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Pakistanis of Greeks and Macedonians Descent

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Genome-wide genotype and sequence-based reconstruction of the 140,000 year history of modern human ancestry

Scientific Reports


doi:10.1038/srep06055
Received

05 March 2014
Accepted

28 July 2014
Published

13 August 2014

We investigated ancestry of 3,528 modern humans from 163 samples. We identified 19 ancestral components, with 94.4% of individuals showing mixed ancestry. After using whole genome sequences to correct for ascertainment biases in genome-wide genotype data, we dated the oldest divergence event to 140,000 years ago. We detected an Out-of-Africa migration 100,000–87,000 years ago, leading to peoples of the Americas, east and north Asia, and Oceania, followed by another migration 61,000–44,000 years ago, leading to peoples of the Caucasus, Europe, the Middle East, and south Asia. We dated eight divergence events to 33,000–20,000 years ago, coincident with the Last Glacial Maximum. We refined understanding of the ancestry of several ethno-linguistic groups, including African Americans, Ethiopians, the Kalash, Latin Americans, Mozabites, Pygmies, and Uygurs, as well as the CEU sample. Ubiquity of mixed ancestry emphasizes the importance of accounting for ancestry in history, forensics, and health.

Methodology
:


We collected genome-wide genotype data on autosomal single nucleotide polymorphisms (SNPs) from publicly available human genomic diversity projects. The global data set included 916 individuals from the Human Genome Diversity Project, 1,092 individuals from the 1000 Genomes Project, 222 individuals from east Africa, 268 individuals from the Singapore Genome Variation Project, 75 individuals from Lebanon, 145 individuals from north Africa and the Basque Country, 323 individuals from south Africa, 18 Arabs from Qatar, 106 individuals from west and central Africa, 133 Maasai from the International HapMap Project, and 462 individuals from a study of the Jewish Diaspora. Data management and quality control were performed using PLINK version 1.07. Graphics were generated using R. Maps were drawn using the R libraries maps and plotrix.


Individuals or markers with genotyping call rates < 95% were excluded. We also removed individuals identified as identical samples, 1st degree relatives, or 2nd degree relatives. After quality control, the global data set comprised 3,528 individuals from 163 samples. The mutual intersection of all data sets yielded 19,372 diallelic, autosomal SNPs with experimentally determined genotypes (i.e., no imputation of missing genotypes was performed). The genotyping call rate in the remaining individuals was 99.8%. The average distance between markers was 142.8 kb (135.4 kb excluding centromeres). Due to very small sample sizes for some samples, no additional pruning of markers based on linkage disequilibrium was performed.

Results:

srep06055-f5.jpg



This was published in nature last year...

The study did statistical analysis on already available data.....


Another statistical analysis published in Science this year

The Kalash Genetic Isolate: Ancient Divergence, Drift, and Selection





    • 1 Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1SA, UK
    • 2 Institute for Maternal and Child Health, IRCCS Burlo Garofolo, University of Trieste, 34137 Trieste, Italy
    • 3 Division of Biological Anthropology, University of Cambridge, Cambridge CB2 1QH, UK
    • 4 Section of Biochemistry, Shifa College of Medicine, Shifa Tameer-e-Millat University, Sector H-8/4, Islamabad 44000, Pakistan
    • 5 Department of Human Genetics & Molecular Biology, University of Health Sciences, Lahore 54000, Pakistan
    • 6 Centre for Human Genetics and Molecular Medicine, Sindh Institute of Urology and Transplantation, Karachi, 74200, Pakistan
Received 9 February 2015, Accepted 26 March 2015, Available online 30 April 2015
The Kalash represent an enigmatic isolated population of Indo-European speakers who have been living for centuries in the Hindu Kush mountain ranges of present-day Pakistan. Previous Y chromosome and mitochondrial DNA markers provided no support for their claimed Greek descent following Alexander III of Macedon's invasion of this region, and analysis of autosomal loci provided evidence of a strong genetic bottleneck. To understand their origins and demography further, we genotyped 23 unrelated Kalash samples on the Illumina HumanOmni2.5M-8 BeadChip and sequenced one male individual at high coverage on an Illumina HiSeq 2000. Comparison with published data from ancient hunter-gatherers and European farmers showed that the Kalash share genetic drift with the Paleolithic Siberian hunter-gatherers and might represent an extremely drifted ancient northern Eurasian population that also contributed to European and Near Eastern ancestry. Since the split from other South Asian populations, the Kalash have maintained a low long-term effective population size (2,319–2,603) and experienced no detectable gene flow from their geographic neighbors in Pakistan or from other extant Eurasian populations. The mean time of divergence between the Kalash and other populations currently residing in this region was estimated to be 11,800 (95% confidence interval = 10,600−12,600) years ago, and thus they represent present-day descendants of some of the earliest migrants into the Indian sub-continent from West Asia.


