AMERICAN JOURNAL OF HUMAN BIOLOGY 18:532–539 (2006)

Original Research Article

Gene Flow in the Iberian Peninsula Determined from

Y-Chromosome STR Loci
JOSÉ A. PEÑA,1* SUSANA GARCIA-OBREGON,1 ANA M. PEREZ-MIRANDA,1
MARIAN M. DE PANCORBO,2 AND MIGUEL A. ALFONSO-SANCHEZ1
1
Departamento de Genética, Antropologı́a Fı́sica y Fisiologı́a Animal, Universidad del Paı́s Vasco,
48080 Bilbao, Spain
2
Servicio de Genómica, Banco de ADN y Departamento de Zoologı́a y Biologı́a Celular Animal,
Facultad de Farmacia, Universidad del Paı́s Vasco, 48080 Bilbao, Spain

ABSTRACT In this work, seven multiallelic short-tandem repeat (STR) loci from the nonre-
combining region of the human Y-chromosome (DYS19, DYS389 I, DYS389 II, DYS390, DYS391,
DYS392, and DYS393) were typed in a sample of residents in the Basque Country (RBAS). In all,
40 different Y-STR haplotypic combinations were identified, resulting in a value of haplotypic di-
versity of 0.979. Y-STR data compiled from previous works were used for studying Y-chromosome
diversity in the Iberian Peninsula and for assessing the effects of migratory movements on the
genetic background of the population living currently in territories traditionally occupied by
native (autochthonous) Basques. An analysis of the spatial distribution of allelic frequencies of
the Y-STRs revealed a geographic pattern characterized by variation gradients (frequency clines)
oriented for the most part in the direction southwest-northeast. Accordingly, a neighbor-joining
analysis showed a relative polarization between populations located in the northeast and center
of the Iberian Peninsula, and the rest of the samples considered. The study sample (RBAS) occu-
pied an intermediate position in the population tree between the autochthonous Basques (BASQ)
and the remaining samples. Interestingly, the RBAS collection only showed genetic heterogeneity
with that of native Basques (FST ¼ 0.013, P < 0.05). Estimates of admixture proportions in the
gene pool of RBAS indicated a high level of hybridization with Basque (56%) and non-Basque
(44%) genes, which could explain the genetic differentiation observed between BASQ and RBAS.
Am. J. Hum. Biol. 18:532–539, 2006. ' 2006 Wiley-Liss, Inc.

There are four known forces involved in evo-
lutionary changes: mutation, genetic drift,
gene flow, and natural selection. Specifically,
gene flow is the movement and/or exchange of
genes by the migration of breeding individuals
(or their gametes) among populations. It is
argued that gene flow or migration has two
paradoxical effects. On the one hand, gene
flow can increase the genetic variation within
a population by introducing novel alleles pro-
duced by mutation in another population, or
maintain existing variation by continually
introducing alleles that are not favored by
selective pressure. On the other hand, contin-
ued gene flow decreases diversity among pop-
ulations, causing gene pools to become simi-
lar; in other words, gene flow tends to homoge-
nize allele frequencies between two or more
populations (Hartl and Clark, 1997).
The degree of mobility of populations is usu-
ally seen as one major factor affecting the in-
tensity of gene flow. In human industrialized
and urbanized societies, the modern lifestyle
has implied a remarkable increase in mobility,
which has often led to the phenomenon known
as ‘‘breakdown of isolation.’’ The intense gene
flow typical of modernized societies is a vari-
able to be considered in both forensic and pop-
ulation genetic studies, since it can introduce
substantial biases in the results. Thus, it is
extremely important to carry out careful sam-
pling processes, trying to investigate accu-
rately the origins of donors and corresponding
Grant sponsor: Ministerio de Ciencia y Tecnologı́a, Spain;
Grant number: BOS 2002-01677; Grant sponsor: Universidad
del Paı́s Vasco/Euskal Herriko Unibertsitatea, Spain; Grant
number: GIU 05/51; Grant sponsor: Programa de Formación de
Investigadores, Departamento de Educación, Universidades e
Investigación, Basque Government.
*Correspondence to: Dr. José A. Peña, Departamento de
Genética, Antropologı́a Fı́sica y Fisiologı́a Animal, Univer-
sidad del Paı́s Vasco, Apartado 644, 48080 Bilbao, Spain.
