The results of studies in Quaternary geology, archeology, paleoanthropology and human genetics demonstrate that the ancestors of Native Americans arrived in mid-latitude North America mainly along the Pacific Northwest Coast, but had previously inhabited the Arctic and during the last glacial maximum were in a refugium in Beringia, a land bridge connecting Eurasia and North America. The gene pool of Native Americans is represented by unique haplogroups of mitochondrial DNA and the Y chromosome, the evolutionary age of which rangesfrom 13 to 22 thousand years. The results of a paleogenomic analysis also show that during the last glacial maximum Beringia was populated by human groups that had arisen as a result of interaction between the most ancient Upper Paleolithic populations of Northern Eurasia and newcomer groups from East Asia. Approximately 20 thousand years ago the Beringian populations began to form, and the duration of their existence in relative isolation is estimated at about 5 thousand years. Thus, the adaptation of the Beringians to the Arctic conditions could have taken several millennia. The adaptation of Amerindian ancestors to high latitudes and cold climates is supported by genomic data showing that adaptive genetic variants in Native Americans are associated with various metabolic pathways: melanin production processes in the skin, hair and eyes, the functioning of the cardiovascular system, energy metabolism and immune response characteristics. Meanwhile, the analysis of the existing hypotheses about the selection of some genetic variants in the Beringian ancestors of the Amerindians in connection with adaptation to the Arctic conditions  shows the ambiguity of the testing results, which may be due to the loss of some traces of the “Beringian” adaptation in the gene pools of modern Native Americans. The most optimal strategy for further research seems to be the search for adaptive variant Interdisciplinary research is important in examining issues relatedto the role of Beringia as an Arctic land bridge in the settlementof the Western Hemisphere. Unfortunately, the floodingof the lowlands of Beringia by rising sea levels 12,000 yearsago led to the loss of a large number of archaeological sites,including possible human settlements. As a result, the mainhypotheses associated with the history of the peopling of theAmericas are based primarily on a synthesis of archaeological,geological, and biological data from the more accessibleregions of Asia and the Americas . According to one model ofAmerican settlement, ancient people from Northeast Asiamigrated to the Americas via Beringia along an ice-freecorridor in what is now Western Canada, which was createdby melting glaciers at the end of the Pleistocene. Accordingto another scenario, Native American ancestors activelydeveloped the coastal zone and migrated to northwest NorthAmerica along the southern coast of Beringia .However, both of these scenarios appear to underestimatethe more complex role that Beringia played in the settlementof the Americas by human populations .Results of new geochronological studies suggest that the“Canadian Corridor” was blocked by ice during presumedhuman migrations ~15–16 thousand years ago and was onlyfreed ~13.8 thousand years ago . Therefore,it seems more likely that the migration route is the northwestcoast of America . Since the melting ofthe glaciers in these territories could have occurred as earlyas 18,000 years ago, the coastal migration route could havebeen developed by the first Americans ~17,000 years ago.At the same time, the results of genomic studies indicate thatthe ancestors of the Native Americans appear to have lived inthe Beringian Arctic for quite a long time and survived the lastglacial maximum there, which terminated ~19,000 years ago. It is assumed that thesepopulations adapted to the harsh conditions of the Far Northwhile living in Beringia, and as the climate warmed, Beringiandescendants migrated to Northwest AmericaDue to the rapid development of DNA sequencing techniques,the genomic approach is widely used in modern research inarchaeology and paleoanthropology. Genetic studies of theindigenous populations of Siberia have shown that the mostancient genetic components, which were present as early as30–40 thousand years ago in the most ancient inhabitantsof both southern and northern Siberia, were later lost as aresult of genetic drift and population replacement periods.Thus, genomic analysis of the ancient populations of Siberia hasshown that in Siberia there were several periods of almostcomplete population replacements . Themost ancient Late Paleolithic population of Siberia was replaced20–11 thousand years ago by newcomers from EastAsia, resulting in the formation of Siberian populations thatgave rise to the ancestors of both Paleosiberians and NativeAmericans. In the time interval