AMERICAN JOURNAL OF HUMAN BIOLOGY 21:407–409 (2009)

Short Report

Y-chromosome Variation in South Iberia: Insights into the North

African Contribution
LUIS ALVAREZ, * CRISTINA SANTOS, RAFAEL MONTIEL, BLAZQUEZ CAEIRO, ABDELLATIF BAALI, JEAN-MICHEL DUGOUJON,
AND MARIA PILAR ALUJA
Unitat d’Antropologia Biològica, Departament Biologia Animal, Biologia Vegetal i Ecologia, Universitat Autònoma de Barcelona,
08193 Cerdanyola del Vallès, Barcelona, Spain

ABSTRACT Population of Pedroches Valley, a hypothetical Berber settlement, located in the northwest portion of
Córdoba province (Andalusia, Spain), had been analyzed for its Y-chromosome diversity. Moreover, to contextualize this
population, 127 Y-chromosomes from a general Andalusia sample and a North African Berber community (Marrakech,
Morocco) were also typed. For all samples, 24 single nucleotide polymorphisms of the non-recombining portion of the Y-
chromosome (NRY) were analyzed and those samples described as belonging to E3b1b-M81 haplogroup were also typed
for 16 Y-chromosome short tandem repeats. Our Analysis showed low levels of North African E3b1b-M81 haplogroup in
the Pedroches Valley population (1.5%), which is a lower contribution than would be expected. This result rejects the
hypothesis of a gradual genetic assimilation of Berber settlers during the Islamic period. Am. J. Hum. Biol. 21:407–409,
2009. ' 2009 Wiley-Liss, Inc.

The current genetic pool of the Iberian Peninsula
results from genetic admixture events concerning various
populations from different origins at different times. One
of those events was 8 centuries of settlement of different
Arabs and Berber populations in the Iberian Peninsula.
This Islamic period involved numerous population move-
ments, mainly Berbers from North Africa into Iberia.
Although toponymic studies showed a high presence of
Berber-origin toponymies in the region between the west-
ern mountains of Andalusia and Toledo, distribution of
Arabs and Berbers in the conquered part of Iberia is still a
controversial issue. Today, some historians argue that the
Berber settlements were dispersed in small villages along
the aforementioned area while maintaining their Magh-
reb tribal structures. Nevertheless, to date, there are no
differentiating elements in the archaeological record
(ceramics, villages structure, etc.) that could represent
evidence of integration of the Berber conquerors in the
settlements (Manzano-Moreno, 2006).
To find more clues to these settlements, the study of the
Y-chromosome represents an adequate tool to access the
possible genetic contribution of the Berber conquerors to
the present-day Iberian genetic pool. The studies per-
formed so far reveal that Berber-speaking populations
present a distribution of E3b1b-M81 Y-chromosome hap-
logroup that closely matches their current presence in
North Africa (Cruciani et al., 2004). In Iberia, E-M81 hap-
logroup showed low levels, 5% in Andalusia (Bosch et al.,
2001), and within a range of 0–11.5% in the occidental
provinces of Andalusia (Flores et al., 2004); however, with
the small sample size of the previously cited works, no
further extrapolations can be performed.
MATERIALS AND METHODS
Samples
Total DNA was extracted from blood or buccal swabs of
195 unrelated males from three populations: 68 from
Pedroches Valley, a region located in the northwest por-
tion of Córdoba province (Andalusia, Spain) that
presents evidences of Berber settlements during the
Islamic occupation; 94 from a representative Andalusian
sample (Spain), considered for comparative purposes; and
33 from different hamlets in Azgour Valley from the Amiz-
miz region (Marrakech, Morocco), which represents a typi-
cally Moroccan Berber population. For all participants,
appropriate informed consent and information about the
geographical origin of their four grandparents was
obtained.
Y-Chromosome biallelic markers
Twenty-four previously published polymorphic sites
were analyzed. Markers M1, M2, M9, M20, M26, M40,
M52, M74, M94, M167, M170, M173, M201, M207, M213,
12f2a, and LLy22g were typed under conditions summar-
ized by Montiel et al. (2005); markers M78, M81, M123,
and M215 were typed following Neto et al. (2007) condi-
tions; the rest of the markers, M215, 223, M253, and
M281, were typed under original conditions. The phylog-
eny of the markers and the names of haplogroups were
used following standardized nomenclature guidelines.
Samples resolved belonging to E3b1b subgroup were
typed for Y-chromosome short tandem repeat (STR) loci
DYS19, DYS389I and II, DYS385, DYS390, DYS391,
DYS392, DYS393, DYS456, DYS458, DYS437, DYS438,
DYS439, DYS448, DYS635, and Y GATA H4; markers
1
were co-amplified with the AmpFlSTR Y-filerTM PCR kit
(Applied Biosystems), under conditions recommended by
the manufacturer, and Polymerase Chain Reactions (PCR)
Contract grant sponsor: MEC; Contract grant numbers: BOS 2002-
00724, and CGL2006-07374.
*Correspondence to: Luis Alvarez, Unitat d’Antropologia Biològica,
Departament Biologia Animal, Biologia Vegetal i Ecologia, Universitat
Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Barcelona, Spain.
E-mail: luis.alvarez@uab.es
Received 7 October 2008; Accepted 19 December 2008
DOI 10.1002/ajhb.20888
Published online 11 February 2009 in Wiley InterScience (www.interscience.
wiley.com).

