ANTHROPOLOGICAL SCIENCE Vol. 119(3), 259–264, 2011

Genetic sketch of the six population groups of Rajasthan: a study based on 12 autosomal loci Rima DADA1, Kallur Nava SARASWATHY2*, Khangembam Somibabu MEITEI2, Prakash Ranjan MONDAL2, Harpreet KAUR3, Kiran KUCHERIA1, Seema BHARDWAJ4, Gaznavi IDRIS1 1Department of Anatomy, All India Institute of Medical Sciences, Delhi 110029, India 2Department of Anthropology, University of Delhi, Delhi 110007, India 3INCLEN Trust International India, Delhi 110020, India 4University of Banswara, Banswara 327001, India

Received 26 August 2010; accepted 11 January 2011

Abstract Rajasthan lies on the northwest border of India, and has acted as a major route for human movements since prehistoric and historic times. The present study was conducted to gain an insight into inter- and intrapopulation affinities or variations among the six population groups of Rajasthan. DNA samples from 221 unrelated individuals belonging to six endogamous population groups of Rajasthan, including both tribal (Bhil, Damaria, Garasia, Mina and Saharia) and caste populations (Rajput) were screened for 12 DNA (seven Alu indel and five RFLP) markers. All the loci were found to be polymorphic in all the studied populations. The GST values which determine the genetic differ- entiation between the population ranges from 1.5% in LPL to 8.7% in APO. This study suggests that the population groups of Rajasthan are genetically close to other Indian populations, reflecting a com- mon genetic unity among the Indian populations. The study also indicates European gene flow into the populations studied in this paper.

Key words: Autosomal markers, common linguistic group, genetic affinity, genetic differentiation

Introduction the Indus Valley civilization that is approximately 4500 years old, but disappeared about 3500 BP. The largest con- Rajasthan lies on the northwest border of India and has centration of Indus Valley civilization sites appear in an area varied topography ranging from Aravalli Hills (the oldest encompassing modern-day Rajasthan, Gujarat and Punjab. hill range in the world) to the Thar Desert, which covers In addition, the linguistic homogeneity of the region, includ- 61% of the land area of Rajasthan. The climate of Rajasthan ing both the caste and tribal groups of Rajasthan, is also ranges from scorching summers to chilly winters. This high- unique as they all fall within the Indo-European linguistic ly heterogeneous landscape could have geographically iso- family, which is mainly confined to the caste populations of lated the populations of Rajasthan and helped to maintain India. However, traces of the Dravidian linguistic group are their endogamous nature. Due to the harsh climatic condi- also found in the literature with respect to the Indus Valley tions, differential selective pressures are also likely to have civilization (Marshall, 1979; Knorozov et al., 1981). It is be- been present. The Indian subcontinent is considered as a lieved that speakers of Dravidian languages were wide- major corridor for the dispersal of modern humans as they spread throughout India before the arrival of Indo-European emerged out of Africa about 100000 years ago (Cavalli- speakers (Thapar, 1966). Subsequently, Indo-European Sforza et al., 1994; Cann, 2001). Since it is located at the speakers exercised dominance over the pre-existing pop- crossroads of Africa and West as well as East Eurasia ulations (Dravidian) and Dravidian speakers are currently (Cordaux et al., 2003), Rajasthan in India probably served as confined primarily to the southern part of India. These cul- one of the major land routes for human migrations. More- tural contacts and invasions have resulted in a high degree of over, from the fourth century BC onwards, India, particular- genetic and cultural differentiation among the people of ly in the north, was subjected for 2000 years to repeated Rajasthan. waves of penetration by alien peoples (Danielou, 2003; Indian populations are basically categorized into tribes Bhasin, 2006). The archaeological evidence for this is and castes. The tribes of India constitute only 8.2% of the present in the form of many prehistoric sites which include total Indian population and are more or less confined to rural India. The tribal populations of Rajasthan studied in this paper speak Indo-European languages as against the South * Correspondence to: Kallur Nava Saraswathy, Biochemical and Indian (Dravidian speakers) and East Indian (Austro-Asiatic Molecular Anthropological Laboratory, Department of Anthropol- ogy, University of Delhi, Delhi 110007, India. and Tibeto-Burman speakers) tribes of India. There are E-mail: [email protected] around 573 tribal populations in India, of which 75 are Published online 5 April 2011 categorized as primitive tribes by the Indian government. in J-STAGE (www.jstage.jst.go.jp) DOI: 10.1537/ase.100826 The caste group Rajput belongs to the major hierarchical