from 11 to 4 thousand yearsago, this population was partially replaced by other groups ofEast Asian origin, which gave rise to the modern populationsof Siberia. Additional gene flows in both directions alongthe Bering Strait were also reported at this time. These flowsaffected the genetic structure of Eskimo and Na-Dene populations.The results of paleogenomic studies have shown that theformation of ancestral populations to ancient peoples ofnorthern Siberia and America could have occurred in a widearea from the Trans-Baikal region to Beringia . Paleogenomic dataindicate that during the Holocene the populations of NortheastAsia  werevery dynamic in contrast to the Trans-Baikal populations,which demonstrate genetic continuity from the Mesolithicto the Bronze Age . Evidence was alsoobtained that bearers of the Belkachin archaeological culturein Yakutia can be considered ancestors of the Paleo-Eskimos,who migrated to the Americas about 5–6 thousand years ago.Population studies of polymorphism of mitochondrial DNA(mtDNA) and Y-chromosome, which are inherited through thematernal and paternal lineages, respectively, have developed greatly over the past 40 years. The results of molecular datingof the evolutionary age of the mtDNA and Y-chromosomehaplogroups distributed among the modern indigenous populationsof Siberia demonstrate that the most ancient geneticlineages in the south of Siberia belong to the postglacialperiod , while in the northeast ofSiberia their evolutionary age does not exceed 10,000 years.Genetic data for the present-day populations of Siberia arethus inconsistent with archaeological and paleoanthropologicaldata showing the presence of Homo sapiens both in thesouth and in the north of Siberia as far back as 40–45 thousandyears ago ,but suggest that due to genetic drift, the most ancient geneticlineages were lost in the modern Siberian population . This assumptionwas completely confirmed by the results of the paleogenomicstudy of the Siberians, which showed, as noted above, that inSiberia there have been several periods of almost completereplacements of the earlier inhabitants .The genetic data obtained thus do not contradict the hypothesisthat, during the last glacial maximum, the Arctic partof Asia and America  could have beeninhabited by humansBased on the mtDNA variability data, it has been establishedthat the divergence between the ancestors of the Siberian andNative American populations occurred ~25,000 years ago,the evolutionary age of Native American founding mtDNAlineages is approximately 18.5 thousand years, and theirspread across Americas began ~16,000 years ago . The mitochondrial gene pool of Native Americansis represented by only seven mtDNA haplogroups , the evolutionary age of whichranges from 13 to 20 thousand years ago . All of the Native AmericanmtDNA haplogroups are unique, and for six of them (exceptD4h3a) there are still no analogues found anywhere else inthe world, which means that these haplogroups were formedin deep isolation and, apparently, while living in BeringiaFor the haplogroup D4h3a, a related D4h3b-haplotype wasfound in modern Chinese, which diverged from the commonD4h3 ancestor by ~18.3 thousand years, and the evolutionaryage of D4h3a itself is ~13 thousand years .It is also noteworthy that in all cases (except X2a) the NativeAmerican mtDNA haplogroups are derived from East Asianancestors within haplogroups A, B, C, and D, predominantlydistributed in East Asia. Meanwhile, the area of haplogroup Xis located in Western Eurasia, and therefore the appearance ofhaplogroup X2a in the Beringians (and thus in the ancestorsof the Amerindians) could have happened on the basis of thegene pool of their Upper Paleolithic ancestors, related to thepopulation of Western Eurasia. This follows from paleogenomicdata showing that the Upper Paleolithic populationsof Siberia and Eastern Europe (in the time range from 18 to45.5 thousand years ago) were characterized by approximatelythe same set of mtDNA haplogroups of West Eurasian origin . Thus, it is notexcluded that the mtDNA X-haplotypes, which emerged onthe basis of haplogroup X2a, were also common in the genepool of this ancient population. The age of haplogroup X2ais approximately 13,000 years , and itsrange is limited to northwestern North America only .Haplogroup C4c has a similar geographic distribution andevolutionary age (~14 thousand years) . This appears to indicate thatthe carriers of haplogroups X2a and C4c were among thelast Beringians who populated North America. Around thesame time, but along the Pacific coast, haplogroup D4h3apresumably spread to the south of America . Thus, the results of mtDNA polymorphism studiesindicate that both pathways (both the “Canadian Corridor”and the Pacific Coast of America) may have been used in thesettlement of the Americas.The results of studies of Y-chromosome variability alsoshowed