doi: 10.1111/j.1469-1809.2007.00421.

A Perspective on the History of the Iberian Gypsies

Provided by Phylogeographic Analysis of Y-Chromosome
Lineages
A. Gusmão1 , L. Gusmão1, ∗ , V. Gomes1 , C. Alves1 , F. Calafell2 , A. Amorim1,3 and M. J. Prata1,3
1
Ipatimup, Instituto de Patologia e Imunologia da Universidade do Porto. R. Dr. Roberto Frias, s/n. 4200-465 Porto. Portugal
2
Unitat de Biologia Evolutiva, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra. 08003 Barcelona,
Catalonia, Spain
3
Faculty of Sciences, University of Porto, Pr. Gomes Teixeira, 4099-002 Porto. Portugal

Summary
The European Gypsies, commonly referred to as Roma, are represented by a vast number of groups spread across many
countries. Although sharing a common origin, the Gypsy groups are highly heterogeneous as a consequence of genetic
drift and different levels of admixture with surrounding populations. With this study we aimed at contributing to the
knowledge of the Roma history by studying 17 Y-STR and 34 Y-SNP loci in a sample of 126 Portuguese Gypsies.
Distinct genetic hallmarks of their past and migration route were detected, namely: an ancestral component, shared by
all Roma groups, that reflects their origin in India (H1a-M82; ∼ 17%); an influence from their long permanence in the
Balkans/Middle-East region (J2a1b-M67, J2a1b1-M92, I-M170, Q-M242; ∼ 31%); traces of contacts with European
populations preceding the entrance in the Iberian Peninsula (R1b1c-M269, J2b1a-M241; ∼ 10%); and a high proportion
of admixture with the non-Gypsy population from Iberia (R1b1c-M269, R1-M173/del.M269, J2a-M410, I1b1b-M26,
E3b1b-M81; ∼ 37%). Among the Portuguese Gypsies the proportion of introgression from host populations is higher than
observed in other groups, a fact which is somewhat unexpected since the arrival of the Roma to Portugal is documented
to be more recent than in Central or East Europe.

Keywords: Portuguese Gypsies, Gypsy diaspora, Roma, Y-chromosome lineages, Y-SNP haplogroups, Y-STR haplo-
types

Introduction
Portugal is the westernmost region reached by the Gypsy
diaspora after the Roma people arrived in Europe 600–
700 years ago (Liégeois, 1989; Fraser, 1998; Kendrick &
Puxon, 1998). The establishment of Gypsy groups in Por-
tugal is recorded since the second half of the 15th century
and at present they are estimated to amount to 30–50 thou-
sand individuals. The Portuguese Gypsies are likely to be
a branch of the group that crossed the Pyrenees as early
as the first quarter of the 15th century and rapidly spread
throughout Spain and Portugal. Contrary to many other
Roma, the Iberian Gypsies, known as Gitanos in Spain
and Ciganos in Portugal, are non-Romani-speakers. Af-
∗ Corresponding Author: Leonor Gusmão, Ipatimup Rua Dr.
Roberto Frias, s/n 4200-465 Porto, Portugal. Tel: +351 22 5570700;
Fax: +351 22 5570799; E-mail address: lgusmao@ipatimup.pt
ter entering Iberia they progressively lost the original lan-
guage, and while nowadays many Gitanos still speak Calo,
which is basically Spanish with a large amount of Romani
loan words, the Ciganos from Portugal speak Portuguese
with the Calo just being a reminiscent reference language
(Fraser, 1998).
The Roma in Portugal, as indeed the Roma elsewhere,
are a transnational genetic isolate which fulfil the prop-
erties that make genetic isolates an interesting resource in
genetic epidemiology; namely, they have reduced genetic
diversity and increased linkage disequilibrium (González-
Neira et al. personal communication). The Roma therefore
present a particular genetic disease spectrum, with some
prevalent diseases almost absent, others specific to Gypsies,
and others with private Roma mutations (for a review,
see Kalaydjieva et al. 2001b). Presently in Portugal, Gypsy
communities are spread all over the country and represent
indeed a conspicuous component of the Portuguese social


C 2008 The Authors Annals of Human Genetics (2008) 72,215–227 215
Journal compilation 
C 2008 University College London
 A. Gusmão et al.
and demographic landscape. Despite that, little is known
about the Portuguese Gypsies. How are they related with
other Gypsy groups? To what extent interaction with other
Portuguese contributed to change the founder gene pool?
What signs do they still retain from the complex history
that pre-dated their entrance in Iberia?
Such questions cannot be satisfactorily answered by
applying conventional historical approaches, because the
proper sources are almost non-existent. As happens with
other Roma groups, the Ciganos from Portugal lack a writ-
ten history of their own and the documentary sources from
non-Gypsies (Coelho, 1996) are limited to episodic refer-
ences.
The absence of records constitutes a major complication
in Gypsy studies and explains why the most instructive
evidences about the original homeland of the proto-
Gypsies have resulted from linguistics and genetics. Lin-
guistics has provided compelling evidence of an ancestral
origin in the Indian subcontinent (Liégeois, 1989; Fraser
1998; Kendrick & Puxon, 1998). Evidence for this hypoth-
esis has been consistently found in genetic analyses (e.g.
Gresham et al. 2001; Morar et al. 2004; Malyarchuk et al.
2006), which, although essentially centred in non-Iberian
Gypsies, have further revealed a particular high level of
sub-structuring among Gypsy groups, this is in sharp con-
trast to the surrounding relatively uniform European pop-
ulations. Small population size, strong drift effects, limited
inter-group gene flow and differential admixture with the
host populations, seem to have acted together leading to
strong genetic differentiation between Gypsy groups. This
means that no group of Gypsies can be representative of
other groups, which consequently makes the group the
basic unit of genetic research, as it is of social organisation
(Fraser, 1998). In view of this and in order to address the
questions raised above about the Portuguese Ciganos, in
this study we firstly examined a sample of 126 unrelated
Gypsy males with a high-resolution Y-chromosome STR
and SNP typing strategy; secondly we exploited the results
in the context of previously published data on other Gypsy
groups and respective host populations, and finally we tried
to unfold layers of genetic male lineages in the Portuguese
Gypsies to retrieve information about major events in their
demographic history.
Material and Methods
Sample Collection
Blood samples were taken under informed consent from 126
unrelated Gypsy males, from 18 different communities in 11 dif-
ferent Portuguese districts. Personal inquiries were made to each
individual in order to avoid close kinship. Genomic DNA was
extracted according to standard phenol-chloroform method.
216 Annals of Human Genetics (2008) 72,215–227
Y-STR Typing
Seventeen Y-STR loci (DYS19, DYS385, DYS389I, DYS389II,
DYS390, DYS391, DYS392, DYS393, DYS437, DYS438,
DYS439, DYS448, DYS456, DYS458, DYS635 and GATA
H4.1) were amplified using the AmpFlSTR Yfiler PCR Am-
plification Kit (AB Applied Biosystems), following the manufac-
turers’ instructions. Genotypes were produced with a 310 Ge-
netic Analyser (AB Applied Biosystems), and by comparison to
reference sequenced ladders provided with the kit. The nomen-
clature was according to the ISFG recommendations (Gusmão
et al. 2006) and alleles for GATA H4.1 locus were named by
adding nine repeats to the allele numbering of the typing kit
(Mulero et al. 2006).
Y-SNP Typing
Thirty-four Y-SNP markers were hierarchically typed, in order to
define the major male lineages (Figure 1). All samples were typed
for the biallelic markers SRY1532, M213, M9, M70, M22, Tat,
92R7, M173 and P25, as previously described by Brión et al.
(2005). For the remaining SNPs, the following typing scheme
was used:
- M201, M170, 12f2, M26, M62 and M172 were typed in samples
carrying the M213 mutation and lacking M9, as in Brión et al.
(2005);
- M96, M35, M78, M81, M123 and M34 were typed in samples
carrying only the SRY1532 mutation, as previously described by
Brión et al. (2005);
- M242 was tested in one individual that fell within haplogroup
P (xR1). This mutation was typed by SNaPshot according to
Blanco-Verea et al. (2006);
- M269, M18 and M73 were screened in all individuals within
R1∗ haplogroup. M18 and M73 were typed by SNaPshot using
the primers described in Brión et al. (2005). M269 was typed by
RFLP (Supplementary Table S1);
- F∗ (xGIJK) individuals were screened for M69, M52, M82
and Apt mutations in a newly implemented multiplex SNaPshot
reaction (Supplementary Table S1);
- Samples carrying M172 were tested for M410, M67, M92,
M12 and M241 using also SNaPshot methods (Supplementary
Table S1).
The YCC (2002) and Jobling & Tyler-Smith (2003) nomen-
clature was updated according to Cruciani et al. (2004); Sengupta
et al. (2006); Semino et al. (2004) and Cinnioğlu et al. (2004).
Data Analysis
Haplotype diversity, mean number of pairwise differences and
pairwise genetic distances (Rst, for STR-haplotypes and Fst for
SNP-haplogroups) were calculated using the Arlequin software
ver. 3.01 (Excoffier & Scheinder, 2005). In genetic distance
analyses, DYS385 was not considered, and the number of re-
peats in DYS389I was subtracted from DYS389II. Haplogroup
frequencies were estimated by direct counting.

