Global Phylogeography of the Loggerhead Turtle (Caretta caretta) as Indicated by Mitochondrial

DNA Haplotypes
Author(s): Brian W. Bowen, Naoki Kamezaki, Colin J. Limpus, George R. Hughes, Anne B.
Meylan and John C. Avise
Source: Evolution, Vol. 48, No. 6 (Dec., 1994), pp. 1820-1828
Published by: Society for the Study of Evolution
Stable URL: http://www.jstor.org/stable/2410511
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Evolution,48(6), 1994, 1820-1828

GLOBAL PHYLOGEOGRAPHY OF THE LOGGERHEAD TURTLE

(CARETTA CARETTA) AS INDICATED BY
MITOCHONDRIAL DNA HAPLOTYPES

BRiANW. BOWEN',NAOKIKAMEZAKI2,COLINJ.LIMPUS3,
GEORGER. HUGHES4,ANNEB. MEYLAN5,AND
JoHNC. AvISE6
1BEECS Genetic AnalysisCore,P.O. Box 110699,
UniversityofFlorida,Gainesville,
Florida32611 and
ArchieCarrCenter forSea TurtleResearch,223 BartramHall,
UniversityofFlorida,Gainesville,
Florida32611
2TheGraduateSchoolofHumanand Environmental Studies,KyotoUniversity,
YoshidaNihonmatsu-cho, Sakyo,Kyoto606,Japan
3Queensland
DepartmentofEnvironment and Heritage,P.O. Box 155,Brisbane,4002, Queensland, Australia
4NatalParksBoard,P.O. Box 662,Pietermaritzburg, 3200 SouthAfrica
100 EighthAvenue,
5FloridaMarineResearchInstitute, S.E., St. Petersburg,
Florida33701-5095
6DepartmentofGenetics,University
ofGeorgia, Athens, Georgia30602

Abstract.-Restriction-siteanalysesofmitochondrialDNA (mtDNA) fromtheloggerheadsea turtle

(Caretta caretta)reveal substantialphylogeographicstructureamong major nestingpopulationsin
the Atlantic,Indian, and Pacific oceans and the Mediterraneansea. Based on 176 samples from
eightnestingpopulations,most breedingcolonies were distinguishedfromotherassayed nesting
locations by diagnosticand oftenfixedrestriction-site differences,indicatinga strongpropensity
fornatal homingby nestingfemales.Phylogeneticanalyses revealed two distinctivematrilinesin
theloggerheadturtlethatdiffer by a mean estimatedsequence divergencep = 0.009, a value similar
in magnitudeto the deepest intraspecificmtDNA node (p = 0.007) reportedin a global surveyof
the greensea turtleChelonia mydas. In contrastto the greenturtle,where a fundamentalphylo-
geneticsplit distinguishedturtlesin the AtlanticOcean and the MediterraneanSea fromthose in
theIndian and Pacificoceans, genotypesrepresenting the two primaryloggerheadmtDNA lineages
wereobservedin bothAtlantic-Mediterranean and Indian-Pacificsamples.We attributethisaspect
ofphylogeographic structurein Carettacaretta to recentinteroceanicgene flow,probablymediated
by the abilityof this temperate-adaptedspecies to utilize habitatsaround southernAfrica.These
resultsdemonstratehow differences in the ecologyand geographicrangesof marine turtlespecies
can influencetheircomparativeglobal population structures.

-Biogeography, Carettacaretta,
Keywords. conservationgenetics,marine turtles,mitochondrial
DNA, phylogeography.

Received July9, 1993. Accepted November 30, 1993.