E-mail: joseangel.pena@ehu.es
Received 7 December 2005; Revision received 10 February
2006; Accepted 13 February 2006
Published online in Wiley InterScience (www.interscience.
wiley.com). DOI 10.1002/ajhb.20525

V
C 2006 Wiley-Liss, Inc.
 Y-CHROMOSOME STR LOCI IN IBERIAN PENINSULA
ancestors, to avoid as many inaccuracies as
possible.
One of the most striking changes in the Late
Holocene’s human populations has been their
trend of accelerating mobility. Among Euro-
pean populations who have been isolated for a
long time and have experienced dramatic de-
mographic changes in the last two centuries
because of the intensification of migratory
movements, the Spanish Basque territory is
paramount. For instance, industrialization in
the Spanish Basque Country began to take
place by the middle of the 19th century, and
reached its height in the 1950s, coinciding
with the industrial modernization of Spain
(Montero, 1995). Its effects on population size
become clear when the timing of population
increase is considered. Since the first general
census of Spain in 1787, the population of the
Basque territories has shown a continued,
steady growth, and therefore, since 1950 this
has been the second most densely populated
community in Spain, with approximately 290
inhabitants/km2. It is estimated that around
40% of the current population of the Basque
Country is the result of continuous large-scale
immigration, which took place in Spain over
the period 1950–1975 (Alfonso-Sánchez et al.,
2001, 2005).
The aim of this work was the genetic charac-
terization of the population living currently in
territories traditionally occupied by native (au-
tochthonous) Basques. To that end, seven
short-tandem repeats (DYS19, DYS389 I,
DYS389 II, DYS390, DYS391, DYS392, and
DYS393) of the nonrecombining region of the
human Y-chromosome (Y-STRs) were analyzed
in a sample of the general (resident) population
of the Spanish Basque territory. In addition,
we compiled Y-STR data from the relevant lit-
erature to assess the impact of gene flow pro-
ceeding from other regions of the Iberian Pen-
insula on the gene pool of the contemporary
resident population of the Basque territories.
Analyses of Y-STRs were used in several previ-
ous studies to trace human migratory move-
ments (Quintana-Murci et al., 2001; Kayser
et al., 2001, 2003; Saha et al., 2003, 2005).
MATERIALS AND METHODS
Whole blood samples were collected from 60
unrelated, healthy men in EDTA vacuum
tubes. Ethical guidelines were adhered to, as
stipulated by each of the institutions involved
in the study. All blood donors gave their
533
informed consent prior to inclusion in the
sample.
The voluntary donors included in the sam-
ple were taken at random among the resident
population from both rural and urban zones of
the Basque Country. Consequently, it can be
assumed that it is a representative sample of
the population nowadays residing in the his-
torical territories of the Basque Country. For a
thorough characterization of the sample (tak-
ing into consideration that in population stud-
ies on Basques, it is usual to differentiate
between natives and residents), all donors
were interviewed to obtain information about
the geographical origins of their parents and
grandparents. The genealogical data usually
recorded in population genetic studies do not
go beyond the grandparents, so they are in-
formative only about very recent immigration
events. However, the mobility of human popu-
lations started to show a steady growth with
the industrialization and urbanization proces-
sess that occurred in the transition between
the 18th and 19th centuries, resulting in the
phenomenon known as ‘‘breakdown of iso-
lates.’’ For this reason, the impact of recent
migrations on the genetic background of the
native population may theoretically be mini-
mized if the Basque ancestry of individuals
included in the sample is assessed, thus avoid-
ing as many inaccuracies as possible in the clas-
sification of an individual’s origin. In our study,
Basque surnames and birthplaces of individu-
als and ancestors (recorded back to the third
generation) were the criteria employed to define
local autochthony. In many cases, donors’
grandparents were natives of the Basque Coun-
try (50.0%), but a substantial proportion of
them were also hybrids (33.3%) or offspring of
people born elsewhere on the Iberian Peninsula
(16.7%). In all, 68.7% of donors’ grandparents
were born in the Basque Country.
DNA was extracted by the standard phenol-
chloroform extraction procedure and ethanol
precipitation (Sambrook et al., 1989). Amplifi-
cation was carried out in a GeneAmp PCR Sys-
tem 9700 (PE Applied Biosystems, Foster City,
CA). Each locus was amplified individually,
except for DYS389 I/DYS389 II and DYS390/
DYS391 loci. The primer sequences and cycling
conditions for DYS19, DYS389 I, DYS389 II,
DYS390, DYS391, DYS392, and DYS393 were
as described in Kayser et al. (1997).
