Forensic Science International: Genetics Supplement Series
Volume 2, Issue 1 , Pages 331-333, December 2009

Mitochondrial DNA control region sequence analysis of Mataco–Guaicurú speaking tribes from Argentina

  • Andrea Sala

      Affiliations

    • Servicio de Huellas Digitales Genéticas, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina
    • Corresponding Author InformationCorresponding author. Tel.: +54 11 4964 8281/8282; fax: +54 11 4964 8281/8282.
    • ,
  • Evguenia Alechine

      Affiliations

    • Servicio de Huellas Digitales Genéticas, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina
    • ,
  • Cecilia Bobillo

      Affiliations

    • Servicio de Huellas Digitales Genéticas, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina
    • ,
  • Luciano J. Merini

      Affiliations

    • Cátedra de Microbiología Industrial y Biotecnología, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Argentina
    • ,
  • Carmen G. Ayala

      Affiliations

    • Laboratorio de Biología Molecular-Servicio de Histocompatibilidad e Inmunogenética, Hospital de Alta Complejidad Pte Juan D. Peron, Formosa, Argentina
    • ,
  • Juan C. Acosta Ferreira

      Affiliations

    • Cuerpo Médico Forense, Poder Judicial de Formosa, Argentina
    • ,
  • Daniel Corach

      Affiliations

    • Servicio de Huellas Digitales Genéticas, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina

Received 11 August 2009; accepted 14 August 2009. published online 27 September 2009.

Article Outline

Abstract 

In this work we analyze three Mataco–Guaicurú speaking Amerindian tribes that inhabit the Northern Argentinean portion of Gran Chaco region. One hundred and sixty-eight samples of unrelated males belonging to two linguistic families: Guaycurú (Toba from Chaco, n=27; Toba from Formosa, n=37 and Pilaga from Formosa, n=56) and Mataco (Wichi from Formosa, n=48) were investigated. The entire Control Region of the mtDNA from position 16024 to 576 was sequenced. EMPOP sequencing strategy was employed including the use of 10 primers for each sample. Specific haplotypes were found in these populations with very high frequency. These findings could provide clues to address the ethnicity of a sample in routine forensic casework.

Keywords: Argentinian Mataco–Guaicurú speakers, mtDNA

 

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1. Introduction 

The investigation of the polymorphisms located in the mitochondrial DNA (mtDNA) genome allows tracing back the evolutionary process in the history of the populations. Restriction fragment analysis in the mtDNA coding region, in first term, and then the analysis of the region called “Hypervariable Region I” (HVRI) in the non-coding region or D-Loop, allowed to detect specific characteristic in Native American populations [1], [2], [3], [4]. More recently, the incorporation to the Hypervariable Region II and III analysis increased the power of the mitochondrial information in anthropological and forensic studies. Based on the complete mtDNA genome sequence it was possible to define the phylogeny of the four pan-American haplogroups A2, B2, C1 and D1 [5] as well as other Native American subhaplogroups (e.g. C4c, D2a, D3, D4h3 and X2a).

Northern Argentina is inhabited by several communities that belongs to different ethnic groups. In particular, Formosa and Chaco provinces lodges the following ethnicities: Toba, Chulupi, Mocovíes, Wichí and Pilagá (the last group is restricted exclusively to Formosa province). These two provinces are part of the so-called Gran Chaco Region and have been the focus of several anthropological and genetic studies in the last years [6], [7], [8], [9], [10].

Aiming to increase the knowledge of the mtDNA sequence data available, we analyzed the mitochondrial sequence variations in three ethnic groups: Pilagá, Wichí and Toba. Previous investigation in these communities was based on restriction fragment polymorphism and HVRI sequencing in order to determine the mitochondrial haplogroups distribution and haplotype variation [7], [9]. In that sense we expanded the investigation including the analysis of the mtDNA HVRI, HVRII, HVRIII, VR1 and VR2. This work constitutes the first investigation of the entire mitochondrial D-Loop following the EMPOP guidelines to obtain high quality sequences in aboriginal individuals from Argentina.

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2. Material and methods 

2.1. Samples 

This study included the analysis of 168 unrelated individuals belonging to Mataco and Guaycurú speaking tribes inhabiting in the north Argentina. Guaycurú-speakers (Toba from Chaco province, n=27; Toba from Formosa province, n=37 and Pilagá from Formosa, n=56) and Mataco-speakers (Wichí from Formosa, n=48). All the samples were obtained after signing a written consent statement by the donors and the investigation was approved by the Ethical Commission of the Faculty of Pharmacy and Biochemistry, University of Buenos Aires. Sampling was carried out with the help of local authorities. DNA was extracted from saliva samples according to Quinque et al. [11].

