The first farmers, with a focus on Anatolia

Background on the Sites and Samples

We first learned about the demographics of the first farmers across the Levant and Zagros from Lazaridis et al. (2016). From that work, we saw that there were at least two independent developments towards agriculture, with two distinct populations (Lazaridis et al., 2016). One was located in the Levant (Natufians), while the other was at Ganj Dareh, in Iran (Lazaridis et al., 2016). We learned from the paper that Natufians and Ganj Dareh were both nearly equal in Basal Eurasian, yet Ganj Dareh had a lot of ancestry from a lineage related to ANE, and Natufians had little to none of this ancestry (Lazaridis et al., 2016).  The Levant Neolithic samples from PPNB to PPNC were a mix of something related to Natufians, and another lineage related to Anatolian farmers from Barcin and Mentese (Mathieson et al., 2015; Lazaridis et al., 2016).

Also in 2016, we caught our first glimpse of the Early Neolithic in Central Anatolia (Kilinc et al., 2016), on the Konya Plain. The Boncuklu group was actually a very early Neolithic group that did have small-scale agriculture (Baird et al., 2012), but was not “aceramic” as many believe. There were very early styles of pottery production and limited use of ceramics at this site; some of the earliest in West Asia (Fletcher et al., 2017). The Boncuklu farmers were a group with long runs of homozygosity, comparable to Western Hunter Gatherers (WHG) (Kilinc et al., 2016). Despite this, the Boncuklu samples are actually quite similar to the later farmers at Barcin, Mentese, and early European farmers (Kilinc et al., 2016), pointing to Anatolia as a potentially third place of independent farming production, along with the Levant and Zagros.

Broushaki et al. (2016) also brought us more samples from the Zagros, that clustered with those samples from Ganj Dareh. These samples were from Tepe Abdul Hosein and Wezmeh Cave (Broushaki et al., 2016).

When it came to actually looking at the ancestral breakdown of Anatolians, Lazaridis et al. (2016) came up with a very solid model where Anatolians were a mix of lineages related to Ganj Dareh, Levant Neolithic, and WHG, with mixture proportions of 0.387, 0.339, and 0.274, respectively.

For the purpose of this post, I am going to focus on Boncuklu and their role in the formation of the Anatolian Neolithic. Since Boncuklu capture method involved shotgun data, I am also using shotgun sequenced genomes from Broushaki et al. (2016) to use as my Iranian farmers (Tepe Abdul Hosein), and the Barcin farmers (Bar8, Bar31). Also, Kostenki14 and MA1 are being used as hunters, as these two form two poles of variation, and are important in the formation of WHG and EHG. Lineages related to both are also important to the formation of these farmers.

In the analysis of the Boncuklu and Barcin farmers, Kilinc et al. (2016) stated that diversity from mixing with other similar groups and/or admixture from more southern groups is possible in the transition from Boncuklu to the later Neolithic of Anatolia. Taking a look at various methods of analysis, it does seem quite possible that a more southern source did admix into the Anatolian farmers of Barcin Hoyuk.

D-stats

First, looking at D-stats, there isn’t anything that is very significant, in determining if Barcin or Boncuklu share more with any particular population. There are near significant results for Natufians and less for the Levant Neolithic samples.

Outgroup	Test	Pop1	Pop2	f4	Z-score	SNPs
Mbuti_DG	Ust_Ishim	Boncuklu	Anatolia_N1	-0.000282	-0.628	1081252
Chimp	Ust_Ishim	Boncuklu	Anatolia_N1	-0.000416	-0.876	1036168
Mbuti_DG	Natufian	Boncuklu	Anatolia_N1	0.00148	2.878	494327
Chimp	Natufian	Boncuklu	Anatolia_N1	0.001535	2.843	472808
Mbuti_DG	Levant_N	Boncuklu	Anatolia_N1	0.000891	2.171	804093
Chimp	Levant_N	Boncuklu	Anatolia_N1	0.000694	1.556	769663
Mbuti_DG	Tepe_Abdul	Boncuklu	Anatolia_N1	-0.000556	-1.448	1003898
Chimp	Tepe_Abdul	Boncuklu	Anatolia_N1	-0.000645	-1.559	962310
Mbuti_DG	Wezmeh	Boncuklu	Anatolia_N1	-0.00025	-0.562	1082448
Chimp	Wezmeh	Boncuklu	Anatolia_N1	-0.000415	-0.882	1037321
Mbuti_DG	CHG	Boncuklu	Anatolia_N1	-0.000361	-0.867	1083101
Chimp	CHG	Boncuklu	Anatolia_N1	-0.000445	-1.031	1037961
Mbuti_DG	Iron_Gates	Boncuklu	Anatolia_N1	-0.000623	-1.69	1076193
Chimp	Iron_Gates	Boncuklu	Anatolia_N1	-0.000712	-1.803	1031346
Mbuti_DG	EHG	Boncuklu	Anatolia_N1	-0.000393	-0.989	995924
Chimp	EHG	Boncuklu	Anatolia_N1	-0.000544	-1.281	954789
Mbuti_DG	W_Siberia	Boncuklu	Anatolia_N1	0.000176	0.404	818559
Chimp	W_Siberia	Boncuklu	Anatolia_N1	0.000181	0.397	783377
Mbuti_DG	Ami	Boncuklu	Anatolia_N1	-0.001048	-1.936	734487
Chimp	Ami	Boncuklu	Anatolia_N1	-0.001122	-1.945	702374