The amplified products were detected and
separated by capillary electrophoresis, using
an ABI PRISM 310 Genetic Analyzer (PE
Applied Biosystems). Fragment sizes were
American Journal of Human Biology DOI 10.1002/ajhb
534 J.A. PEÑA ET AL.
Fig. 1. Map showing locations of populations in-
cluded in analysis. ANDL, Andalusia (N ¼ 163); ASTU,
Asturias (N ¼ 90); BASQ, autochtonous Basques (N ¼
168); CACR, Cáceres (N ¼ 91); CANT, Cantabria (N ¼
101); CATL, Catalonia (N ¼ 224); CPOR, Central Portu-
gal (N ¼ 185); GALC, Galicia (N ¼ 103); MADR, Madrid
(N ¼ 148); NPOR, North Portugal (N ¼ 182); RBAS, resi-
dent Basques (N ¼ 60); SARG, Saragossa (N ¼ 120);
SPOR, South Portugal (N ¼ 112); VALN, Valencia (N ¼
140). N is sample size.
determined automatically using Genescan
Analysis software version 3.1, and by compari-
son with the supplied allelic ladders.
Allelic and haplotypic frequencies of Y-STR
markers were obtained by the simple gene-
counting method. In order to unveil patterns of
genetic affinities among geographically tar-
geted groups of the Iberian Peninsula (Fig. 1),
based on Y-STR allelic and haplotypic frequen-
cies, data previously compiled were used.
Except for data on the native Basque collection
(Garcı́a et al., 2004), all samples were logged in
the Y-STR haplotype reference database
(YHRD), maintained at the Institute of Legal
Medicine, Humboldt University (Berlin, Ger-
many), and made public via the Internet
(http://www.yhrd.org) (Roewer et al., 2005). To
assess genetic relationships among popula-
tions, we computed FST, an analogue of
Wright’s FST that takes the evolutionary dis-
tance between individual haplotypes into
account (Excoffier et al., 1992; Excoffier and
Smouse, 1994). Estimates of FST were obtained
using Arlequin software (Schneider et al.,
2000), and tested for statistical significance by
means of randomization (1,000 replicates per
comparison). Allelic frequencies were used to
calculate FST unbiased genetic distances (Reyn-
olds et al., 1983) between all pairs of popula-
tions. From the resultant FST genetic distance
American Journal of Human Biology DOI 10.1002/ajhb
matrix, a phylogenetic tree based on the neigh-
bor-joining (NJ) method (Saitou and Nei, 1987)
was constructed using the PHYLIP program,
version 3.2 (Felsenstein, 1989). The reliability
of the dendrogram was tested by bootstrap
resampling (Felsenstein, 1985), with 1,000 iter-
ations. In addition, we investigated the exis-
tence of frequency clines in a system of moving
coordinates (Pérez-Miranda et al., 2003, 2004).
The analysis of frequency clines is based on the
calculation of the Pearson correlation coeffi-
cient between the frequency of a given allele in
a series of populations and the geographical
coordinates of these populations with regard to
an axis that rotates in successive iterations
from a direction north-south up to a direction
south-north (1808). In those cases where, for
the same allele, more than one statistically sig-
nificant association is obtained between allelic
frequency and spatial distribution, we select
that direction of the axis for which the correla-
tion coefficient produces a maximum value.
Finally, to calculate the genetic contribution of
Basque and non-Basque populations to the
gene pool of Basque residents, a frequency-
based admixture method was used. The esti-
mator of the method is based on weighted least
squares (Long et al., 1991), and takes into
account sampling error and drift. Briefly, this
estimator is the solution to the equation:
X
J
pih ¼ pij
mj
j¼1
where pih is the frequency of the i-th allele in
the hybrid population, pij denotes the fre-
quency of the i-th allele in the j-th reference
population (j ¼ 1,...,J), mj is the proportionate
contribution of the j-th reference
PJ gene pool to
the hybrid population, and j¼1 mj ¼ 1.