© 2011 The Anthropological Society of Nippon 259 260 R. DADA ET AL. ANTHROPOLOGICAL SCIENCE varna system of India, comprising Brahmins, Kshatriyas, Materials and Methods Vaishyas, and Shudras. In this context the present study analyses five tribal populations (Bhils, Damaria, Garasia, Intravenous blood samples (5 ml) were collected from Minas, and Saharia) and one caste group of Rajasthan healthy, unrelated individuals of both sexes from six endog- (Rajputs) with respect to 12 autosomal markers. All the stud- amous groups (Bhils, Minas, Saharia, Garasia, Damaria, and ied populations belong to the Indo-European linguistic fam- Rajputs) of Rajasthan, with proper informed written consent. ily. Except for the Indo-European speaking Rajputs, which Samples were collected from the Banswara district of Rajas- is a caste group, all the other population groups are tribal. than (Figure 1) as this has the highest concentration of tribal Historically, the Bhils (Mann, 1987) and the Minas population (72.3%) (Census of India, 2001). This district (Bhasin, 2007) are taken to be the original inhabitants of shares borders with Gujarat and Madhya Pradesh states Rajasthan, constituting 39.3% and 50%, respectively, of the (Table 1). total tribal population of the state, whereas the Saharia is the only primitive tribe of Rajasthan, constituting 1% of the total Laboratory Analysis tribal population, and bears a resemblance to the Hindu caste populations of the area. Rajputs are thought to have originat- DNA was extracted following the salting-out protocol ed in Rajasthan only, but, according to James Tod (1832), (Miller et al., 1988). Seven human-specific Alu Indel (PV92, Rajputs have a Scythian origin—a stock which moved out of FXIIIB, D1, CD4, APO, ACE, mtNUC) and five restriction Caucasus towards the Indus Valley. The Garasias constitute fragment length polymorphism (RFLP) (NAT, LPL, PSCR, about 2.7% of the total tribal population and are thought to T2, ALB) markers were analyzed from the six population be descendants of Rajputs by Bhil women (Kapoor, 2002). samples. Amplification and analysis of DNA samples was The Damaria constitute 0.62% of the tribal population of the carried out using polymerase chain reaction (PCR), followed state and are taken to be a branch of the Bhils (Mehta, 2004). by restriction digestion as required. The primers and proto- In view of the diverse ethnic background of the popula- cols used for typing Alu Indel and mtNUC markers were as tions of Rajasthan vis-à-vis the numerous waves of migra- given by Stoneking et al. (1997), Majumder et al. (1999), tion in India, the main objective of the present paper is to and Watkins et al. (2001). The details of RFLPs studied are gain some insight into the peopling of India and to assess the given by the Allele Frequency Database (http://alfred.med. inter- and intra-population variation as well as the affinities yale.edu/alfred/index.asp). The PCR product for Alu Indel of the population groups of Rajasthan. markers and the digested PCR product for RFLP markers were run on a 2% agarose gel stained with EtBr (ethidium bromide) and then visualized in UV light for genotyping.

Figure 1. Geographical location of the region (Banswara district) of sample collection. Vol. 119, 2011 GENOMIC STUDY AMONG RAJASTHAN POPULATIONS 261

Table 1. Anthropological information of the study populations Population Number Location of sampling Anthropological information Bhil 47 Banswara district Tribe, Indo-European (Bhili), numerically large, wide geographical distribution, among the earliest inhabitants of Rajasthan, trace their origin from Dravidian word for bow, ‘bilu,’ which is a characteristic weapon of Bhils Mina 31 Banswara district Largest tribal group of Rajasthan, Indo-European, among the earliest inhabitants of Rajasthan, claim to originate from Matsya Avatr. Saharia 30 Banswara district Tribe, Indo-European (Harauti), literal meaning—companion of tiger; clan names according to places of origin Garasia 53 Banswara district Tribe, thought to be descendents of Rajputs by Bhil women, Indo-European (dialect akin to Bhili) Damaria 26 Banswara district Tribe, Indo-European (Wagdi), taken to be a branch of the Bhils, migrated from Gujarat Rajputs 34 Banswara district Upper caste, Indo-European (Hindi), name derived from Sanskrit word ‘rajputra’ meaning son of a ruler