2.2. Methods 

The entire D-Loop was amplified using L15971-H649 flanking primers. Sequences were obtained with the following 10 primers: L15971, L15989, H00016, H00270, L00015, L00029, H00639, H00599, L00314 and H00159. Electrophoresis was conducted in an ABI 3100 Avant, Applied Biosystems automated sequencer. The software Sequencher v4.8 was employed for sequence analysis. Sequences were aligned with Alignment v1.1.30. Molecular diversity indices as well as Fst distances were obtained with the software Arlequin v3.1. Haplotype diversity was calculated according to Nei [12]. For statistical analysis both Toba communities (Chaco and Formosa) were pooled and consider as Toba group.

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3. Results and discussion 

Sequences from 168 individuals were performed following the EMPOP guidelines, obtaining unambiguous haplotypes. The entire Control Region of the mtDNA spanning position 16024 to 576 was sequenced. Table 1 summarizes haplogroup distribution, molecular diversity indices and haplotype diversity calculated in the three groups investigated. The four pan-American haplogroups (hgs) were observed in the groups investigated, except for hg C1 in Wichí group. As expected transition are the most common substitution observed. Insertions were observed in HVR II and HVRIII. Unusual transversions to G or A purine bases was observed in position 16111 in the most frequents haplotypes belonging to hg D1 in Toba group. Two heteroplasmic positions were observed: in 150 (C/T) in Toba group and in 234 (G/A) in Wichí group.

Table 1. Haplogroups distribution and standard diversity indices in the three ethnic groups analyzed.
HgsA2 (%)B2 (%)C1 (%)D (%)No. transicNo. transvNo. indelsNo. different haplotypesaNucleotide diversitybHaplotype diversityb
Pilagá (n=56)10.742.916.130.3393915/56 (26.8)0.0118±0.00590.8889±0.0170
Wichí (n=48)22.933.3043.8332422/48 (45.8)0.0115±0.00580.9140±0.0169
Toba (n=64)21.932.84.740.65451030/64 (46.8)0.0109±0.00550.9419±0.0113

aNumber of different haplotypes is indicated respect to the sample size and in percentage between brackets.

bValues of nucleotide and haplotype diversity±standard deviation.

Nucleotide diversities were similar in the three groups whereas differences were observed in the haplotype diversities. The lowest haplotype diversity was observed in the Pilagá group, where only 15 different haplotypes were observed in 56 samples analyzed. Between these, only five haplotypes were unique and a very frequent D haplotype (88.23% of the D samples) were shared by 15 individuals. The HVRI sequence of these samples present identity with the HVRI of the most frequent D haplotype described by Cabana et al. [9] in a sample of 38 Pilagá. The particularity of this haplotype is that the transition 16325 that characterize D1 haplogroup [5] is absent. On the other hand, transitions 152 and 16291 allows to suppose that these samples could belong to hg D4. Further investigations involving the coding region will permit to determine the subhaplogroup of this particular haplotype.

In addition with the homopolymeric C-strech region between 16182 and 16193 positions observed in hg B2 samples in the three groups, a particular insertion in 463.1 together with a transition at 460 generated another homopolymeric C-strech in HVRIII. This feature was observed in 3/56 individuals from Pilagá group, all of them belong to hg B2 and shared identical haplotype. This motif was not observed in a set of 388 samples from different regions of Argentina [13]. On the other hand, scarce information in the third Hypervariable Region of the mtDNA does not permit to compare this finding with other ethnic groups.

Significant Fst values were obtained for the comparison between Pilagá and Wichí (Fst=0.10328, p=0.0000), Pilagá and Toba (Fst=0.0646, p=0.0000), Wichí vs Toba (Fst=0.07375, p=0.00195). Only the Fst between Toba from Chaco and Toba from Formosa was no significant (Fst=0.03087, p=0.07812). These finding reinforce previously described relationship between the three communities based on Y-chromosome and autosomal markers [14], [15].

This preliminary analysis of the mitochondrial D-Loop variations in three communities allowed to detect some peculiarities in the groups analyzed. Further investigations in the coding region will allow obtaining a complete information about the mitochondrial characteristics of these groups. These findings could provide clues to address the ethnicity of a sample in routine forensic casework and will constitute a valuable research in the anthropological field.

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Conflict of interest 

None.

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Acknowledgements 

This work was supported, in part, by grant UBACyT.B-047, to DC. DC and AS are members of the Carrera del Investigador Científico y Tecnológico-CONICET, Argentina.

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References 

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PII: S1875-1768(09)00059-6

doi:10.1016/j.fsigss.2009.08.085

Forensic Science International: Genetics Supplement Series
Volume 2, Issue 1 , Pages 331-333, December 2009

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