RESULTS
Among 60 individuals typed in the sample
of residents of the Basque Country, 40 differ-
ent haplotypes were identified through the
combination of allelic states of the seven Y-
STR loci analyzed (Table 1). The overall hap-
lotype diversity was 0.979. Only 5 of the 40 Y-
STR haplotypes detected showed a frequency
5% (3 individuals). Gene diversity values per
loci were, in decreasing order of magnitude,
0.691 (DYS389 II), 0.603 (DYS389 I), 0.563
(DYS390), 0.544 (DYS391), 0.489 (DYS19),
0.363 (DYS392), and 0.347 (DYS393).
The genetic heterogeneity among popula-
tions was assessed by means of the coefficient
 Y-CHROMOSOME STR LOCI IN IBERIAN PENINSULA 535
TABLE 1. Y-STR haplotypes detected in 60 individuals residents in Basque Country (Spain)

DYS19 DYS389 I DYS389 II DYS390 DYS391 DYS392 DYS393 N

13 12 29 24 11 13 15 1
13 13 29 24 11 13 13 2
13 13 29 25 10 13 12 1
13 13 29 25 10 13 13 1
13 13 30 23 10 11 13 1
13 13 30 24 11 13 13 1
13 14 30 24 9 11 13 1
14 12 28 22 10 11 13 1
14 12 28 23 10 11 13 1
14 12 28 23 11 14 13 1
14 12 28 24 11 13 13 3
14 13 29 23 10 13 13 1
14 13 29 23 11 13 12 1
14 13 29 24 10 13 12 2
14 13 29 24 10 13 13 4
14 13 29 24 10 13 14 1
14 13 29 24 11 13 13 5
14 13 29 25 11 13 13 1
14 13 29 26 11 13 14 1
14 13 30 24 11 13 13 2
14 13 30 25 11 13 13 1
14 14 30 23 10 13 13 1
14 14 30 23 11 13 13 2
14 14 30 23 11 13 14 2
14 14 30 24 10 13 13 1
14 14 30 24 11 13 13 5
14 14 30 24 11 13 14 1
14 14 30 25 11 14 14 1
14 14 31 23 11 13 13 1
14 14 31 24 10 13 13 3
15 12 28 24 10 13 13 1
15 12 30 22 10 11 14 1
15 13 28 23 10 11 13 1
15 13 30 23 11 13 13 1
15 13 30 24 10 13 13 1
15 14 30 24 11 12 13 1
16 13 28 25 9 11 13 1
16 13 30 24 10 11 13 1
17 13 28 23 10 11 13 1
18 13 28 24 9 11 13 1

FST, which was computed from haplotypic fre-
quencies. Estimates of FST between pairs of
the 14 populations considered in the analyses
(Fig. 1) indicated statistically significant dif-
ferences between the sample of autochthonous
Basques (BASQ) and the rest of the popula-
tions included (data not shown). Thus, the res-
ident Basque study population (RBAS) dif-
fered significantly only from native Basques
(FST ¼ 0.013, P < 0.05), and not from other
Iberian populations.
The genetic relationships derived from hap-
lotypic frequencies were corroborated through
distance methods. To that end, we computed
FST unbiased genetic distances (Reynolds
et al., 1983) between all pairs of populations,
based on the allelic frequencies. Figure 2
depicts the dendrogram obtained using the NJ
method. In agreement with the results des-
cribed above, the most notable finding was the
extreme position displayed by the sample of
autochthonous Basques (BASQ). Interest-
ingly, resident Basques (RBAS) appeared seg-
regated in an intermediate position in the NJ
tree, between the autochthonous Basques
(BASQ) and the main cluster of non-Basque
populations. Considering this fact, we esti-
mated the degree of genetic admixture of
RBAS, based on the allelic frequencies of Y-
STRs. The estimates of admixture proportions
give information about the relative contribu-
tion of Basque and non-Basque genes to the
shaping of the genetic background of RBAS.