C 2008 The Authors
Journal compilation 
C 2008 University College London
 A Perspective on the History of the Iberian Gypsies

Figure 1 Phylogenetic network of Y-SNP haplogroups, indicating the biallelic markers typed in the present
work and their absolute frequencies in a sample of 126 Portuguese Gypsies. See Adams et al. (2006) for details
on P25 recurrence.

The YHRD - Y Chromosome Haplotype Reference Database
(Willuweit & Roewer, 2007), URL: www.ystr.org, release
22 from August 10, 2007, was used for haplotype match
analysis.
Phylogenetic median-joining networks of Y-STR haplotypes
inside haplogroups were constructed using Network 4.2.0.0
(www.fluxus-technology.com).
Results and Discussion
Y-STR Haplotype Diversity
The analysis of 17 Y-STR loci in the Portuguese Gypsies
revealed 52 different haplotypes yielding a proportion of
42% of distinct lineages (Supplementary Table S2). Out of
these, 38 were found only once (73.1% of unique haplo-
types) while the remaining 14 were shared by two or more
males. Three of the shared haplotypes were very well rep-
resented (H18-7.9%; H21-15.9% and H24-13.5%) reach-
ing unusually high values in comparison to those found
in most populations for lineages defined by such a high
number of Y-STRs (e.g. Alves et al. 2007; Turrina et al.
2006; Berger et al. 2005; Soltyszewski et al. 2007). In most
European populations haplotype diversity is higher than
99.9%, in contrast with the low value now detected in the
Portuguese Gypsies (0.9437 ± 0.0107).
Previous studies (Gresham et al. 2001; Zaharova et al.
2001; Ploski et al. 2002; Nagy et al. 2007; Füredi et al.


C 2008 The Authors Annals of Human Genetics (2008) 72,215–227 217
Journal compilation 
C 2008 University College London
 A. Gusmão et al.

Pairs of Roma populations Pairs of non-Roma populations

3500 3500

3000 3000
Geographic distance (Km)

Geographic distance (Km)

2500 2500

2000 2000
R2 = 0,6493
1500 2
R = 0,0499 1500 P = 0,0002
P = 0,2529
1000 1000

500 500

0 0
0,000 0,050 0,100 0,150 0,200 0,250 0,000 0,050 0,100 0,150 0,200 0,250
Genetic distance (Rst) Genetic distance (Rst)

Figure 2 Correlation between genetic and geographic distances in Roma and non-Roma populations.

1999; Peričić et al. 2005a; Kalaydjieva et al. 2001a) al- from the distant Lithuanian or Slovakian groups. Accord-
ready had assessed Y-STR diversity in other Roma groups ingly, while genetic distances in the European host popula-
(see Table 1) using, however, very distinct sets of loci. Only tions were found to be correlated with geographic distance
seven Y-STRs were common to all studies, namely DYS19, (Figure 2), in agreement with the general picture widely
DYS389 I, DYS389 II, DYS390, DYS391, DYS392 and reported for Europe (Rosser, 2000), the same was not ob-
DYS393. In view of that, comparisons with Roma and served in the Gypsy groups (r2 = 0.0499; P = 0.2529;
their host populations (which were available for all except Figure 2) as previously reported by Gresham et al. (2001)
Slovakia) were based exclusively upon those seven loci. in a study restricted to Gypsies from Bulgaria, Spain and
Haplotype diversity in the Portuguese Gypsies, using these Lithuania.
markers, was now 0.9097 ± 0.0121, fitting well in the The overall structure of the distance matrix can be
range of values observed in other Roma groups, which visualised by means of multidimensional scaling (MDS,
varied between a minimum of 0.8430 ± 0.0218 in Bul- Fig. 3), in which Roma and non-Roma groups are clearly
garian Gypsies and a maximum of 0.9200 ± 0.0373 in separated. Such separation is statistically significant: the dif-
Spanish Gypsies, both studied by Gresham et al. (2001). Y- ference between groups accounted for 14.8% of the to-
STR diversity in each Gypsy group was remarkably lower tal genetic variance (as computed with AMOVA; p =
than in any non-Gypsy host population where diversities 0.001), and was larger than that within groups (8.0%,
were always higher than 0.98. p < 0.001). Differentiation among Roma (Fst = 8.71%,
The reduced diversities in all Gypsy groups are signs of p < 0.000) was larger than among non-Roma (Fst =
drift effects acting in their pool of Y-STR lineages and 7.70%, p < 0.000). The high genetic distances observed be-
such effects can be explained by the small population size tween Roma groups pinpoints the fast differentiation that
of the Roma groups, together with the deeply rooted en- Gypsy populations experienced during their dispersion in
dogamy that characterises the Gypsy people (Kalaydjieva et Europe.
al. 2001b; Gresham et al. 2001).
Y-SNP Haplogroup Variation
Pairwise Genetic Distance Analysis
In the Portuguese Gypsies, 17 different Y-SNP haplogroups
Pairwise genetic affinities between Gypsy and non-Gypsy were detected (Figure 1) from which five accounted for
host populations were evaluated through Rst distances. 88% of their chromosomes, namely R1b1c-M269, J2a1b-
On average, the highest genetic distances were observed M67, J2a-M410, H1a and I-M170.
in comparisons involving a Gypsy and a non-Gypsy Comparing the Y-SNP profile of Gypsy and non-Gypsy
population (Table 1), with all values reaching statistical Portuguese populations (Beleza et al. 2003) highly signifi-
significance. Among the non-Gypsy pairs, absence of dif- cant differences were observed (Fst = 0.152; P < 0.001).
ferentiation was only detected between Spain/Portugal, What mainly differentiates them is the low prevalence of
Bulgaria/Macedonia and Bulgaria/Hungary, which de- R1b1c in non-Gypsies (∼ 60% against 27%), the much
notes an Y-STR sub-structuring pattern visibly influenced higher frequency of J lineages (∼ 40% versus ∼ 10%) as well
by geographical distance. Geography does not seem to play as the absence of H haplogroup in non-Gypsies, whereas
such a linear role in the inter-group differentiation be- in the Gypsy sample it reached a proportion nearing 17%.
tween Gypsies. In fact the Spanish Gypsies neither differed Furthermore, although the number of different hap-
from their Iberian neighbours, the Portuguese Gypsies, nor logroups in both samples is similar, the frequency