Marine turtlesof the familyCheloniidae en- marine turtlespecies? In the greenturtle(Che-

compass an ecological diversitythat contrasts
withtheirapparentmorphologicalconservatism.
In terms of feedingecology, the spongivorous
hawksbill (Eretmochelysimbricata) consumes
sessile poriferidsthat are toxic to most verte-
brates(Meylan 1988),theherbivorousgreenturtle
(Chelonia mydas) grazes on sea grass and algal
pastures (Bjorndal 1985), and the carnivorous
loggerhead(Carettacaretta)feedson crustaceans
and mollusks(Mortimer 1982). With respectto
reproductiveecology,greenturtlesand hawksbill
turtlesnestprimarilyin thetropics,whereaslog-
gerheadturtlesnest almost exclusivelyin warm
temperateregions(Pritchardand Trebbau 1984).
To what extentmightthese and related eco-
logical factorsinfluencethe phylogeographyof
1820
? 1994The SocietyfortheStudyofEvolution.All rights
reserved.
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lonia mydas),continentalbarriershave been of
overridingimportancein partitioningphyloge-
netic lineages (Bowen et al. 1992). Since green
turtlesare primarilytropicalin distribution,the
southernextensionsofAfricaand SouthAmerica
representprominentbarriersto contemporary
dispersal. By comparison,the loggerheadhas a
moretemperatedistribution, includingan Indian
Ocean rookery(Natal, SouthAfrica)within1000
km of the South Atlantic Ocean (Hughes
1974a,b). Given thistemperatehabitat,southern
Africamayhave been less formidableas a barrier
to interoceanicgene flowin Carettacarettathan
in the more tropicalmarineturtlespecies, a hy-
pothesisthatcan be testedwithmoleculargenetic
data.
 LOGGERHEAD TURTLE PHYLOGEOGRAPHY 1821

Adult loggerheadsundertakereproductivemi-
grationsthat range from tens to thousands of
kilometers(Meylan et al. 1983; Hughes 1989;
Limpus et al. 1992). Females typicallynest on
continentalcoastlinesadjacent to warm temper-
ate currents,and tag-recapturestudies indicate
thatnearlyall femalesreturnto the same nesting
beach in successivenestingseasons (Dodd 1988).
Major reproductiveassemblagesare knownfrom
the Mediterraneansea as well as the Atlantic,
Indian, and westernPacific oceans, but nesting Collectionlocales forCarettacaretta.The
FIG. 1.
is rareor absentin thecentraland easternPacific numbers
referto locationsin table1.
(Ross 1982; Frazier 1985). Deraniyagala (1945)
described putative subspecies based on subtle jor nesting concentrations for Caretta caretta
morphologicaldifferences betweenAtlantic(Ca- (Pritchardand Trebbau 1984; Dodd 1988). The
retta caretta caretta) and Indian-Pacific forms nations representedin this surveyconstitutea
(C. c. gigas), but recentreviewshave questioned subsetof targetlocations forwhichpermitagen-
theseassignments(Pritchardand Trebbau 1984; cies were accessible and receptiveto biological
Dodd 1988). Estimatesof age at maturityrange research.
from22-26 yr in the westernAtlantic(Klinger Each sample consistedof two eggsfroma nest
and Musick 1995) to 30+ yrin easternAustralia (to offsetpotentialmortalityduringtransporta-
(Limpus 1985). tion-loggerhead eggsare highlysensitiveto mo-
In recentreports,analysesof maternallytrans- tion duringthe firstfew weeks of development
mitted mitochondrial DNA (mtDNA) have [Limpus et al. 1979]), or a singlehatchling.Eggs
provenusefulforresolvingquestionsabout nest- wereremovedfromthenestduringlaying,trans-
ingbehaviorand populationdemographyin ma- portedto the laboratory,and incubatedfor4 to
rine turtles.MitochondrialDNA data can yield 6 wk beforeprocessing.Hatchlingswereeuthan-
informationabout gene flowover both contem- ized and processed in the laboratory.Because
poraryand evolutionarytimescales,therebyper- both full-and half-siblingsare expected to be
mittingappraisal of present-dayphilopatryto identicalwithrespectto mtDNA genotype,field
natal site(Meylan et al. 1990; Allard et al. 1994),
collectionsweredesignedto ensurethatno more
historicalpatternsofglobal colonization(Bowen than one nest per femalewas sampled.
et al. 1991, 1992), and deeper evolutionaryhis- Closed-circular mtDNA was isolated from
tory(Bowen et al. 1993a). In a surveyofregional whole embryos(eggs) or softtissues(hatchlings)
population structure,the distributionof logger- by CsCl-ethidiumbromidedensity-gradient cen-
head mtDNA lineages among fourlocations in trifugation(Lansman et al. 1981). Aliquots of
the southeasternUnited States revealed two ge- purifiedmtDNA were digestedwith the 17 in-
netic population units,correspondingto nesting formativerestrictionenzymes (includingfour-,
beaches in (1) easternand westernFlorida and five-,and six-base recognitionsequences) listed
(2) Georgia and South Carolina (Bowen et al. in table 2. In addition, representativesamples
1993b). These data are consistent with natal were digestedwithBamHI, Clal, EcoRI, KpnI,
homing on a regional scale. Here we assess NsiI, Sacl, Sall, and SmaI, but these enzymes
mtDNA haplotyperelationshipsamong nesting provedto be uninformative, producingeitherzero
colonies on a global scale to resolve recentevo- or one cut in preliminarytests.Digestion frag-
lutionaryhistoryand patternsof historicaldis- mentswereend-labeledwith35S nucleotidesand
persal forthis temperatemarine reptile. separatedon 1.0%/o-1.7% agarosegels.Restriction
fragmentswere visualized by autoradiography
MATERIALS AND METHODS and assigned molecular weightsthroughcom-
Between 1987 and 1992, 176 Caretta caretta parison to a 1-kbladder.
nestsweresampled fromeightnestingaggregates, Restriction-fragment profileswere compiled
includingrookeriesin Greece, Brazil, South Af- into composite lettercodes thatrepresentedthe
rica, Oman, Japan, Australia,and two popula- observed mtDNA haplotypes.Estimates of nu-
tions in the southeasternUnited States (table 1; cleotidesequence divergence(p values) werecal-
fig.1). These locationsrepresentmostofthe ma- culatedbythe"site" approach(Nei and Li 1979).

