The fateful triangle

Not long ago Lazaridis et al. proposed that most present-day Europeans were derived from three distinct ancestral populations: Ancient North Eurasians (ANE), Early European Farmers (EEF) and Western European Hunter-Gatherers (WHG).

However, this is essentially a stop-gap model, which will in all likelihood be replaced by a partly revised and more robust model once someone manages to sequence a genome or two from the Neolithic Near East. That's because EEF is clearly a hybrid component, largely made up of ancient Near Eastern ancestry and something very WHG-like, sometimes in very different proportions depending on the location and archeological context of the EEF genomes being analyzed.

So what will this new model look like, you might ask? Probably like this, where EEF is replaced by an Early Neolithic Farmer (ENF) component from the ancient Near East, or something very similar:

The diagram above is basically a Principal Component Analysis (PCA) based on output from my new West Eurasia K8 test (see here), in which the Near Eastern component is synonymous with ENF.

I'm quite certain that these results are very close to the truth. However, just in case the Near Eastern ancestry proportions are a little bit too high (and we won't know until we see those ancient genomes from the Near East), I've got another version that offers lower bound Near Eastern estimates.

It might be useful to keep in mind that I rotated the plots to fit geography. As a result, Component 1, which packs around 85% of the variance on both plots, appears smaller than Component 2, which only carries around 10% of the variance.

A spreadsheet with West Eurasia K8 results for a wide variety of populations is available here. Please note that there are two sheets, with the second sheet showing the lower bound Near Eastern ancestry proportions.

We'll probably learn of more ancient European meta-populations as many more genomes are sequenced from across Eurasia. Nevertheless, I doubt this will affect the model outlined above. That's because I'm expecting all such meta-populations to be mixtures of ANE, ENF and/or WHG, as well as, in some cases, extra-West Eurasian components.

However, I suspect that West Eurasia will have to be modeled in a different way from Europe, with, amongst other things, the so called Basal Eurasian component replacing ENF. But for this to happen we'll need at least one ancient genome that is in large-part of Basal Eurasian origin. In any case, that's a whole different subject.

See also...

4mix: four-way mixture modeling in R

Gokhem2 + Motala12 =/= present-day Swedes

I've seen quite a few comments on this blog suggesting that most of the Ancient North Eurasian (ANE) admixture found in Northern Europe today might come from Scandinavian hunter-gatherers like Motala12 and Ajvide58. It's probably obvious to most that this is not realistic, because the Scandinavian forager genomes sequenced to date show very high ratios of Western European Hunter-Gatherer (WHG) ancestry (>80%), so basically the math doesn't add up.

Nevertheless, I thought it might be useful to drive the point home using this Principal Component Analysis (PCA) based on my new West Eurasia K8 test. The datasheet is available here. You can view a spreadsheet of the results with extra samples here.

Please note that neither Motala12 nor Gokhem2, a late Neolithic farmer from south Sweden belonging to the Funnelbeaker culture, can pass for present-day Swedes. Moreover, mixing Gokhem2 with Motala12, in any proportions, will not produce a result even vaguely similar to present-day Swedes (ie. the outcome will fall somewhere along the dotted line).

I'd say one of the most obvious ways to get the right result would be to blend the Scandinavian forager and farmer with at least one other sample from somewhere below (ie. geographically speaking, east or southeast) of the Swedish cluster.

It might be possible to come up with a more precise plot location, and thus perhaps geographic origin, for this putative third source of Swedish ancestry by running some complex tests with the PCA datasheet. If anyone wants to have a go at that, and you actually manage to come up with a coherent outcome, then feel free to post your findings in the comments below.

I've decided not to bother, because as far as I can see, the options are infinite. What we really need are more genomes from the Swedish late Neolithic/early Bronze Age (LN/EBA), preferably belonging to one of the local spin-offs of the Corded Ware culture, which is thought to have originated in Eastern Europe, to provide more datapoints and help narrow down the options.

On a related note, I'm catching up on some reading this holiday season, and currently going through this book chapter which discusses the upheavals during the LN/EBA in south Scandinavia as seen through its archeology.

Rune Iversen, Beyond the Neolithic transition - the "de-Neolithisation" of south Scandinavia

See also...

Bell Beaker, Corded Ware, EHG and Yamnaya genomes in the fateful triangle

ANE is the primary cause of west to east genetic differentiation within West Eurasia

Here's a Principal Component Analysis (PCA) and an accompanying biplot based on output from an improved version of my ANE K7 ancestry test. Let's call it the West Eurasia K8. This one gives more accurate estimates of Western European Hunter-Gatherer (WHG) and Near Eastern admixture proportions, thanks to the use of new ancient samples.

When rotated accordingly (like here), the results are basically indistinguishable from those I get with genotype data (for instance, see here and here), which suggests that they're correct and based on ancestry proportions that are close to the truth. The Past3 data sheet used to create the PCA is available here. You can view a spreadsheet of the results with extra samples here.

Clearly, ANE is the main agent causing the west to east differentiation in dimension 2. Note that even a small rise in ANE, say, 4-5%, creates significant distance between samples on the PCA plot.

East and South Eurasian admixture has a similar effect, but must be more considerable to make an impact on a West Eurasian-specific PCA like this (and it does with the obvious Volga-Ural outliers, who come from Chuvashia and Tatarstan).

On the other hand, Near Eastern admixture without ANE creates almost the opposite effect. Note, for instance, that Neolithic genomes Stuttgart and NE1 show much higher levels of Near Eastern ancestry than most Europeans, and yet they're amongst the most western samples on the plot.

This suggests that the Near East, and in particular the Caucasus, experienced a significant rush of ANE admixture after early Neolithic farmers left the region for Europe. Alternatively, Caucasus populations may have carried even higher levels of ANE than they do today, before newcomers from the Near East mixed with them. But either way, a lot of ANE arrived in the Near East at some point.

It also suggests that, overall, the populations that moved west across northern Europe after the Neolithic, and shifted northern European genetic structure to the east, did not carry high ratios of Near Eastern ancestry. Instead, they harbored high ratios of ANE and WHG. What these ratios were exactly I haven't a clue, but ancient DNA should tell us that soon.

Below are the ancestry proportions for the five ancient genomes in this analysis, in chronological order. It's interesting to note (yet again) the rising and falling Near Eastern admixture, from the Mesolithic to Neolithic and then from the Neolithic to Bronze Age, respectively, as well as the steady rise of ANE from the Bronze Age to the Iron Age.

Loschbour (Mesolithic)

ANE 0
South_Eurasian 0
Near_Eastern 0
East_Eurasian 0
WHG 99.5
Oceanian 0.5
Pygmy 0
Sub-Saharan 0

Stuttgart (Neolithic)

ANE 0
South_Eurasian 0
Near_Eastern 72.19
East_Eurasian 0
WHG 27.8
Oceanian 0
Pygmy 0
Sub-Saharan 0

NE1 (Neolithic)

ANE 0
South_Eurasian 0
Near_Eastern 69.82
East_Eurasian 0
WHG 30.17
Oceanian 0
Pygmy 0
Sub-Saharan 0

BR2 (Bronze Age)

ANE 9.62
South_Eurasian 0.08
Near_Eastern 43.96
East_Eurasian 0
WHG 45.44
Oceanian 0.48
Pygmy 0.23
Sub-Saharan 0.19

Hinxton4 (Iron Age)

ANE 15.08
South_Eurasian 0.06
Near_Eastern 35.44
East_Eurasian 0.46
WHG 48.5
Oceanian 0
Pygmy 0
Sub-Saharan 0.46

See also...