To do these calculations, we took as allelic fre-
quencies of the non-Basque populations the
average values estimated from the different
Iberian samples considered. The Y-STR data
are highly revealing of the admixture process

American Journal of Human Biology DOI 10.1002/ajhb

536
Fig. 2. Neighbor-joining tree for 14 Iberian popula-
tions, based on Reynolds FST genetic distance calcu-
lated from Y-STR allelic frequencies. Figures are boot-
strap values (1,000 replicates). Population abbrevia-
tions as in Figure 1.
that has taken place in the Basque Country,
which was probably caused by recent migra-
tory flows promoted by large-scale industriali-
zation, among other factors. As expected, a
major contribution to the gene pool of RBAS
comes from natives of the Basque Country
(BASQ), since Basque genes represented a
56% (m1 ¼ 0.562 6 0.049, mean 6 SD through-
out). Similarly, the contribution of Iberian
non-Basque populations is also substantial,
reaching approximately 45% of the gene pool
of RBAS (m2 ¼ 0.438 6 0.056). The proportion
of ‘‘indigenous’’ genes (56.7%) is lower than
the proportion of donors’ grandparents born in
the Basque Country (68.7%). This absolute
difference of 12% between molecular and de-
mographic evidence might reflect the effect of
ancient gene flows (at least previous to the
19th century). According to the results of the
NJ tree (Fig. 2), the populations less distanced
(from the genetic viewpoint) from the samples
of autochthonous (BASQ) and resident (RBAS)
Basques were those of Saragossa, Cantabria,
Catalonia, Madrid, and Valencia. This popula-
tion group shows a certain geographical con-
sistency, since it is concentrated in the center
and northeast of the Iberian Peninsula. The
rest of the samples included in our study
(Andalusia, Galicia, Asturias, Cáceres, and
Central, North, and South Portugal) appeared
closely associated in the same population clus-
ter, positioned at the opposite end of autoch-
thonous Basques (BASQ).
An analysis of the spatial distribution of Y-
STR allelic frequencies revealed some geo-
graphic patterning. In Figure 3, gradients are
depicted such that lines indicate the direction
of maximum correlation between allelic fre-
quency and geographical position, whereas
arrows indicate the direction of the increase in
frequencies. In addition, line length repre-
sents the relative weight of the targeted allele
American Journal of Human Biology DOI 10.1002/ajhb
J.A. PEÑA ET AL.
Fig. 3. Y-chromosomal STR allelic frequency clines
on Iberian Peninsula. Lines indicate direction of maxi-
mum correlation between allelic frequencies and geo-
graphical coordinates. Arrows indicate direction of
increase in frequencies. In addition, line length re-
presents relative weight of targeted allele within corre-
sponding locus. Thus, maximum line length (delimited
by outer circumference) implies that represented allele
is, on average, most frequent of its locus.
within the corresponding locus. Thus, a maxi-
mum line length (delimited by the outer cir-
cumference) implies that the allele repre-
sented is, on average, the most frequent of its
locus. In all, we found six gene frequency
clines, most of them characterized mainly by a
tendency toward an increase in frequencies
with a southwest-northeast orientation (Fig.
3). In this way, with the exception of allele 17
of the DYS19 locus, whose frequencies
increased along the west-east axis (r ¼ 0.785,
P < 0.001), the rest of the detected frequency
gradients tended to increase in the direction
southwest-northeast (DYS393*13, r ¼ 0.756,
P < 0.01, and DYS389 I*14, r ¼ 0.689, P <
0.01), or exactly in the opposite direction, i.e.,
northeast-southwest, as occurred in the cases
of allele 10 of DYS391 (r ¼ 0.663, P < 0.01), al-
lele 11 of DYS392 (r ¼ 0.667, P < 0.01), and al-
lele 12 of DYS393 (r ¼ 0.700, P < 0.01).
DISCUSSION
In the present study, a variety of analyses
based on Y-STR haplotypic and allelic frequen-
cies in a group of populations from the Iberian
Peninsula revealed both a certain degree of
genetic heterogeneity, and the existence of
spatial patterning for allelic frequencies. This
latter finding was confirmed by statistically
 Y-CHROMOSOME STR LOCI IN IBERIAN PENINSULA
significant correlations between allele frequen-
cies and the geographic position of populations.
With respect to the study population (resi-
dents in the Basque Country, RBAS), the anal-
yses reflected a conspicuous lack of genetic dif-
ferentiation when they were compared to
other samples from the Iberian Peninsula.
This is probably an outcome of intense migra-
tory waves toward the Basque Country, which
took place mainly through the 19th and 20th
centuries.
