REP。RTs PeeDee Belemnite(VPDB)standard and for normalized properly, each run also versus NBS-19 for 8C= 1.95% versus VPDB and 0.0%o versus air nitrogen. The first anaylsis(sealed- quots of a air nitrogen. Precision in organic st of±0.1% o of its correct as measured by including repeats of A curacy sample of known isotopic ratio in each run, was pectrometry(EA/CFIRMS) alue. Each autosampler run contained seven org ±0.10%.Ato Carlo Erba NC2500 interfaced through a Finnigan acid standards, four natural sample standards, and 40 notation, or part CONFLO lI to a Finnigan Delta XL mass spectromet of the sample amount. Samples were prepared VPDB. where 8 [=C/C)sample/(C/ organic standards and homogenous natural samples. he EA/CFIRMS analyses were calibrated as follow iler boats (measuring s m by mm) and aidf. 12.S. Carter Mem ished reut o 2 0 smpling g ersus a pulse of 99.999% pure standard gas injected rernight at 50C, the sample boats were sealed and Kennecott Point. to the mass spectrometer source immediately be neasured as follows: Isotope analyses were per- 14 M J. Orchard, PJ. Forster,Geol.Surv.Can. Pap ormed by EA/CFIRMS, using a Carlo Erba NC2500 90-10.453(1991) Because the isotope ratios obtained were dependent terfaced through a Finnigan CONFLO l to a Finni- by a grant from NSF (H. Lane, program on mixing ratios of carrier gas and dilutant in the an Delta XL mass spectrometer. Sampl dministrator) to P.D. W and by Geological Survey of Canada Project number 870070 organic standard run with the samples under the same conditions. To verify that the corrections were tope ratios calibrated in sealed-tube combustions 22 December 2000, accepted 28 March 2001 African Origin of Modern (C to T mutation, also called RPS4Y)(Fig. 1) (21, 22). Therefore, these three markers can be Humans in East Asia: A Tale of used to test the completeness of the replacement of modern humans of African origin in East Asia. An observation of a male individual not 12,000 Y Chromosomes carrying one of the three polymorphisms would be indicative of a potential ancient origin and Yuehai Ke, *Bing Su,2,1. 3* Xiufeng Song, 'Daru Lu, could possibly lead to the rejection of such Lifeng Chen, Hongyu Li, Chunjian Qi, Sangkot Marzuki completeness. Ranjan Deka, Peter Underhill, Chunjie Xiao, Mark Shriver, Each of the 12, 127 samples typed carried jeff Lell, Douglas Wallace,R Spencer Wells, one of the three polymorphisms (YAP+, Mark Seielstad, 1 Peter Oefner, 6 Dingliang Zhu, 2 Jianzhong Jin, M89T, or M130T)(Table 1). In other words, they all fall into the lineage of M168T that was Wei Huang, 2,13 Ranajit Chakraborty, Zhu Chen, 123Li Jin3,13t originally derived from Africa. Hence, no an- cient non-African y chromosome was found in To test the hypotheses of modern human origin in East Asia, we sampled 12, 127 the extant East Asian populations(P= 5.4 X male individuals from 163 populations and typed for three Y chromosome 10-0 assuming a frequency of 1/1000 of local biallelic markers(YAP, M89, and M130). All the individuals carried a mutation contribution in the extant populations), suggest- at one of the three sites. These three mutations(YAP+, M89T, and M130T) ing an absence of either an independent origin coalesce to another mutation(M168T), which originated in Africa about 35,000 or a 1,000, 000-year shared global evolution. to 89,000 years ago. Therefore, the data do not support even a minimal in situ This result indicates that modern humans of hominid contribution in the origin of anatomically modern humans in East Asia. African origin completely replaced earlier pop- ulations in East asia. TheOut-of-Africa"hypothesis suggests that typed for three Y chromosome biallelic markers atomically modern humans originated in (YAP, M89, and M130)(7, 18)(Table D). ' State Key Laboratory of Genetic Engineering Insti- Africa about 100,000 years ago and then Being a single-locus multiple-site (i.e haplo e of genetics, School of Life sciences. Fudan Uni- spread outward and completely replaced local type) system, the Y chromosome is one of the versity, 220 Handan Road, Shanghai, China 200443, archaic populations outside Africa (1, 2). most powerful molecular tools for tracing hu- and Morgan-Tan International Center for Life Scienc- This proposition has been supported by ge- man evolutionary history(, 9, 19-21). In pre- the Chinese Academy of Sciences, Kunming, Chin netic evidence and archaeological findings vious Y chromosome studies, an extreme geo- Human Genetics center 9). The replacement in Et was sup- graphic structure was revealed in global n, 1200 Herman Pressler E547, Houston, TX 77030 recent ancient DNA analyses, lations in which the oldest clade represen hich ruled out the contribution of Neander- Africans and the younger ones represent some artment of envir thals to