The fateful triangle

Bell Beaker, Corded Ware, EHG and Yamnaya genomes in the fateful triangle

Milk consumption in late Neolithic/Bronze Age West Eurasia

The map below is based on data from Warinner et al. 2014. It shows the consumption of milk, or lack of, among Late Neolithic/Bronze Age (LN/BA) individuals from across West Eurasia. Admittedly, the sampling is very sparse, but like I've said before on these blogs, the LN/BA was a time of profound changes in Europe, so every scrap of data from this period is very valuable.

Note the lack of milk consumption among the samples from north of the Alps, where today the vast majority of people consume milk as adults, and can do so because they carry the Lactase Persistence Allele (T-13910). This doesn't look like a coincidence, considering the mounting evidence of a major population turnover across much of Europe during the LN/BA, mostly as a result of migrations from the east.

Citation...

Warinner, C. et al. Direct evidence of milk consumption from ancient human dental calculus. Sci. Rep. 4, 7104; DOI:10.1038/srep07104 (2014).

See also...

Lactase persistence and ancient DNA

Ancient genomes from the Great Hungarian Plain

The Y-chromosome tree bursts into leaf

Update 20/05/2015: Large-scale recent expansion of European patrilineages

...

I wonder what the hardcore Y-DNA genetic genealogists will say about this effort? I know that many of those guys have been working with full Y-chromosome sequences for a while now. It's open access with lots of supplementary info.

Abstract: Many studies of human populations have used the male-specific region of the Y chromosome (MSY) as a marker, but MSY sequence variants have traditionally been subject to ascertainment bias. Also, dating of haplogroups has relied on Y-specific short tandem repeats (STRs), involving problems of mutation rate choice, and possible long-term mutation saturation. Next-generation sequencing can ascertain single nucleotide polymorphisms (SNPs) in an unbiased way, leading to phylogenies in which branch-lengths are proportional to time, and allowing the times-to-most-recent-common-ancestor (TMRCAs) of nodes to be estimated directly. Here we describe the sequencing of 3.7 Mb of MSY in each of 448 human males at a mean coverage of 51x, yielding 13,261 high-confidence SNPs, 65.9% of which are previously unreported. The resulting phylogeny covers the majority of the known clades, provides date estimates of nodes, and constitutes a robust evolutionary framework for analysing the history of other classes of mutation. Different clades within the tree show subtle but significant differences in branch lengths to the root. We also apply a set of 23 Y-STRs to the same samples, allowing SNP- and STR-based diversity and TMRCA estimates to be systematically compared. Ongoing purifying selection is suggested by our analysis of the phylogenetic distribution of non-synonymous variants in 15 MSY single-copy genes.

Here are a couple of interesting quotes. You can see the samples they're talking about on the tree below. As per the second paragraph, it seems there's a paper about to be published at Nature Communications on European Y-chromosome haplogroups based on some heavy resequencing data (see Batini et al. in the references list). Can't wait for that.

(viii) Rare deep-rooting hg Q lineages in NW Europe: Hg Q has been most widely investigated in terms of the peopling of the Americas from NE Asia (Karafet et al. 1999). Here, as well as an example of the common native American Q-M3 lineage, we included examples of rare European hg Q chromosomes. One of the English chromosomes belongs to the deepest-rooting lineage within Q (Q-M378) and may reflect the Jewish diaspora (Hammer et al. 2009); the other is distantly related, shares a deep node with the Mexican Q-M3 chromosome, and has an STR-haplotype closely related to those of scarce Scandinavian hg Q chromosomes (unpublished data).

(ix) Structure within the west Eurasian hg R: The TMRCA of hg R is 19 KYA, and within it both hgs R1a and R1b comprise young, star-like expansions discussed extensively elsewhere (Batini et al. submitted). The addition of Central Asian chromosomes here contributes a sequence to the deepest subclade of R1b-M269, while another, in a Bhutanese individual, forms an outgroup almost as old as the R1a/R1b split.

Citation...

Hallast et al., The Y-chromosome tree bursts into leaf: 13,000 high-confidence SNPs covering the majority of known clades, Molecular Biology & Evolution, published online December 2, 2014, doi: 10.1093/molbev/msu327

Hinxton ancient genomes roundup

Most visitors here are probably aware by now that the Iron Age genomes from Hinxton are the two male samples 1 and 4 (ERS389795 and ERS389798, respectively). You can find confirmation of this at the link below.

The researchers were surprised to find that the older Iron Age men were genetically more similar to people living in Britain today than the Anglo-Saxon women were. Stephan Schiffels of the Wellcome Trust Sanger Institute reported the results October 20 at the annual meeting of the American Society of Human Genetics.

“It doesn’t look like these Anglo-Saxon immigrants left a big impact on the genetic makeup of modern-day Britain,” Schiffels said.

The finding raises an intriguing possibility that indigenous people in Britain may have repelled the Anglo-Saxons but adopted the invaders’ language and culture, says Eimear Kenny, a population geneticist at the Icahn School of Medicine at Mount Sinai in New York City, who was not involved in the work. More ancient samples from other times and parts of Britain should give a clearer picture of that episode of history, she said.

Anglo-Saxons left language, but maybe not genes to modern Britons

In regards to the main thrust of the article above, I'm not sure if there's much point discussing whether the British today are mostly of Celtic or Anglo-Saxon stock based on just five ancient genomes from a single location in England. However, if I was told that Hinxton4, the only high coverage genome in this collection, was a modern sample, I'd say it belonged to an Irishman from western Ireland, rather than an Englishman from eastern England.

Thus, unless Hinxton4 was an ancient migrant from Ireland, then it does seem to me as if there was a fairly significant admixture event in England between the indigenous Irish-like Celts and newcomers from the east, which eventually resulted in the present-day English population.

In any case, there are indeed some noticeable differences between the two sets of samples, and these can be visualized by plotting their f3 shared drift statistics.

For instance, plotting the f3-statistics of Hinxton2, which actually looks like the genome of someone straight off the boat from the Jutland Peninsula, against those of Hinxtons 1 and 4, we see that the former shares most drift with the Danes. Moreover, the Danes, Swedes and Germans, all Germanic speakers of course, deviate strongly on both graphs from the lines of slope that run from the Erzya to the Irish. The reason they deviate from these lines is because they don't share enough drift with Hinxtons 1 and 4 compared to the other reference populations from Northwestern Europe, especially the Irish.

A similar pattern can be seen when plotting the average results of Hinxtons 1 and 4 against those of 2, 3 and 5. However, the effect isn't nearly as pronounced, possibly because Hinxtons 3 and 5 are of mixed Celtic/Germanic origin.

See also...

Analysis of an ancient genome from Hinxton

Analysis of Hinxton2 - ERS389796

Analysis of Hinxton3 - ERS389797

Analysis of Hinxton4 - ERS389798

Analysis of Hinxton5 - ERS389799

Analysis of Hinxton5 - ERS389799

Hinxton5, or ERS389799, is one of five ancient English genomes stored at the Sequence Read Archive under accession number ERP003900. However, this analysis is based on the genotype file of Hinxton5 available at Genetic Genealogy Tools. For more information and some speculation about these genomes see my earlier blog post here.