218 Annals of Human Genetics (2008) 72,215–227 

C 2008 The Authors
Journal compilation 
C 2008 University College London
 A Perspective on the History of the Iberian Gypsies


C 2008 The Authors Annals of Human Genetics (2008) 72,215–227 219
Journal compilation 
C 2008 University College London
 A. Gusmão et al.
0,8
RoHun
RoBul1 0,6
0,4
RoBul2 RoSlo
0,2
Dimension 2
0
-1,5 -1 RoSpa -0,5 0 0,5
-0,2
RoLit
RoPor
-0,4
-0,6
RoMac
-0,8
-1
Dimension 1
Figure 3 Multidimensional scaling plot of Rst pairwise values between Roma (grey
circles) and non-Roma (black circles) populations. Codes for populations as in
Table 1.
distribution in Gypsies was much less monotonous than in
non-Gypsies, as indicated by haplogroup diversities that are
0.8367 ± 0.0138 and 0.6520 ± 0.0206, respectively. The
high haplogroup diversity in Gypsies can be explained by
admixture during the history of the group, which can be
simplistically outlined as encompassing three main phases:
the period beyond the early establishment and fragmenta-
tion of founder groups in Europe, possibly in the Balkan
region (since it represented the fons et origo of Roma in Eu-
rope; Fraser 1998), the fast journey towards Western Eu-
rope, and the arrival in Iberia and subsequent settlement in
Portugal.
However, haplogroup diversity does not reflect other
relevant demographic processes, such as founder effects,
although they are quite evident when comparing levels
of diversity within haplogroups assessed from STR vari-
ability. Dramatically low diversities characterise most of
the haplogroups present more than once in the sample
(Supplementary Table S2), being particularly clear within
(i) J2a∗ (xJ2a1b)-M410, in which 20 out of 21 chromo-
somes share exactly the same 16-STR haplotype; (ii) the
25 J2a1b∗ (xJ2a1b1)-M67 lineages, where 17 individuals
belong to the same STR background and 5 belong to a
different one; or (iii) I∗ (xI1b1b)-M170, with 10 out of 12
individuals sharing a given haplotype.
Phylogeographical Analyses
Haplogroup H
Haplogroup H, defined by the M69 mutation (Underhill
et al. 2001), possibly arose in the boundaries of present-
220 Annals of Human Genetics (2008) 72,215–227
Mac
Lit
Bul
Hun
1 1,5 2
Spa
Por
day India (Sengupta et al. 2006) where it is widespread
throughout the entire subcontinent, reaching the highest
frequencies in Central and Southern India. Being a major
haplogroup in all European Roma but absent in their host
populations, it provides clear genetic evidence of the an-
cestral geographical origin of the proto-Gypsies (Gresham
et al. 2001; Kalaydjieva et al. 2001a).
Haplogroup H was one of the most represented in
our sample (16.7%). All individuals inside this haplogroup
were H1a, carrying both the M52 and M82 mutations.
Figure 4A depicts the network constructed after recover-
ing the STR information currently available on H1 chro-
mosomes of different Gypsy groups (Gresham et al. 2001;
Kalaydjieva et al. 2001a). It shows a star-shaped network
where an outstandingly high number of chromosomes lie
in a modal haplotype, shared by all Gypsy groups, and the
remaining differing from the modal by no more than two
molecular steps. Taking together all Gypsy H1 chromo-
somes, the internal STR diversity was remarkably reduced
with a haplotype diversity of 0.398 ± 0.049 and a mean
variance across the seven STR loci of 0.033. This pattern
of STR diversity within H1 is a clear indication that a
profound bottleneck happened in the most distant past of
the Gypsy people, suggesting that, in full accordance with
Gresham et al. (2001), a remarkably small number of
founders must have been present in the ancestral group
from which new Roma populations emerged.
When searching for matches with the Gypsy H1 modal
haplotype (DYS19∗ 15, DYS389I∗ 14, DYS389II∗ 30,
DYS390∗ 22, DYS391∗ 10, DYS392∗ 11 and DYS393∗ 12)
it was possible to observe that it was shared by two tribal
groups from Southern India, being rather common among

C 2008 The Authors
Journal compilation 
C 2008 University College London
 A Perspective on the History of the Iberian Gypsies

Figure 4 Median-joining networks of Y STR haplotypes within haplogroups H1, J2a1b∗ (xJ2a1b1), I and R1∗ of Gypsy
populations. For each haplogroup the area of the circles is proportional to the number of individuals; the smallest area is
equivalent to one individual. Within the pie-charts, orange refers to Portuguese, blue to Bulgarian (Gresham et al. 2001 and
Kalaydjieva et al. 2001a), yellow to Spanish, and red to Lithuanians (Gresham et al. 2001). Arrows assign lineages possessing
the Dinaric modal haplotype (DMH) or the Nordic modal haplotype (NMH). ∗ identifies the Portuguese individual
belonging to I1b12.

one of them (Kivisild et al. 2003). Additionally, four exact
matches for the Gypsy H1 haplotype extended to three
more markers (DYS437∗ 14, DYS438∗ 9 and DYS439∗ 11)
were also found among individuals belonging to castes
of Indian warriors, two of them from the North (Zerjal
et al. 2007). These findings illustrate how difficult it is to
address the questions: which specific Indian region and
social stratum did the first Gypsies arise? The widespread,
although uneven, distribution of the Gypsy H1 modal
haplotype in different Indian populations (either castes or
tribes from North or South), means that it is not possible
to exclude any scenario for the Indian source population
of the first wave of migration.
Portuguese Gypsies preserved a low proportion of H
lineages in comparison to other Roma from Europe (17%
against ∼ 80% in the Bulgarians, for instance).
Haplogroup J
Over one third of the Portuguese Gypsy males be-
long to haplogroup J2, being distributed in five different


