaenrdeyberitdgael.s, monly detected from populations in the Caucasus and Iranian ril 20
Plateau. Mismatch tests further indicated a history of sudden 1
2004; Magyari et al., 2008) and, for some temperate animal 9
range expansion for populations encompassing the diagonal
species there is clear evidence of inter-continental gene flow
and the adjacent eastern Anatolia. We note that for several
(Dubey et al., 2007; Furman et al., 2009; Stamatis et al.,
temperate animal species the diagonal does function as a
2009). In contrast, for alpine plant species, the current and
barrier to lineage mixing (Rokas, et al., 2003; Dubey et al.,
past ecological conditions around the Sea of Marmara
2006; Bilgin et al., 2009), and the present contrasting results
potentially represent a substantial geographic/ecological
for an alpine species highlight the importance of individual
barrier for inter-continental gene flow. Surprisingly, related
ecologieswheninterpretingmountainsasbarriersorcorridors.
B-haplotypes were found distributed in both western
Anatolia and the southern Balkans and there is negligible
genetic differentiation between these two areas (F ¼
0.095). Hence we are forced to conclude that the SeSaTof Conclusions
Marmara region has not been a historical ecological barrier Despite the dominating influence of mountain systems on
for A. alpina. Inter-continental genetic exchange most likely the biogeographic structure of Anatolia, the phylogeographic
occurred during favourable glacial conditions, when the histories of mountain species are poorly known in this
250 Ansell et al. — Anatolian mountain plant phylogeography
biodiversityhotspot.Theprsentstudydemonstratestheimpor- synthase intron (chsi) sequences and chloroplast DNA varation.
tanceoftheAnatoliamountainsasthecentreforglobaldiver- MolecularEcology15:4065–4083.
BeilsteinMA,Al-ShehbazIA,MathewsS,KelloggEA.2008.Brassicaceae
sification for arctic–alpine A. alpina, that the local Anatolian
phylogenyinferredfromphytochromeAandndhFsequencedata:tribes
diversity pattern mirrors the global pattern, and that local
andtrichomesrevisited.AnnalsofBotany95:1307–1327.
Pleistocene population history has left a genetic imprint on BilginR,Karatas¸A,C¸oramanE,PandurskiI,PapadatouE,MoralesJC.
the global population structure. Relatively dense and even 2006. Molecular taxonomyand phylogeography of Miniopterus schrei-
sampling was crucial for recovering critical genotypes in bersii (Kuhl,1817)(Chiroptera:Vespertilionidae),intheEurasiantran-
sition.BiologicalJournaloftheLinneanSociety87:577–582.
southern Anatolia to substantiate these findings. Currently
BilginR,FurmanA,C¸oramanE,Karatas¸A.2008.Phylogeographyofthe
therearefewdetailedstudiesofintra-specificgeneticdiversity
Mediterranean horseshoe bat, Rhinolophus euryale (Chiroptera:
from other plants to reliably assess Anatolia’s Pleistocene Rhinolophidae), in southeastern Europe and Anatolia. Acta
history as a centre of gene pool amalgamation or centre of Chiropterologica10:41–49.
diversity (Kucˇera et al., 2006; Go¨mo¨ry et al., 2007; BilginR,KaratasA,C¸oramanE,DisotellT,MoralesJC.2009.Regionally
D
Nthaeydleanrgoev ehtearbl.a,ri2u0m07)l.egWaecyweorfe ftohretunFaloteratoThuarvkeeyaccperossjetcot aiVnnedspacelritmmiliiaogtinrcaiatdolalryey).rebBsatMtricCtsepdeEcpvieoastlt,uetrinMosnianorifyopdtBiesritoirsilbougstyciohnr8e:iobf2er0gs9ei.inedt(ioCcih:1dir0iov.1per1tes8irt6ay/: ownlo
(Davis, 1965–85) as a molecular resource to discriminate 1471-2148-8-209. ad
e
between these alternative interpretations for A. alpina. Challis RJ, Mutun S, Nieves-Aldrey J-L, et al. 2007. Longitudinal range d
Consequently our study also illustrates the importance of expansionandcrypticeasternspeciesinthewesternPalaearcticoakgall- fro
wasp,Andricuscoriarius.MolecularEcology16:2103–2114. m
sampling and scaling effects when conducting phylogeo- h
graphic analysis, especially in complex regions of lineage C¸iplak B. 2003. Distribution of Tettigoniinae (Orthoptera, Tettigoniidae) ttp
bush-crickerts in Turkey: the importance of the Anatolian Taurus s
and flora mixing. Mountains in biodiversity and implications for conservation. ://a
BiodiversityandConservation12:47–64. ca
ClementM,PosadaD,CrandallKA.2000.TCS:acomputerprogramtoesti- de
m
SUPPLEMENTARY DATA mategenegenealogies.MolecularEcology9:1657–1660. ic
ComesHP,KaderitJW.2003.Spatialandtemporalpatternsintheevolution .o