modern Europeans (10, 11). Howev- Africans and all non-African populations(21) 5267,Us4 er, it has been argued that the abundant hom- One Y chromosome polymorphism (C to T ford, CA 94305, USA"Department of Biology Yunn inid fossils found in China and other regions mutation) at the M168 locus is shared by all University, Kunming, China. Department of Anthro in East Asia(e.g, Peking man and Java man) non-African populations and was originally de- logical characters but also in spatial and tem- 1062 globally representative male individuals 30322. use sow elcome Trust cente poral distributions(12-16). In this report, we (21). The age of M168 was estimated at 44, 000 Genetics, University of Oxford, UK. Asian human origins using Y chromosome 89,000 years), marking the recent Out-of-Africa Boston, MA 02115. UsA 12Shanghai test the competing hypotheses of modern years (95% confidence interval: 35, 000to P migrations(21). Under the M168T lineage niversity, Shanghai, China. National Human G Center at Shanghai, China. We sampled 12, 127 male individuals from there are three major derived sublineages de- contributed equally to this work. 3 populations across Southeast Asia, Oce- fined by polymorphisms at loci YAP (Alu in- espondence should be addressed. E- ania, East Asia, Siberia, and Central Asia and sertion)(), M89(C to T mutation), and M130 www.sciencemag.orgSciEnceVol29211May2001 1151PeeDee Belemnite (VPDB) standard and for 15N 5 0.0‰ versus air nitrogen. The first anaylsis (sealed￾tube combustion with subsequent measurement on a Finnigan MAT 251 mass spectrometer) has a repro￾ducibility of 6 0.08‰ for organic standards. The second analysis [elemental analyzer–continuous-flow isotope ratio mass spectrometry (EA/CFIRMS)] used a Carlo Erba NC2500 interfaced through a Finnigan CONFLO II to a Finnigan Delta XL mass spectrometer. Reproducibility in this system averages 60.12‰ for organic standards and homogenous natural samples. The EA/CFIRMS analyses were calibrated as follows: For each sample, d15N and d13C were measured versus a pulse of 99.999% pure standard gas injected into the mass spectrometer source immediately be￾fore or after the sample pulse eluted from the EA. Because the isotope ratios obtained were dependent on mixing ratios of carrier gas and dilutant in the CONFLO, the ratios were normalized to a known organic standard run with the samples under the same conditions. To verify that the corrections were normalized properly, each run also contained four aliquots of a natural sample whose ratios were known from sealed-tube combustion. This natural sample had a precision identical to the organic stan￾dard (60.12‰) and the average of four determina￾tions was within a range of 60.1‰ of its correct value. Each autosampler run contained seven organic acid standards, four natural sample standards, and 40 samples and blanks. Blanks typically were less than 1% of the sample amount. Samples were prepared for analysis by grinding, followed by weighing into silver boats (measuring 5 mm by 9 mm) and acidifi- cation with 20 ml of 50% HCl. After air drying overnight at 50°C, the sample boats were sealed and measured as follows: Isotope analyses were per￾formed by EA/CFIRMS, using a Carlo Erba NC2500 interfaced through a Finnigan CONFLO II to a Finni￾gan Delta XL mass spectrometer. Sample isotope ratios were normalized in each run to the values obtained for an organic standard with known iso￾tope ratios calibrated in sealed-tube combustions versus NBS-19 for d13C 5 1.95‰ versus VPDB and for d15N 5 0.0‰ versus air nitrogen. Precision in this system averages 60.12‰ for organic stan￾dards and homogenous natural samples. Accuracy, as measured by including repeats of a natural sample of known isotopic ratio in each run, was 60.10‰. All isotope ratios are expressed in delta notation, or parts per thousand deviation from VPDB, where d13C 5 {[(13C/12C)sample/(13C/ 12C)VPDB] 2 1} 3 1000. 12. E. S. Carter, Mem. Geol. (Lausanne) 11, 175 (1993). 13. E. S. Carter, unpublished results of 2000 sampling at Kennecott Point. 14. M. J. Orchard, P. J. L. Forster, Geol. Surv. Can. Pap. 90-10, 453 (1991). 15. Supported by a grant from NSF (H. Lane, program administrator) to P.D.W. and by Geological Survey of Canada Project number 870070. 