V
C 2009 Wiley-Liss, Inc.
408 L. ALVAREZ ET AL.

TABLE 1. NRY haplogroup frequencies in the populations analysed that generally show different levels of this haplogroup
(Arredi et al., 2004).
N (%)
Haplogroup Haplogroup E3b-M215 shows high levels of frequency
definition Andalusian Pedroches Valley Amizmiz Valley in Northwest Africa, East Africa, and the Middle East
(Cruciani et al., 2004), whereas in Iberia this haplogroup
A 1 (1.1) 0 (0.0) 1 (3.0) presents a widespread distribution with an overall fre-
E(xE3a, E3b) 1 (1.1) 0 (0.0) 0 (0.0)
E3a 0 (0.0) 0 (0.0) 1 (3.0) quency of 8.46% (Arroyo-Pardo et al., 2007); inside this
E3b(xE3b1a–c) 0 (0.0) 0 (0.0) 1 (3.0) haplogroup, the subgroup E3b1b-M81 presented a fre-
E3b1a 3 (3.2) 3 (4.4) 1 (3.0) quency of 2.1% in general Andalusia and 1.5% in
E3b1b 2 (2.1) 1 (1.5) 28 (84.8) Pedroches Valley. These proportions were similar to that
E3b1c 0 (0.0) 0 (0.0) 1 (3.0)
F 1 (1.1) 2 (2.9) 0 (0.0) observed in other Iberian populations (Alonso et al., 2005;
G 2 (2.1) 2 (2.9) 0 (0.0) Bosch et al., 2001; Flores et al., 2004), although the value
I(xI1a, I1b2, I1c) 3 (3.2) 3 (4.4) 0 (0.0) detected in the Pedroches Valley was lower than the a pri-
I1a 0 (0.0) 2 (2.9) 0 (0.0) ori expected because this subgroup is associated with
I1b2 2 (2.1) 0 (0.0) 0 (0.0)
J 8 (8.5) 5 (7.3) 0 (0.0) North African Berber communities. In this sense, the
K, M, O(xL, N, P) 7 (7.4) 1 (1.5) 0 (0.0) Azgour Valley population presented a high value (84.8%)
P(xR) 1 (1.1) 3 (4.4) 0 (0.0) of chromosomes belonging to subgroup E3b1b-M81. This
R(xR1) 1 (1.1) 3 (4.4) 0 (0.0) frequency value agrees with previous studies that show
R1(xR1b3f) 55 (58.5) 41 (60.3) 0 (0.0)
R1b3f 7 (7.4) 2 (2.9) 0 (0.0) an average value of 80% in Berber populations (Arredi
TOTAL 94 68 33 et al., 2004; Bosch et al., 2001; Cruciani et al., 2004).
To evaluate the diversity levels of the three analyzed
populations, Nei’s gene diversity was calculated from the
haplogroups frequency distribution. The Pedroches Valley
and the Andalusia samples presented no significant differ-
were run in a ABI 3130XL sequencer (Servei de Genòm- ences when their values were compared; moreover, both
ica, Universitat Autònoma de Barcelona). populations showed values that fit within the variability
observed in the Iberian populations that were considered
Statistical analysis for comparison. The Azgour Valley Berber sample showed
the lowest level of genetic diversity (0.284 6 0.102)
The haplogroup frequencies detected in the studied pop- detected for all North African populations because of the
ulations (Table 1) were recorded together with the fre- substantial presence of the subgroup E3b1b-M81.
quency of the populations used for comparisons, namely
the Iberian Peninsula (Alonso et al., 2005; Flores et al.,
2004; Goncalves et al., 2005), Balkans (Bosch et al., 2006),
PCA and network analysis
North Africa (Alonso et al., 2005; Arredi et al., 2004; Bosch
et al., 2001), and East Africa (Fernandes et al., 2008). To better access the relationships among populations, a
Arlequin v3.11 was run to estimate Nei’s gene diversity PCA was performed (Figure 1a). The first two components
(Nei, 1987) for each population and to perform the exact of the analysis account for 43.77% of the genetic variance.
test of population differentiation (Rousset and Raymond, The South Iberian populations analyzed stand with the
1995) used to compare the haplogroup distribution rest of Iberian populations in the negative quadrant of the
between populations. Moreover, a principal components first component because of the frequency of M173-R1. A
analysis (PCA) using frequencies of haplogroups of popu- geographical or country-based substructure is observed
lations reported was conducted using SPSS 15.0 (SPSS, for the North African populations (Eastern, Central, and
1989–2006). Western), which is determined by differences in the sec-
A median-joining network of STR haplotypes in M81- ond component that is mainly influenced by the frequency
E3b1b haplogroup was constructed and drawn with the of E3b1b-M81.
Network 4.5.0.0. Program (Fluxus Technology, 2004– Furthermore, the E3b1b-M81 Network was built to
2007). The locus-specific weight, used in the construction detect possible shared STRs haplotypes between Iberians,
of the networks, was proportional to the mutation rate per Berbers, and other populations for which both single nu-
loci (Gusmao et al., 2005). cleotide polymorphism (SNPs) and STRs were typed. The
network diagram (Figure 1b) showed a central cluster
RESULTS AND DISCUSSION representing the founder haplotype. In this cluster, Portu-
Y-SNP haplogroup variation guese and one Andalusia sample shared the central haplo-
type with North African populations including the Ber-
Haplogroup frequencies obtained for the analyzed popu- bers analyzed in this work. This can represent the prehis-
lations are shown in Table 1. Eighteen different hap- toric links between the Iberians and North Africans
logroups were found in the 195 analyzed chromosomes. In (Flores et al., 2004) or recent admixture phenomena as a
both the samples analyzed from the Pedroches Valley and consequence of the Islamic occupation of the peninsula
Andalusia, haplogroup M173-R1 was the most frequent Y- (Bosch et al., 2001). The same can be considered with the
chromosome lineage (60%). This haplogroup had been unique E3b1b-M81 Y-chromosome observed in the
described as the most frequent in Iberia (Alonso et al., Pedroches Valley that stands in a mainly European
2005; Bosch et al., 2001; Flores et al., 2004). On the other branch. Moreover, a haplotype from the Andalusia ana-
hand, in the Berber sample analyzed from the Azgour Val- lyzed populations, located in a terminal position of a com-
ley, no M173-R1 lineages were reported. The lack of plete North African branch, could be considered to be a
M173-R1 chromosomes is not usual in Berber populations recent contribution to the Iberian gene pool.

