letters to nature 22. Begun, D& Walker, A in The Narickotome Homo erectus Stedetom(eds Walker, A Leakey, R) researchers in many field culture is thus of great interest to 21. Riscutia, C in Paleoanthropology, Morphology and Paleoecology (ed. Tuttle, R. H)373-375(Mouton, expansion process of H According to the historical records, the Hans were descended R. LHuman paleontological evidence relevant to language behavior. Hum. Neurobiol 2, from the ancient Huaxia tribes of northern China, and the Han culture(that is, the language and its associated cultures)expanded 24. Holloway, R. L The Indonesian Homo erectus brain endocasts revisited. Am. /. Phys Anthropol 55, into southern China-the region originally inhabited by the 25 Sartono, S&Tyler, D E A New Homo erectus Skll from Sangiran, Arvar: am Annoumcement 1-4 southern natives, including those speaking Daic, Austro-Asiatic (Intern. Conf. Human Paleoecol LIPPI, Jakarta, 1993). nd Hmong-Mien languages-in the past two millennia". Studies 26. Rak Y.& Rensburg. B. Kebara 2 neanderthal pelvis: first look at a complete inlet. Am. J. Phys. on classical genetic markers and microsatellites show that the a.73,227-231(1987) [ cortical bone and dental Han people, like East Asians, are divided into two genetically namel by computed tomography: applications and problems. Am I. Phys. Anthropol. 91, 469-484 differentiated groups, northern Han and southern Han, separated 28. Hohne, K.H. et al. A new representation of knowledge concerning human anatomy and function. approximately by the Yangtze river. Differences between thes Natere Med. 1, 506-511(19 groups in terms of dialect and customs have also been noted o npanzee, with special reference to growth of brain, eruption of Such observations seem to support a mechanism involving rimarily cultural diffusion and assimilation(the cultural diffusion 30. Schultz, A H in Contributions to Embryology no 170(ed, Schultz, A.H. )1-63(Camegie Institution of model)in Han expansion towards the south. However, the sub Washington Publication no 518, Washington DC, 1940). stantial sharing of Y-chromosome and mitochondrial lineages SupplementaryInformationaccompaniesthepaperonwww.nature.com/nature between the two groups.and the historical records describing the expansion of Han people' contradict the cultural diffusion Acknowledgements We are grateful to the following individuals for their assistance in accessing model hypothesis of Han expansion. In this study, we aim to collections and their advice and comments during the preparation of this paper: S. Anton, examine the alternative hypothesis; that is, that substantial popu L P Bocquet-Appel, J. Braga, G Brauer, M. Braun, P Darlu, M. Haas, M. von Harling, C. He er,S Paabo, F. Renoult, M. Richards, Ph. Rightmire, F. Schrenk, ation movements occurred during the expansion of Han culture HSick,ESpoor,TStriano, ITreil, w.Wendelen and V Zeitoun. This research was supported by ( the demic diffusion model) grants from CNRS and by the Max Planck Society. To test this hypothesis, we compared the genetic profiles of southern Hans with their two parental population groups: northern Competing interests statement The authors deare that they have no competing financial Hans and southern natives, which include the samples of Daic, Hmong-Mien and Austro-Asiatic speaking populations currently Correspondence and requests for materials should be addressed to J.J.H. (hublineeva.mpg. de residing in China, and in some cases its neighbour Ing countries. Genetic variation in both the non-recombining region of the Y chromosome(NRY) and mitochondrial DNA (mtDNA)S- were surveyed in 28 Han populations from most of the China(see Fig. I and Supplementary Table 1 for details On the paternal side, southern Hans and northern Hans share similar frequencies of Y-chromosome haplogroups(Supplementary Genetic evidence supports demic Table 2), which are characterized by two haplogroups M122-C mutations(O3-M122 and O3e-M134)that are prevalent diffusion of han culture in