Despite its relatively low North Sea score in the Eurogenes K15, and pronounced western shift on the Principal Component Analysis (PCA) plots, this genome appears mostly Germanic. In my opinion, the shared drift stats and also oracle results are quite convincing in this regard. If this were a modern sample it could probably pass for 3/4 north Dutch and 1/4 Irish. By the way, the Sub-Saharan admixture just looks like noise; this is, after all, a low coverage genome.

Shared drift stats of the form f3(Mbuti;Hinxton5,Test) - Eurogenes dataset

Shared drift stats of the form f3(Mbuti;Hinxton5,Test) - Human Origins dataset

Eurogenes K15 4 Ancestors Oracle results

See also...

Analysis of Hinxton2 - ERS389796

Analysis of Hinxton3 - ERS389797

Analysis of Hinxton4 - ERS389798

Hinxton ancient genomes roundup

Ust'-Ishim belongs to K-M526

Not long ago I predicted that Ust'-Ishim belonged to a basal form of Y-chromosome haplogroup P (see here). As it turns out, the 45,000 year-old western Siberian genome belongs to K(xLT) or K-M526, which is actually pretty close to my guess. The Ust'-Ishim paper was published today and is sitting behind a paywall here, but the extensive supp info is free.

Here's a map to help visualize the information, featuring Ust'-Ishim as well as Mal'ta boy, another North Eurasian Upper Paleolithic genome published recently.

The Ust'-Ishim genome was sequenced from the fossil of a femur bone found on the right bank of the Irtysh River. This area is very close to the Urals, and almost in the middle of the former Mammoth steppe that once stretched across North Eurasia from Iberia to Alaska. Interestingly, M526 is an ancestral mutation to the markers that define Y-chromosome haplogroups N, Q and R, which possibly dominated North Eurasia since the Upper Paleolithic (note that the 24,000 year-old Mal'ta boy belongs to a basal form of R).

Moreover, R1a and R1b are the most frequent haplogroups in Europe today. Thus, it would seem that most European males derive their paternal ancestry from North Eurasian hunter-gatherers whose ancestors spread out across Eurasia from the Middle East over 45,000 years ago.

I know that a lot of people have been arguing recently that K-M526 and the derived P-M45 originated and diversified in Southeast Asia, and then migrated north well within the last 45,000 years (for instance, see here). However, considering that K-M526 was already in reindeer country 45,000 years ago, as well as the Denisovan (ancient Siberian hominin) admixture among Southeast Asians, that might well turn out to be the equivalent of arguing that up is down and down is up.

By the way, Ust'-Ishim also belongs to pan-Eurasian mitochondrial (mtDNA) haplogroup R*, and in terms of genome-wide genetic structure appears roughly intermediate between West and East Eurasians. These outcomes fit very nicely with its Y-haplogroup.

However, it's slightly closer to Mesolithic Iberian genome La Brana-1, Upper Paleolithic Siberian MA-1 (or Mal'ta boy), and present-day East Asians, than to present-day West Eurasians, including Europeans. That's because it lacks "ancestry from a population that did not participate in the initial dispersals of modern humans into Europe and Asia". This is obviously the so called Basal Eurasian admixture discussed in Lazaridis et al. (see here), which is probably associated with early Neolithic farmers.

Also worth mentioning is that Ust'-Ishim harbors longer stretches of Neanderthal chromosomal segments than present-day Eurasians, which suggests that admixture between modern humans and Neanderthals took place in the Middle East not long before the ancestors of Ust-Ishim moved into Siberia (50-60,000 years ago). But this was already covered months ago, and you'll find lots of links on the topic on Google.

Citation...

Qiaomei Fu et al., Genome sequence of a 45,000-year-old modern human from western Siberia, Nature 514, 445–449 (23 October 2014) doi:10.1038/nature13810

See also...

Y-haplogroup P1 in Pleistocene Siberia (Sikora et al. 2018 preprint)

Analysis of Hinxton4 - ERS389798

Hinxton4, or ERS389798, is one of five ancient English genomes stored at the Sequence Read Archive under accession number ERP003900. However, this analysis is based on the genotype file of Hinxton4 available at Genetic Genealogy Tools. For more information and some speculation about these genomes see my earlier blog post here.

I still don't know who these samples represent exactly, but in all likelihood, this is one of the two Iron Age sequences from the collection, and probably belongs to a Briton of Celtic stock. Note, for instance, its high affinity to the present-day Irish, relatively low North Sea score in the Eurogenes K15, and pronounced western shift on the second Principal Component Analysis (PCA) plot below.

Interestingly, Lithuanians top its shared drift list based on the Human Origins dataset and more than 360K SNPs. I'm not entirely sure what this means, but it's probably related in some way to the unusually high level (>45%) of indigenous European hunter-gatherer ancestry carried by Lithuanians.

Shared drift stats of the form f3(Mbuti;Hinxton4,Test) - Eurogenes dataset

Shared drift stats of the form f3(Mbuti;Hinxton4,Test) - Human Origins dataset

Eurogenes K15 4 Ancestors Oracle results

See also...

Analysis of Hinxton2 - ERS389796

Analysis of Hinxton3 - ERS389797

Analysis of Hinxton5 - ERS389799

Hinxton ancient genomes roundup

Analysis of Hinxton3 - ERS389797

Hinxton3, or ERS389797, is one of five ancient English genomes stored at the Sequence Read Archive under accession number ERP003900. However, this analysis is based on the genotype file of Hinxton3 available at Genetic Genealogy Tools. For more information and some speculation about these genomes see my earlier blog post here.

Despite the exaggerated North Sea score in the Eurogenes K15, Hinxton3 could easily pass for a present-day Briton from the eastern coast of England or Scotland, albeit with a stronger than usual pull towards Scandinavia. Indeed, the f3-statistics show that it shares most genetic drift with the British and Icelanders from Eurogenes and Human Origins, respectively.

Shared drift stats of the form f3(Mbuti;Hinxton3,Test) - Eurogenes dataset

Shared drift stats of the form f3(Mbuti;Hinxton3,Test) - Human Origins dataset

Eurogenes K15 4 Ancestors Oracle results

See also...

Analysis of Hinxton2 - ERS389796

Analysis of Hinxton4 - ERS389798

Analysis of Hinxton5 - ERS389799

Hinxton ancient genomes roundup

Analysis of Hinxton2 - ERS389796

Hinxton2, or ERS389796, is one of five ancient English genomes stored at the Sequence Read Archive under accession number ERP003900. However, this analysis is based on the genotype file of Hinxton2 available at Genetic Genealogy Tools. For more information and some speculation about these genomes see my earlier blog post here.

Interestingly, f3-statistics in the form f3(Mbuti;Hinxton2,Test) show that Hinxton2 shares most genetic drift with present-day Danes and Norwegians. Please refer to the relevant spreadsheets below.

Shared drift stats of the form f3(Mbuti;Hinxton2,Test) - Eurogenes dataset

Shared drift stats of the form f3(Mbuti;Hinxton2,Test) - Human Origins dataset

Eurogenes K15 4 Ancestors Oracle results

See also...

Analysis of Hinxton3 - ERS389797

Analysis of Hinxton4 - ERS389798

Analysis of Hinxton5 - ERS389799

Hinxton ancient genomes roundup

Ancient genomes and the calculator effect

Several ancient genomes have been posted online as text files and uploaded to GEDmatch over the last couple of weeks, and many more are likely to follow in the future. A lot of people have already taken this opportunity to analyze these files with various online ancestry tools, usually DIY calculators.

That's actually not a bad way of doing things, as long as everyone's aware that almost all of these calculators produce biased results. They produce biased results because they violate a very basic rule of science, which is this:

Do not test more than one variable at a time.