Material and Methods

DNA Samples and Genotyping
The Kalash samples were collected from three valleys in the Hindu Kush mountain ranges in northwest Pakistan (Figure 1A). In accordance with the Declaration of Helsinki, the samples were collected after informed consent was obtained, and the study was approved by all relevant institutional ethics committees. Lymphoblastoid cell lines were established from all collected blood samples, and some (n = 25) were deposited with CEPH; these latter samples form part of the South and Central Asian collection of the HGDP-CEPH cell-line panel. We used 10 of these and an additional 13 samples that are not in the collection for our analysis. All of these unrelated (n = 23) Kalash males were genotyped on the Illumina HumanOmni2.5M-8 BeadChip with 200 ng of DNA (26 ng/μl) prepared from these lymphoblastoid cell lines. Genotyping calls and quality control (QC) were performed with GenoSNP9 by the Sanger Institute’s core genotyping facility. Genotypes were called only for samples passing Sequenom genetic fingerprinting and gender concordance. These were run through the standard QC pipeline. All 23 samples passed a call-rate threshold of 95% and were used in the downstream analysis. Genotyping quality was assessed by comparison of 178,072 SNPs that overlapped the Illumina 650,000 K SNP chip. Ten of the Kalash samples analyzed in this study had also been genotyped on this platform, and the sample genotype concordance was 99.999%. Comparative data were obtained from 35 populations representing Africa, Europe, Caucasus, and West, Central, East, and South Asia (Table S1)

1-s2.0-S0002929715001378-gr1.jpg


Results

The Kalash Are a Genetic Isolate
PCA using only Eurasian and South Asian populations separated the populations from Europe, Caucasus, and West Asia from East Asians in the first component and from South and Central Asians in the second component; Central Asians lay closer to the Sherpa from Nepal and CHB from East Asia. The Kalash samples clustered together as an outlier population to the other South Asian samples from India and Pakistan (Figure 1B). The Kalash genetic isolation was also supported by the ADMIXTURE plot (Figure 1C), in which the lowest cross-validation error was achieved with seven ancestry components. In this analysis, the Kalash were characterized mainly by a unique genetic component (dark green), although many samples shared a proportion of their ancestry with their neighbors in Pakistan (light orange and light blue). This light-blue component was also shared among many diverse populations from West, Central, and South Asia.


May I ask, in that branch diagram what is the significance of it, if you can explain. Thanks!
 