that the male gene pool of Native Americans is characterizedby a small set of genetic lineages – these are haplogroupsQ-M3, Q-M848 and Q-Z780, widespread among NativeAmericans, as well as the more rare haplogroups C-P39and Q-Y4276, typical of Native North Americans, and haplogroupC-MPB373, found in Native South Americans . The evolutionary age ofthese Y-haplogroups ranges from 15.0 to 21.6 thousand years,indicating that they originated in Beringia. This hypothesis issupported by the discovery of the rare haplogroup Q-L804,related to the Native American haplogroup Q-M3, in populationsof Northwestern Europe . Since the divergence time between these haplogroupsis ~17.3 thousand years, it is assumed that migrations fromBeringia took place not only in the direction of America, butalso in the opposite direction to Europe .In addition, it should be noted that according to mtDNAstudies, very rare haplogroups C1e and C1f, related to NativeAmerican C1 haplogroups , were found inIcelanders as well as in the Mesolithic population of Karelia(Yuzhnyi Oleniy Island) . Thus, these data allow us to believethat in Beringia there was not a brief stop of human populationsthat migrated from the south, but indeed the Beringianpopulations, with their own genetic characteristics and uniquemtDNA and Y-chromosome haplogroups, were formed thereover a long time.The question about the duration of the Beringian Standstillis also of great interest and is regularly studied. One ofthe first genetic studies of this problem concluded that theBeringian Standstill can be estimated at ~15,000 years, if westart from the first discoveries of Homo sapiens remains inthe Asian part of the Arctic ~30,000 years ago and until themoment when the Beringian expansion to America took place,i. e. ~15,000 years ago according to genetic data . M. Raghavan et al. (2015) showed based on theresults of a paleogenomic study that the upper boundary ofdivergence between the ancestors of the Native Americansand East Asians should be considered as ~23 thousand years In that case, the isolation period in Beringia  was ~8,000 years. Estimatesof the duration of the Beringian Standstill obtained by usingthe analysis of ancient mitogenomes demonstrated that, givenvarious kinds of uncertainties, the isolation period of the Beringianpopulation could have ranged from 2.4 to 9.0 thousandyears .Such a long stay in Beringia suggests that its population wasquite large in number and subdivided within Beringia . It is assumedthat the western part of Beringia (Chukotka) was inhabited byPaleosiberian populations, while the eastern part of Beringia(Alaska and Yukon) was inhabited in the south by the NativeAmerican ancestors and in the north by a separate group ofancient Beringians who disappeared or were assimilated bynorthern Native American tribes . Both of thesegroups of the ancient Eastern Beringians were very closelyrelated, since they showed the C1b and B2 mitochondrialhaplogroups typical of the Native Americans . Moreover, at present thesehaplogroups are more characteristic of the southern NativeAmerican populations, but in the past they were present inthe north of Eastern Beringia, thus confirming the BeringianStandstill hypothesis .It is likely that during the glacial maximum, the southernpart of Beringia was a refugium, characterized by a fairly mildclimate, rich biota, and therefore quite suitable for settlementof ancient people .According to the results of paleogenomic data modeling,the divergence between the Paleosiberians and the populationthat gave rise to the Beringians occurred approximately24 thousand years ago, but the Upper Paleolithic geneticcomponent was obtained by both populations approximately20 thousand years ago (in the ancient Beringians its fractionwas ~18 %) . Thus, it is assumed thatthe most ancient Upper Paleolithic peoples were actuallypresent in Beringia during the last glacial maximum, andthat the formation of Beringians, including Native Americanancestors, began through interaction with the newcomer EastAsia-related peoples ~20 thousand years ago . In this scenario, the time of existence of the relativelyisolated Beringian population is estimated to be ~5,000 years.A study of Y-chromosome polymorphism in Native Americanpopulations also showed that the duration of the BeringianStandstill may not have exceeded 4,600 years . Thus, the results of genomic studies suggest that theadaptation to the Arctic conditions of the ancient populationsthat took refuge in Beringia may have taken several millennia.According to some genetic studies, it is proposed that the genomesof the Native Americans may contain signals of adaptationto the Arctic conditions, which emerged during theBeringian stage of the Native American ancestors formation,which, as noted above, may have taken a fairly long period oftime . A recentstudy of genomic polymorphism in various