Obviously, the variable we want to test with these calculators is ancestry. However, when the reference samples are tested in a different way to the test samples, which is what usually happens, then this adds another variable to the proceedings. As a result, we simply can't compare the results of the reference samples to those of the test samples.

I know that a lot of people find this difficult to grasp, and many just seem hell bent on not grasping it. However, anyone who isn't completely insane, and takes five minutes out of their day to try and understand the concepts involved, has to agree that this is a real problem. It can be proven empirically, like I did over two years ago (see here).

I suspect that a lot of confusion has been caused by the fact that the people who were used as reference samples in the making of the various DIY calculators saw highly accurate results when running them, and so assumed everything was fine. The accuracy of the DIY calculators for such people is indeed impressive, and I show that at the link above, but unfortunately the story is very different for everyone else.

Here's the good news: the Eurogenes calculators don't suffer from the calculator effect. That's because the reference samples are treated in the same way as the test samples, so there's only one variable: ancestry. What this means is that when you run a modern or ancient genome with a Eurogenes calculator you can confidently compare the result to those of the reference samples (provided enough SNPs are used), and then be able to make sensible inferences about its genetic origins.

Analysis of an ancient genome from Hinxton

I've just added an ancient sample from Hinxton, England, to my burgeoning ancient genomes collection. It's a pre-publication release freely available here as ERS389795. Thanks to Felix C. for breaking the news. We've both called this sample Hinxton1.

Unfortunately, its archeological context is a mystery to me, but it's possibly one of the ancient genomes mentioned in the recent Schiffels et al. ASHG abstract (see here).

In terms of genome-wide genetic structure, Hinxton1 is most similar to present-day Orcadians, Irish, western Scots, Icelanders and western Norwegians, more or less in that order. However, it's fairly distinct from the modern inhabitants of England, or at least those in my datasets, who mostly come from Kent and Cornwall.

Please note, this analysis features two different datasets: Eurogenes and Human Origins. Eurogenes, which is my own dataset, includes more populations than Human Origins, and is based on SNPs used in commercial ancestry and medical work. On the other hand, Human Origins shows a more varied sampling strategy, and is based on SNPs specifically chosen for population genetics.

Shared drift stats in the form f3(Mbuti;Hinxton1,Test) - Eurogenes dataset

Shared drift stats in the form f3(Mbuti;Hinxton1,Test) - Human Origins dataset

Eurogenes K15 4 Ancestors Oracle results

See also...

Analysis of Hinxton2 - ERS389796

Analysis of Hinxton3 - ERS389797

Analysis of Hinxton4 - ERS389798

Analysis of Hinxton5 - ERS389799

Hinxton ancient genomes roundup

Genetic structure in the Western Balkans

PLoS ONE has a new paper by Kovacevic et al. on the genetic structure of Western Balkan populations. Here's the abstract:

Contemporary inhabitants of the Balkan Peninsula belong to several ethnic groups of diverse cultural background. In this study, three ethnic groups from Bosnia and Herzegovina - Bosniacs, Bosnian Croats and Bosnian Serbs - as well as the populations of Serbians, Croatians, Macedonians from the former Yugoslav Republic of Macedonia, Montenegrins and Kosovars have been characterized for the genetic variation of 660 000 genome-wide autosomal single nucleotide polymorphisms and for haploid markers. New autosomal data of the 70 individuals together with previously published data of 20 individuals from the populations of the Western Balkan region in a context of 695 samples of global range have been analysed. Comparison of the variation data of autosomal and haploid lineages of the studied Western Balkan populations reveals a concordance of the data in both sets and the genetic uniformity of the studied populations, especially of Western South-Slavic speakers. The genetic variation of Western Balkan populations reveals the continuity between the Middle East and Europe via the Balkan region and supports the scenario that one of the major routes of ancient gene flows and admixture went through the Balkan Peninsula.

Among the most eye catching figures from the study is this TreeMix graph with ten migration edges or admixture events. Note the 44% migration edge running from the base of the Eastern European branch to the French. Is this perhaps a legacy of the Proto-Celts and early Germanics? In any case, something similar can be seen on this TreeMix graph from the supplementary PDF to Skoglund et al. 2014, where a French genome is modeled as a clade closely related to Upper Paleolithic Siberian forager MA-1, but with considerable Sardinian admixture.

Also, the position of the Poles at the tip of the tree, and thus near the North Russians, is somewhat curious. However, I know that several of these individuals are ethnic Poles from Estonia, so that might be the problem.

Update 25/08/2014: Here's a typical Eurogenes Principal Component Analysis (PCA) of West Eurasia with the new samples from this paper (Bosnians, Kosovars, Macedonians, Montenegrins and Serbs).

Citation...

Kovacevic L, Tambets K, Ilumäe A-M, Kushniarevich A, Yunusbayev B, et al. (2014) Standing at the Gateway to Europe - The Genetic Structure of Western Balkan Populations Based on Autosomal and Haploid Markers. PLoS ONE 9(8): e105090. doi:10.1371/journal.pone.0105090

Haplotype-based PCA of West Eurasia and Europe

The Principal Component Analyses (PCA) below are based on pairwise Identity-by-Descent (IBD) sharing inferred with fastIBD. My aim was to create PCA that took into account haplotype information to see how they might differ from similar plots based on unlinked loci (such as here).

Clearly, they're less reflective of geography and isolation-by-distance, and instead more profoundly influenced by relatively recent isolation, founder effects and/or rapid expansions, especially in Northern and Eastern Europe, and in particular among the Finns, Balts and East Slavs. Unfortunately, I don't have time to say much more about these results. But feel free to post any questions or observations in the comments below. I have done something very similar in the past, but with far fewer samples (see here).

Please note, to ensure that the PCA were as informative as possible I was forced to drop several populations that produced unusual results, probably because of extreme founder effects. This is why, for instance, there are no Ashkenazi Jews on any of the plots, and the only Finns you'll find come from western Finland.

I'll try this again on a much larger dataset when more samples come in, and also include populations from Central and South Asia.

Update 7/8/2014: Apparently some people are wondering what the plots with Finns and Jews look like. Here you go...

Coming soon: genome-wide data from more than forty 3-9K year-old humans from the ancient Russian steppe

Below is a presentation abstract from the upcoming SMBE 2014 conference. I simply can't wait to see the paper, which I'm guessing will be published very soon.

A central challenge in ancient DNA research is that for many bones that contain genuine DNA, the great majority of molecules in sequencing libraries are microbial. Thus, it has been impractical to carry out whole genome analyses of substantial numbers of ancient individuals. We report a strategy for in-solution capture of ancient DNA from approximately 390,000 single nucleotide polymorphism (SNP) targets, adapting a method of Fu et al. PNAS 2013 who enriched a 40,000 year old DNA sample for the entire chromosome 21. Of the SNPs targets, the vast majority overlap the Affymetrix Human Origins array, allowing us to compare the ancient samples to a database of more than 2,700 present-day humans from 250 groups.

We applied the SNP capture as well as mitochondrial genome enrichment to a series of 65 bones dating to between 3,000-9,000 years ago from the Samara district of Russia in the far east of Europe, a region that has been suggested to be part of the Proto-Indo-European homeland. We successfully extracted nuclear data from 10-90% of targeted SNPs for more than 40 of the samples, and for all of these samples also obtained complete mitochondrial genomes. We report three key findings:

- Samples from the Samara region possess Ancient North Eurasian (ANE) admixture related to a recently published 24,000 year old Upper Paleolithic Siberian genome. This contrasts with both European agriculturalists and with European hunter-gatherers from Luxembourg and Iberia who had little such ancestry (Lazaridis et al. arXiv.org 2013). This suggests that European steppe groups may be implicated in the dispersal of ANE ancestry across Europe where it is currently pervasive.