American Journal of Human Biology

 Y-CHROMOSOME VARIATION IN SOUTH IBERIA
Fig. 1. a, Principal component plot of Y-chromosome Hg frequency
data (accounting for 43.6% of the detected variance). Triangles sym-
bolize Iberian populations: White triangles represent populations of
this study (1-Pedroches Valley, 2-Andalusia), dark grey triangles cor-
respond to other Andalusia populations (3- Cadiz, 4- Cordoba, 5-
Huelva, 6- Malaga, and 7- Seville), grey triangles stand for other Ibe-
rian population (8- Biscay Basques, 9- Cantabria, 10- Castile, 11- Cat-
alonia, 12- Galicia, 13- Gipuzkoa Basques, 14- Leon, 15- Northern
Portugal, 16- Central Portugal, 17- Southern Portugal, and 18- Valen-
cia); Pentagons symbolize Northern African populations: White pen-
tagons correspond to the population of this study (19- Azgour Valley),
black pentagons stand for Moroccans populations (20- Arabs, 21-
Northern Berbers, 22- Sahara, and 23- Southern Berbers), dark grey
pentagons represent central north African populations (24- Algerian
Arabs, 25- Algerian Berbers, 26- Tunisia), and grey pentagons corre-
spond to Egyptians populations (27- Northern, 28- Southern). b,
Microsatellite networks of M81-E3b1b haplogroup. Code colors for
populations in the networks: Red 5 North Africa; Blue 5 West Africa;
Green 5 Portugal; Orange 5 Balkans; Gray 5 Azgour Valley; Yellow 5
Pedroches Valley; White 5 Andalusia.
Overall, the Pedroches Valley and the Andalusian sam-
ples typed in this study show a perfect affinity to other
Iberian populations. Our data indicate that the Berber
presence in the Pedroches Valley is less than would be
409
expected when historical or toponymic records are taking
into account, thus rejecting the hypothesis of integration,
at least at the genetic level, of the Berbers settlers in the
conquered areas. A more detailed revision of the people
movements for the last centuries is necessary to clarify
the present Y-chromosome genetic composition in the
analyzed area.
LITERATURE CITED
Alonso S, Flores C, Cabrera V, Alonso A, Martin P, Albarran C, Izagirre
N, de la Rua C, Garcia O. 2005. The place of the Basques in the Euro-
pean Y-chromosome diversity landscape. Eur J Hum Genet 13:1293–
1302.
Arredi B, Poloni ES, Paracchini S, Zerjal T, Fathallah DM, Makrelouf M,
Pascali VL, Novelletto A, Tyler-Smith C. 2004. A predominantly neo-
lithic origin for Y-chromosomal DNA variation in North Africa. Am
J Hum Genet 75:338–345.
Arroyo-Pardo E, Baeza C, Fernández E, López-Parra AM. 2007. Genetic
history of the Iberian Peninsula. In: Santos C, Lima M, editors. Recent
advances in molecular biology and evolution: applications to biological
anthropology. Kerala, India: Research Singpost. p 389–411.
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 North-
western Africa and the Iberian Peninsula. Am J Hum Genet 68:1019–
1029.