C 2008 The Authors Annals of Human Genetics (2008) 72,215–227 221
Journal compilation 
C 2008 University College London
 A. Gusmão et al.
subgroups. Of the two subgroups defined by mutation
M67, J2a1b∗ (xJ2a1b1) was very well represented account-
ing for ∼ 20% of the Gypsy males. J2a1b∗ was also com-
monly observed in the Roma from Bulgaria, Lithuania and
Spain (Gresham et al. 2001).
The network produced with the STR haplotypes within
J2a1b∗ lineages (Figure 4B) was very similar to the one
obtained for haplogroup H1. However, it is worth notic-
ing the almost opposite gradient of distribution of H1
and J2a1b∗ between the Bulgarian Gypsies, on one side,
and the Portuguese, Spanish and Lithuanian, on the other.
The frequency of J2a1b∗ in the latter three groups varies
between approximately 21% and 33%, whereas in the
Bulgarian Roma it just attains 9%. Adding to these findings
the non-detection of J2a1b∗ (xJ2a1b1) among the Macedo-
nian Roma (Peričić et al. 2005b), it seems likely that the
haplogroup was not integrated in the ancestral gene pool
of the first proto-Roma migrating from India, and that
later on, during the fragmentation of the Roma settled
in the Balkan region (which gave rise to the groups that
continued to move west and northward in Europe), strong
drift effects operated successively, enhancing the frequency
of J2a1b∗ (xJ2a1b1) in a group that would turn out to be
a common founder of both Iberian and North European
Roma.
The spatial distribution pattern of the haplogroup in
Eurasia makes it likely that indeed the proto-Roma already
carried J2a1b∗ (xJ2a1b1) when they first arrived in Europe.
Like other subgroups of the J2a-M410 cluster, its presence
in Eurasia is widely recognised to be associated with the de-
mographic spread of the Neolithic farmers. In India, M67
lineages were found to be almost virtually absent (Zerjal
et al. 2007; Sengupta et al. 2006) although two isolated
occurrences have been reported (Kivisild et al. 2003). The
region where their frequency peaks is South Caucasus, but
they have been observed from the Middle East through
Pakistan and also diffused throughout Europe. Moreover,
since higher frequencies and variances have been registered
in Europe and Turkey than in the Middle East, it has been
suggested that M67 chromosomes arrived in Europe from
Turkey (Semino et al. 2004) probably following an Aegean
route (Di Giacomo et al. 2004).
Having this in mind and knowing from historical sources
that Turkey and Greece represented open doors for the in-
flux of Roma into the Balkan region, it seems quite admis-
sible that the introduction of J2a1b∗ (xJ2a1b1) in the Gypsy
gene pool occurred prior to their arrival in the Balkans,
maybe in the Turkish region where the Gypsies settled for
a long time before entering Europe, either crossing the
Bosphorus or following an Aegean route.
Besides M67, one isolated chromosome further har-
boured M92, thus falling into J2a1b1. This haplogroup
222 Annals of Human Genetics (2008) 72,215–227
was also observed at low frequencies in the Bulgarian and
Spanish Gypsies (Gresham et al. 2001) with the STR back-
ground of two of the Bulgarian males differing just one
molecular step from the Portuguese J2a1b1 lineage. J2a1b1
has been predominantly found in the Northern Mediter-
ranean from Turkey westward (Di Giacomo, 2004), having
an overall geographical distribution and history of disper-
sion quite similar to J2a1b∗ (xJ2a1b1). Thus, an identical
explanation can be argued for the introduction of both
lineages in the Gypsies, though not followed in the case of
J2a1b1 by the strong drift effects that drove the distribution
of J2a1b∗ (xJ2a1b1).
Another very common J lineage in the Portuguese Gyp-
sies was J2a∗ (xJ2a1b), reaching 16.7%. It is characterised by
the absence of M67 and the presence of the very recently
discovered mutation M410 (Sengupta et al. 2006). It seems
extremely unlikely that such lineages were present among
the Romani groups studied by Gresham et al. (2001), since
with their typing strategy only five out of 252 samples were
not assigned to a specific haplogroup and these five sam-
ples had microsatellite backgrounds very distinct from the
J2a∗ (xJ2a1b) chromosomes now detected. As the pattern
of geographical distribution of J2a∗ (xJ2a1b) is still incipi-
ently known, in order to address the question of its origin
among the Portuguese Gypsies we recruited the STR in-
formation anchored in those lineages and we conducted a
haplotype match analysis using the YHRD. Based on the
‘minimal haplotype+SWGDAM core set’ of the database
(11 STR loci) we only observed a full match in a world-
wide population sample of 23,979 haplotypes, in a set of
221 populations, and the match was with a Portuguese
sample. This led us to presume that admixture with Por-
tuguese non-Gypsies was responsible for the introduction
of the haplogroup in the Gypsy gene pool and again a strong
founder effect extraordinarily raised its frequency. Interest-
ingly, 20 of the 21 J2a∗(xJ2a1b) chromosomes shared the
same 16 STR haplotype which was only one molecular
step distant from the first. Given that the founder haplo-
type appeared in Gypsy communities spread throughout
Portugal, the introduction event must have occurred soon
after the arrival of the first Gypsies in the country.
Each of the remaining two J lineages detected, J2b-M12
and J2b1a-M241, were observed only once in the sample.
Consulting the YHRD, we found eleven full matches with
the STR haplotype harboured by the J2b chromosome
in individuals from Romania, Macedonia, Bosnia, Albania
and Italy. However, the same haplotype did not match any
of the Romani samples contained in the database, namely
from Hungary (N = 224), Bulgaria (N = 81), Turkey
(N = 111), and Slovakia (N = 63). We can admit that
some J2b-M12 lineages were assimilated in the Balkan
region.

C 2008 The Authors
Journal compilation 
C 2008 University College London

Concerning J2b1a-M241, several haplotypes molecu-
larly related with the Portuguese Gypsy J2b1a were ob-
served disseminated in Western European populations and
a Pyrenean sample showed a full match to it. Therefore, this
might have been incorporated during their passage through
Western Europe.
Haplogroup I
The M170 mutation defining haplogroup I was carried
by 10.3% of Portuguese Gypsy males. The clade contains
a variety of lineages restricted to Europe and almost ab-
sent elsewhere including in the Near-East and Middle-East.
In Europe, where the haplogroup arose in middle Upper
Palaeolithic times (Semino et al. 2000; Rootsi et al. 2004),
two distinct areas of very high incidence are observed,
one located around Scandinavia and the other around the
Dinaric Alps in the Balkan region. These two foci of fre-
quency are due to different I sub-clades, namely I1a and
I1b∗ , whose defining mutations were not assessed in this
study. Besides M170, only M26 was additionally tested and
the sub-clade it defines, I1b1b, was found in one out the 13
I chromosomes. The Portuguese Gypsy lineage classified
as I1b1b had a STR background quite distinct from others
but differing by just two steps from a chromosome of a
Spanish Gypsy, which may also be I1b1b. Western Europe
is the region where I1b1b peaks and where the defining
mutation, M26, most possibly arose in a population from
Iberia/South France (Rootsi et al. 2004). A match anal-
ysis with the extended haplotype revealed that the Por-
tuguese I1b1b chromosome fully matched six records in
the YHRD, all from Portugal or Spain. We can safely con-
clude that the introgression of I1b1b in the Iberian Gypsies
occurred after the Pyrenees crossing.
Gresham et al. (2001) showed that M170 lineages were
present in Roma from Bulgaria (∼ 25%), Spain (∼ 15%) and
Lithuania (∼ 5%), Peričić et al. (2005b) also found them
among the Macedonian Romani (∼ 5%).
In the YHRD, for the most frequent haplotype inside
I∗ (xI1b1b), we found 20 identical haplotypes. Fifteen were
in Eastern Europe, eight of them in Gypsy populations
from Slovakia and Hungary, supporting their possible in-
corporation in the Balkan region.
The phylogenetic network reconstructed with the STR
data within I chromosomes from Gypsy individuals is pre-
sented in Figure 4C and shows that the most common of
the two Portuguese I∗ (xI1b1b) haplotypes is also the most
frequent among the Bulgarian Roma. Lineages molecu-
larly close to this modal haplotype are found in Gypsies
from Bulgaria, Portugal and Spain. A substantial propor-
tion of Bulgarian haplotypes fall in a second molecular
cluster to which the single Lithuanian I chromosome also
belongs. Interestingly, while the haplotypes within this last