- The mtDNA composition of the steppe population is primarily West Eurasian, in contrast with northwest Russian samples of this period (Der Sarkissian et al. PLoS Genetics 2013) where an East Eurasian presence is evident.

- Samara experienced major population turnovers over time: early samples (>6000 years) belong primarily to mtDNA haplogroups U4 and U5, typical of European hunter-gatherers but later ones include haplogroups W, H, T, I, K, J.

We report modeling analyses showing how the steppe samples may relate to ancient and present-day DNA samples from the rest of Europe, the Caucasus, and South Asia, thereby clarifying the relationship of steppe groups to the genetic, archaeological and linguistic transformations of the late Neolithic and Bronze ages.

David Reich et al., Genotyping of 390,000 SNPs in more than forty 3,000-9,000 year old humans from the ancient Russian steppe, SMBE 2014 abstract.

The other really interesting abstract from this conference concerns the Ust-Ishim genome from Upper Paleolithic western Siberia (see here). I'm betting its Y-chromosome haplogroup will be P*, but that's pure speculation on my part.


Update 11/02/2015: Massive migration from the steppe is a source for Indo-European languages in Europe (Haak et al. 2015 preprint) .

PCA projection bias in ancient DNA studies

Many Principal Component Analyses (PCA) in papers on ancient genomes clearly suffer from projection bias. However, most people don't seem to understand this problem and the impact it can have on the interpretation of the data.

Here's a demonstration of this effect using two PCA. In the first PCA, La Brana-1, a Mesolithic genome from Iberia, was projected onto the PC eigenvectors computed with modern individuals from the HGDP. However, in the second PCA the ancient genome was run together with these samples. Note the clear difference between the two outcomes.

The second outcome does look a bit strange, but it's actually the correct one, because it's now an established fact that Mesolithic hunter-gatherers, like La Brana-1, were clearly outside the range of modern European, and indeed West Eurasian, genetic variation.

For a technical discussion of this problem, which is also sometimes known as "shrinkage", refer to Lee et al. 2012. To get an idea of the confusion that it can cause, see the discussion in the comments section under my last blog post:

More info on two Thracian genomes from Iron Age Bulgaria + a complaint

The above experiment with La Brana-1 was run with PLINK 2, which is freely available here, using just over 16K SNPs. Only markers with a read depth of 4x or higher were considered, and the marker set was further pruned to account for no-calls (--geno 0.005), LD (--indep-pairwise 200 25 0.4), and minor allele frequency (--maf 0.05).

More info on two Thracian genomes from Iron Age Bulgaria + a complaint

PLoS Genetics has just published a new paper on the genetic affinities of Oetzi the Iceman (see here). As far as I can tell, it simply affirms what we've already learned about Oetzi from previous studies, but it does feature interesting new insights into a couple of genomes from Iron Age Bulgaria, aka. Thrace:

The first individual (P192-1) was excavated from a pit sanctuary near Svilengrad, Bulgaria, dated to 800–500 BCE. The other individual (K8) was found in the Yakimova Mogila Tumulus in southeastern Bulgaria, dated to 450–400 BCE.

...

For the Thracian individuals from Bulgaria, no clear pattern emerges. While P192-1 still shows the highest proportion of Sardinian ancestry, K8 more resembles the HG individuals, with a high fraction of Russian ancestry.

...

Interestingly, this individual [K8] was excavated from an aristocratic inhumation burial containing rich grave goods, indicating a high social standing, as opposed to the other individual, who was found in a pit [15]. However, the DNA damage pattern of this individual does not appear to be typical of ancient samples (Table S4 in [15]), indicating a potentially higher level of modern DNA contamination.

K8 might well be contaminated with modern DNA to some degree, but I'd say there's a much better explanation for these signals of non-trivial genetic substructures within the Thracian population.

Archeology suggests that during the Bronze Age the Balkans were invaded from the east by nomads associated with the Yamnaya culture of the Pontic-Caspian Steppe. These invaders, possibly of early Indo-European stock, liked to build Tumuli mounds for their important dead, which were essentially copies of the Kurgan mounds built by the Yamnaya and related peoples.

Moreover, we now know that indigenous European hunter-gatherer (HG) ancestry survived best in Eastern Europe (see here), so it's very likely that the aforementioned invaders from the steppe were significantly HG-like in terms of genetic structure.

Therefore, the fact that K8 was buried in a richly furnished Tumulus (essentially a Kurgan), and genetically more similar to indigenous Europeans than P192-1, who was genetically more Near Eastern-like, and basically thrown into a ditch after he died, doesn't appear to be a coincidence.

In other words, perhaps K8 belonged to a ruling class of steppe origin, while P192-1 was largely of native Balkan stock, whose ancestors were conquered centuries earlier by the steppe nomads and forced to live as an underclass? If so, it wouldn't be the only time in history that this sort of thing has happened, especially within Indo-European societies.

By the way, unfortunately I have to add that the Principal Component Analyses (PCA) in this paper featuring the two HG genomes, ajv70 and La Brana-1, are simply woeful (PDF link). These genomes should be clearly outside the range of modern European genetic variation, but here they land among the Orcadian and French samples. Where was the peer review I wonder?

Citation...

Sikora M, Carpenter ML, Moreno-Estrada A, Henn BM, Underhill PA, et al. (2014) Population Genomic Analysis of Ancient and Modern Genomes Yields New Insights into the Genetic Ancestry of the Tyrolean Iceman and the Genetic Structure of Europe. PLoS Genet 10(5): e1004353. doi:10.1371/journal.pgen.1004353

See also...

Ancient DNA from prehistoric Bulgaria and Denmark

PCA projection bias in ancient DNA studies

The really old Europe is mostly in Eastern Europe

A new version of the Lazaridis et al. ancient genomes preprint has just appeared at arXiv (see here). It includes several new Principal Component Analyses (PCA), TreeMix graphs, a ChromoPainter/fineSTRUCTURE co-ancestry matrix, and an updated ADMIXTURE analysis. The revised text underlines the relatively close genetic relationship between indigenous European hunter-gatherers and present-day Eastern Europeans:

The co-ancestry matrix (Fig. S19.3) confirms the ability of this method to meaningfully cluster individuals. We highlight two clusters: Stuttgart joins all Sardinian individuals in cluster A and Loschbour joins a cluster B that encompasses all Belarusian, Ukrainian, Mordovian, Russian, Estonian, Finnish, and Lithuanian individuals. These results confirm Sardinia as a refuge area where ancestry related to Early European Farmers has been best preserved, and also the greater persistence of WHG-related ancestry in present-day Eastern European populations. The latter finding suggests that West European Hunter-Gatherers (so-named because of the prevalence of Loschbour and La Braña) or populations related to them have contributed to the ancestry of present-day Eastern European groups. Additional research is needed to determine the distribution of WHG-related populations in ancient Europe.

Fig. S10.5 suggests that the main axis of differentiation in Europe when the subcontinent is considered as a whole may tend to Northeastern Europe rather than SSE/NNW (8). This is consistent with our analysis of ancestry proportions in European populations (Fig. 2B, Extended Data Table 3) which indicate a cline of reduced EEF (and increasing WHG) ancestry along that direction.