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1822 B. W. BOWEN ET AL.

TABLE 1. Sample locations,rookerysizes, and population informationforCaretta caretta.

Sample Rookery size

Rookerylocation size (females/yr) Comments
AtlanticOcean
1. East and west Florida 29 10,000-25,000 Feeding groundsinclude the east coast
of the U.S., the Bahamas, Gulf of
Mexico, and Caribbean Sea
2. Georgia and South Carolina 63 1000-3000 Feeding groundsalong east coast of
U.S.
3. Bahia, Brazil 11 -400
Mediterranean
4. Kiparissia Bay, Peloponnesus, 21 -300 Feeding groundsinclude Italy,Malta,
Greece and Tunisia
Indian Ocean
5. Tongaland, Natal, South Africa 15 300-600 Feeding groundsinclude Tanzania,
Madagascar, Kenya, South Africa,
and Mozambique
6. Masirah Island, Oman 8 30,000 Largestrookeryin the world; feeding
groundsextendto the Horn of Afri-
ca, Red Sea, and Gulf of Arabia
PacificOcean
7. Senrigahama Beach, Minabe, Wa- 15 100-300 Feeding groundsinclude the eastern
kayama Prefecture,
Japan China Sea
8. Mon Repos, Queensland, Australia 14 300-600 Feeding groundsinclude New South
Wales, Solomon Islands, Indonesia,
Papua New Guinea, and New Cale-
donia
References:
1, Meylanet al. 1983;Murphyand Hopkins1984;Conleyand Hoffman 1987;Ehrhart
and Raymond1987;2,
Richardson1982;Murphy andHopkins-Murphy 1989;3,M. Marcovaldi
pers.comm.1993;4,Margaritoulis
1988a,b;5,Hughes
1974a,b,1989,unpubl.data;6, Rossand Barwani1982;7, Iwamotoetal. 1985;K. Gotopers.comm.1993;8, Limpus1985;
Limpusetal. 1992.