C 2008 The Authors
Journal compilation 
C 2008 University College London
A Perspective on the History of the Iberian Gypsies
cluster fit in or are molecularly related to the known Di-
naric modal haplotype (16–24-11–11-13 at DYS19, 390,
391, 392, 393), which is an STR motif strongly associated
with the sub-cluster I1b (Barać et al. 2003; Rootsi et al.
2004), the haplotypes within the most represented cluster
belong to, or are molecularly related to, the Nordic modal
haplotype (14–23-10-11-13 at the same STR loci), which
in turn has been connected to the sub-cluster I1a (this is
only moderately observed in the Balkans). Admitting that
both clusters were incorporated into the Roma gene pool
by admixture events occurring in the Balkan region, it is
noteworthy that in Romani people the two clusters occur
at inverted proportions compared to their host populations,
a finding that can easily be explained by drift effects, but
which also testifies how these events can lead to misinter-
pretation when addressing the origin of specific lineages.
Haplogroup R1b
The second most common Y clade in the Portuguese Gyp-
sies was R1, defined by M173, and detected in 36 indi-
viduals (∼ 29%), 35 of them holding the derivative muta-
tion M269 that characterises the sub-clade R1b1c. In one
sample it was not possible to type M269, because
of a large deletion of at least 3.5 Mb affecting this
∗
chromosome (see haplotype H33, classified as R1 -
M173/del.M9/del.M269; Supplementary Table S2). Nev-
ertheless, this sample haplotype profile is identical to the
most frequent haplotype inside R1b1c haplogroup.
R1 was found in all Roma groups with variable fre-
quency: 3.5% in Macedonians (Peričić et al. 2005b),
4.4% in Bulgarians; 10% in Lithuanians, 22.2% in Spanish
(Gresham et al. 2001). The gradient of distribution within
Gypsy groups overlaps quite well with the global clinal
pattern of R1 across Europe where the Palaeolithic R1b
subgroup of this paragroup occurs significantly more fre-
quently in Western populations, reaching 60% frequency
in Portugal (Beleza et al. 2006). The Iberian Peninsula is
thought to be the region of origin of the major deriva-
tive R1b1c as well as of its expansion after the Last Glacial
Maximum (Semino et al. 2001; Cinnioğlu et al. 2004). In
agreement with the R1 distribution pattern, the Gypsies
from Spain and Portugal present the highest frequencies
of these chromosomes, and most likely a large proportion
of them were essentially acquired by admixture with the
Iberian surrounding populations. Two lines of evidence
lead to this interpretation. Firstly, the absence of founder
lineages being transversal to all Roma groups, as shown in
Figure 4D, where the network of the STR haplotypes an-
chored in Gypsy R1 is depicted. We can observe a cluster
of haplotypes molecularly related that contains chromo-
somes from Portuguese, Spanish and Lithuanian Roma.
The remaining haplotypes are very divergent and occur
Annals of Human Genetics (2008) 72,215–227 223
 A. Gusmão et al.
erratically in distinct Roma populations. This suggests that
R1 chromosomes were incorporated independently in dif-
ferent Roma and that diversification in loco within each
group was of minor relevance in the context of the Gypsy
R1 diversity.
The other evidence comes from the haplotype match
analysis. Out of 35 STR haplotypes within R1b1c chro-
mosomes, 28 showed complete coincidence within Euro-
pean populations in the YHRD, in a total of 183 matches.
From those, only 14 matches were with individuals from
Eastern European populations and none was with any of
the 479 haplotypes from Gypsy populations contained in
the database. The other 169 coincidences were dissemi-
nated in Western Europe but nearly 38% were with Iberian
haplotypes, 57% of which inside Portugal.
These results led us to conclude that the majority of
R1b1c lineages were assimilated after the arrival in Iberia.
Other Lineages
Three chromosomes were positive for M35, a mutation
that defines haplogroup E3b. This haplogroup was not de-
tected among the Lithuanian or Spanish Gypsies, but was
present in 4.4% of Bulgarian Roma and reached 29.8% in
the Macedonian Romani. At least the two M35 chromo-
somes from the Portuguese Gypsies that additionally had
M81 (E3b1b), do not seem to belong to the array of E3b
lineages of the Roma from the Balkans. M81 was absent in
the Macedonian Romani, and although not typed in the
Bulgarian Gypsies, their E3b chromosomes had STR hap-
lotypes quite different from the Portuguese E3b1b. This
haplogroup was likely introduced in Iberia during the Is-
lamic occupation (Bosch et al. 2001; Cruciani et al. 2004)
being present in ∼ 6% of the Portuguese non-Gypsy pop-
ulation (Beleza et al. 2006). Searching in YHRD with
the extended haplotype, one of the chromosomes fully
matched 12 records out of which eight were from Portugal
and one from Spain, seeming to imply that the lineages
were introduced into the Gypsies already in Iberia.
Concerning the third E3b chromosome, it belonged to
E3b1a-M78, with a distribution that encompasses North
and Eastern Africa, all of Europe, and the Middle East.
In YHRD only two full matches were observed with both
chromosomes from non-Gypsies, one from Poland and an-
other from Germany.
A single male was found carrying the M242 mutation,
belonging to haplogroup Q. This mutation originated in
central Asia, although it is also been detected at low fre-
quencies in India, Pakistan (Sengupta et al. 2006), Iran
(Regueiro et al. 2006) and Turkey (Cinnioğlu et al. 2004).
Apart from its presence in Turkey, this mutation is virtu-
ally absent in Europe, which supports its acquisition by the
224 Annals of Human Genetics (2008) 72,215–227
Gypsies before entrance in Europe. This mutation was not
detected in a sample of 57 Macedonian Romani, and the
lack of resolution in other studies on Gypsy groups does
not allow confirmation of its presence among them.
E∗ (xE3b1)-M96, G-M201, and K2-M70 were observed
once each in the Portuguese Gypsy sample. They were not
found in other Roma and all are relatively uncommon in
Europe. No matches were observed for these lineages. Like
in other European populations, they are present in Portugal
at low frequencies (Beleza et al. 2006).
Conclusions
In this work we studied the Y-chromosome diversity of
Portuguese Gypsies, aiming at deepening previous knowl-
edge about the population structure of the Roma in
Europe.
Combining a large amount of both SNP and STR in-
formation, it was possible to unfold layers of information
kept in the Portuguese Gypsy male gene pool, enabling
an extensive reconstruction of the major episodes in their
demographic history.
The Ancestral Component Shared by the
European Roma
In the Portuguese Ciganos four lineages were detected, H1a,
I∗ (xI1b1b), J2a1b∗ (xJ2a1b1), and J2a1b1, which are widely
distributed in Roma groups. The sharing of this set of lin-
eages, usually uncommon in European non-Gypsy popu-
lations, is a genetic link among Gypsies that clearly bear
witness to the common origin of all Roma in Europe.
Two other lineages, J2b∗ (xJ2b1a) and Q, could have also
been incorporated into the Gypsies prior to their entrance
into Europe. On the whole, the proportion retained from
an ancient gene pool already present in the Balkan Roma
(before the fragmentation event and westward dispersion
that occurred during the 15th century) amounted to about
48.4% of the Portuguese Gypsy chromosomes.
The Passage from the Balkans to Iberia
During the short period spanning the exodus from the
Balkans to the entrance into Iberia, the frequency of lin-
eage R1b1c might have been considerably enriched. R1∗
was present at 28.6% frequency in the Portuguese Gyp-
sies, but only at 2%, 4% and 10% in Macedonian, Bulgar-
ian and Lithuanian Roma, respectively, which implies that
about 18.6% of R1b1c lineages can be attributed to Iberian
influx. Actually, out of the 35 STR-haplotypes within the
R1b1c Portuguese Gypsy chromosomes, 23 had previously

