Since the 1990s, human diversity studies have increasingly utilized DNA-based analyses. Y-chromosome and mtDNA markers provide complementary perspectives. Interpretive problems were apparent in the definition of a number of the ethnic study populations.
Since the 1990s, human diversity studies have increasingly utilized DNA-based analyses. Y-chromosome and mtDNA markers provide complementary perspectives. Interpretive problems were apparent in the definition of a number of the ethnic study populations.
Since the 1990s, human diversity studies have increasingly utilized DNA-based analyses. Y-chromosome and mtDNA markers provide complementary perspectives. Interpretive problems were apparent in the definition of a number of the ethnic study populations.
Abstract Initial physical anthropology studies into ethnic
diversity were largely dependent on comparative whole body
and craniometric measurements, and through time assessments of ethnic diversity based on these measures exhibited increasing statistical sophistication. Since the 1990s, in Asia as elsewhere in the world, human diversity studies have increasingly utilized DNA-based analyses, with Y-chromosome and mtDNA markers providing complementary perspectives on the origins and gene pool structures of different ethnic groups. This approach is illustrated in a study of population genetic structure in PR China, in which DNA samples from the Han majority and eight ethnic minorities were analyzed. The Y- chromosome and mtDNA data showed multiple paternal geographical and ethnic origins but restricted maternal ancestries. However, interpretive problems were apparent in the denition of a number of the ethnic study populations, which appear to reect political as well as genetic inuences. In all anthropological studies, whether based on anthropometry or genomic analysis, unambiguous and appropriate community identication is a prerequisite. J Physiol Anthropol 26(2): 77 82, 2007 http://www.jstage.jst.go.jp/browse/jpa2 [DOI: 10.2114/jpa2.26.77] Keywords: Anthropometry, Y-chromosome, mtDNA, India, China Introduction The impetus for much of the early research in physical anthropology was based on the premise that characteristic differences between human races could be demonstrated using anthropometric and craniometric measurements. An early attempt at such an approach was made by Francis Galton, who derived numerical formulae to describe facial proles, with the three-fold aim of measuring: i) hereditary likenesses or differences, ii) racial likenesses or differences, and iii) ascertaining whether special types of physiognomy were correlated with denite mental or moral characteristics (Pearson, 1924). The methodology initiated by Galton was adopted and extended by Pearson and colleagues in the Galton Laboratory at University College, London. This culminated in the derivation of the Coefcient of racial likeness (Pearson, 1926, 1928), which was applied to the quantication of physical differences between racial groups, e.g., using craniometric analysis to produce a classication of Asiatic races (Woo and Morant, 1932). The Coefcient of racial likeness gained considerable popularity and, for example, it was employed in the 1931 Census of India, with the 29 anthropometric and craniometric measures listed in Table 1, plus data on skin, hair, and eye colour, used to compare and categorize individuals drawn from a range of Indian populations. The application of the method to individuals sampled from the four southern states of India, modern-day Andhra Pradesh (Telegu), Karnataka (Kanarese), Kerala (Malayali), and Tamil Nadu (Tamil) is illustrated is Figure 1, with the calculated Coefcients of racial likeness given as mean values (Guha, 1935). Despite stringent critiques of the methodology by eminent contemporaries, e.g., Fisher (1936) and Seltzer (1937), the Coefcient of racial likeness continued to be employed as the method of choice in differentiating between the races of man although with a decline in usage following the abuses of the discriminatory racial identication legislation and practices in Germany in the 1930s and early 1940s. Today, comparative anthropometric and craniometric analysis remains in use as an investigative tool, e.g., in studies on caste and tribal groups in India (Sirajuddin et al., 1994; Bharati et al., 2004), and the Coefcient of racial likeness was adopted in a recent study of body mass index (Adak et al., 2006). During the 1940s and 1950s, blood group assays, mainly based on serotyping techniques, were introduced for inter- ethnic comparisons, and by the late 1950s and 1960s protein electrophoresis was in widespread use for the study of polymorphic variation. The introduction of genomic analysis in the 1990s has not quelled the controversy as to whether or Physical Anthropology and Ethnicity in Asia: The Transition from Anthropometry to Genome-based Studies A. H. Bittles 1,3) , M. L. Black 1,2) and W. Wang 2,1) 1) Centre for Human Genetics, Edith Cowan