Bosch E, Calafell F, Gonzalez-Neira A, Flaiz C, Mateu E, Scheil HG, Huck-
enbeck W, Efremovska L, Mikerezi I, Xirotiris N, Grasa C, Schmidt H,
Comas D. 2006. Paternal and maternal lineages in the Balkans show a
homogeneous landscape over linguistic barriers, except for the isolated
Aromuns. Ann Hum Genet 70(Pt 4):459–487.
Cruciani F, La Fratta R, Santolamazza P, Sellitto D, Pascone R, Moral P,
Watson E, Guida V, Colomb EB, Zaharova B, Lavinha J, Vona G, Aman
R, Cali F, Akar N, Richards M, Torroni A, Novelletto A, Scozzari R. 2004.
Phylogeographic analysis of haplogroup E3b (E-M215) y chromosomes
reveals multiple migratory events within and out of Africa. Am J Hum
Genet 74:1014–1022.
Fernandes AT, Rosa A, Goncalves R, Jesus J, Brehm A. 2008. The Y-chro-
mosome short tandem repeats variation within haplogroup E3b: evi-
dence of recurrent mutation in SNP. Am J Hum Biol 20:185–190.
Flores C, Maca-Meyer N, Gonzalez AM, Oefner PJ, Shen P, Perez JA, Rojas
A, Larruga JM, Underhill PA. 2004. Reduced genetic structure of the
Iberian peninsula revealed by Y-chromosome analysis: implications for
population demography. Eur J Hum Genet 12:855–863.
Goncalves R, Freitas A, Branco M, Rosa A, Fernandes AT, Zhivotovsky LA,
Underhill PA, Kivisild T, Brehm A. 2005. Y-chromosome lineages from
Portugal, Madeira and Acores record elements of Sephardim and Berber
ancestry. Ann Hum Genet 69 (Part 4):443–454.
Gusmao L, Sanchez-Diz P, Calafell F, Martin P, Alonso CA, Alvarez-Fer-
nandez F, Alves C, Borjas-Fajardo L, Bozzo WR, Bravo ML, Builes JJ,
Capilla J, Carvalho M, Castillo C, Catanesi CI, Corach D, Di Lonardo
AM, Espinheira R, Fagundes de Carvalho E, Farfan MJ, Figueiredo HP,
Gomes I, Lojo MM, Marino M, Pinheiro MF, Pontes ML, Prieto V,
Ramos-Luis E, Riancho JA, Souza Goes AC, Santapa OA, Sumita DR,
Vallejo G, Vidal Rioja L, Vide MC, Vieira da Silva CI, Whittle MR, Zabala
W, Zarrabeitia MT, Alonso A, Carracedo A, Amorim A. 2005. Mutation
rates at Y chromosome specific microsatellites. Hum Mutat 26:520–528
Manzano-Moreno E. 2006. Conquistadores, emires y califas: los Omeyas y
la formación de Al-Andalus. Crı́tica, editor. Barcelona: Editorial Crı́tica
S. A. 620 p.
Montiel R, Bettencourt C, Silva C, Santos C, Prata MJ, Lima M. 2005.
Analysis of Y-chromosome variability and its comparison with mtDNA
variability reveals different demographic histories between islands in
the Azores Archipelago (Portugal). Ann Hum Genet 69 (Part 2):135–144.
Nei M. 1987. Molecular evolutionary genetics. New York, NY: Columbia
University Press.
Neto D, Montiel R, Bettencourt C, Santos C, Prata MJ, Lima M. 2007. The
African contribution to the present-day population of the Azores Islands
(Portugal): analysis of the Y chromosome haplogroup E. Am J Hum Biol
19:854–860.
Rousset F, Raymond M. 1995. Testing heterozygote excess and deficiency.
Genetics 140:1413–1419.
American Journal of Human Biology