f3-ratio

Also, looking at f3, there isn’t a lot to really support much in the way of admixture from Levant or Natufians into the Barcin Hoyuk samples.

Pop1	Pop2	Target	f3	std error	Z-score	SNPs
Boncuklu	Tepe_Abdul	Anatolia_N1	0.004	0.002013	1.987	494742
Boncuklu	Wezmeh	Anatolia_N1	0.002506	0.002218	1.13	486870
Boncuklu	Levant_N	Anatolia_N1	-0.000089	0.001916	-0.047	382114
Boncuklu	Tepecik	Anatolia_N1	0.004217	0.00194	2.174	361064
Boncuklu	Natufian	Anatolia_N1	-0.001456	0.00235	-0.619	222845

qpAdm Models

Switching over to qpAdm, the first thing to check is what Boncuklu is best modeled as. For the right populations the simplest and seemingly best for using Kostenki, Natufians, and Tepe_Abdul_Hosein_N as the left pops turned out to be Mbuti_DG, Ust_Ishim, GoyetQ116-1, Vestonice16, MA1, Ami, Onge, and Karitiana.

probability	Natufian	Tepe_Abdul	Kostenki14	std error	std error	std error
0.236	0.232	0.449	0.32	0.142	0.107	0.074

While the probability and standard errors are a little high, it is hard to keep them any lower. While adding something like Ganj_Dareh_N or Levant_N will reduce the standard error, the probability also dips significantly.

Moving onto Anatolia_N1 (Bar31, Bar8), the right pops, or outgroups became Mbuti_DG, Ust_Ishim, Kostenki14, MA1, Natufian, Tepe_Abdul_Hosein_N, Ami, Onge, Karitiana.

probability	Boncuklu	Levant_N	std error	std error
0.815	0.687	0.313	0.064	0.064

Also, swapping Natufian and Levant_N between left and right populations:

probability	Boncuklu	Natufian	std error	std error
0.302	0.771	0.229	0.071	0.071

qpGraph Models

After seeing some decent fits in qpAdm, I tried to use qpGraph and see how easily they could be replicated. To being with, I started with a simple tree that contained just the Natufians and Tepe_Abdul_Hosein_N. The Interestingly, CHG comes out as a mix of Natufian, Tepe_Abdul_Hosein, and MA1. first tree should what would be expected, with Natufians and Tepe_Abdul_Hosein_N having similar amounts of Basal Eurasian, and with Iranian farmers having ancestry from a lineage related to MA1.

6/4/18 New Update: Simple qpGraphs

The purpose of these simple graphs is to avoid a lot of admixture edges and to show the steps in finding the ancestry of Boncuklu, Levant, and Barcin samples. Samples that are not absolutely needed, such as Ust_Ishim, MA1, CHG, and any ENA population will be left out. This also shows the work process in how I find where to look at for admixture events.

The first graph here shows that by branching Boncuklu off of Natufian, there is a significant Z-score with Iranian farmers. The next step will be to branch from Iranians to Boncuklu.

 

 

The next output had a worst Z-score that involved Kostenki, meaning that an admixture line from the Kostenki branch will be the next admixture source into Boncuklu.

After adding the admixture edge from near Kostenki, Boncuklu comes out mostly Kostenki-related, with the rest being a near even mix of Tepe Abdul Hosein and Natufian.

This created a pretty solid fit, so, the next step would be to see how the graph works with Adding Levant_N. Since the Levant samples harbor a lot of Natufian-related ancestry, that is where they will first be branched off of.

This output shows that an admixture edge is needed from a node on the Boncuklu line to Levant_N in order to drop the Z-score. A shared drift edge with Boncuklu was needed to move the worst Z-score closer to 2.