Obviously, such events of gene flow would
have introduced a certain homogeneity into
the Iberian gene pool. This is mirrored in the
NJ tree, where the residents’ sample is located
in an intermediate position between autoch-
thonous Basques (BASQ) and other Iberian
populations. One of the most notable findings
of the present study is the statistically signifi-
cant genetic heterogeneity observed between
RBAS and BASQ. In fact, the non-Basque
genes seem to make a contribution similar to
that of the Basque ones (44% and 56%, respec-
tively) to the genetic makeup of the contempo-
rary population of RBAS. Genetic differentia-
tion between RBAS and BASQ was previously
described in studies on polymorphic Alu inser-
tions and on autosomal short-tandem repeats
(de Pancorbo et al., 2001; Pérez-Miranda
et al., 2005).
It is possible that the degree of genetic
admixture of RBAS was slightly overesti-
mated, however, bearing in mind the fact that
it was calculated from Y-chromosome microsa-
tellites (Y-STR). In effect, the industrial boom,
involving a displacement of male individuals
(Montero, 1995), caused most of the migratory
movements to the Basque Country. From the
beginning of the 19th century up to the ends
of the 20th century, mining activities and,
later, the metallurgical industry attracted a
great number of immigrants, mostly men
searching for a job. On the other hand, it is
very probable that the process of genetic
admixture has not been homogeneous across
the Basque territory. Most likely, the hybrid-
ization process has varied among different
Basque regions, depending on the degree of in-
dustrialization and urbanization, and even
among different generations of the same
region. The impact of recent migrations on
marital structure and inbreeding levels in the
Spanish Basque territories, according to
degree of industrial development, urban-rural
status, population size, and the geography of
peopling (concentrated or dispersed popula-
tions), was analyzed thoroughly in some re-
537
cent works with a biodemographic perspective
(Alfonso-Sánchez et al., 2001, 2004, 2005).
Another interesting result, albeit of more
difficult interpretation, is the gene-frequency
gradient that emerged when the whole set of
Iberian populations was analyzed. An impor-
tant contribution to the understanding of
the Iberian Peninsula’s demographic history
came from a study of Y-STR variability (Bosch
et al., 2001). Those authors stated that, in
spite of allelic frequencies of STR markers are
strongly affected by genetic drift (and the sto-
chastic nature of drift is likely to accumulate
differences through time), eight centuries of
Muslim colonization of the Iberian Peninsula
left only a minimum imprint on Y-chromosome
Iberian lineages. These results were further
confirmed by using Y-chromosome haplotypes
from biallelic markers, STRs, and the minisa-
tellite MSY1 (Brion et al., 2003). Yet all these
authors coincided in pointing out that the
interpretation of STR variability is often com-
plicated, probably because of the high mobility
associated with modern lifestyles, so that the
origin of contemporary human populations
cannot be interpreted as a single event, but as
a much more complex process.
The existence of a certain degree of genetic
dissimilarity between the populations of the
northeast and center of the Iberian Peninsula
with regard to the rest of the populations (sup-
ported by NJ data and by the finding of sev-
eral frequency gradients with an orientation
northeast-southwest) nonetheless suggests the
occurrence of some events of remarkable de-
mographic impact that could have affected the
spatial distribution pattern of Y-STR markers
in Spain and Portugal.
The observed allelic frequency clines could
have originated simply through isolation-by-
distance. Yet, bearing in mind the reduced
geographical scope analyzed and the lack of
notable physical barriers, it seems reasonable
to look for an explanation related to some past
demographic events that could have triggered
intense movements of settling and/or repopu-
lation. A hypothetical scenario could be the
Reconquista (Reconquest), a historical episode
by means of which the Christian kingdoms
acquired the control of the whole Iberian Pen-
insula after almost eight centuries of war with
the different Arabic kingdoms established in
the south. After the Moslem invasion, which
took place in 711 AD, a sizable number of Cen-
tral Spain’s regions remained practically un-
inhabited. Both Central Spain and other
southern regions were repopulated mainly by
American Journal of Human Biology DOI 10.1002/ajhb
538 J.A. PEÑA ET AL.
people who came from the kingdoms of Castile
and Leon, located in the north and northwest
of the peninsula. On the contrary, some cen-
tral and mostly eastern regions were recon-
quered and, up to a certain point, repopulated
by individuals from the kingdom of Aragon,
which was located in the northeast (Garcı́a de
Cortázar, 2005). These medieval demographic
conflicts, together with the presence of the
Basques (who even nowadays show some sin-
gularities in their genetic background), might
explain the existence of the allelic frequency
clines cited above.