Citation...

Iosif Lazaridis, Nick Patterson, Alissa Mittnik, et al., Ancient human genomes suggest three ancestral populations for present-day Europeans, arXiv, April 2, 2014, arXiv:1312.6639v2

Extreme positive selection for light skin, hair and eyes on the Pontic-Caspian steppe...or not

Unusually strong positive selection over the past 5,000 years, rather than population replacement or even admixture, is responsible for the high frequencies of light skin, hair and eyes among present-day Eastern Europeans, according to a new paper by Wilde et al. at PNAS.

The authors were able to infer pigmentation traits from ancient DNA for 63 Eneolithic and Bronze Age samples, mostly from Kurgan mounds from the Pontic-Caspian steppe of Ukraine and surrounds. The results suggest that the ancient individuals were overall much darker than present-day Ukrainians, who, nevertheless, appear to be their direct descendants based on mitochondrial DNA (mtDNA) sequences. Quoting the paper:

To this end we compared the 60 mtDNA HVR1 sequences obtained from our ancient sample to 246 homologous modern sequences (29–31) from the same geographic region and found low genetic differentiation (FST = 0.00551; P = 0.0663) (32). Coalescent simulations based on the mtDNA data, accommodating uncertainty in the ancient sample age, failed to reject population continuity under a wide range of assumed ancestral population size combinations (Fig. 1).

Conversely, continuity between early central European farmers and modern Europeans has been rejected in a previous study (33). However, the Eneolithic and Bronze Age sequences presented here are ∼500–2,000 y younger than the early Neolithic and belong to lineages identified both in early farmers and late hunter–gatherers from central Europe (33).

...

In sum, a combination of selective pressures associated with living in northern latitudes, the adoption of an agriculturalist diet, and assortative mating may sufficiently explain the observed change from a darker phenotype during the Eneolithic/Early Bronze age to a generally lighter one in modern Eastern Europeans, although other selective factors cannot be discounted. The selection coefficients inferred directly from serially sampled data at these pigmentation loci range from 2 to 10% and are among the strongest signals of recent selection in humans.

Well, either this is indeed a remarkable finding, or something's not quite right. I think it's the latter.

The argument for genetic continuity from the Eneolithic/Bronze Age to the present on the Pontic-Caspian steppe based on mtDNA sequences is actually very weak. The results could simply mean that the ancient samples shared deep maternal ancestry with modern Ukrainians and most other Europeans.

Indeed, we know for a fact that much of the Pontic-Caspian steppe was occupied by Turkic groups of Asian origin from the early Middle Ages until only a couple of hundred years ago. They were eventually cleared out by Tsarist Russia, and mainly replaced by East Slavic settlers from just northwest of the steppe. This process might not be easy to see by comparing low resolution mtDNA data, even between European populations separated by 5,000 years, but it's likely to be obvious when looking at full mtDNA genomes, high-density genome-wide data, and/or Y-chromosome haplogroups.

Surprisingly, the article doesn't mention Keyser et al. 2009, a very important study which showed that a sample of Kurgan nomads from Bronze and Iron Age South Siberia had frequencies of light hair and eyes comparable to those of present-day Northern and Eastern Europeans (see here). Also worth noting is that the most common Y-chromosome haplogroup among these individuals was R1a, which is today the most frequent haplogroup in Eastern Europe, including Ukraine.

What this suggests to me is that the Kurgan cultural horizon was not genetically homogeneous. I suspect that Kurgan groups closer to the Balkans carried significantly higher levels of Near Eastern Neolithic farmer ancestry, and were thus much darker than those in the more temperate northerly regions. However, it seems that at some point, the Neolithic farmer DNA was diluted enough by continuous movements of light pigmented groups from the north and east, possibly made up mostly of males, that there was a major shift in pigmentation traits from Near Eastern-like to North European-like across most of Eastern Europe. This scenario actually fits very nicely with the latest on the genetic origins of Europeans (see here).

We won't know what really happened until we see at least a few complete ancient genomes from Eastern Europe. But for now, I'd have to suspend my disbelief to accept that present-day Eastern Europeans are, by and large, descendants of these exceedingly brunet prehistoric people of the Pontic-Caspian steppe.

Citation...

Wilde et al., Direct evidence for positive selection of skin, hair, and eye pigmentation in Europeans during the last 5,000 y, PNAS, Published online before print on March 10, 2014, DO:I10.1073/pnas.1316513111

See also...

PCA of ancient European mtDNA

Khazar shmazar

Human Biology recently posted several open access manuscripts dealing with the topic of Jewish origins (see submissions from 2013 here). One of these preprints is essentially a rebuttal to an Eran Elhaik paper from a couple of years ago, which argued that a substantial part of Ashkenazi Jewish ancestry was derived from within the Khazar Empire. The leading author of the new preprint is Doron M. Behar, but thirty people in all, many of them well known scientists, have put their names on it. Here's the abstract:

The origin and history of the Ashkenazi Jewish population have long been of great interest, and advances in high-throughput genetic analysis have recently provided a new approach for investigating these topics. We and others have argued on the basis of genome-wide data that the Ashkenazi Jewish population derives its ancestry from a combination of sources tracing to both Europe and the Middle East. It has been claimed, however, through a reanalysis of some of our data, that a large part of the ancestry of the Ashkenazi population originates with the Khazars, a Turkic-speaking group that lived to the north of the Caucasus region ~1,000 years ago. Because the Khazar population has left no obvious modern descendants that could enable a clear test for a contribution to Ashkenazi Jewish ancestry, the Khazar hypothesis has been difficult to examine using genetics. Furthermore, because only limited genetic data have been available from the Caucasus region, and because these data have been concentrated in populations that are genetically close to populations from the Middle East, the attribution of any signal of Ashkenazi-Caucasus genetic similarity to Khazar ancestry rather than shared ancestral Middle Eastern ancestry has been problematic. Here, through integration of genotypes on newly collected samples with data from several of our past studies, we have assembled the largest data set available to date for assessment of Ashkenazi Jewish genetic origins. This data set contains genome-wide single-nucleotide polymorphisms in 1,774 samples from 106 Jewish and non- Jewish populations that span the possible regions of potential Ashkenazi ancestry: Europe, the Middle East, and the region historically associated with the Khazar Khaganate. The data set includes 261 samples from 15 populations from the Caucasus region and the region directly to its north, samples that have not previously been included alongside Ashkenazi Jewish samples in genomic studies. Employing a variety of standard techniques for the analysis of populationgenetic structure, we find that Ashkenazi Jews share the greatest genetic ancestry with other Jewish populations, and among non-Jewish populations, with groups from Europe and the Middle East. No particular similarity of Ashkenazi Jews with populations from the Caucasus is evident, particularly with the populations that most closely represent the Khazar region. Thus, analysis of Ashkenazi Jews together with a large sample from the region of the Khazar Khaganate corroborates the earlier results that Ashkenazi Jews derive their ancestry primarily from populations of the Middle East and Europe, that they possess considerable shared ancestry with other Jewish populations, and that there is no indication of a significant genetic contribution either from within or from north of the Caucasus region.

I'm really not sure what to make of all of this attention that the Khazar hypothesis is still getting? It's been obvious for a while now that in terms of genetic structure Ashkenazi Jews are basically a group of East Mediterranean origin. But Elhaik's paper did get a fair bit of media coverage, so I suppose after that a rebuttal was to be expected.