C 2008 The Authors
Journal compilation 
C 2008 University College London

been detected in non-Gypsy Iberians. Assuming that these
23 lineages were integrated into the Gypsy gene pool al-
ready in Iberia, we can estimate that ∼ 18.3% (23 in 126)
of chromosomes in Gypsies resulted from R1b1c admix-
ture in Iberia, in full conformity with that obtained when
the Lithuania Gypsies were used as reference. Admitting
this, the remaining proportion of the stock of Portuguese
Gypsies’ R1b1c, together with J2b1a, roughly 10.3%, may
have been acquired in Europe, mainly during the journey
towards Iberia. Despite the supposedly short lasting passage
from the Balkans to Iberia, interaction with surrounding
populations was nonetheless enough to leave visible im-
prints in the genetic composition of the Portuguese Gyp-
sies.
Acquisitions after Arrival in Iberia
R1b1c and J2a∗ (xJ2a1b) are the two lineages among the
Portuguese Ciganos that provide the best clues to the ex-
tent of admixture that occurred in Iberia. Within Iberia
it is impossible to discriminate between the Portuguese
and Spanish contributions since the Iberian Y-chromosome
landscape is quite homogeneous.
We have estimated the proportion of R1b chromosomes
of Iberian descendent to be ∼ 18.3%.
We also hypothesized an Iberian origin for J2a∗ (xJ2a1b),
encompassing 16.7% of the Portuguese Ciganos, and for
I1b1b and E3b1b, observed at the residual frequency of
2.4%.
Summing up all contributions of probable Iberian ori-
gin, we suggest 37.4% as the minimum estimate of the
admixture proportion between Gypsies and non-Gypsy in-
dividuals in Iberia.
A number of other lineages detected in the Portuguese
Gypsies, namely E∗ (xE3b1), E3b1a, G, J2b1a and K2, to-
taling ∼ 4.0%, probably introgressed during their passage
from the Balkans to Iberia or after arrival in Iberia. Whilst
fully absent in the Balkan Roma, all are found dispersed
at low or moderate frequencies throughout Europe, it is
therefore difficult to infer where they were absorbed into
the Gypsy gene pool.
We have previously estimated that 23 R1b1c chromo-
somes were of Iberian origin, and that they are represented
by ten different 17-STR haplotypes. Pairwise comparisons
between these ten haplotypes revealed that (with a single
exception) out of 45 possible comparisons the observed
differences were always higher than four (between four to
13 repeats). Therefore, we can say that at least nine R1b1c
Iberian chromosomes have been incorporated in Gypsies.
Because this haplogroup exists in 60% of the non Gypsy
Iberian males (Beleza et al. 2006), we can extrapolate that,
from the remaining 40% of the Iberian non-Gypsy gene