Table 2. Allele frequencies for 12 autosomal markers in the six Statistical Analysis populations of Rajasthan Allele frequencies and heterozygosities at individual loci Populations as well as overall heterozygosity were calculated by using Marker the software POPGENE version 1.32 (Yeh and Yang, 1999). Bhil Rajput Saharia Mina Garasia Damaria Hardy–Weinberg equilibrium was tested using the chi Pv92 (+) 0.422 0.405 0.433 0.523 0.403 0.296 square (χ2) goodness of fit. Bonferroni’s correction for FXIIIB (+) 0.715 0.709 0.629 0.583 0.519 0.421 multiple comparisons was also applied. In order to evaluate D1 (+) 0.234 0.336 0.312 0.326 0.461 0.310 genetic differentiation among the studied groups, the locus- APO (+) 0.880 0.894 0.800 0.629 0.923 0.948 wise measure of genetic diversity (Nei, 1973) was calculated ACE (+) 0.510 0.613 0.578 0.416 0.730 0.544 using the software DISPAN (Ota, 1993). The neighbour CD4 (−) 0.358 0.038 0.709 0.017 0.080 0.232 joining tree was constructed based on the DA matrix (Saitou mtNUC (+) 0.538 0.539 0.413 0.407 0.500 0.653 and Nei, 1987). In order to determine the population cluster- NAT (+) 0.659 0.613 0.703 0.466 0.423 0.532 ing of the populations studied, principal coordinate analysis T2 (+) 0.619 0.650 0.560 0.466 0.423 0.514 was carried out using available data on world populations LPL (+) 0.563 0.622 0.485 0.450 0.538 0.602 (Stoneking et al., 1997; Majumder et al., 1999; Mukherjee et PSCR (+) 0.475 0.424 0.469 0.285 0.437 0.450 al., 2000; Nasidze et al., 2001; Antunez-de-Mayolo et al., ALB (+) 0.639 0.556 0.548 0.464 0.615 0.431 2002; Vishwanathan et al., 2003; Comas et al., 2004; +=insertion; −=deletion. Saraswathy et al., 2008). Table 3. Gene diversity analysis for individual loci and all loci Results considered jointly

All the loci were found to be polymorphic in all popula- Marker GST HT HS tions. Allele frequencies are summarized in Table 2. In most PV92 0.018 0.485 0.476 of the markers the maximum attainable value (0.5) of het- F13B 0.044 0.044 0.459 erozygosity for a biallelic marker is actually attained. The D1 0.020 0.442 0.433 average heterozygosity ranges from 0.21 to 0.5. After apply- APO 0.087 0.260 0.237 ing Bonferronin’s correction, significant departure from ACE 0.037 0.491 0.472 Hardy–Weinberg equilibrium was observed for the Minas CD4 0.321 0.364 0.247 with respect to three markers (D1, CD4, and PSCR), where- MtNUC 0.028 0.499 0.485 as the Damaria deviated for two markers (D1 and NAT) and NAT 0.041 0.491 0.470 the Bhils (CD4), Saharia (ACE), Garasia (NAT) and Rajputs T2 0.026 0.496 0.483 (D1) for one marker each. In terms of the markers, maxi- LPL 0.015 0.496 0.488 mum deviation was observed for the D1 marker among the PSCR 0.016 0.488 0.480 Mina, Damaria and Rajputs; for the CD4 marker among the ALB 0.022 0.496 0.485 Bhil and Mina; for the NAT marker among the Garasia and All loci 0.049 0.457 0.435 Damaria; for the ACE marker among theSaharia; and for the PSCR marker among the Mina. The GST value (Table 3) calculated for each locus to deter- the total genomic diversity is attributable to inter-population mine the degree of gene differentiation between the popula- variation. The total genomic diversity (HT) among the sub- tions varies between 1.5% at the LPL locus to 32.1% at the populations is quite high, ranging from 0.260 (APO) to CD4 locus. When all loci are considered together, 4.9% of 0.499 (MtNUC). The HT value over all loci is 0.457, whereas 262 R. DADA ET AL. ANTHROPOLOGICAL SCIENCE

Figure 2. Neighbour joining tree depicting genomic affinities among six ethnic populations of Rajasthan based on six Alu markers.

the HS values, which determine the genomic diversity be- tween individuals within populations, ranges from 0.237 (APO) to 0.488 (LPL). The HS value over all loci is 0.435. The Rajput and Mina make one cluster, while the Garasia Figure 3. Neighbour joining tree depicting genomic affinities and Bhil are separated in the neighbour joining tree among six ethnic populations of Rajasthan and tribes of South India (Figure 2). based on six Alu markers.