Haplotypediversitieswereestimatedwithmeth- Slatkin(1989). An estimateofaveragemigration

ods described by Nei and Tajima (1981) and
nucleotidediversitieswithmethodsdescribedby
Nei (1987). Relationships among mtDNA ge-
notypes were assessed by UPGMA clustering
(Sneath and Sokal 1973) using a programin the
PHYLIP computersoftwarepackage(version3.4;
Felsenstein 1989) and by an exhaustive search
of branchingnetworksusing parsimonycriteria
in PAUP (version 3.1.1; Swoffordand Olsen
1990; Swofford1993). The bootstrappingoption
in PAUP (with200 replicates)was used to assess
statisticalsupport for nodes in the parsimony
network(Felsenstein 1985).
Nesting populations were tested in pairwise
comparisons for significantdifferencesin hap-
lotype frequencyusing the G test with Yates'
correction(Sokal and Rohlf 1981). Estimatesof
maternalgene flow among nestingpopulations
(Nm values) were obtained using the cladistic
approach (Slatkinand Maddison 1989). In cases
where no haplotypes were shared between lo-
cations,an upper bound on the estimateof Nm
was calculated with the approach described by
rate among assayed nestingcolonies was calcu-
lated with the private-allele method (Slatkin
1985), usingthe equation in Slatkinand Barton
(1989).
RESULTS
Variation in mtDNA genome size was indi-
cated by concordantdifferencesin the relative
mobilityof mtDNA fragments across severalre-
striction endonucleaseprofiles.For example,SpeI
digests revealed length heterogeneity in an
mtDNA fragmentof approximately3.0 kilobas-
es (kb), withdifferentindividuals exhibitingho-
mologous fragmentsrangingfrom2.8 to 3.2 kb
(fig.2). Lengthvariantswerelocalized to a single
regionof the molecule,probablythe controlre-
gion (Berminghamet al. 1986). Because the in-
heritanceof these size variantsis uncertain,we
excluded size variants fromphylogeneticanal-
yses. All subsequentdiscussion will concernre-
striction-sitevariationonly.
Based on a mean of 82 restrictionsites scored
per individual, eight differentmtDNA haplo-

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 LOGGERHEAD TURTLE PHYLOGEOGRAPHY 1823

TABLE 2. DescriptionanddistributionofmtDNAgenotypes observedin Carettacaretta.Italicizedletters refer

to mtDNArestricton-fragment profiles
producedby(fromleftto right): AvaII, BclI, BglI, BgIII, BstEII, BstNI,
DraII, EcoRV, Hindll, HindIII, MspI, NdeI, PvuII, SpeI, SstII, StuI, and XbaI. For each enzyme,adjacent
lettersinthealphabetindicatethatfragmentprofilesdifferedbya singlerestriction sitegainorloss;nonadjacent
lettersdiffered
byat leasttwosites.

Compos- No. of
itecode mtDNAgenotype Rookery
location nests
A DCCCCCCCCACCCCCBC Georgia-SouthCarolina, USA 2
B DCCCCCCCCBCCCCCBC Carolina,USA
Georgia-South 60
Florida, USA 9
C DCCCCBCCCBCCCCCBC Bahia, Brazil 11
Florida,USA I
D ACCCCDCCCBBCCCCCC Florida, USA 19
Zakynthos,Greece 21
Natal, South Africa 15
E ACCCCDCBCBBCCCCCC Georgia-SouthCarolina, USA I
F DCCCCBCCCBBCCCCBC Masirah Island, Oman 8
G BCCBCDCCCCBCCCCCC Queensland, Australia 14
H BCCBCDCDCCBCCCCCC Wakayama, Japan 15

typesweredetectedamongassayednesting pop- suspected evolutionaryrates for marine turtle