.
Genome-wide genotype and sequence-based reconstruction of the 140,000 year history of modern human ancestry

Scientific Reports


doi:10.1038/srep06055
Received

05 March 2014
Accepted

28 July 2014
Published

13 August 2014

We investigated ancestry of 3,528 modern humans from 163 samples. We identified 19 ancestral components, with 94.4% of individuals showing mixed ancestry. After using whole genome sequences to correct for ascertainment biases in genome-wide genotype data, we dated the oldest divergence event to 140,000 years ago. We detected an Out-of-Africa migration 100,000–87,000 years ago, leading to peoples of the Americas, east and north Asia, and Oceania, followed by another migration 61,000–44,000 years ago, leading to peoples of the Caucasus, Europe, the Middle East, and south Asia. We dated eight divergence events to 33,000–20,000 years ago, coincident with the Last Glacial Maximum. We refined understanding of the ancestry of several ethno-linguistic groups, including African Americans, Ethiopians, the Kalash, Latin Americans, Mozabites, Pygmies, and Uygurs, as well as the CEU sample. Ubiquity of mixed ancestry emphasizes the importance of accounting for ancestry in history, forensics, and health.

Methodology
:


We collected genome-wide genotype data on autosomal single nucleotide polymorphisms (SNPs) from publicly available human genomic diversity projects. The global data set included 916 individuals from the Human Genome Diversity Project, 1,092 individuals from the 1000 Genomes Project, 222 individuals from east Africa, 268 individuals from the Singapore Genome Variation Project, 75 individuals from Lebanon, 145 individuals from north Africa and the Basque Country, 323 individuals from south Africa, 18 Arabs from Qatar, 106 individuals from west and central Africa, 133 Maasai from the International HapMap Project, and 462 individuals from a study of the Jewish Diaspora. Data management and quality control were performed using PLINK version 1.07. Graphics were generated using R. Maps were drawn using the R libraries maps and plotrix.


Individuals or markers with genotyping call rates < 95% were excluded. We also removed individuals identified as identical samples, 1st degree relatives, or 2nd degree relatives. After quality control, the global data set comprised 3,528 individuals from 163 samples. The mutual intersection of all data sets yielded 19,372 diallelic, autosomal SNPs with experimentally determined genotypes (i.e., no imputation of missing genotypes was performed). The genotyping call rate in the remaining individuals was 99.8%. The average distance between markers was 142.8 kb (135.4 kb excluding centromeres). Due to very small sample sizes for some samples, no additional pruning of markers based on linkage disequilibrium was performed.

Results:

srep06055-f5.jpg



This was published in nature last year...

The study did statistical analysis on already available data.....


Another statistical analysis published in Science this year

The Kalash Genetic Isolate: Ancient Divergence, Drift, and Selection





    • 1 Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1SA, UK
    • 2 Institute for Maternal and Child Health, IRCCS Burlo Garofolo, University of Trieste, 34137 Trieste, Italy
    • 3 Division of Biological Anthropology, University of Cambridge, Cambridge CB2 1QH, UK
    • 4 Section of Biochemistry, Shifa College of Medicine, Shifa Tameer-e-Millat University, Sector H-8/4, Islamabad 44000, Pakistan
    • 5 Department of Human Genetics & Molecular Biology, University of Health Sciences, Lahore 54000, Pakistan
    • 6 Centre for Human Genetics and Molecular Medicine, Sindh Institute of Urology and Transplantation, Karachi, 74200, Pakistan
Received 9 February 2015, Accepted 26 March 2015, Available online 30 April 2015
The Kalash represent an enigmatic isolated population of Indo-European speakers who have been living for centuries in the Hindu Kush mountain ranges of present-day Pakistan. Previous Y chromosome and mitochondrial DNA markers provided no support for their claimed Greek descent following Alexander III of Macedon's invasion of this region, and analysis of autosomal loci provided evidence of a strong genetic bottleneck. To understand their origins and demography further, we genotyped 23 unrelated Kalash samples on the Illumina HumanOmni2.5M-8 BeadChip and sequenced one male individual at high coverage on an Illumina HiSeq 2000. Comparison with published data from ancient hunter-gatherers and European farmers showed that the Kalash share genetic drift with the Paleolithic Siberian hunter-gatherers and might represent an extremely drifted ancient northern Eurasian population that also contributed to European and Near Eastern ancestry. Since the split from other South Asian populations, the Kalash have maintained a low long-term effective population size (2,319–2,603) and experienced no detectable gene flow from their geographic neighbors in Pakistan or from other extant Eurasian populations. The mean time of divergence between the Kalash and other populations currently residing in this region was estimated to be 11,800 (95% confidence interval = 10,600−12,600) years ago, and thus they represent present-day descendants of some of the earliest migrants into the Indian sub-continent from West Asia.