Discussion Damaria cluster with tribals from the Nilgiri Hills—it has The allele frequency distribution pattern of the pop- been reported previously that Nilgiri Hills tribals have a ulations studied in this paper is quite similar to those ob- diverse genetic origin (Saraswathy et al., 2008); and the served in other Indian populations (Majumder et al., 1999; Damaria too trace their ancestry to two different sources Mukherjee et al., 2000; Veerraju et al., 2001; Vishwanathan (Bhils and Rajputs), which perhaps justifies their grouping et al., 2003; Saraswathy et al., 2008). The relatively higher with Nilgiri Hill tribals. The Rajputs and Minas are grouped heterozygosity (0.21–0.5) and GST (4.9%) values reflect the together, whereas as satellite tribes of the Gonds, the Thotis genetic heterogeneity in the studied populations. Earlier and Kolams form a cluster. The Garasias and Damaria form studies have reported GST values ranging from as low as independent branches, whereas the Bhils are grouped with 2.5% (Mukherjee et al., 2000) among Madhya Pradesh pop- south Indian tribes—this latter finding may well be ex- ulations to as high as 8.3% among Southern Indian popula- plained by the influence of proto-Dravidian culture on the tions (Vishwanathan et al., 2003). Bhils, going back to Indus Valley civilization culture (the It is accepted that, after Africa, the history of India has Bhils derive their name from the Dravidian word ‘Bilu’). played a key role in human evolution. Further admixture fol- Thus, the present study suggests that the population groups lowed by geographical, cultural, linguistic isolation has led of Rajasthan are genetically diverse and also reflect some to relatively homogeneous gene pools in the Indian context. Dravidian genetic background. Rajasthan also displays huge genetic diversity attributable to The principal coordinate analysis (PCO) scatter plot the region’s biological, social, and geographic heterogene- (Figure 4) drawn using the available data on six Alu markers ity. In archaeologically terms, too, Rajasthan is very rich, from the various tribal and non-tribal groups along with having Paleolithic (Didwana), Mesolithic (Bagor), Neo- European populations reveals a very scattered distribution of Chalcolithic (Ahar, Balathal, and Gilund) and even Indus Indian populations, suggesting diversified sources of gene Valley civilization sites (Kalibangan). flow into the existing gene pools. All the studied six popula- Comparison of the six population groups of Rajasthan tions, including both tribal and caste populations, are rela- using a neighbour joining tree shows that the Rajputs and tively close to one another (Figure 4). Almost all the tribal Minas form a group (Figure 2). Ethnically, it has also been population group of south India are placed far apart from reported that the Minas share several clan names with the one another, which is consistent with a previous report on Rajputs, whereas the Damaria, Saharias, Bhils, and Garasias south Indian populations (Guha, 1944; Sastri, 1955; Cavalli- all form separate branches which may be attributed to their Sforza et al., 1994; Cordaux et al., 2003; Saraswathy et al., diverse origins. 2008), indicating founder effects with relative isolation In order to assess the ethnolinguistic affiliations of the among the south Indian tribal population groups due to Rajasthan populations with other Indian populations, and endogamy. However, genetic influx is seen among the also to understand the influence of proto-Dravidian genes on Andhra Pradesh Brahmin and Nayakpod population groups the populations of Rajasthan, a separate neighbour joining with the exception of the Badaga tribe. All the European tree was constructed using similar data sets for south Indian populations are relatively clustered together in the PCO plot populations for six Alu InDel markers (Figure 3). The (Figure 4) and their genetic continuity is seen with the Badaga, Kurumba, Irula, and Kota are the important tribal studied populations (Damaria and Mina) and other Indian groups of the Tamil Nadu, speaking Dravidian languages. population, which coincides with archaeological and his- The Totis, Kolams (primitive tribe), and Nayakpods (tribe) torical findings. The Indus Valley civilization was spread come from Andhra Pradesh and also speak Dravidian over Pakistan and western India, parts of Afghanistan and languages. In this analysis, the Rajputs cluster with the Turkmenistan, and was linked to a huge area including por- Minas, whereas the Bhils cluster with the Nayakpods; all the tions of Afghanistan, coastal Persia, north and central India, other populations considered were found to be scattered with and Mesopotamia by trade links that followed the major other south Indian population groups. It can be seen that the rivers, which might have attracted people from these places Vol. 119, 2011 GENOMIC STUDY AMONG RAJASTHAN POPULATIONS 263