ulations(table2). Digestionprofiles areavailable mtDNA, the deepest bifurcationin the logger-
fromB.W.B.Overallhaplotypic diversity among head mtDNA phylogenywould be roughly2-4
surveyedloggerheads was 0.732, slightly lower my old.
thanthevalue of 0.874 reportedfora compa- Withinthe Indian Ocean, samples fromSouth
rableglobalmtDNAsurvey ofgreen turtles(Bowen Africa and Oman were fixedfor separate hap-
etal. 1992).Nucleotide diversityamongsurveyed lotypes(D and F in table 2) that belong to the
specimens was 0.002,essentially identicalto the two distinctmtDNA lineagesdescribedin figure
valuereported forgreenturtles. Theseestimates 3. In the Pacific,collections fromnortheastern
ofmtDNAvariationarenearthelowendofthe Australiaand Japanwerefixedforalternatehap-
spectrum ofvaluesreported forconspecific com- lotypes(G and H in table 2) thatdifferbya single
parisonsin vertebrates (Aviseetal. 1987,1989). restriction-site change. In the Atlantic-Mediter-
Low levelsofgeneticvariability also havebeen ranean,our sample fromGreece was fixedfora
reportedin a protein-electrophoretic surveyof mtDNA haplotype(D) thatalso was observedat
loggerhead turtles (Gyurisand Limpus1988). 66% frequencyin Florida samples (but notably
AlldifferencesamongmtDNArestriction pro-
filescouldbe interpreted as specific sitegainsor
losses.The mostprominent topologicalfeature Spe I -- loggerheads
of the mtDNA phylogeny is a relatively deep

(N
bifurcation definingtwo distinctevolutionary
lineages(haplotypes A, B, C, and F vs. D, E, G,
and H) separatedby a meanlevel of sequence length
divergence p = 0.009 (fig.3). Thisdivergence is variation ( - 3 kb

similarin magnitudeto the deepestforkob-

servedwithinan mtDNAphylogeny ofthegreen
turtle(p = 0.007; Bowenet al. 1992).Represen-
tativesof the two primaryloggerhead lineages
wereobservedin boththeAtlanticand Indian
oceans.
On thebasisofseveralrestriction-site studies FiG.2. SpeI digestsof mtDNAfrom18 loggerhead
ofmtDNA,a molecularclockformarineturtles turtles collectedin Georgia.Notethefragment at ap-
hasbeententatively calibrated at 0.20/o-0.4% per proximately 3.0 kb whichvariesin size amongindi-
viduals.Thespecimen inthesecondlanefrom theright
my(Aviseetal. 1992),a pace thatis severalfold appearstobe heteroplasmic fortwomtDNAsdiffering
slowerthanconventional estimates forotherver- byabout50 bp. The rightlaneis a molecularweight
tebrategroups(Brownet al. 1979). Fromthese standard(1-kbladder).

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1824 B. W. BOWEN ET AL.

G (QLD 100% dicate thatgene flowis sufficient to preventfix-

ation of alternatemtDNA genotypesin assayed
H (JAPAN 100%
populations,whereas lower values indicate that
E (GA-SC 2% gene flowis insufficient to preventdivergenceof
D
(GREECE 100%
S. AFRICA100%
isolated gene pools by geneticdrift(Birkyet al.
FLORIDA66% 1983; Slatkin 1987; but see Allendorf 1983).
A (GA-SC 3%
However,Nm values based on mtDNA data must
__ GA-SC 95%
z B FLORIDA31% be interpreted withcaution,because of a poten-
CC BRAZIL 100% tially high variance in migrationestimatesde-
FLORIDA3%