contemporarycontinental groups showed that Native Americans harbor atleast 20,424 genetic variants suspected to have originated inBeringia . This is comparable tothe number of genetic variants that distinguish Africans fromnon-Africans. However, there are tens of times less specificgenetic variants in Eurasia. At the same time, group-specificpolymorphisms have not been found at all in Europeans, thusindicating rather intensive inter-ethnic contacts within Eurasia.A study of the influence of adaptive selection on the geneticprofile of Native Americans has shown that the appearance ofAmerican-specific genetic variants is associated with variousmetabolic pathways, but the most important ones are associatedwith the production of melanin in the skin, hair and eyes,as well as cardiovascular function .It should be noted, however, that the most distinct traces ofadaptation to the severe conditions of the Far North have beenrevealed in the gene pools of the Eskimo and Paleo-Asiaticpeoples rather than in those of the Native Americans. Eskimoand Paleo-Asiatic populations also originated in the Asian andNorth American Arctic during the last 5–6 thousand years and thus,their periods of adaptation are quite comparable in time to theBeringian population. However, in the next 15 thousand yearsafter their isolation, the Beringians began to occupy moresouthern American territories, and therefore it is unknownto what extent the traces of their ancestors’ adaptation tothe Arctic are preserved in the gene pools of modern NativeAmericans .A number of studies of modern and ancient populations ofthe Far North of Asia and America have found evidence ofgenetic and physiological adaptation of Eskimo and Paleo-Asiatic ancestors to low temperatures and an “Arctic” dietbased primarily on the consumption of seafood rich in polyunsaturatedfatty acids (PUFAs) .One of the most striking examples of diet-related geneticmarkers is the “Arctic” variant of the CPT1A gene(rs80356779-A) .It is known that the “Arctic” variant is widely spread in modernpopulations of the Eskimos, Chukchi, Koryaks, andother peoples of the Sea of Okhotsk region, whose economicstyle is connected with sea hunting .According to paleogenomic data, the earliest records of the“Arctic” variant of the CPT1A gene were found among theGreenland and Canadian Paleo-Eskimos ,representatives of the Tokarev culture of the North of the Seaof Okhotsk region  and carriers of the LateJomon culture of Hokkaido Island .The appearance of the “Arctic” variant of the CPT1A geneis most likely associated with the adaptive response of theFar North indigenous peoples to the “Arctic” diet, which ischaracterized by a marked excess of lipids and proteins anda deficit of carbohydratesIn addition, genetic variants associated with carbohydratemetabolism are very frequent in indigenous populations of Northeast Asia. For example, the maximum frequency (52 %)of deletion of the pancreatic amylase gene AMY2A, necessaryfor starch digestion, was found in the Eskimo, Chukchi, andKoryak populations; ~30 % of the indigenous peoples ofNortheast Asia lack this gene . The highfrequency of AMY2A gene deletion in Northeast Asia may beexplained by the deficiency of starch and disaccharides in thetraditional diet of indigenous peoples in the past. A deficiencyin oligosaccharides can also explain the high incidence ofthe inactive sucrose-isomaltase gene (gene SI ) among theindigenous populations of the Far North of Asia and America. In GreenlandicEskimos, glucose homeostasis-related mutations in theTBC1D4 and ADCY3 genes have also been identified, whichincrease the risk of obesity and type 2 diabetes .Thus, genetic changes that occurred in the past due toadaptationto extreme environments, in the current conditions(when there is no carbohydrate deficiency in the diets of theFar North indigenous populations) become harmful, becausethey increase the risk of metabolic and other diseases.Interestingly, in a number of cases, adaptive changes inthe gene pools of Eskimo and Paleo-Asiatic populations arecaused by nonsynonymous substitutions with high pathogenicityindices , nonsense mutationsleading to stop codons or splicing disruption , and whole gene deletions (as inthe case of the AMY2A gene) . It seems likely thatthis kind of fairly profound genetic changes could have beenpreserved in the gene pools of Native Americans if they hadadapted to the Arctic environment in a similar way, involvinga very specific “Arctic” diet.There is a suggestion that since the ancient inhabitants ofBeringia migrated first to the sub-Arctic Pacific coast and thenon to Northwest America, they may have practiced a maritimetype of economy at the sub-Arctic stage . But,apparently, this stage of migration history was not reflectedin the gene pools of the Native Americans, or their ancestorsdid not hunt sea mammals, but hunted mainly megafauna:mammoths, bison, horses