Figure 4. PCO scatter plot analysis of 38 populations using five common autosomal markers. to explore India, entering India probably through present- of Indian populations and diversified sources of genetic con- day Rajasthan and Gujarat. Also, from the fourth century BC tribution followed by distinctive isolation through endoga- onwards for 2000 years, India, particularly in the north, was mous forms of marriage among the various geographical subjected to waves of penetration: history testifies that population groups of India. Greeks, Parthans, Sakas (Scythians), and Pahalvas, includ- ing the Kushans, were the first to arrive, after the Indo- Aryan civilization had entered its settled course (Bhasin, Conclusion 2006). The impact of these Indo-European speakers can be The present study reveals a high genetic heterogeneity of accounted from the fact that even the tribal people of Rajas- the Rajasthan populations with respect to 12 autosomal loci. than speak dialects belonging to the Indo-European family, By and large, the caste and tribal groups have a close genetic which is unique as this linguistic group mainly encompasses relation. The major inference from the populations studied in caste groups. However, Sir John Marshall (1979) was the this paper is the existence of an inflow of Eurasian genes first to suggest that the language of the Indus civilization into already existing gene pools, i.e. probably proto- was Dravidian, and most scholars have taken the ‘Dravidian Australoid, which was reported to have a Dravidian linguis- hypothesis’ seriously. Even Parpola (1994), after much care- tic lineage. ful and detailed sifting of the evidence, opines “that the Harappan language is most likely to have belonged to the Dravidian family.” Hence, it may be said that the Indus Acknowledgments Valley civilization culture is proto-Dravidian. The early in- We are grateful to the Department of Biotechnology, fluence of Dravidian culture in Rajasthan may be seen from Government of India, for providing the financial support to the fact that the earliest inhabitants of Rajasthan, the Bhils, conduct the present study. We are also grateful to the people trace their name from the Dravidian word ‘Bilu.’ Intra- of Rajasthan for providing the blood samples. We also ac- population variation, reflected by high heterozygosity val- knowledge Prof. P.P. Majumder for providing support and ues, is found to be high in Indian populations including both help in standardization and analyzing the laboratory works. caste and tribal populations from north and south India. However, the inter-population difference is found to be less in the populations studied in this paper as they cluster to- References gether in the PCO plot (Figure 4). In contrast, in south Indi- Antunez-de-Mayolo G., Antunez-de-Mayolo A., Antunez-de- an populations the inter-population variation is higher as Mayolo P., Papiha S.S., Hammer M., Yunis J.J., Yunis E.J., reflected by their scattered distribution in the PCO plot Damodaran C., Martinez-de-Pancorbo M., Luis Caeiro J.L., (Figure 4). It is interesting to note here that almost all the Puzyrev V.P., and Herrera R.J. (2002) Phylogenetics of worldwide human populations as determined by polymorphic studied populations cluster with the north Indian non-tribal Alu insertions. Electrophoresis, 23: 3346–3356. (Uttar Pradesh Muslim, Brahmin and Rajput) and south Bhasin M.K. (2006) Genetics of castes and tribes of India: Indian Indian tribal (Nayakpods) population, indicating genetic population milieu. International Journal of Human Genetics, closeness. This probably indicates ancestral commonalities 6: 233–274. 264 R. DADA ET AL. ANTHROPOLOGICAL SCIENCE

