C. R. Biologies 335 (2012) 698–707 Contents lists available at SciVerse ScienceDirect Comptes Rendus Biologies w ww.sciencedirect.com Anthropology/Anthropologie The Hmong Diaspora: Preserved South-East Asian genetic ancestry in French Guianese Asians La Diaspora Hmong : un patrimoine ge´ne´tique conserve´ chez la population asiatique de Guyane franc¸aise a,b c, a d Nicolas Brucato , Ste´phane Mazie`res *, Evelyne Guitard , Pierre-Henri Giscard , a,y a a E´ tienne Bois , Georges Larrouy , Jean-Michel Dugoujon a UMR 5288 CNRS, Laboratoire d’Anthropologie Mole´culaire et Imagerie de Synthe`se (AMIS), Universite´ Paul-Sabatier Toulouse III, Toulouse, France b Language and Genetics Department, Max Planck Institute for Psycholinguistics, Nijmegen, The Netherlands c CNRS, EFS-AM, ADES UMR 7268, Faculte´ de Me´decine, Aix Marseille Universite´, Secteur Nord, baˆtiment A - CS80011, 51, boulevard Pierre-Dramard, 13344 Marseille cedex 15, France d Institut des De´serts et des Steppes–Muse´um, Paris, France A R T I C L E I N F O A B S T R A C T Article history: The Hmong Diaspora is one of the widest modern human migrations. Mainly localised in Received 4 July 2012 South-East Asia, the United States of America, and metropolitan France, a small Accepted after revision 8 October 2012 community has also settled the Amazonian forest of French Guiana. We have biologically Available online 7 November 2012 analysed 62 individuals of this unique Guianese population through three complementary genetic markers: mitochondrial DNA (HVS-I/II and coding region SNPs), Y-chromosome Keywords: (SNPs and STRs), and the Gm allotypic system. All genetic systems showed a high Anthropology conservation of the Asian gene pool (Asian ancestry: mtDNA = 100.0%; NRY = 99.1%; Genetic markers Gm = 96.6%), without a trace of founder effect. When compared across various Asian Asian populations populations, the highest correlations were observed with Hmong-Mien groups still living Gene diversity in South-East Asia (Fst < 0.05; P-value < 0.05). Despite a long history punctuated by Diaspora exodus, the French Guianese Hmong have maintained their original genetic diversity. ß 2012 Acade´mie des sciences. Published by Elsevier Masson SAS. All rights reserved. 1. Introduction the 16th century, forced the Hmong to leave their heartland and to settle the Yunnan highlands [2]. There, they The Hmong Diaspora is widely scattered in Asia, but also developed an economy based on a shifting agriculture of in America, Europe and Oceania. This human migration crops like maize and millet, together with husbandry and would have originated from the South Chinese province of hunting. These strategies permitted the Hmong to prosper Kweichow [1]. Associated with Neolithic cultures of Daxi among the other ethnic minorities of China, where they are (5300–6400 BP) and Qujialing (4600–5000 BP), the Hmong also known as Miao. The Hmong share common cultural is an ancient and an important component of the human traits particularly with the Mien, including language, and diversity of South East Asia [2]. The Han expansion, during genetic characteristics that cluster the Hmong and Mien together in the so-called Hmong-Mien group [3,4]. Following a period of calm, since the 18th century the Hmong have conflicted at various times with the Chinese * Corresponding author. government. They took refuge in northern Indochina E-mail address: [email protected] (S. Mazie`res). y Deceased author. where their skills in shifted cultivation rapidly integrated 1631-0691/$ – see front matter ß 2012 Acade´mie des sciences. Published by Elsevier Masson SAS. All rights reserved. http://dx.doi.org/10.1016/j.crvi.2012.10.003 N. Brucato et al. / C. R. Biologies 335 (2012) 698–707 699 them into the regional economic network, and progres- available to identify the ancestry of each haplotype sively gave the Hmong a relative institutional power, observed in this study [11,12]. The third system was the under the French colonial influence. However, the down- biparental Gm allotypic system. This system comprises side of the strategic role of the Hmong was the direct polymorphic antigenic markers located on the constant implication in the First Indochina War (1946–1954), the regions of the heavy chains of three subclasses of Vietnam War (1959–1975) and the Secret War in Laos immunoglobulin G (IgG1, IgG2, and IgG3) [13,14]. The (1964–1975). The latter triggered a massive exodus of Gm system has been frequently studied in human Hmong from Laos toward northern Thailand, but also populations and has shown heterogeneous haplotype toward other continents. Almost 200,000 individuals went frequencies among populations [15,16]. Our study pro- into exile to the United States and 15,000 to metropolitan poses the first genetic insight into the Hmong Diaspora and France. Most of these migrants preferred to live in its particular example in French Guiana. The present study occidental cities but others were steered toward an will aim to answer two questions in particular: unusual destination: French Guiana [5]. This South American territory, localised between has the original Hmong genetic pattern been shaped by Surinam and Brazil, has been a French colony since the founder effect and admixture during their long exodus? 