University, Perth, Australia 2) Capital Medical University, Beijing, PR China 3) Centre for Comparative Genomics, Murdoch University, Perth, Australia not inter-ethnic genetic differences are meaningful, although in recent years the debate has largely concentrated on the question of ethnicity in biomedical studies and clinical practice (Cooper et al., 2003; Gonslez Burchard et al., 2003; Jorde and Wooding, 2004). However, there is considerable disquiet surrounding the relevance and applicability of ethnic differences in disciplines such as human biology and biological anthropology. Genome-based Studies on the Relationship between Chinese Ethnic Minorities To study genetic differences between the majority Han population of PR China and eight representative ethnic minority populations, nger-prick blood samples or DNA specimens were obtained from unrelated males belonging to the majority Han population and the Muslim Hui community of Liaoning Province; the Muslim Bonan, Dongxiang, and Sala communities of Gansu Province; the Kucong, Miao, and Yao communities of Yunnan Province; and Tibetans resident in the Tibetan Autonomous Province. The samples were analysed for eight Y-chromosome binary markers (M1 (YAP), M216/M130 (RPS4Y), M89/M213, M9, M175/M214, M122, M45, and M17) and ve Y-chromosome microsatellite markers ( DYS19, DYS388, DYS 389a, DYS389b, DYS393), via PCR-RFLP and uorescent genotyping respectively (Black et al., 2006). The mtDNA hypervariable region I (HV-I) was amplied and sequenced using ABI Prism dye primer kits running on 6% denaturing polyacrylamide gels, with sequence data analysed using ABI DNA analysis software and SEQUENCER (Gene Codes). The subjects also were screened for the mtDNA 9-bp deletion in the COII/tRNA Lys intergenic region. To identify the origins and genetic relatedness of the nine study communities, the Y-chromosome and mtDNA data were separately analyzed by summary statistic and phylogenetic methods (Wang et al., 2003; Black et al., 2006). Broad haplogroups were inferred from Y-chromosome binary polymorphisms according to the Y Chromosome Consortium (YCC 2002), while mtDNA haplogroups were inferred from HV1 sequences (Yao and Zhang, 2002). Genetic distances were calculated from Y-chromosome microsatellite data (dm 2 distance) with the software package MICROSAT, while mtDNA genetic distances (TamuraNei distance) were calculated with MEGA3 software. Neighbour-joining trees were then generated from these distance matrices using MEGA3. The Y-chromosome haplogroup frequencies indicated a range of dispersed male origins (Table 2). The Muslim Bonan, Dongxiang, and Sala displayed a combination of Middle Eastern, Central Asian, and East Asian male ancestries, indicated by the presence of haplogroups P*(xR1), R1a1*, and O3 respectively. The Muslim Hui additionally showed a signicant frequency of haplogroup C, indicative of a North Asian, and possibly Mongolian, male ancestral component. The Han and Miao had predominantly post-Neolithic East Asian male ancestries, comprising haplogroups O* and O3, whereas the Tibetan and Yao populations showed older pre- Neolithic East Asian origins with a high frequency of haplogroup D ancestries, a relatively rare haplogroup indicated by the presence of the YAP insertion which also is found at high frequency in Japanese populations (Hammer et al., 2006). The Kucong contained a pre-Neolithic male ancestry of southern Asian origin, which was possibly older than the other study populations (Black et al., 2006). The neighbourjoining tree derived from the Y-chromosome microsatellite data (Fig. 2a) showed no immediately obvious grouping of populations equivalent to the genetic ancestries found with the binary data, and the genetic distance measure conrmed the high levels of male diversity. Examples of seemingly incongruous relationships include the Hui grouping closely with the Yao, and the co-clustering of the Bonan and the Miao. On initial inspection these groupings could be 78 Anthropology and ethnicity in Asia Table 1 Anthropometric and craniometric measurements used to calculate the Coefcient of racial likeness. Stature Nasal length Breadth height ratio Auricular height Nasal breadth Trans fronto-parietal index Maximum head length Nasal height/depth Orbitonasal index Maximum head breadth Upper facial length Nasal index Minimum frontal breadth Total facial length Nasal elevation index Maximum bizygomatic breadth Horizontal head circumference Upper facial index Bigonial breadth Sagittal arc Total facial index Inter-orbital breadth Transverse arc Trans cephalo-facial index Orbitonasal breadth Length breadth index Vertical cephalo-facial index Orbitonasal arc Length height index Fig. 1 Coefcients of racial likeness for four South Indian populations. interpreted as inferring close ancestral ties, but the histories of both pairs of populations are notably different (Du and Yip, 1993), and would not support any such relationships. It, therefore, seems probable that