In conclusion, all these findings stress the
need to clarify the complex genetic relation-
ships of the Iberian groups. With this purpose
in mind, population analyses require multiple
unlinked genetic markers, taking into account
that the amount of genetic information increa-
ses with the number of independent loci sur-
veyed (Rosenberg and Nordborg, 2002). It is
also essential to carry out thorough and ex-
haustive sampling processes, emphasizing ac-
curate knowledge of the place of origin of DNA
donors and their ancestors. The careful selec-
tion of the sample is key in both forensic and
evolutionary studies.
ACKNOWLEDGMENTS
S.G.-O. and M.A.A.-S. were supported by
the Programa de Formación de Investiga-
dores, Departamento de Educación, Universi-
dades e Investigación (Basque Government).
We are particularly grateful to all the volun-
tary donors, who cooperated generously in the
development of this study.
LITERATURE CITED
Alfonso-Sánchez MA, Peña JA, Aresti U, Calderón R. 2001.
An insight into recent consanguinity within the Basque
area in Spain. Effects of autochthony, industrialization
and demographic changes. Ann Hum Biol 28:505–521.
Alfonso-Sánchez MA, Calderón R, Peña JA. 2004. Opportu-
nity for natural selection in a Basque population and its
secular trend: evolutionary implications of epidemic mor-
tality. Hum Biol 76:361–381.
Alfonso-Sánchez MA, Aresti U, Peña JA, Calderón R. 2005.
Inbreeding levels and consanguinity structure in the
Basque province of Guipúzcoa (1862–1980). Am J Phys
Anthropol 127:240–252.
Bosch E, Calafell F, Comas D, Oefner PJ, Underhill PA,
Bertranpetit J. 2001. High-resolution analysis of human
Y-chromosome variation shows a sharp discontinuity and
limited gene flow between northwestern Africa and the
Iberian Peninsula. Am J Hum Genet 68:1019–1029.
Brion M, Salas A, González-Neira A, Lareu MV, Carracedo
A. 2003. Insights into Iberian population origins through
the construction of highly informative Y-chromosome
haplotypes using biallelic markers, STRs, and the MSY1
minisatellite. Am J Phys Anthropol 122:147–161.
American Journal of Human Biology DOI 10.1002/ajhb
de Pancorbo MM, López-Martı́nez M, Martı́nez-Bouzas C,
Castro A, Fernández-Fernández I, Antúnez de Mayolo G,
Antúnez de Mayolo A, Antúnez de Mayolo P, Rowold DJ,
Herrera RJ. 2001. The Basques according to polymorphic
Alu insertions. Hum Genet 109:224–233.
Excoffier L, Smouse PE. 1994. Using allele frequencies and
geographic subdivision to reconstruct gene trees within
a species: molecular variance parsimony. Genetics 136:
343–359.
Excoffier L, Smouse PE, Quattro JM. 1992. Analysis of mo-
lecular variance inferred from metric distances among
mtDNA haplotypes: application to human mitochondrial
DNA restriction data. Genetics 131:479–491.
Felsenstein J. 1985. Confidence limits on phylogenies: an
approach using the bootstrap. Evolution 39:783–791.
Felsenstein J. 1989. PHYLIP: phylogeny inference package
(version 3.2). Cladistics 5:164–166.
Garcı́a O, Martı́n P, Gusmão L, Albarrán C, Alonso S, De
La Rua C, Flores C, Izagirre N, Peñas R, Pérez JA,
Uriarte I, Yurrebaso I, Alonso A. 2004. A Basque Country
autochthonous population study of 11 Y-chromosome
STR loci. Forensic Sci Int 145:65–68.
Garcı́a de Cortázar F. 2005. Atlas de historia de España,
2nd ed. Barcelona: Editorial Planeta, S.A.
Hartl DL, Clark AG. 1997. Principles of population genet-
ics, 3rd ed. Sunderland, MA: Sinauer Associates, Inc.
Kayser M, Caglià A, Corach D, Fretwell N, Gehrig C, Gra-
ziosi G, Heidorn F, Herrmann S, Herzog B, Hidding M,
Honda K, Jobling M, Krawczak M, Leim K, Meuser S,
Meyer E, Oesterreich W, Pandya A, Parson W, Penacino
G, Perez-Lezaun A, Piccinini A, Prinz M, Schmitt C,
Schneider PM, Szibor R, Teifel-Greding J, Weichhold G,
De Knijff P, Roewer L. 1997. Evaluation of Y-chromo-
somal STRs: a multicenter study. Int J Legal Med 110:
125–149.