F ( OMAN100%
1.0 0.8 0.6 0.4 0.2 0.0
SEQUENCEDIVERGENCE(%)
FIG. 3. UPGMA phenogram summarizing
tionships among the eight observed haplotypes
scribed in table 2. The same branchingorder is ob- recentevolutionarytimescales.
served in a parsimonyanalysis withminorrearrange-
ments of genotypesA, B, C, and F. The distinction
betweenthe lineagecontaininggenotypesA, B, C, and
F versusthelineagecontaininggenotypesD, E, G, and
H is supportedat a bootstrappinglevel of 100% in a
parsimonyanalysisofrestriction-site presence/absence
data using an exhaustive search in the computerpro-
gramPAUP. By the same criteria,the groupingof ge-
notypesG and H is supportedat 86%, and thegrouping
of genotypesD and E is supported at 60%, and no
branchingorderforA, B, C, and F was supportedabove
50%. The geographiclocation(s) whereeach genotype
was observedand thefrequencyofeach genotypewith-
in these locations are indicated to the right.Abbrevi-
ations: GA-SC, Georgia and South Carolina; QLD,
Queensland, Australia.
absent fromthe adjacent nestingpopulation in
Georgia and South Carolina) and at 100% fre-
quency in the sample fromSouth Africa.Thus,
this haplotypewas shared among nestingpop-
ulations in threedifferentocean basins. Finally,
samples fromBrazil were fixedfora haplotype
(C) thatalso was foundat low frequency(3%) in
Florida.
Notwithstanding theselatterinstancesof hap-
lotypesharing,the surveyednestingpopulations
were distinguishedby significantdifferencesin
haplotypefrequencyin 27 of 28 pairwise com-
parisons (table 3). Pairwise estimates of inter-
rookerygene flow(Nm values; table 3) are low.
Based on the frequencyof endemic or private
haplotypes(i.e., those confinedto a singlepop-
ulation;Slatkin198 5), averagegeneflowbetween
assayed nestingpopulations is Nm 0. 1. This
value is qualitativelysimilar to migrationesti-
mates fromgreenturtles:Nm 0.3 forthe At-
lantic-Mediterraneanand Nm 0.2 forthe In-
dian-Pacific(Bowen et al. 1992).
In general,Nm values greaterthan 1 to 4 in-
rived fromsinglehaploid genealogies,and pos-
sible violations of the equilibriumassumptions
thatsupporttheseestimates(see Slatkinand Bar-
ton 1989). For these reasons, Nm values pre-
therela- sented here should be viewed as general indi-
de- catorsofthemagnitudeofgeneticexchangeover
DISCUSSION
Population-GeneticStructure
If female loggerheadsreturnto natal sites for
nesting,thenrookeriesshould tendto show pro-
nounceddifferences withrespectto female-trans-
mitted genetic markerssuch as mtDNA (irre-
spectiveof the magnitudeof male-mediatednu-
clear gene flow,possibly via interrookery mat-
ings; see Karl et al. 1992). Alternatively,in the
absence ofnatal homing,rookerieswould be sub-
ject to thehomogenizinginfluenceoffemale-me-
diated gene flow,and accordinglyshould exhibit
littlegeographicpartitioningof mtDNA haplo-
types.In the Indian-Pacificbasin, samples from
all four surveyed rookeries exhibited fixed
mtDNA haplotypedifferences fromone another.
These resultsare entirelyconsistentwitha strong
behavioral disposition for natal homing by fe-
male loggerheadturtles,at least on a regional
scale, and extendthe geneticevidence fornatal
homingpresentedpreviouslyforloggerheadsin
the Atlanticand Mediterraneanbasins (Bowen
et al. 1993b).
- Natal homingin loggerheadturtlescannot be
absolute, because new nestingbeaches must be
colonized at some reasonable frequencyby tur-
tles hatched elsewhere(Carr et al. 1978). Over
evolutionarytimescales the availabilityand lo-
cations of appropriatenestinghabitat no doubt
change in responseto alterationsin climate,sea
level, and geography.This may be reflectedin
theintraoceanicmtDNA phylogenies:geneticdif-
ferentiation betweennestingcolonieswithineach
ocean basin is generallyshallow, a findingthat
indicates recentconnectionsamong colonies in
a historical,phylogeneticsense. We suspectthat

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 LOGGERHEAD TURTLE PHYLOGEOGRAPHY 1825

changes in climate and coastal geographydrive

an ongoing process of rookeryextinctionsand
I..
colonizations.This process,perhapscoupled with 001

rare lapses in strictfemale natal homing,has a

homogenizingeffectwherebythe accumulation 0o
oV)0
of greatermutationalseparationamong nesting
populations is prevented.Overall, the mtDNA
data suggest that loggerhead turtle rookeries e e t
-sQ cj liv
withinan ocean basin tendto be stronglyisolated
fromone anotherover ecologicaltimescales,but
tightlyconnectedover evolutionarytimeframes. 00 ~ ~ ~ ~ ~ ~ 0*
IVlv
0 o