In any case, I'm not complaining. This paper includes a very interesting genotype dataset of many previously unpublished samples, which I tested last week with PCA (see here).

Citations...

Behar, Doron M.; Metspalu, Mait; Baran, Yael; Kopelman, Naama M.; Yunusbayev, Bayazit; Gladstein, Ariella; Tzur, Shay; Sahakyan, Havhannes; Bahmanimehr, Ardeshir; Yepiskoposyan, Levon; Tambets, Kristiina; Khusnutdinova, Elza K.; Kusniarevich, Aljona; Balanovsky, Oleg; Balanovsky, Elena; Kovacevic, Lejla; Marjanovic, Damir; Mihailov, Evelin; Kouvatsi, Anastasia; Traintaphyllidis, Costas; King, Roy J.; Semino, Ornella; Torroni, Anotonio; Hammer, Michael F.; Metspalu, Ene; Skorecki, Karl; Rosset, Saharon; Halperin, Eran; Villems, Richard; and Rosenberg, Noah A., No Evidence from Genome-Wide Data of a Khazar Origin for the Ashkenazi Jews (2013). Human Biology Open Access Pre-Prints. Paper 41.

Elhaik E. The missing link of Jewish European Ancestry: contrasting the Rhineland and Khazarian hypotheses. Genome Biol Evol. 2012. doi:10.1093/gbe/evs119, Advance Access publication December 14, 2012.

See also...

Near Eastern origin of Ashkenazi Levite R1a

A Mesolithic genome from Spain

Nature today published a paper on the complete genome of La Brana 1, a Mesolithic hunter-gatherer from Iberia: Olalde et al. 2014. Based on genetic variants associated with pigmentation traits, it's likely that this individual had blue eyes, dark hair and deep brown skin.

Moreover, he was probably lactose intolerant (in other words, unlike most Europeans today, he couldn't drink milk as an adult), and his Y-chromosome belonged to the European-specific, but today extremely rare, haplogroup C6 (aka. C-V20), and mtDNA to haplogroup U5b2c1, which again is a European-specific marker. Below is an artist's impression of his mug (courtesy of CSIC), and below that the paper abstract.

Ancient genomic sequences have started to reveal the origin and the demographic impact of farmers from the Neolithic period spreading into Europe1, 2, 3. The adoption of farming, stock breeding and sedentary societies during the Neolithic may have resulted in adaptive changes in genes associated with immunity and diet4. However, the limited data available from earlier hunter-gatherers preclude an understanding of the selective processes associated with this crucial transition to agriculture in recent human evolution. Here we sequence an approximately 7,000-year-old Mesolithic skeleton discovered at the La Braña-Arintero site in León, Spain, to retrieve a complete pre-agricultural European human genome. Analysis of this genome in the context of other ancient samples suggests the existence of a common ancient genomic signature across western and central Eurasia from the Upper Paleolithic to the Mesolithic. The La Braña individual carries ancestral alleles in several skin pigmentation genes, suggesting that the light skin of modern Europeans was not yet ubiquitous in Mesolithic times. Moreover, we provide evidence that a significant number of derived, putatively adaptive variants associated with pathogen resistance in modern Europeans were already present in this hunter-gatherer.

Indeed, the pigmentation traits are basically the same as those of Loschbour, a Mesolithic genome from Luxembourg, featured recently in the groundbreaking Lazaridis et al. preprint (see here). So we can already speculate with some confidence that this was a common, and perhaps dominant, trait combination among European hunter-gatherers.

However, early European farmers, whose ancestors almost certainly migrated to Europe from the Near East during the Neolithic, probably had somewhat different pigmentation traits. We know this because a 7,500 year-old Linearbandkeramik (LBK) farmer genome from Stuttgart, Germany, also featured in Lazaridis et al., showed markers for brown eyes, dark hair, and relatively light skin.

So as things stand, it appears that Europeans only acquired their present coloring, including pale skin and a high incidence of light eyes, relatively recently, well after the hunter-gatherers and farmers began mixing, and their hybrid DNA had time to go through some really powerful selective sweeps. These sweeps were possibly in part a reaction to the Neolithic diet, rich in carbohydrates but poor in vitamin D, amongst other things. Vitamin D doesn't have to be acquired from food because the body can synthesize it from the sun, but this is done more effectively by people with fair skin, giving them an advantage, especially in places like Europe, which has fairly long winters and lots of cloud cover.

But perhaps this isn't the full story, and present-day European pigmentation traits are also sourced from a late migration into Europe of a prevailingly blond people from somewhere in what is now Russia?

This might sound far fetched, but during the middle Bronze Age the Eurasian steppe was home to the Andronovo culture, with archeological links to earlier cultures in what is now southern Russia. Based on the DNA of Andronovo nomads from Kurgans in South Siberia, it seems they had fair skin and a lot of blue eyes and blond hair (see here). They also overwhelmingly belonged to Y-chromosome haplogroup R1a1a, which is very common today in Central and Eastern Europe and also parts of Scandinavia. So it'll be interesting to see the pigmentation markers of Mesolithic Eastern Europeans and Central Asians when their genomes become available, probably in the not too distant future, and if they contributed any ancestry to present-day Europeans. Early indications are that they did, and I discussed that in my previous blog entry here.

La Brana 1 and Loschbour were both classified as part of the West European Hunter-Gatherer (WHG) mata-population by Lazaridis et al., even though only a partial sequence from La Brana 1 was available at the time. As far as I can see, the results in Olalde et al. based on the complete genome don't contradict this classification, because they show that La Brana 1 is most similar to present-day Europeans from around the Baltic Sea, just like Loschbour. Note, for instance, the position of Swedes (SE) and Poles (PL) on the far right of these graphs, indicating inflated allele sharing between them and La Brana 1 relative to other Europeans.

Unfortunately, I have to say that the main Principal Component Analysis (PCA) from the paper isn't as informative as it could have been, due to the large number of Finnish individuals included in the analysis. It's mostly a reflection of the recent population growth, founder effect and genetic drift among Finns, particularly those from eastern Finland.

Nevertheless, note that all of the non-Finnish Europeans more or less fall along the cline that runs from La Brana 1 to present-day Cypriots. This suggests that Europeans today are mostly the product of mixture, in varying degrees, between indigenous European hunter-gatherers, like La Brana 1 and Loschbour, and immigrant Neolithic farmers from the East Mediterranean. So it's a result that basically agrees with the findings of Lazaridis et al.

Interestingly, Loschbour and four other Mesolithic samples from Lazaridis et al. belonged to Y-chromosome haplogroup I, which is not at all closely related to C6. This hints at the presence of a diverse Y-chromosome gene pool in pre-Neolithic Europe, and indeed I'm still confident of seeing R1 and/or R1a among Mesolithic remains from Eastern Europe.

Even though the vast majority of haplogroup C clades are today specific to Eastern Asia, Oceania and the Americas, C6 has only been found among a handful of individuals from across Southern, Western and Central Europe, many of whom are listed at the FTDNA haplogroup C project (look for the V20+ results here). It's difficult to say when this marker or its ancestral lineage migrated to Europe, but C is one of the most basal human Y-chromosome clades, so it could represent the very first Anatomically Modern Human (AMH) wave into Europe, which actually isn't a new concept (see Scozzari et al. 2012).

The Olalde et al. paper includes a lot more information than I'm willing to cover in this blog entry. If you don't have access to the main report, please note that the extended and supplementary data are very detailed and open access.