Kayser M, Krawczak M, Excoffier L, Dieltjes P, Corach D,
Pascali V, Gehrig C, Bernini LF, Jespersen J, Bakker E,
Roewer L, De Knijff P. 2001. An extensive analysis of Y-
chromosomal microsatellite haplotypes in globally dis-
persed human populations. Am J Hum Genet 68:990–
1018.
Kayser M, Brauer S, Weiss G, Schiefenhovel W, Underhill
P, Shen P, Oefner P, Tommaseo-Ponzetts M, Stoneking
M. 2003. Reduced Y-chromosome, but not mitochondrial
DNA, diversity in human populations from west New
Guinea. Am J Hum Genet 72:281–302.
Long JC, Williams RC, McAuley JE, Medis R, Partel R, Tre-
gellas M, South SF, Rea AE, McCormick B, Iwaniec U.
1991. Genetic variation in Arizona Mexican Americans:
estimation and interpretation of admixture proportions.
Am J Phys Anthropol 84:141–157.
Montero M. 1995. Historia del Paı́s Vasco (de los orı́genes a
nuestros dı́as). San Sebastián: Ediciones Txertoa.
Pérez-Miranda AM, Alfonso-Sánchez MA, Peña JA, Cal-
derón R. 2003. HLA-DQA1 polymorphism in autochtho-
nous Basques from Navarre (Spain): genetic position
within European and Mediterranean scopes. Tissue Anti-
gens 61:465–474.
Pérez-Miranda AM, Alfonso-Sánchez MA, Vidales MC,
Calderón R, Peña JA. 2004. Genetic polymorphism
and linkage disequilibrium of the HLA-DP region in
Basques from Navarre (Spain). Tissue Antigens 64:264–
275.
Pérez-Miranda AM, Alfonso-Sánchez MA, Kalantar A, Gar-
cı́a-Obregón S, de Pancorbo MM, Peña JA, Herrera RJ.
2005. Microsatellite data support subpopulation struc-
turing among Basques. J Hum Genet 50:403–414.
Quintana-Murci L, Krausz C, Zerjal T, Sayar SH, Hammer
MF, Mehdi SQ, Ayub Q, Qamar R, Mohyuddin A, Rad-
hakrishna U, Jobling MA, Tyler-Smith C, McElreavey K.
2001. Y-chromosome lineages trace diffusion of people
 Y-CHROMOSOME STR LOCI IN IBERIAN PENINSULA
and languages in southwestern Asia. Am J Hum Genet
68:537–542.
Reynolds J, Weir BS, Cockerham CC. 1983. Estimation of
the coancestry coefficient: basis for a short-term genetic
distance. Genetics 105:767–779.
Roewer L, Croucher PJP, Willuweit S, Lu TT, Kayser M,
Lessig R, de Knijff P, Jobling MA, Tyler-Smith C, Krawc-
zak M. 2005. Signature of recent historical events in the
European Y-chromosomal STR haplotype distribution.
Hum Genet 116:279–291.
Rosenberg N, Nordborg M. 2002. Genealogical trees, coales-
cent theory and the analysis of genetic polymorphisms.
Nat Rev Genet 3:380–390.
Saha A, Udhayasuriyan PT, Bhat KV, Bamezai R. 2003.
Analysis of Indian population based on Y-STRs reveals
539
existence of male gene flow across different language
groups. DNA Cell Biol 22:707–719.
Saha A, Sharma S, Bhat A, Pandit A, Bamezai R. 2005. Genetic
affinity among five different population groups in India
reflecting a Y-chromosome gene flow. J Hum Genet 50:49–51.
Saitou N, Nei M. 1987. The neighbor-joining method: a new
method for reconstructing phylogenetic trees. Mol Biol
Evol 4:406–425
Sambrook J, Fritsch E, Maniatis T. 1989. Molecular clon-
ing: a laboratory manual. Cold Spring Harbor: Cold
Spring Harbor Laboratory Press.
Schneider S, Roessli D, Excoffier L. 2000. Arlequin: a soft-
ware for population genetics data analysis, version 2.000.
Geneva: Genetics and Biometry Laboratory, Department
of Anthropology, University of Geneva.
American Journal of Human Biology DOI 10.1002/ajhb