C 2008 The Authors
Journal compilation 
C 2008 University College London
A Perspective on the History of the Iberian Gypsies
pool, influxes of roughly six additional non R1b1c lineages
might also have occurred. From these, four were already
considered as Iberian influx (J2a, I1b1b and E3b1b). The
other expected lineages can belong to any of the tails of
haplogroups, E∗ (xE3b1), G, J2b1a or K2, referred to above,
since they are also present in Iberia.
Summarizing, the emergence of the Portuguese Gyp-
sies was the result of a complex process stretching over a
vaguely known history of migration, multiple fragmenta-
tion and interaction with other populations. In the current
study some progress was made towards disentangling a little
more of their past. Emphasising a few, it seems now possi-
ble to refute the hypothetical relevance of a second route
of entrance of Gypsies into Europe through North Africa,
at least from the male point of view, as we did not find ge-
netic evidence supporting any considerable North African
influence.
We can also contest the common idea that the Por-
tuguese Gypsies (and Gypsies in general) are an extremely
closed group. The minimum estimate of around 37% as the
proportion of male admixture that had already occurred in
Iberia is in clear contradiction with the stereotype. This
high rate of lineage assimilation means that through adop-
tion of a socio-cultural style of life, many people became
and are becoming Gypsy (and vice-versa). More than re-
flecting a biological/linguistic entity, the term “Gypsy”
must be perceived as a social construct in a continually
changing environment.
Despite the high rate of gene pool renewal, the process
did not erase signatures of the strong endogamous nature
of the Portuguese Gypsy society. All Gypsy groups share
this structural tradition and all of them experienced iden-
tical and peculiar population histories. Along time, both
demography and culture acted together providing the con-
ditions for profound and continuous reshuffling of previ-
ous genetic compositions. Eventually, new founder effects
inside old founder effects under a continuous corridor of
gene influx will progressively attenuate the existing genetic
signs of their origin. As this and previous works demon-
strate, at the moment, genetic studies can still contribute
to partially recover that past.
Acknowledgements
We thank all DNA donors and those, which made possible the
contacts with Gypsy communities in Portugal, including ACIME
and Cruz Vermelha Internacional. This work was partially sup-
ported by Fundação para a Ciência e a Tecnologia, through POCI
(Programa Operacional Ciência e Inovação 2010) and project
PTDC/ANT/70413/2006. The primers used to type M242 mu-
tation were kindly provided by Maria Brión (Institute of Legal
Medicine, Santiago de Compostela, Spain).
Annals of Human Genetics (2008) 72,215–227 225
 A. Gusmão et al.
References
Adams, S. M., King, T. E., Bosch, E. & Jobling, M. A. (2006)
The case of the unreliable SNP: recurrent back-mutation of Y-
chromosomal marker P25 through gene conversion. Forensic Sci
Int 159, 14–20.
Alves, C., Gomes, V., Prata, M. J., Amorim, A. & Gusmão, L.
(2007) Population data for Y-chromosome haplotypes defined by
17 STRs (AmpFlSTR YFiler) in Portugal. Forensic Sci Int 171,
250–255.
Barać, L., Pericić, M., Klarić, I. M., Rootsi, S., Janićijević, B.,
Kivisild, T., Parik, J., Rudan, I., Villems, R. & Rudan, P. (2003)
Y chromosomal heritage of Croatian population and its island
isolates. Eur J Hum Genet 11, 535–542.
Beleza, S., Alves, C., Gonzalez-Neira, A., Lareu, M., Amorim, A.,
Carracedo, A. & Gusmão, L. (2003) Extending STR markers in
Y chromosome haplotypes. Int J Legal Med 117, 27–33.
Beleza, S., Gusmão, L., Lopes, A., Alves, C., Gomes, I., Giouzeli,
M., Calafell, F., Carracedo, A. & Amorim, A. (2006) Micro-
Phylogeographic and Demographic History of Portuguese Male
Lineages. Ann Hum Genet 70, 181–194.
Berger, B., Lindinger, A., Niederstätter, H., Grubwieser, P. &
Parson, W. (2005) YSTR typing of an Austrian population sample
using a 17-loci multiplex PCR assay. Int J Legal Med 119, 241–246.
Blanco-Verea, A., Brión, M., Sánchez-Diz, P., Jaime, J. C., Lareu, M.
V. & Carracedo, A. (2006) Analysis of Y chromosome lineages in
native South American population. Progress in Forensic Genetics
11. International Congress Series 1288, 222–224.
Bosch, E., Calafell, F., Comas, D., Oefner, P. J., Underhill, P. A.
& 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.
Brión, M., Sobrino, B., Blanco-Verea, A., Lareu, M. V. & Carracedo,
A. (2005) Hierarchical analysis of 30 Y-chromosome SNPs in Eu-
ropean populations. Int J Legal Med 119, 10–15.
Cinnioğlu, C., King, R., Kivisild, T., Kalfoğlu, E., Atasoy, S.,
Cavalleri, G. L., Lillie, A. S., Roseman, C. C., Lin, A. A., Prince,
K., Oefner, P. J., Shen, P., Semino, O., Cavalli-Sforza, L. L. &
Underhill, P. A. (2004) Excavating Y-chromosome haplotype strata
in Anatolia. Hum Genet 114, 127–148.
Coelho, F. A. (1996) Os ciganos de Portugal, com um estudo do
calão. Dom Quixote. Lisboa
Cruciani, F., La Fratta, R., Santolamazza, P., Sellitto, D., Pascone,
R., Moral, P., Watson, E., Guida, V., Colomb, E. B., Zaharova,
B., Lavinha, J., Vona, G., Aman, R., Cali, F., Akar, N., Richards,
M., Torroni, A., Novelletto, A. & Scozzari, R. (2004) Phylogeo-
graphic analysis of haplogroup E3b (E-M215) Y chromosomes
reveals multiple migratory events within and out of Africa. Am J
Hum Genet 74, 1014–1022.
Di Giacomo, F., Luca, F., Popa, L. O., Akar, N., Anagnou, N.,
Banyko, J., Brdicka, R., Barbujani, G., Papola, F., Ciavarella,
G., Cucci, F., Di Stasi, L., Gavrila, L., Kerimova, M. G.,
Kovatchev, D., Kozlov, A. I., Loutradis, A., Mandarino, V.,
Mammi, C., Michalodimitrakis, E. N., Paoli, G., Pappa, K. I.,
Pedicini, G., Terrenato, L., Tofanelli, S., Malaspina, P. & Novel-
letto, A. (2004) Y chromosomal haplogroup J as a signature of the
post-neolithic colonization of Europe. Hum Genet 115, 357–371.
Excoffier, L. G. & Scheinder, S. (2005) Arlequin ver.3.0: An in-
tegrated software package for population genetics data analysis.
Evolutionary Bioinformatics Online 1, 47–50.
Fraser, A. (1998) História do Povo Cigano. Tradução de Costa T.
Editorial Teorema, LDA
226 Annals of Human Genetics (2008) 72,215–227
Füredi, S., Woller, J., Padar, Z. & Angyal, M. (1999) Y-STR haplo-
typing in two Hungarian populations. Int J Legal Med 113, 38–42.
Gresham, D., Morar, B., Underhill, P. A., Passarino, G., Lin, A.
A., Wise, C., Angelicheva, D., Calafell, F., Oefner, P. J., Shen,
P., Tournev, I., de Pablo, R., Kucinskas, V., Perez-Lezaun, A.,
Marushiakova, E., Popov, V. & Kalaydjieva, L. (2001) Origins and
divergence of the Roma (gypsies). Am J Hum Genet 69, 1314–
1331.
Gusmão, L., Butler, J. M., Carracedo, A., Gill, P., Kayser, M., Mayr,
W. R., Morling, N., Prinz, M., Roewer, L., Tyler-Smith. C. &
Schneider, P. M. (2006) DNA Commission of the International
Society of Forensic Genetics (ISFG): An update of the recom-
mendations on the use of Y-STRs in forensic analysis. Forensic Sci
Int 157, 187–197.
Jobling, M. A. & Tyler-Smith, C. (2003) The Human Y Chromo-
some: An evolutionary Marker Comes of Age. Nature Reviews
Genetics 4, 598–612.
Kalaydjieva, L., Calafell, F., Jobling, M. A., Angelicheva, D., de Kni-
jff, P., Rosser, Z. H., Hurles, M. E., Underhill, P., Tournev, I.,
Marushiakova, E. & Popov V (2001a) Patterns of inter- and intra-
group genetic diversity in the Vlax Roma as revealed by Y chro-
mosome and mitochondrial DNA lineages. Eur J Hum Genet 9,
97–104.
Kalaydjieva, L., Gresham, D. & Calafell, F. (2001b) Genetic studies
of the Roma (Gypsies): a review. BMC Medical Genetics 2, 5.
Kendrick, D. & Puxon, G. (1998) Ciganos: Da Índia ao
Mediterrâneo. Colecção Interface 3.
Kivisild, T., Rootsi, S., Metspalum, M., Mastanam, S., Kaldmam,
K., Parik, J., Metspalu, E., Adojaan, M., Tolk, H. V., Stepanov,
V., Golge, M., Usanga, E., Papiha, S. S., Cinnioğlu, C., King,
R., Cavalli-Sforza, L., Underhill, P. A. & Villems, R. (2003) The
genetic heritage of the earliest settlers persists both in Indian tribal
and caste populations. Am J Hum Genet 72, 313–332.
Liégeois, J. P. (1989) Ciganos e Itinerantes: Dados socioculturais. Da-
dos sociopolı́ticos. Santa Casa da Misericórdia de Lisboa. 1Edição
Malayarchuk, B. A., Grzybowski, T., Deremko, M. V., Czarny, J. &
Miscicka-Sliwka, D. (2006) Mitochondrial DNA diversity in the
Polish Roma. Ann Hum Genet 70, 195–206.
Martı́n, P., Garcia-Hirchfeld, J., Garcia, O., Gusmão, L., Albarran,
C., Sancho, M. & Alonso, A. (2004) A Spanish population study
of 17 Y chromosome STR loci. Forensic Sci Int 139, 231–235.
Morar, B., Gresham, D., Angelicheva, D., Tournev, I., Gooding,
R., Guergueltcheva, V., Schmidt, C., Abicht, A., Lochmüller, H.,
Tordai, A., Kalmár, L., Nagy, M., Karcagi, V., Jeanpierre, M.,
Herczegfalvi, A., Beeson, D., Venkataraman, V., Carter, K. W.,
Reeve, J., de Pablo, R., Kučinskas, V. & Kalaydjieva, L. (2004)
Mutation history of the Roma/Gypsies. Am J Hum Genet 75,
596–609.
Mulero, J. J., Budowle, B., Butler, J. M. & Gusmão, L. (2006) Letter
to the editor – nomenclature and allele repeat structure update for
the Y-STR locus GATA H4. J Forensic Sci 51, 694.
Nagy, M., Henke, L., Henke, J., Chatthopadhyay, P. K., Volgyi, A.,
Zalan, A., Peterman, O., Bernasovska, J. & Pamjav, H. (2007)
Searching for the origin of Romanies: Slovakian Romani, Jats of
Haryana and Jat Sikhs Y-STR data in comparison with different
Romani populations. Forensic Sci Int 69, 19–26.
Peričić, M., Klarić, I. M., Lauc, L. B., Janićijević, B., Dordević, D.,
Efremovska, L. & Rudan, P. (2005a) Population genetics of 8 Y
chromosome STR loci in Macedonians and Macedonian Romani
(Gypsy). Forensic Sci Int 154, 257–261.
Peričić, M., Lauc, L. B., Klarić, I. M., Rootsi, S., Janićijević, B.,
Rudan, I., Terzić, R., Čolak, I., Kvesić, A., Popović, D., Šijački,