Material and Methods

DNA Samples and Genotyping
The Kalash samples were collected from three valleys in the Hindu Kush mountain ranges in northwest Pakistan (Figure 1A). In accordance with the Declaration of Helsinki, the samples were collected after informed consent was obtained, and the study was approved by all relevant institutional ethics committees. Lymphoblastoid cell lines were established from all collected blood samples, and some (n = 25) were deposited with CEPH; these latter samples form part of the South and Central Asian collection of the HGDP-CEPH cell-line panel. We used 10 of these and an additional 13 samples that are not in the collection for our analysis. All of these unrelated (n = 23) Kalash males were genotyped on the Illumina HumanOmni2.5M-8 BeadChip with 200 ng of DNA (26 ng/μl) prepared from these lymphoblastoid cell lines. Genotyping calls and quality control (QC) were performed with GenoSNP9 by the Sanger Institute’s core genotyping facility. Genotypes were called only for samples passing Sequenom genetic fingerprinting and gender concordance. These were run through the standard QC pipeline. All 23 samples passed a call-rate threshold of 95% and were used in the downstream analysis. Genotyping quality was assessed by comparison of 178,072 SNPs that overlapped the Illumina 650,000 K SNP chip. Ten of the Kalash samples analyzed in this study had also been genotyped on this platform, and the sample genotype concordance was 99.999%. Comparative data were obtained from 35 populations representing Africa, Europe, Caucasus, and West, Central, East, and South Asia (Table S1)

1-s2.0-S0002929715001378-gr1.jpg


Results

The Kalash Are a Genetic Isolate
PCA using only Eurasian and South Asian populations separated the populations from Europe, Caucasus, and West Asia from East Asians in the first component and from South and Central Asians in the second component; Central Asians lay closer to the Sherpa from Nepal and CHB from East Asia. The Kalash samples clustered together as an outlier population to the other South Asian samples from India and Pakistan (Figure 1B). The Kalash genetic isolation was also supported by the ADMIXTURE plot (Figure 1C), in which the lowest cross-validation error was achieved with seven ancestry components. In this analysis, the Kalash were characterized mainly by a unique genetic component (dark green), although many samples shared a proportion of their ancestry with their neighbors in Pakistan (light orange and light blue). This light-blue component was also shared among many diverse populations from West, Central, and South Asia.

Your one of the brightest people here. I am sure most will agree PDF would be poorer with you. Thank god your a Pakistani or else like in that other thread you would have me stumped ...

Trust me that does not happen often ..
 
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This is what I was thinking but I didn't say it. :lol:
This is just BS.. Now one can explain the beliefs of Kalash people (brainwashing by bunch of Greek oldies living in the region n the Greeks suckin upto them.

But people of Gilgit Baltistan? Hunza ? WTF!!

Those ppl have more in common with Central Asia n Tibet n Pakistan than greeks!! Those are proud ppl.

Although now I even see Greeks commenting on Pak pics with their retarded Greek bloodline BS!
 
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All the sources which are found on internet are pro greek and macedonian which people call quality source or valid or neutral. This is known as creating inferior complex through print media and information war. Porus (Purshottam) would have either died or given his territory to Mecedonians if was defeated. Who the hell is Alexander to judge the bravery. Well Alexander fought with bravery hence Porus(Purshottam) granted him life by allowing him safe escape. On his return.