the clustering displayed by the neighbouringjoining tree linked populations on the basis of their similar internal population structures, rather than their ancestral origins. By comparison, the equivalent mtDNA haplogroup frequency (Table 2) and the neighbour-joining tree (Fig. 2b) plotted from the mtDNA data strongly suggested that the major Bittles, AH et al. J Physiol Anthropol, 26: 7782, 2007 79 Table 2 Y-chromosome and mtDNA haplogroup frequencies of the nine Chinese study populations. Province/region Liaoning Yunnan Xizang Gansu Population Han Hui Kucong Miao Yao Tibet Bonan Dongxiang Sala Y-chromosome haplogroup frequency Haplogroup DE 0.190 0.571 0.550 0.056 0.065 0.062 C 0.049 0.227 0.053 0.028 0.062 F*(xK) 0.146 0.227 0.524 0.132 0.071 0.100 0.167 0.152 0.094 O 0.227 0.090 0.282 0.071 0.250 0.056 0.065 0.218 O3 0.480 0.136 0.286 0.533 0.286 0.250 0.196 0.312 K*(xO,P) 0.098 0.136 0.100 0.111 0.109 R1a1 0.181 0.25 0.321 0.218 P*(xR) 0.056 0.092 0.032 mtDNA haplogroup frequency Haplogroup D 0.500 0.385 0.111 0.273 0.533 0.091 0.111 0.455 M* 0.167 0.231 0.083 0.111 0.091 0.200 0.111 0.182 C 0.083 0.167 0.273 Z 0.231 0.222 G 0.077 A 0.077 0.167 0.133 0.455 W 0.111 B 0.167 0.250 0.222 0.091 0.067 0.222 0.273 F 0.083 0.333 0.182 0.182 0.111 R* 0.083 0.25 0.222 0.273 0.067 0.111 0.091 Adapted from Black et al. (2006) Fig. 2 (a) Unrooted neighbourjoining tree based on the genetic distances from Y-chromosome microsatellite haplotypes of nine Chinese ethnic populations. The scale bar indicates branch length in dm 2 genetic distance units. (b) Unrooted neighbourjoining tree based on the genetic distances from mtDNA HV1 sequences of nine Chinese ethnic populations. The scale bar indicates branch length in TamuraNei genetic distance units. female contribution in each of the ethnic communities was provided by East Asian women, with considerable homogeneity indicated by the TamuraNei genetic distance measure of just 0.002 units. Despite the fact that, since the foundation of communities such as the Muslim Bonan, Dongxiang, Hui, and Sala from the 8 th century onwards (Lipman, 1997; Gladney, 1998), each community has been endogamous (Du and Yip, 1993; Black et al., 2006). The two complementary genomic data sets clearly indicate that, while the male origins of the non-Han ethnic minorities were drawn from multiple Chinese and non-Chinese populations, the females of all of the ethnic minorities showed an almost exclusively Chinese ancestry, ndings which are in accord with historical records. Assessing the Results of Anthropogenetic Studies There are obvious problems in seeking to compare results obtained via anthropometric measurements to those determined on the basis of blood group or protein polymorphisms, and to an even greater extent to studies based on genomic analysis. Anthropometric measures, such as the Coefcient of racial likeness, are based on variation that derives from combinations of genes variously expressed from conception to adulthood and interacting with environmental factors that inuence their expression. In contrast, blood group and protein polymorphisms are mainly single gene measures which may be selective or neutral in effect, with partial neutrality the most commonly observed outcome. The single tandem repeat Y-chromosome loci and the HV-1 mtDNA markers used in the present study of the Chinese ethnic communities (Table 2) provide a selectively neutral perspective on former generations, which can be used to identify the geographical origins and approximate dates of past migrational events. As such, the level and precision of the genetic information they provide is much greater than that available from blood group or protein polymorphisms, and incomparably more specic than the genetic content of the anthropometric analyses. However, what each of these measures has in common is absolute reliance on: i) adequate and representative sample sizes; ii) the accuracy and relevance of the denitional information available for their study populations; and iii) the application of appropriate statistical methods. In a large number of studies, the sample sizes are unrealistically small to provide a representative picture of the population. Thus, in the Census of India of 1931 (Guha, 1935), the all-India Coefcient of racial likeness was calculated from a total of 2,163 males and 348 females, at a time when the population of the country totalled 279 million (Dyson, 2001). Of equal importance, in the South Indian comparative study (Fig. 1), the four different ethnicities were based on individuals drawn from the Brahmin caste (varna). Yet the Brahmin varna is notably sub- divided and, for example, in Karnakata, the home state of the Kanarese, 101 different and mainly endogamous Brahmin sub-communities were identied and enumerated in the 1901 Census (Ananda Row, 1901). In this example, the question arises as to whether each of the four ethnic groups tested was sampled from a single sub-community or from multiple endogamous