Supplementary data are available online at www.aob.oxford ofthefloraoftheEuropeanalpinesystem.Taxon52:451–462. up
journals.org and consist of the following. Table S1: Cooper SJB, Ibrahim KM, Hewitt GM. 1995. Postglacial expansion and .co
genomesubdivisionintheEuropeanGrasshopperChorthippusparallelus. m
Sampling information. Table S2: Haplotype alignments and MolecularEcology4:49–60. /ao
cplraosgsriafimcatainoanlsy.sTeasb.le S3: Prior values used for the various IM CouvMtrioeunumramnTdeLnpPhr,oinfFcfirpKale.nsz2k0oe1f0eA.vM,oloAultlei-ocSnuhlaeinrhbptahhzeylomIgAue,snteaBtridacksf,kaetmermilyFpoT(rB,alrKadsoisvciceharscieMfiaceAa)-,. b/article
MolecularBiology&Evolution27:55–71. -a
b
ACKNOWLEDGEMENTS DavisEdPiHnb.u1r9g6h5:–U8n5i.veFrlsoirtyaPofreTsusrEkedyinabnudrgthh.eeastAegeanislands,Vols1–9. stra
c
We thank Mary Gibby and Andreas Hemp for collecting DavisPH.1971.DistributionpatternsinAnatoliawithparticularreferenceto t/1
material in Yemen and eastern Africa, Cathy Smith for endemism.In:DavisPH,HarperPC,HedgeIC.eds.Plantlifeofsouth- 08
geo-referencing, Bob Press for comments on the manuscript DubewyesSt,AZsaiait.sAevbeKrd,eCeno:ssUonnivJe-rFs,itAybPdreusksaAdibeerrdAee,nV,o1g5e–l2P8..2006.Pliocene /2/2
development, Maria Albani and George Coupland for andPleistocenediversificationandmultiplerefugiainaEurasianshrew 41
support during fieldwork in Greece, and Julia Llewellyn- (Crocidura suaveolens group). Molecular Phylogenetics and Evolution /15
2
Hughes and her team for the NHM DNA sequencing 38:635–647. 1
1
sBeMrv,icCeO. WanedaErefogrraatlelofuwlintog uthsetocuursaetolersafosfamthpelehsefrrboamriaheBr-, DubeEaynvdiSd,tehCnecoNesseooafnrPEJla-esFits,:totVhcoeehncreaasble´ıikdoifVreth,cetKiobrniycasˇoltluofafueurkendaBls,herxDeciwhkae(nrCgrEeo,cbiVedtouwgraeeellnePu.Ecu2or0do0op7ne,. 8 by g
bariasheetsinourstudy.Theprojectwasfinanciallysupported Soricidae).JournalofEvolutionaryBiology20:1799–1808. ue
s
by the Department Botany at the NHM London, and by the Ehrich D, Gaudel M, Assefa A, et al. 2007. Genetic consequences of t o
Swedish and Norwegian Research Councils to H.K.S. Pleistocene range shifts: contrast between the Arctic, the Alps and the n 0
EastAfricanmountains.MolecularEcology16:2542–2559. 4
El Mousadik A, Petit RJ. 1996. High level of genetic differentiation for Ap
LITERATURE CITED a(Lll.e)liSckreieclhsn]eesnsdaemmoicngtopMopourolactcioon.sThoefotrheeticAarlgaanndtrAepep[lAierdgaGneinaestipcisno9s2a: ril 20
AbbottRJ,BrochmannC.2003.Historyandevolutionofthearcticflora:in 832–839. 19
thefootstepsofEricHulte´n.MolecularEcology12:299–313. EkimT,Gu¨nerA.1986.TheAnatolianDiagonal:factorfiction?Proceedings
AnsellSW,SchneiderH,PedersenN,GrundmannM,RussellSJ,Vogel oftheRoyalSocietyofEdinburgh89B:69–77.
JC. 2007. Recombination diversifies chloroplast trnF pseudogenes in Erinc¸S.1978.ChangesinthephysicalenvironmentinTurkeysincetheendof
Arabidopsislyrata.JournalofEvolutionaryBiology29:2400–2411. thelastglacial.In:BriceWC.ed.TheenvironmentalhistoryoftheNear
Ansell SW, Grundmann M, Russell SJ, Schneider H, Vogel JC. 2008. and Middle East since the last Ice Age. London: Academic Press,
Genetic discontinuity, breeding system change and population history 87–110.
of Arabis alpina in the Italian Peninsula and adjacent Alps. Molecular Excoffier L, Smouse P, Quattro J. 1992. Analysis of molecular variance
Ecology17:2245–2257. inferred from metric distances among DNA haplotypes: applicationsto
Assefa A, Ehrich D, Taberlet P, Nemomissa S, Brochmann C. 2007. humanmitochondrialDNArestrictiondata.Genetics131:479–491.
Pleistocenecolonizationofafro-alpine‘skyislands’bythearctic-alpine ExcoffierL,LavalG,SchneiderS.2005.ARLEQUIN(version3.0):anintre-
Arabisalpina.Heredity99:133–142. gatedsoftwarepackageforpopulationgeneticdataanalysis.Evolutionary
Atalay I. 1996. Palaeosoils as indicators of the climatic changes during BioinformaticsOnline1:47–50.
Quaternary period in S. Anatolia. Journal of Arid Environments 32: Fady B, Lefe`vre F, Vendramin GG, AmbertA, Re´gnierC, BariteauM.
23–35. 2008. Genetic consequences of past climate and human impact on
Bartish IV, Kadereit JW, Comes HP. 2006. Late Quaternary history of easternMediterraneanCedruslibaniforests:impactsfortheirconserva-
Hippophae¨ rhamnoides L. (Elaeagnaceae) inferred from chalcone tion.ConservationGenetics9:85–95.