almost all Han populations studied(mean and range: 53.8%6 -71%0; 54.2%, 35-74%6, for northern and southern Hans, respect ively). Haplogroups carrying MI19-C(O1* and Olb)and/or Bo Wen, Hui ui, Daru Lu, Xiufeng Song, Feng Zhang, Yungang He, M95-T(O2a* and O2a1)(following the nomenclature of the Y Jianzhong Jin', Wei Huang, Ranjan Deka, Bing Su, Jingze Tan Chromosome Consortium)which are prevalent in southern Feng Li, Yang Gao, Xianyun Mao, Liang Zhang,, Ji Qia natives, are more frequent in southern Hans(19%,3-42%)than Ranajit Chakraborty &LiJin in northern Hans (5%, 1-10%). In addition, haplogroups O1b-M110, O2al-M88 and O3d-M7, which are prevalent in tg and Center for Anthropological southern natives " 7, were only observed in some southern Hans Studies, School of life Sciences and Morgan-Tan International Center for Li Sciences, Fudan University, Shanghai 200433, China 4%on average), but not in northern Hans. Therefore, the contri- bution of southern natives in southern hans is limited if we assume Chinese National Human Genome Center, Shanghai 201203, China Center for Genome Information, Department of Environmental Heai that the frequency distribution of Y lineages in southern natives niversity of Cincinnati, Cincinnati, Ohio 45267, USA presents that before the expansion of Han culture that started "Key Laboratory of Cellular and Molecular Evolution, Kunming Institute of 000 yr ago. The results of analysis of molecular variance Zoology, the Chinese Academy of Sciences, Kunming 650223, China (AMOVA) further indicate that northern Hans and southern Hans are not significantly different in their Y haplogroups The spread of culture and language in human populations is (FST=0.006, P>0.05), demonstrating that southern Hans bear explained by two alternative models: the demic diffusion model, a high resemblance to northern Hans in their male lineages which involves mass movement of people; and the cultural On the maternal side, however, the mtDNA haplogroup distri- diffusion model, which refers to cultural impact between popu- bution showed substantial differentiation between northern Hans lations and involves limited genetic exchange between them. The and southern Hans(Supplementary Table 3). The overall frequen mechanism of the peopling of Europe has long been debated, a cies of the northern East Asian-dominating haplogroups(A, C,D, key issue being whether the diffusion of agriculture and language G, M8a, Yand Z)are much higher in northern Hans(55%, 49-64%) from the Near East was concomitant with a large movement of than are those in southern Hans(36%, 19-52%) In contrast, the farmers. Here we show, by systematically analysing Y-chromo- frequency of the haplogroups that are dominant lineages(B, F, R9a, some and mitochondrial DNA variation in Han populations, that R9b and N9a)in southern natives 4Is is much higher in southern the pattern of the southward expansion of Han culture is (55%, 36-72%)than it is in northern Hans(33%, 18-42%) consistent with the demic diffusion model, and that males played Northern and southern Hans are significantly different in their larger role than females in this expansion. The Han people, who mtDNA lineages(FsT=0.006, P< 10 ). Although the Fsr values all share the same culture and language, exceed 1.16 billion(2000 between northern and southern Hans are similar for mtDNA census), and are by far the largest ethnic group in the world. The and the Y chromosome, Fsr accounts for 56% of the total among e2004NaturePublishingGroupNatuReIvoL431116SePtEmbEr2004www.nature.com/nature© 2004 NaturePublishingGroup 21. Riscutia, C. in Paleoanthropology, Morphology and Paleoecology (ed. Tuttle, R. H.) 373–375 (Mouton, The Hague, 1975). 22. Begun, D. & Walker, A. in The Nariokotome Homo erectus Skeleton (eds Walker, A. & Leakey, R.) 326–358 (Springer, Berlin, 1993). 23. Holloway, R. L. Human paleontological evidence relevant to language behavior. Hum. Neurobiol. 2, 105–114 (1983). 24. Holloway, R. L. The Indonesian Homo erectus brain endocasts revisited. Am. J. Phys. Anthropol. 