Brahmin communities. Further, the 1931 Census of India drew specic attention to the fact that, In India, due to the social prestige and privileges enjoyed by upper castes, there is always a tendency for individuals to surreptitiously pass off as members of castes higher than the ones to which they rightly belong (Guha, 1935, p. iv). Under these circumstances, unless the selection of each study population has been conducted with absolute rigour, the results obtained may be of questionable validity or value. Despite the demonstrated differences between the Han and the eight ethnic minority communities (Table 1), as already indicated, the apparent genetic relationships suggested by the neighbour-joining trees derived from the Y-chromosome data (Figs. 2a, 2b) merit some scepticism. During the course of the study it also became apparent that problems existed in the identication of the Chinese study communities (Black et al., 2006). The starting-point for the study was to select representative ethnic communities, and for this purpose a number of specic minzu (ethnic populations) were chosen. In China, 56 minzu received governmental recognition following the establishment of the Peoples Republic and, besides the Han, seven of the communities sampled, the Bonan, Dongxiang, Sala, Hui, Miao, Yao, and Tibetans were recognized minzu. At the time of the establishment of the Peoples Republic, the Kucong followed a hunter-gatherer lifestyle in the tropical forests on the Yunnan-Burma border and they were not accorded ofcial recognition as a minzu. On subsequent investigation, it became apparent that at least two of the ofcially recognized minzu could have been notably heterogeneous. The Yao are resident in six different provinces of PR China, and over thirty different zhixi (sub-groups) have been identied within the ofcial Yao minzu structure (Litzinger, 1995). Their cultural diversity is further demonstrated by the worship of separate deities in different Yao communities, and by particular sub-community beliefs in mythological origins and religious festivities, all of which were allegedly unknown to other Yao communities before the establishment of the Yao minzu in 1949 (Litzinger, 1995). The Miao also present classicatory difculties, and it has been claimed that early use of the term Miao referred to most of the disparate indigenous peoples across southwest China rather than to a single ethnic group (Diamond, 1995). The present- day Miao can be sub-divided into three broad groups, the Hmong in Yunnan, the Hmu in Guizhou, and the Xioob in Guizhou and Hunan, which exhibit cultural and linguistic differences (Diamond, 1995). From an anthropogenetic perspective, historical sources would strongly suggest that it would be more realistic and prudent to conduct subject selection in the Yao and the Miao on a sub-community by sub-community basis. This contention also applies to the 9.8 million Hui Muslims who are resident across all of the provinces of PR China (National Bureau of 80 Anthropology and ethnicity in Asia Statistics of China, 2002). Apart from their shared religion, the Hui do not appear to have any other unifying cultural or language criteria that would convincingly permit their analysis as a single ethnic minority. The population of Asia is now estimated at 3,921 million, 60.5% of the world total (PRB 2006), and in a large country such as India the population of 1,104 million is sub-divided into 50,00060,000 discrete endogamous communities (Bittles, 2005). Since some 93% of the population of PR China are Han, the level of genetic differentiation is unlikely to match that of India. But it still seems improbable that 1,304 million people can be convincingly sub-divided into just 56 discrete ethnicities, or that 1,213 million Han are genetically homogeneous. The original decision to create recognized ethnic minzu, therefore, appears to have been as much a matter of political convenience as a recognition of longstanding anthropological divisions, e.g., based on cultural, religious, and language diversity (Black et al., 2006). Most Asian countries, large and small, are characterized by high levels of clan and tribal sub-division. While community knowledge of family lineages and clan afliations usually appears to be credible, genomic studies in Central Asia have shown that the wider tribal origins claimed by communities may more mythical than factual (Chaix et al., 2004). Whatever the method of analysis adopted, in all anthropological studies, appropriate care is essential in the sampling of study populations. Otherwise, in the absence of stringent selectional criteria, the outcomes obtained and hence the conclusions drawn may be seriously impaired, misleading, or simply erroneous. 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Bittles, Room 19.387, Edith Cowan University, 100 Joondalup Drive, Perth WA 6015, Australia Phone: 61863045623 Fax: 61863045851 e-mail: a.bittles@ecu.edu.au 82 Anthropology and ethnicity in Asia