C 2008 The Authors
Journal compilation 
C 2008 University College London

A., Behluli, I., Dordević, D., Efremovska, L., Bajec, D. D., Ste-
fanović, B. D., Villems, R. & Rudan, P. (2005b) High-resolution
phylogenetic analysis of southeastern Europe traces major episodes
of paternal gene flow among Slavic populations. Mol Biol Evol 22,
1964–1975.
Ploski, R., Wozniak, M., Pawlowski, R., Monies, D. M., Branicki,
W., Kupiec, T., Kloosterman, A., Dobosz, T., Bosch, E., Nowak,
M., Lessig, R., Jobling, M. A., Roewer, L. & Kayser, M. (2002)
Homogeneity and distinctiveness of Polish paternal lineages re-
vealed by Y chromosome microsatellite haplotype analysis. Hum
Genet 110, 592–600.
Regueiro, M., Cadenas, A. M., Gayden, T., Underhill, P. A. & Her-
rera, R. J. (2006) Iran: Tricontinental nexus for Y-chromosome
driven migration. Hum Hered 61, 132–143.
Rosser, Z. H., Zerjal, T., Hurles, M. E., Adojaan, M., Alavantic,
D., Amorim, A., Amos, W., Armenteros, M., Arroyo, E., Bar-
bujani, G., Beckman, G., Beckman, L., Bertranpetit, J., Bosch,
E., Bradley, D. G., Brede, G., Cooper, G., Côrte-Real, H.B, de
Knijff, P., Decorte, R., Dubrova, Y. E., Evgrafov, O., Gilissen,
A., Glisic, S., Gölge, M., Hill, E. W., Jeziorowska, A., Kalayd-
jieva, L., Kayser, M., Kivisild, T., Kravchenko, S. A., Krumina,
A., Kucinskas, V., Lavinha, J., Livshits, L. A., Malaspina, P., Maria,
S., McElreavey, K., Meitinger, T. A., Mikelsaar, A. V., Mitchell,
R. J., Nafa, K., Nicholson, J., Nørby, S., Pandya, A., Parik, J.,
Patsalis, P. C., Pereira, L., Peterlin, B., Pielberg, G., Prata, M. J.,
Previderé, C., Roewer, L., Rootsi, S., Rubinsztein, D. C., Saillard,
J., Santos, F. R., Stefanescu, G., Sykes, B. C., Tolun, A., Villems,
R., Tyler-Smith, C. & Jobling, M. A. (2000) Y-chromosomal di-
versity in Europe is clinal and influenced primarily by geography,
rather than by language. Am J Hum Genet 67, 1526–1543.
Rootsi, S., Magri, C., Kivisild, T., Benuzzi, G., Help, H., Bermi-
sheva, M., Kutuev, I., Barac, L., Peričić, M., Balanovsky, O.,
Pshenichnov, A., Dion, D., Grobei, M., Zhivotovsky, L. A.,
Battaglia, V., Achilli, A., Al-Zahery, N., Parik, J., King, R., Cin-
nioğlu, C., Khusnutdinova, E., Rudan, P., Balanovska, E., Schef-
frahn, W., Simonescu, M., Brehm, A., Goncalves, R., Rosa, A.,
Moisan, J. P., Chaventre, A., Ferak, V., Füredi, S., Oefner, P. J.,
Shen, P., Beckman, L., Mikerezi, I., Terzic, R., Primorac, D.,
Cambon-Thomsen, A., Krumina, A., Torroni, A., Underhill, P.
A., Santachiara-Benerecetti, A. S., Villems, R. & Semino, O.
(2004) Phylogeography of Y-chromosome haplogroup I reveals
distinct domains of prehistoric gene flow in Europe. Am J Hum
Genet 75, 128–137.
Semino, O., Passarino, G., Oefner, P. J., Lin, A. A., Arbuzova, S.,
Beckman, L. E., De Benedictis, G., Francalacci, P., Kouvatsi, A.,
Limborska, S., Marcikiae, M., Mika, A., Mika, B., Primorac, D.,
Santachiara-Benerecetti, A. S., Cavalli-Sforza, L. L. & Under-
hill, P. A. (2000) The genetic legacy of paleolithic homo sapiens
in extant europeans: A Y-chromosome prespective. Science 290,
1155–1159.
Semino, O., Magri, C., Benuzzi, G., Lin, A. A., Al-Zahery, N.,
Battaglia, V., Maccioni, L., Triantaphyllidis, C., Shen, P., Oefner, P.
J., Zhivotovsky, L. A., King, R., Torroni, A., Cavalli-Sforza, L. L.,
Underhill, P. A. & Santachiara-Benerecetti, A. S. (2004) Origin,
diffusion, and differentiation of Y-chromosome haplogroups E and
J: inferences on the neolithization of Europe and later migratory
events in the Mediterranean area. Am J Hum Genet 74, 1023–1034.
Sengupta, S., Zhivotovsky, L. A., King, R., Mehdi, S. Q., Edmonds,
C. A., Chow, C. T., Lin, A. A., Mitra, M., Sil, S. K., Ramesh,
A., Rani, M. V. U., Thakur, C. M., Cavalli-Sforza, L. L., Ma-
jumder, P. P. & Underhill, P. A. (2006) Polarity and temporality

C 2008 The Authors
Journal compilation 
C 2008 University College London
A Perspective on the History of the Iberian Gypsies
of high-resolution Y-chromosome distribution in India identify
both indigenous and exogenous expansions and reveal minor ge-
netic influence of Central Asian Pastoralists. Am J Hum Genet 78,
202–221.
Soltyszewski, I., Pepinski, W., Spolnicka, M., Kartasinska, E.,
Konarzewska, M. & Janica, J. (2007) Y-chromosomal haplotypes
for the AmpFlSTR Yfiler PCR Amplification Kit in a population
sample from Central Poland. Forensic Sci Int 168, 61–7.
Turrina, S., Atzei, R. & De Leo, D. (2006) Y-chromosomal STR
haplotypes in a Northeast Italian population sample using 17plex
loci PCR assay. Int J Legal Med 120, 56–59.
Underhill, P. A., Passarino, G., Lin, A. A., Shen, P., Mirazon-Lahar,
M., Foley, R. A., Oefner, P. J. & Cavalli-Sforza, L. L. (2001)
The phylogeography of Y-chromosome binary haplotypes and the
origins of modern human populations. Ann Hum Genet 65, 43–62.
Willuweit, S., Roewer, L., on behalf of the International Forensic
Y Chromosome User Group (2007) Y chromosome haplotype
reference database (YHRD): update. Forensic Sci Int: Genetics 1,
83–87.
Y Chromosome Consortium (2002) A nomenclature system for the
tree of human Y-chromosomal binary haplogroups. Genome Res
12, 339–348.
Zaharova, B., Andonova, S., Gilissen, A., Cassiman, J. J., Decorte,
R. & Kremensky, I. (2001) Y-chromosomal STR haplotypes in
three major population groups in Bulgaria. Forensic Sci Int 124,
182–186.
Zerjal, T., Pandya, A., Thangaraj, K., Ling, E. Y., Kearley, J.,
Bertoneri, S., Paracchini, S., Singh, L. & Tyler-Smith, C. (2007)
Y-chromosomal insights into the genetic impact of the caste system
in India. Hum Genet 121, 137–144.
Supplementary Material
The following material is available for this article online:
Table S1. Multiplex H, J2 and M269. Details on sequences
of PCR and SBE primers used, and typing conditions
Table S2. Y-chromosome haplotypes in 126 Gypsies from
Portugal. District codes are: Aveiro (Av); Lisboa (Li); Viana
do Castelo (VC); Braga (B); Porto (P); Bragança (Bn);
Vila Real (VR); Viseu (Vi); Coimbra (Co); Guarda (Gu);
Castelo Branco (CB); Leiria (Le); Santarém (Sa); Setúbal
(Se); Portalegre (Po); Évora (Ev); Beja (Be); Faro (Fa); Fun-
chal (Fu).
This material is available as part of the online article from:
http://www.blackwell-synergy.com/doi/abs/10.1111/
j.1469-1809.2007.00421.x
(This link will take you to the article abstract).
Please note: Blackwell Publishing are not responsible
for the content or functionality of any supplementary
materials supplied by the authors. Any queries (other than
missing material) should be directed to the corresponding
author for the article.
Received: 15 October 2007
Accepted: 20 October 2007
Annals of Human Genetics (2008) 72,215–227 227