For the next run, I added the two shotgun samples from Barcin (Bar8, Bar31) to the tree.This left a worst Z > 3, so, the next step will be an edge from a node related to Natufians into Anatolia_N1.

For the last graph, the edge from a Natufian-related node admixes into the Barcin Hoyuk samples. There remains a worst Z > 2.5, that appears to be asking for Iranian admixture into the Levant_N population. While it isn’t necessary, I did look at adding an admixture edge from the Iranian farmers and it only added 2% admixture. This graph should be sufficient enough for a simple tree that incorporates all of the first farmers.

Below are more complex graphs, with an Eastern non-African source (Onge), Ust-Ishim and also MA-1.

For the next step, I also added CHG to the tree, just to see where it was placed and to add another admixture source in West Asia. As other stats show, CHG was less basal than Iranian farmers, but also shifted towards Natufians, potentially sharing a formation story with Boncuklu.

After achieving a good fit with CHG, I then went on to see just how Boncuklu fit into the picture. As expected, Boncuklu is best fit as a mix of Natufian, Iranian, and a lineage related to Kostenki14, with admixture also from CHG. Trying to do just Iranian or just CHG led to significantly higher Z-scores. Both were needed. It appears there is additional ANE in Boncuklu that is not accounted for in Iranians.

After applying a good fit for Boncuklu, I next wanted to see just how the farmers at Barcin Hoyuk fit into tree. For this, I first created a new simplified tree to use and also co-fit the Levant_N samples to see if the best fit includes additional southern ancestry into the later Northwest Anatolian Neolithic.

I also tried a run with just transversion sites, just to see how that would turn out.

Just as a precaution, I ran another test, with Levant_N receiving admixture from a separate branch related to Boncuklu, that is ancestral to Anatolia_N1, to see if that reduced or eliminates the need for admixture from Levant_N. As it turns out, Anatolia then asked first for admixture from Natufians, rather than Levant_N. So, the question then becomes, was there another population that was related to Levant_N which lacked or had less Iranian farmer ancestry and mixed into the ancestors of the Barcin and European farmers? More samples, specifically from southern and especially southeast Anatolia, and PPNA will provide the answers which are needed.For argument sake, I tried both the Natufian and Levant_N admixture edges to see how they turned out. Having more a higher f4 and f3 with Natufians, rather than Levant, leads me to believe this is possible.

Conclusions

The end result here has the Barcin samples as a mix of Boncuklu and Levant_N (or a more Natufian-like group), in agreement with qpAdm. However, this output is only about two-thirds as much Levantine flow into later Anatolian farmers as qpAdm showed. Transversion sites placed more ancestry related to Iranian farmers into both Boncuklu and Levant_N. Boncuklu became significantly more Iranian than the previous run. The amount of flow from Levant_N into Barcin held pretty steady. While more samples across time and space, with similar capture methods, will help this to be resolved. There aren’t much in the way of D-stats or f3_ratio to suggest there is a lot of gene flow there. qpAdm and qpGraph do provide a strong case of gene-flow into Northwest Anatolia.

In upcoming posts I will look at the formation of European farmers, hunter admixture into the Middle Neolithic, and also at the roots of Epipaleolithic and Mesolithic European hunters.

 

References

Baird D. The Late Epipaleolithic, Neolithic, and Chalcolithic of the Anatolian Plateau, 13,000–4000 BC. In: Potts D., editor. A Companion to the Archaeology of the Ancient Near East. Wiley-Blackwell; 2012. pp. 431–466.

Broushaki F., Thomas M.G., …, Berger J. Early Neolithic genomes from the eastern Fertile Crescent. Science. 2016; 353: pp. 499-503.

Fletcher A., Baird D., Spataro M. & Fairbairn A. Early ceramics in Anatolia: Implications for the production and use of the earliest pottery. The evidence from Boncuklu Hoyuk. Cambridge Archaeological Journal. 2017; 27 (2): pp. 351-369.

Kilinc G.M., Omrak A., …, Gotherstrom A. The demographic development of the first farmers in Anatolia. Current Biology. 2016;  26(19): pp. 2659–2666.

Lazaridis I., Nadel D., …, Reich D. Genomic insights into the origin on farming in the ancient Near East. Nature. 2016;536: pp. 419-424.

Mathieson I., Lazaridis I., Rohland N., Mallick S., Patterson N., Roodenberg S.A., Harney E., Stewardson K., Fernandes D., Novak M. Genome-wide patterns of selection in 230 ancient Eurasians. Nature. 2015;528:499–503. [PubMed]