And I am bloody damn sure, had he won the battle he would have left with nothing to fight with other empires. And his escape would have been made much more tougher.



it's on youtube, you will miss it those who can watch do see.
@18:03
he said these greek kaffirs lineage of selucus the greek general are living in chitral part of kafiristan and left with few days only to survive, etc etc etc

OMG hamare AAJ TAK koh tumne AAP TAK banadiya logo bhi nahi choda
 
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May I ask, in that branch diagram what is the significance of it, if you can explain. Thanks!
What does the dendogram/ phylogenetic tree look like?

Your one of the brightest people here. I am sure most will agree PDF would be poorer with you. Thank god your a Pakistani or else like in that other thread you would have me stumped ...

Trust me that does not happen often ..
No I just happen to be in the field (genetics) and I do get pissed when people misinterpret it :unsure:
 
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This does not make them Greek or European.
I know, i was just asking to see if this was in fact credible.
I was always curious on this topic, i was always joked on being a 'gora' all my life, lolz .
some members on this thread have reached a new low.
 
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What does the dendogram/ phylogenetic tree look like?


No I just happen to be in the field (genetics) and I do get pissed when people misinterpret it :unsure:

I could not understand, how Kalash and Indian are coming out of similar branch and are close with Caucasians? How is that possible?
 
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That is Hindutva theory and rest of the world agree in Ukraine as the homeland of the Aryans. You are not Pakistani remove your false flag.

In wikipedia, the term aryan is described as this.. more I don't know. Indians can put more light. Secondly I am not talking about religion, I am talking about the ancient name of the region, to clarify that world knows ancient India ( India , Pakistan, Bangladesh), later Indians took this name of India. That's why India has different names in different languages.
Aryan - Wikipedia, the free encyclopedia
 
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I could not understand, how Kalash and Indian are coming out of similar branch and are close with Caucasians? How is that possible?
1stly, the article is based on autosomes
2ndly, it is statistical analysis showing generation breakdown....

Statistics is a good tool to play with...You feed it x it will give you x results, you feed it y it will give you y results...You feed it xy it will give you xy results...
Article says:
Here, we meta-analyzed ancestry from 12 global and regional diversity projects....So it is basically large sets of pooled data...

2ndly they "estimated the divergence times of the ancestral components and compared these divergence times to historical records. "

In wikipedia, the term aryan is described as this.
You can edit wikipedia anyone can....it isnt science!
 
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1stly, the article is based on autosomes
2ndly, it is statistical analysis showing generation breakdown....

Statistics is a good tool to play with...You feed it x it will give you x results, you feed it y it will give you y results...You feed it xy it will give you xy results...
Article says:
Here, we meta-analyzed ancestry from 12 global and regional diversity projects....So it is basically large sets of pooled data...

2ndly they "estimated the divergence times of the ancestral components and compared these divergence times to historical records. "

Thanks

so in conclusion, what is the basic difference between, europeans, we Pakistanis(north) and North Indians and Arabs? I seriously want know now after being termed as 'Indian', shamefuly.

You can edit wikipedia anyone can....it isnt science!

I know, that, and this is the reason I do not agree to most of things I read on Internet as they are politically motivated and edited. Specially the History because, it is never sure. hence I do not accept that Porus lost against Alexander as widely accepted.
 
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Thanks

so in conclusion, what is the basic difference between, europeans, we Pakistanis(north) and North Indians? I seriously want know now after being termed as 'Indian', shamefuly.
Read the article! If you can read wiki you can read the article...

They used autosomal DNA meaning maternal and paternal mixed...used statistics meaning probability meaning if you are more similar to something you will be linked together (EVEN if you maybe different)..So go be shameful somewhere else!

now after being termed as 'Indian', shamefuly.
I am surprised you completely IGNORED the 2nd study
 
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I read it :)
the whole word is dependant on each other when it comes to DNA. It's all mixed.
We share 99% DNA with a chimp....but we are considered human...Looking for differences in that 1% that is non chimp DNA is not easy nor do we have absolute computer power to tweak all the confusions!
 
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