17th century. Today it only has three individuals per is the French Guianese Hmong (FGH) gene pool still close square kilometre. Welcoming the Hmong onto its soil to the Hmong pool seen in South East Asia today? represented a double advantage for the French govern- ment: to shelter former cooperating people along with 2. Materials and methods populating areas of low population density where condi- tions of life are similar to those found in Laos [5]. 2.1. Population sampled Approximately 2100 Hmong are living in French Guiana. They reinstated their former rural farming communities A total of 141 individuals were sampled in the Cacao with presenting all the social characteristics of South East and Javouhey communities (Fig. 1) in 1980 under the Asia, and preserved their original Hmong cultural identity. auspices of the Institut National de la Sante´ et de la But more than an ethnic isolate, they are also completely Recherche Me´dicale (Paris, France). All samples were integrated in the French Guianese social network by obtained with the informed consent of the participants. significantly increasing the local agricultural production. DNA was extracted recently by a phenol-chloroform Today, they constitute an important component of the protocol from sera, and stored at À20 8C. During the Cacao ethnic diversity of French Guiana that is primarily and Javouhey sample collection, pedigrees were recorded composed by Amerindians, Europeans, Creoles and Noir and used to select 62 maternal and 33 paternal unrelated Marron populations [6–10]. lineages for the present study. The French Guianese example of the Hmong Diaspora represents a unique human migration that has never been 2.2. Laboratory methods studied genetically. To this aim, three different and complementary genetic systems were analysed in this 2.2.1. mtDNA study. Uniparental lineages were determined through the Maternal lineages were characterised by the sequenc- analysis of mitochondrial DNA (mtDNA) and non-recom- ing of a large part of the D-loop region (16012-263) binant Y-chromosome (NRY) haplotypes. Through the following previously described protocol [17]. All data were large number of populations that have previously been obtained on an ABI PRISM 3730 sequencer (PE, Applied genotyped for these two systems, informative data are Biosystems) and analysed with Sequence Scanner v.1.0 (PE, Fig. 1. Geographic location of the two Hmong communities sampled in French Guiana. 700 N. Brucato et al. / C. R. Biologies 335 (2012) 698–707 Applied Biosystems). For preliminary haplogroup assign- Haplotype networks were generated for two pan-Asiatic ment, each sequence was firstly aligned with rCRS [18] lineages, the mtDNA haplogroup M* and the Y-chromosome using BioEdit v.7.0.9.0, then tested for relevant single haplogroup O3*, using respectively the HVS-I data and the Y- nucleotide polymorphisms (SNPs) of the coding region STR core haplotype (DYS19, DYS389I, DYS389II, DYS390, (positions 10400, 10398, 7028, 5178, 13263, 6392) typed DYS391, DYS392, DYS393) from the FGH data and all Asian by Restriction Fragment Length Polymorphisms with the comparable data, via the median-joining algorithm of following enzymes: AluI, HpaI, and DdeI. The final Network v.4.5.1.6 (www.fluxus-engineering.com). To ob- haplogroup assignment was obtained from the differences tain the most parsimonious networks the reticulation with the rCRS by the most recent mtDNA phylogeny permissivity was set to zero. Data were pre-processed using [12,19], then the haplotypes presented in relation to both the star contraction option in Network v.4.5.1.6 [27]. For the the rCRS and RSRS references sequences [20]. mtDNA data, hypermutable sites were identified by post- processing using the Steiner (MP) algorithm and removed 2.2.2. NRY from the analysis [28]. Weight of each Y-STR loci were Paternal lineages were determined by two types of characterised according to its variance in the O3a* sample, markers. NRY haplotypes are a combination of seventeen as previously described [29]. Tajima’s D and Fu’s Fs tests Short Tandem Repeats (STRs), typed using the AmpFLSTR were calculated with mtDNA data using the ARLEQUIN 3.11 1 Yfiler kit (PE, Applied Biosystems), and informative SNPs software package [26]. (UEPs: SRY 10831, M213, M9, M70, M22, Tat, 92R7, M173, Cross-population comparisons based on HVS-I mtDNAs, P25, M122, M134, M175, P31, M50, M101, M119, SRY465, Y-STR haplotypes and Gm lineages were performed using 1 47z, M88, M95, M216, M174, P197), typed by SNaPshot ARLEQUIN 3.11 [26] and depicted through the Fst minisequencing (PE, Applied Biosystems) using already parameter Significance is given for P-values under 0.05.