55, 503–521 (1981). 25. Sartono, S. & Tyler, D. E. A New Homo erectus Skull from Sangiran, Java: an Announcement 1–4 (Intern. Conf. Human Paleoecol., LIPPI, Jakarta, 1993). 26. Rak, Y. & Arensburg, B. Kebara 2 neanderthal pelvis: first look at a complete inlet. Am. J. Phys. Anthropol. 73, 227–231 (1987). 27. Spoor, C. F., Zonneveld, F. W. & Macho, G. A. Linear measurements of cortical bone and dental enamel by computed tomography: applications and problems. Am. J. Phys. Anthropol. 91, 469–484 (1993). 28. Ho¨hne, K. H. et al. A new representation of knowledge concerning human anatomy and function. Nature Med. 1, 506–511 (1995). 29. Zuckerman, S. Age-changes in the chimpanzee, with special reference to growth of brain, eruption of teeth, and estimation of age; with a note on the Taung ape. Proc. Zool. Soc. Lond. 1, 1–42 (1928). 30. Schultz, A. H. in Contributions to Embryology no 170 (ed. Schultz, A. H.) 1–63 (Carnegie Institution of Washington Publication no 518, Washington DC, 1940). Supplementary Information accompanies the paper on www.nature.com/nature. Acknowledgements We are grateful to the following individuals for their assistance in accessing collections and their advice and comments during the preparation of this paper: S. Anton, J. P. Bocquet-Appel, J. Braga, G. Bra¨uer, M. Braun, P. Darlu, M. Haas, M. von Harling, C. Hemm, J.-L. Kahn, C. Lefe`vre, W. van Neer, S. Pa¨a¨bo, F. Renoult, M. Richards, Ph. Rightmire, F. Schrenk, H. Sick, F. Spoor, T. Striano, J. Treil, W. Wendelen and V. Zeitoun. This research was supported by grants from CNRS and by the Max Planck Society. Competing interests statement The authors declare that they have no competing financial interests. Correspondence and requests for materials should be addressed to J.J.H. (hublin@eva.mpg.de). .............................................................. Genetic evidence supports demic diffusion of Han culture Bo Wen1,2, Hui Li1 , Daru Lu1 , Xiufeng Song1 , Feng Zhang1 , Yungang He1 , Feng Li1 , Yang Gao1 , Xianyun Mao1 , Liang Zhang1 , Ji Qian1 , Jingze Tan1 , Jianzhong Jin1 , Wei Huang2 , Ranjan Deka3 , Bing Su1,3,4, Ranajit Chakraborty3 & Li Jin1,3 1 State Key Laboratory of Genetic Engineering and Center for Anthropological Studies, School of Life Sciences and Morgan-Tan International Center for Life Sciences, Fudan University, Shanghai 200433, China 2 Chinese National Human Genome Center, Shanghai 201203, China 3 Center for Genome Information, Department of Environmental Health, University of Cincinnati, Cincinnati, Ohio 45267, USA 4 Key Laboratory of Cellular and Molecular Evolution, Kunming Institute of Zoology, the Chinese Academy of Sciences, Kunming 650223, China ............................................................................................................................................................................. The spread of culture and language in human populations is explained by two alternative models: the demic diffusion model, which involves mass movement of people; and the cultural diffusion model, which refers to cultural impact between popu￾lations and involves limited genetic exchange between them1 . The mechanism of the peopling of Europe has long been debated, a key issue being whether the diffusion of agriculture and language from the Near East was concomitant with a large movement of farmers1–3. Here we show, by systematically analysing Y-chromo￾some and mitochondrial DNA variation in Han populations, that the pattern of the southward expansion of Han culture is consistent with the demic diffusion model, and that males played a larger role than females in this expansion. The Han people, who all share the same culture and language, exceed 1.16 billion (2000 census), and are by far the largest ethnic group in the world. The expansion process of Han culture is thus of great interest to researchers in many fields. According to the historical records, the Hans were descended from the