Citation...

Olalde et al., Derived immune and ancestral pigmentation alleles in a 7,000-year-old Mesolithic European, Nature (2014), doi:10.1038/nature12960

Another look at the Lazaridis et al. ancient genomes preprint

I've now had a chance to look over the Lazaridis et al. preprint a few times, and also take part in several online discussions about the results, at these blogs and elsewhere. So I thought it might be useful to put together another post on the paper to report what I've learned and reiterate a few points. First of all, to understand the results, it's really important to known what the four main ancestral components in this study represent:

- West European Hunter-Gatherer (WHG), based on an 8,000 year-old genome from Loschbour, Luxembourg

- Ancient North Eurasian (ANE), based on a 24,000 year-old genome from South Siberia (dubbed Mal'ta boy or MA-1)

- Early European Farmer (EEF), based on a 7,500 year-old genome from Stuttgart, Germany, belonging to the Neolithic Linearbandkeramik (LBK) culture

- Eastern non-African (ENA), this basically means East Eurasian, and is based on samples of present-day Onge, Han Chinese and Atayal from Taiwan

Now, from what I've seen online, many people seem to think that ANE is more East Asian than European, and can be considered a signal of pretty much any population expansion from the east into Europe. This is not true. ANE is Amerindian-like, but actually also very similar to WHG. In fact, they're equidistant from ENA:

The results of Table S12.1 provide suggestive evidence that Onge share more common ancestry with hunter-gatherers than with Stuttgart. All statistics involving two hunter-gatherer populations have |Z|<0.9, so ancient Eurasian hunter-gatherers are approximately symmetrically related to Onge, and they are all more closely related to them than is Stuttgart.

We next consider the relationship of ancient samples to East Asia using the set (Ami, Atayal, Han, Naxi, She). East Asians are more closely related to all hunter-gatherers than to Stuttgart, but there are no significant differences between hunter-gatherers (all such statistics have |Z|<1.1) (Table S12.2).

...

We have conveniently labeled MA1-related ancestry “Ancient North Eurasian” because of the provenance of MA1 in Siberia, but at present we cannot be sure whether this type of ancestry originated there or was a recent migrant from some western region.

The various Uralic, Turkic and Mongolian groups expanding into Europe, usually after the Bronze Age, no doubt carried significant ENA, so these groups can't be the source of the fairly high levels of ANE across Europe today, because most Europeans lack ENA. Below is a graph based on two f4 tests, comparing ANE and ENA ancestry among Europeans, this time with the Han Chinese as ENA proxies. Note that most of the samples fall within a cline that runs from the Stuttgart sample to Estonians. The only outliers in the direction of the Han are groups from current or former Uralic and Turkic speaking areas of Europe.

ANE was actually present in Scandinavia during the Mesolithic, because Motala12, the 8,000 year-old hunter-gatherer genome from Sweden, has an ANE ratio of 19%. But this isn't enough to explain the ANE levels carried by most present-day Europeans, so it's very likely there were at least two expansions of ANE into Europe.

Considering that Loschbour and Stuttgart totally lack ANE, it's plausible that a major wave of ANE moved across much of Europe sometime after the early Neolithic, but obviously before the Uralic and Turkic expansions, which, as per above, were rich in ENA. Based on recently published ancient mtDNA evidence from Central Europe (see here), Lazaridis et al. propose that this timeframe was the Copper and/or Bronze Age.

This of course is the generally accepted Proto-Indo-European timeframe. Indeed, the theory I put forward in the previous blog entry (see here) that most of the ANE in Europe today was the result of the Proto-Indo-European expansion, probably from Eastern Europe, looks even better on closer inspection.

Note the elongated cline formed by the European samples running from WHG to EEF on Fig 2B, shown below. It correlates well with latitude, and very likely reflects northward migrations of Neolithic farmers into Europe from the Mediterranean Basin, followed by isolation-by-distance. In other words, this cline probably took thousands of years to form.

On the other hand, there is no cline running from WHG/EEF to ANE, but all of the Indo-European and/or Eastern European samples are fairly evenly lifted up towards ANE relative to a few outliers. These outliers are all southwestern Europeans: Basques, Pais Vasco (Basque Country) Spaniards, southern French and Sardinians.

Of course, southwestern Europe is the most distant part of the continent from the generally accepted Indo-European homeland near the middle Volga. Moreover, Basques don't speak an Indo-European language, while Sardinians were only Indo-Europeanized during historic times.

Indeed, even though a couple of tables in the study report considerable ANE ancestry among Basques and Pais Vasco Spaniards, the authors admit that this need not be the case. For instance:

We next attempted to fit individual West Eurasian populations as a mixture of Loschbour and Stuttgart, as representatives of Early European farmers and West European Hunter Gatherers.

Fig. 1B suggests that this is not possible, as most Europeans form a cline that cannot be reconciled with such a mixture [Davidski's note: I think they actually mean Fig. 2B]. Nonetheless, for Sardinians (Extended Data Table 1), the most negative f3-statistic is of the form f3(Test; Loschbour, Stuttgart), which suggests that at least some Europeans may be consistent with having been formed by such a mixture. We thus fit each European population into the topology of Fig. S12.6. Only Basques, Pais_Vasco, and Sardinians, can be fit successfully with this model. Fig. S12.8 shows a successful fit.

Most European populations cannot be fit as this type of 2-way mixture and, intuitively, this is due to their tendency (Fig. 1B) towards Ancient North Eurasians that is not modeled by such a mixture.

Another intriguing thing about the results shown in Fig 2B is that the expansions of ANE across Europe appear not to have disturbed the presumably Neolithic WHG/EEF cline to any great extent. What this suggests is that ANE was spread largely independently of EEF and even WHG. In other words, the groups that pushed ANE deep into Europe probably had very high ratios of this component. This also seems to be true for the groups that brought ANE to the Near East:

A geographically parsimonious hypothesis would be that a major component of present-day European ancestry was formed in eastern Europe or western Siberia where western and eastern hunter-gatherer groups could plausibly have intermixed. Motala12 has an estimated WHG/(WHG+ANE) ratio of 81% (S12.7), higher than that estimated for the population contributing to modern Europeans (Fig. S12.14). Motala and Mal’ta are separated by 5,000km in space and about 17 thousand years in time, leaving ample room for a genetically intermediate population. The lack of WHG ancestry in the Near East (Extended Data Fig. 6, Fig. 1B) together with the presence of ANE ancestry there (Table S12.12) suggests that the population who contributed ANE ancestry there may have lacked substantial amounts of WHG ancestry, and thus have a much lower (or even zero) WHG/(WHG+ANE) ratio.

So perhaps the 17,000 year-old Afontova Gora 2 (AG2) genome from Central Siberia, classified as part of the ANE meta-population by Lazaridis et al., is genetically the closest sample we have to the Proto-Indo-Europeans? Based on a couple of the PCA from Lazaridis et al. (below) and Raghavan et al. (see here), this genome doesn't appear to be 100% ANE. My very rough estimate is 85/15 ANE/WHG.

If my assumptions are correct here, then it's no wonder that this Bronze Age Danish sample (M4) from the recent Carpenter et al. paper (see here) shows a clear shift towards the Americans on the global PCA. M4 is better known as "the old man" from the giant Borum Eshøj barrow (see here), presumably built by some of the earliest Indo-Europeans in Scandinavia. We can probably expect such Afontova Gora 2-like results from many European samples archeologically linked to the early Indo-Europeans.