Marine TurtlePhylogeography 0~~~

0 ~ ~ c 0

The absence of a clear matrilinealseparation

betweenoceanic basins in the temperatelogger- .C) <
head turtlecontrastswith phylogeographicpat-
ternsrecentlyreportedfortwo tropicalmarine-
turtleassemblages. In the greenturtlecomplex
(Chelonia mydas and the nominal C. agassizi),
a phylogenyfor mtDNA haplotypesis charac- 2 , ~O000 0
terizedby a fundamentalbifurcationthatdistin- = ~~~o. o. o.
guishes all Atlantic-Mediterraneanfromall In-
dian-Pacificsamples (Bowen et al. 1992). In the N
0 00 0
ridleycomplex (Lepidochelyskempi and L. oli- 06
e 0 o
0en e0
vacea), morphologicaland geneticevidence in- .Cl
dicatethatan ancestralpopulationmayhave been
split by the Isthmus of Panama into Atlantic
(proto-L. kempi) and Pacific(proto-L. olivacea)
formsabout 3 to 4 mya (Pritchard1969; Hen- a;; ~~t O
0
O O
0
O 7.OO O

drickson1980; Bowen et al. 1991). Lepidochelys

olivacea may have subsequentlycolonized the
AtlanticOcean via southernAfricaduringrecent
evolutionaryhistory(Pritchard1969), a scenario
supportedby morphologicaldata (Pritchardand
Trebbau 1984), distributional data (Hughes 0 :
1972), and mtDNA analyses(Bowen et al. 1991, 0 0
o0 o
W:000~
^ 0 o o6 o6 N
1993a).
Representativesof the two primarymtDNA
lineagesin Carettacarettawereobservedin both
o ~ ~0 10 O00 o I0o o000oxo0

the Atlantic-Mediterraneanand Indian-Pacific

basins. We conclude that the temperatedistri- 0 cn e NOtO
butionof loggerheadturtlesmay have facilitated
at least two effectivetransfersof matrilinesbe-
tween the Atlantic and Indian oceans by gene
flow around southern Africa. Based on the
mtDNA haplotypedistributionsand phylogeny,
we advance the followingtentativescenario as
one example of how such colonization events
mighthave proceeded duringthe recentevolu- U

tionaryhistoryof C. caretta.
During cooler periods of the Pleistocene,log-
3
EXi 00
gerhead populations probably were isolated by
geographyand climateintoAtlanticand Indian-