Bhasin V. (2007) Health status of tribals of Rajasthan. Ethno- Nasidze I., Risch G.M., Robichaux M., Sherry S.T., Batzer M.A., Medicine, 1: 91–125. and Stoneking M. (2001) Alu insertion polymorphisms and Cann R.L. (2001) Genetic clues to dispersal of human populations: the genetic structure of human populations from the Cauca- retracing the past from the present. Science, 291: 1742–1748. sus. European Journal of Human Genetics, 9: 267–272. Cavalli-Sforza L.L., Menozzi P., and Piazza A. (1994) The History Nei M. (1973) Analysis of gene diversity in sub-divided popula- and Geography of Human Genes. Princeton University Press, tions. Proceedings of the National Academy of Science USA, Princeton, NJ. 70: 3321–3323. Census of India (2001) Register General and Census Commis- Ota T. (1993) Dispan: Genetic Distance and Phylogenetic Analy- sioner, Government of India, New Delhi. sis. Institute of Molecular Evolutionary Genetics, Pennsylva- Comas D., Schmid H., Braeuer S., Flaiz C., Busquets A., Calafell nia State University, University Park, PA. F., Bertranpetit J., Scheil H.G., Huckenbeck W., Efremovska Parpola A. (1994) Deciphering the Indus Script. Cambridge Uni- L., and Schmidt H. (2004) Alu insertion polymorphisms in versity Press, Cambridge. the Balkans and the origins of the Aromuns. Annals of Saitou N. and Nei M. (1987) The neighbor-joining method: a new Human Genetics, 68: 120–127. method for reconstructing phylogenetic trees. Molecular Cordaux R., Saha N., Bentley G., Aunger R., Sirajuddin S.M., and Biology and Evolution, 4: 406–425. Stoneking M. (2003) Mitochondrial DNA analysis reveals Saraswathy K.N., Sachdeva M.P., Mukhopadhyay R., Shukla D., diverse histories of tribal populations from India. European Devi N.K., Rawat S., Rao A.P., and Kalla A.K. (2008) Diver- Journal of Human Genetics, 3: 253–264. sified genomic contribution among south Indian popula- Danielou A. (2003) A Brief History of India. Inner Traditions tions—a study on four endogamous groups of Andhra International, Rochester, VT. Pradesh. Annals of Human Biology, 35: 499–508. Fairservis W.A. (1992) The Harappan Civilization and its Writing: Sastri N.K.A. (1955) A History of South India. Oxford University A Model for the Decipherment of the Indus Script. Oxford Press, New York. and IBH, New Delhi. Singh K.S. (2002) People of India: Introduction National Series. Guha B.S. (1944) Racial Elements of India. Oxford University Anthropological Survey of India. Oxford University Press, Press, London. Delhi. Heras H. (1953) Studies in Proto-Indo-Mediterranean Culture, Stoneking M., Fontius J.J., Cliffotr S.L., Soodyal H., Arcot S.S., Vol. 1. Indian History of the Indian Historical Research Insti- Saha N., Jenkins T., Tahir M.A, Deininger P.L., and Batzer tute, Bombay. M.A. (1997) Alu insertion polymorphisms and human evolu- Kapoor S. (2002) The Indian Encyclopedia, Vol. 19. Cosmo Publi- tion: evidence for a larger population size in Africa. Genome cations, New Delhi. Research, 7: 1061–1071. Knorozov Y.V., Al Bedil M.F., and Volchok B.Y. (1981) Proto- Thapar R. (1966) A History of India, Vol. 1. Penguin, Harmonds- Indica: 1979, Report on the Investigation of the Proto-Indian worth. Texts. Nauka Publishing House, Moscow. Tod J. (1832) Annals and Antiquities of Rajasthan, Vol. II. Oxford Majumder P.P., Roy B., Banerjee S., Chakroborty M., Dey B., University Press, London. Mukherjee N., Roy M., Thakurta P.G., and Sil S.K. (1999) Veerraju P., Rao T.V., Lakshmi N., Reshmi S., Dey B., and Human-specific insertion/deletion polymorphism in India Majumder P.P. (2001) Insertion/deletion DNA polymor- populations and their possible evolutionary implications. phisms in two south Indian tribal populations. International European Journal of Human Genetics, 7: 435–446. Journal of Human Genetics, 1: 129–132. Mann K. (1987) Tribal women in a changing society. Mittal Publi- Vishwanathan H., Deepa E., Usharani M.V., and Majumder P.P. cations, Delhi. (2003) Insertion/deletion polymorphisms in tribal populations Marshall J. (1979) First light on the longer forgotten civilization: of southern India and their possible evolutionary implica- the Illustrated London News (1924). In: Possehl L. (ed.), tions. Human Biology, 75: 873–887. Ancient Cities of the Indus, New Delhi, pp. 105–107. Watkins W.S., Ricker C.E., Bamshad M.J., Carroll M.L., Nguyen Mehta P.C. (2004) Ethnographic Atlas of Indian Tribes. Discovery S.V., Batzer M.A., Harpending H.C., Rogers A.R., and Jorde Publishing House, New Delhi. L.B. (2001) Patterns of ancestral human diversity: an analysis Miller S.A., Dykes D.D., and Polesky H.F. (1988) A simple salting of Alu-insertion and restriction-site polymorphisms. Ameri- out procedure for extracting DNA from human nucleated can Journal of Human Genetics, 68: 738–752. cells. Nucleic Acids Research, 16: 215. Yeh F.C. and Yang R.C. (1999) A joint project development, Mukherjee N., Mitra M., Chakraborty M., and Majumder P.P. POPGENE 1.32. University of Alberta, and Tim Boyle, (2000) Congruence of genomic and ethnolinguistic affinities Centre for International Forestry Research. http://alfred.med. among five tribal populations of Madhya Pradesh. Journal of yale.edu/alfred/index.asp Genetics, 79: 41–46.