ancient Huaxia tribes of northern China, and the Han culture (that is, the language and its associated cultures) expanded into southern China—the region originally inhabited by the southern natives, including those speaking Daic, Austro-Asiatic and Hmong-Mien languages—in the past two millennia4,5. Studies on classical genetic markers and microsatellites show that the Han people, like East Asians, are divided into two genetically differentiated groups, northern Han and southern Han6,8, separated approximately by the Yangtze river9 . Differences between these groups in terms of dialect and customs have also been noted10. Such observations seem to support a mechanism involving primarily cultural diffusion and assimilation (the cultural diffusion model) in Han expansion towards the south. However, the sub￾stantial sharing of Y-chromosome and mitochondrial lineages between the two groups11,12 and the historical records describing the expansion of Han people5 contradict the cultural diffusion model hypothesis of Han expansion. In this study, we aim to examine the alternative hypothesis; that is, that substantial popu￾lation movements occurred during the expansion of Han culture (the demic diffusion model). To test this hypothesis, we compared the genetic profiles of southern Hans with their two parental population groups: northern Hans and southern natives, which include the samples of Daic, Hmong-Mien and Austro-Asiatic speaking populations currently residing in China, and in some cases its neighbouring countries. Genetic variation in both the non-recombining region of the Y chromosome (NRY) and mitochondrial DNA (mtDNA)13–16 were surveyed in 28 Han populations from most of the provinces in China (see Fig. 1 and Supplementary Table 1 for details). On the paternal side, southern Hans and northern Hans share similar frequencies of Y-chromosome haplogroups (Supplementary Table 2), which are characterized by two haplogroups carrying the M122-C mutations (O3-M122 and O3e-M134) that are prevalent in almost all Han populations studied (mean and range: 53.8%, 37–71%; 54.2%, 35–74%, for northern and southern Hans, respect￾ively). Haplogroups carrying M119-C (O1* and O1b) and/or M95-T (O2a* and O2a1) (following the nomenclature of the Y Chromosome Consortium) which are prevalent in southern natives, are more frequent in southern Hans (19%, 3–42%) than in northern Hans (5%, 1–10%). In addition, haplogroups O1b-M110, O2a1-M88 and O3d-M7, which are prevalent in southern natives17, were only observed in some southern Hans (4% on average), but not in northern Hans. Therefore, the contri￾bution of southern natives in southern Hans is limited, if we assume that the frequency distribution of Y lineages in southern natives represents that before the expansion of Han culture that started 2,000 yr ago5 . The results of analysis of molecular variance (AMOVA) further indicate that northern Hans and southern Hans are not significantly different in their Y haplogroups (FST ¼ 0.006, P . 0.05), demonstrating that southern Hans bear a high resemblance to northern Hans in their male lineages. On the maternal side, however, the mtDNA haplogroup distri￾bution showed substantial differentiation between northern Hans and southern Hans (Supplementary Table 3). The overall frequen￾cies of the northern East Asian-dominating haplogroups (A, C, D, G, M8a, Yand Z) are much higher in northern Hans (55%, 49–64%) than are those in southern Hans (36%, 19–52%). In contrast, the frequency of the haplogroups that are dominant lineages (B, F, R9a, R9b and N9a) in southern natives12,14,18 is much higher in southern (55%, 36–72%) than it is in northern Hans (33%, 18–42%). Northern and southern Hans are significantly different in their mtDNA lineages (FST ¼ 0.006, P , 1025 ). Although the FST values between northern and southern Hans are similar for mtDNA and the Y chromosome, FST accounts for 56% of the total among￾letters to nature 302 NATURE | VOL 431 | 16 SEPTEMBER 2004 | www.nature.com/nature