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1826 B. W. BOWEN ET AL.
Pacificbasins. One mtDNA lineage(represented
in our surveyby haplotypesA, B, C, and F) may
have evolvedin theAtlantic,and anothermtDNA
lineage(representedbyD, E, G, and H) mayhave
evolved in the Indian-Pacific. Subsequently,
warmertemperatures associatedwithinterglacial
periodsallowed an expansionofloggerheadhab-
itatto higherlatitudes,openinga temperatecor-
ridor around southernAfrica. During such pe-
riods, an Atlanticlineage (precursorto F) may
have invaded the Indian Ocean, and an Indian-
Pacificlineage (precursorto D and E) may have
invaded the Atlantic.These particularlineages
are indicatedbecause (1) the A, B, C, F lineage
is widely distributedin the Atlanticbut repre-
sentedby onlyone observedhaplotype(F) in the
Indian-Pacific,and (2) the D, E, G, H lineage is
widespreadin theIndian-Pacificbut represented
by onlyone common haplotype(D) and one rare
haplotype (E) in Atlantic-Mediterraneansam-
ples. The low diversityof these lineages in the
putative "invaded" ocean basin indicates that
these transplantationsoccurred relatively re-
cently,perhaps duringthe last 20,000 yr.
An alternativepossibility,that both major
mtDNA lineageshave been retainedin bothocean
basins for several million years, cannot be ex-
cluded. If true,however,recentinteroceanicex-
change of mtDNA haplotypesis stillimplicated
to account for the similarityof haplotypes in
separate oceans. Under any of these or related
scenarios,the coastline around southernAfrica
has provided(and may continueto offer)a step-
ping-stone for transplantationof loggerhead
mtDNA genotypesbetween the Indian-Pacific
and Atlantic-Mediterranean basins. Indeed, a re-
cent investigationof hatchlingmovementfrom
the Tongaland (South Africa)rookeryhas dem-
onstrated"leakage" ofneonatesfromthisIndian
Ocean rookeryinto the South Atlantic (G. R.
Hughes unpubl. data). Perhaps thesehatchlings,
carriedinto the Atlanticthrougha narrowcor-
ridor of warm temperatewater,are a source of
Atlanticcolonizers.This would explainthepres-
ence ofhaplotypeD (observed at 100% frequen-
cy in Tongaland samples) in two surveyedAt-
lantic-Mediterraneannestingpopulations.
The presenceof the two primarymtDNA lin-
eages in both the Atlantic-Mediterraneanand
Indian-Pacific basins (fig. 3) is consistentwith
recenttaxonomicreappraisalsthathave rejected
subspecificdesignationsforAtlanticand Indian-
PacificCarettacaretta(Hughes 1974a; Pritchard
and Trebbau 1984; Dodd 1988).
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ConservationConcerns
Althoughwe have analyzed only a small frac-
tionofthegeneticarchitecture ofloggerheadturtle
populations (the matrilinealcomponent),these
resultshave special implicationsforthe conser-
vation biology of this species. First,the haplo-
types definedin this reportmay aid in recon-
structing themigratory routesand feedingground
demographicsof C. caretta. In particular,four
majornestingaggregatesin theIndian and Pacific
oceans (in South Africa,Oman, easternAustra-
lia, and Japan) are characterizedby fixedgenetic
differences in our assays. The mtDNA sequences
thereforeprovide natural markersto detectthe
contributionsof these populations to habitats
outside the nestingarea. In cases wherefeeding-
ground populations are harvestedor otherwise
impacted by human activities,the conservation
value of this data is readilyapparent.
Secondly,the mtDNA data indicate a strong
propensityfornatalhomingbyfemales,suchthat
each regional nestingpopulation comprises an
independentdemographic unit. Although new
nestingbeaches must be occasionally colonized
over evolutionarytimescales,both taggingstud-
ies and mtDNA data (table 3) indicate that the
frequencyof such events over ecological times-
cales is low. Thus, a rookeryextirpatedbyhuman
encroachmentor naturalphenomenais notlikely
to be reestablishedover a timeframerelevantto
human interests.This conclusion holds regard-
less of the level of interrookery exchangeof nu-
clear genes that mightbe mediated by males.
Accordingly,the protectionof nestinghabitats
should remaina high conservationpriority.
ACKNOWLEDGMENTS
This project was made possible by the out-
standingcontributionsof K. Goto, S. Hopkins-
Murphy,R. Klinger,M. Marcovaldi, D. Mar-
garitoulis,W. Nelson, J. Richardson,J. P. Ross,
and K. Sato. For fieldassistance, we gratefully
recognizeM. Camhi, P. Casteneda, C. Coogan,
L. Ehrhart,L. Fisher,G. Marcovaldi, R. Mezich,
B. Schroeder,C. Sears, and J.Zurita.For logistic
supportwe are indebtedto R. M. Ball, R. Carthy,
R. Ferl, M. Harris,K. Horikoshi,J. A. Huff,E.
Possardt,J.Woody, Florida DepartmentofNat-
ural Resources, Georgia Departmentof Natural
Resources, National Marine Fisheries Service
(U.S.), Projeto Tartaruga Marinha (TAMAR;
Brazil), Queensland Department of Environ-
mentand Heritage(Australia),Ministryof Fish-
 LOGGERHEAD TURTLE PHYLOGEOGRAPHY 1827

eries (Sultanate of Oman), Natal Parks Board vance.Proceedings oftheNationalAcademyofSci-

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