22 December 2000; accepted 28 March 2001 African Origin of Modern Humans in East Asia: A Tale of 12,000 Y Chromosomes Yuehai Ke,1 * Bing Su,2,1,3* Xiufeng Song,1 Daru Lu,1 Lifeng Chen,1 Hongyu Li,1 Chunjian Qi,1 Sangkot Marzuki,4 Ranjan Deka,5 Peter Underhill,6 Chunjie Xiao,7 Mark Shriver,8 Jeff Lell,9 Douglas Wallace,9 R Spencer Wells,10 Mark Seielstad,11 Peter Oefner,6 Dingliang Zhu,12 Jianzhong Jin,1 Wei Huang,12,13 Ranajit Chakraborty,3 Zhu Chen,12,13 Li Jin1,3,13 † To test the hypotheses of modern human origin in East Asia, we sampled 12,127 male individuals from 163 populations and typed for three Y chromosome biallelic markers (YAP, M89, and M130). All the individuals carried a mutation at one of the three sites. These three mutations (YAP1, M89T, and M130T) coalesce to another mutation (M168T), which originated in Africa about 35,000 to 89,000 years ago. Therefore, the data do not support even a minimal in situ hominid contribution in the origin of anatomically modern humans in East Asia. The “Out-of-Africa” hypothesis suggests that anatomically modern humans originated in Africa about 100,000 years ago and then spread outward and completely replaced local archaic populations outside Africa (1, 2). This proposition has been supported by ge￾netic evidence and archaeological findings (3–9). The replacement in Europe was sup￾ported by recent ancient DNA analyses, which ruled out the contribution of Neander￾thals to modern Europeans (10, 11). Howev￾er, it has been argued that the abundant hom￾inid fossils found in China and other regions in East Asia (e.g., Peking man and Java man) demonstrate continuity, not only in morpho￾logical characters but also in spatial and tem￾poral distributions (12–16). In this report, we test the competing hypotheses of modern Asian human origins using Y chromosome polymorphisms. We sampled 12,127 male individuals from 163 populations across Southeast Asia, Oce￾ania, East Asia, Siberia, and Central Asia and typed for three Y chromosome biallelic markers (YAP, M89, and M130) (17, 18) (Table 1). Being a single-locus multiple-site (i.e., haplo￾type) system, the Y chromosome is one of the most powerful molecular tools for tracing hu￾man evolutionary history (5, 9, 19–21). In pre￾vious Y chromosome studies, an extreme geo￾graphic structure was revealed in global popu￾lations in which the oldest clade represents Africans and the younger ones represent some Africans and all non-African populations (21). One Y chromosome polymorphism (C to T mutation) at the M168 locus is shared by all non-African populations and was originally de￾rived from Africa on the basis of a study of 1062 globally representative male individuals (21). The age of M168 was estimated at 44,000 years (95% confidence interval: 35,000 to 89,000 years), marking the recent Out-of-Africa migrations (21). Under the M168T lineage, there are three major derived sublineages de￾fined by polymorphisms at loci YAP (Alu in￾sertion) (5), M89 (C to T mutation), and M130 (C to T mutation, also called RPS4Y) (Fig. 1) (21, 22). Therefore, these three markers can be used to test the completeness of the replacement of modern humans of African origin in East Asia. An observation of a male individual not carrying one of the three polymorphisms would be indicative of a potential ancient origin and could possibly lead to the rejection of such completeness. Each of the 12,127 samples typed carried one of the three polymorphisms (YAP1, M89T, or M130T) (Table 1). In other words, they all fall into the lineage of M168T that was originally derived from Africa. Hence, no an￾cient non-African Y chromosome was found in the extant East Asian populations (P 5 5.4 3 1026 assuming a frequency of 1/1000 of local contribution in the extant populations), suggest￾ing an absence of either an independent origin or a 1,000,000-year shared global evolution. This result indicates that modern humans of African origin completely replaced earlier pop￾ulations in East Asia. 1 State Key Laboratory of Genetic Engineering, Insti￾tute of Genetics, School of Life Sciences, Fudan Uni￾versity, 220 Handan Road, Shanghai, China 200443, and Morgan-Tan International Center for Life Scienc￾es, Shanghai, China. 2 Kunming Institute of Zoology, the Chinese Academy of Sciences, Kunming, China. 3 Human Genetics Center, University of Texas–Hous￾ton, 1200 Herman Pressler E547, Houston, TX 77030, USA. 4 Eijkman Institute for Molecular Biology, Jakarta, Indonesia. 5 Department of Environmental Health, University of Cincinnati, Cincinnati, OH 45267, USA. 6 Department of Genetics, Stanford University, Stan￾ford, CA 94305, USA. 7 Department of Biology, Yunnan University, Kunming, China. 8 Department of Anthro￾pology, Pennsylvania State University, University Park, PA 16802, USA. 9 Center for Molecular Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA. 10Wellcome Trust Center for Human Genetics, University of Oxford, UK. 11Program for Population Genetics, Harvard School of Public Health, Boston, MA 02115, USA. 12Shanghai Second Medical University, Shanghai, China. 13National Human Ge￾nome Center at Shanghai, China. *These authors contributed equally to this work. †To whom correspondence should be addressed. E￾mail: ljin@fudan.edu or ljin@sph.uth.tmc.edu R EPORTS www.sciencemag.org SCIENCE VOL 292 11 MAY 2001 1151