and other terrestrial animals of theGreat Arctic Plain . In that case, it is assumed that Amerindianancestors had to adapt primarily to vitamin D deficiency,because at high latitudes UV radiation levels are insufficientfor annual synthesis of cholecalciferol D3 in the skin, and theintake of ergocalciferol D2 depends on dietGenetic adaptation to vitamin D deficiency suggests thatthe dietary sources of this substance that are available toArctic peoples  were scarceor absent in the Beringian steppe-tundra biome. Vitamin Ddeficiency affects the health of children to a greater extent,and so L.J. Hlusko et al. (2018) suggested that Beringiansadapted to UV deficiency through a V370A substitution inthe ectodyslasin receptor encoded by the EDAR gene. Thismutation is widespread in populations of East and North Asia,as well as in Native Americans. One manifestation of this mutationis associated with an increase in the branching densityof mammary gland ducts, which leads to higher amounts ofvitamin D in breast milk .A recent study of Latin American populations also showedthat genes related to energy metabolism played a crucialrole in population adaptation to the environment of America. The great importance of immuneresponse genes, primarily HLA genes, in adaptive processeshas also been highlighted. Moreover, positive selectionin these genes is recorded in Native Americans at differentperiods of their history, both after the European colonizationof America, which introduced new pathogens, and during theinitial settlement of America at the first contact with endemicpathogens .The hypothesis suggesting the influence of high-latitude UVdeficiency on the genetic features of the ancient Beringiansis based on an additional argument involving the adaptiveselection of the FADS genes variants in Native Americans. The fatty acid desaturase genes FADS1and FADS2 encode enzymes involved in PUFA biosynthesisfrom shorter precursors . It is assumedthat the ancestral A-variants of the FADS genes gave advantagesto human populations that consumed food rich in lipidsand proteins , but later in the Neolithic, after the emergence ofagricultural technology, the D-variants became more common,allowing for a higher rate of synthesis of PUFAs from plantlipids .Studies have shown that FADS genes have been affectedby positive selection for a long time – for example, in Europethere has been an increase in the frequency of haplotype Dfrom less than 10 % 10,000 years ago to 60–75 % at present. However, the exact reasons for theincreased frequencies of the FADS genes variants in certainregions of the world have not yet been established. This is dueto the fact that FADS genes are highly pleiotropic. Associationswere found for FADS1 and FADS2 genetic variants with bloodlipids and other metabolites, various blood cell phenotypes,suggesting a link with cardiovascular function . Besides, metabolic balance with respect to PUFAs andother lipids is very important for brain function, in which theFADS1 gene is actively expressed It has been suggested that the emergence of the D haplotypemay have been due not only to the need to increase theefficiency of PUFA synthesis when there is a deficiency inthe diet , but also contributed to an increasein the brain size . In addition, it was foundthat in Greenlandic Eskimos, FADS genes are involved in theprocesses of adaptation to the cold, as there are links betweengenetic variants and the distribution of body fat and height;and the participation of FADS genes in the regulation of growthhormones is also suggested .Population genetic studies have shown that the ancestralhaplotype A is much more common among the indigenouspopulations of Siberia and America . It has also been found that in Greenlandic Eskimosand Native Americans, the FADS1 and FADS2 genes are underpositive selection, which contributed to an almost completefixation of haplotype A in these populations . Meanwhile, a number of otherstudies on Arctic populations of Siberia  and North America (Eskimos)have not revealed the effect of selection on the FADS1 andFADS2 genes .Nevertheless, there is a hypothesis that the original positiveselection signal in the FADS1 and FADS2 genes arose inthe ancient population of Beringia due to the need to adaptto the cold and limited food resources provided by the Arctic. It is assumed that these adaptive geneticvariants have been preserved in the gene pools of the NativeAmericans and Eskimos. However, different combinationsof the FADS genes variants are responsible for the effect ofselection in Arctic and Native American aborigines and inEuropeans . This can be due to differentreasons for the selection of the FADS genes variants in populations.In Europeans it was a shift towards a plant-based diet inthe Neolithic, due to which the D-haplotypes with increaseddesaturase activity were more favorable, and in Eskimos ancestorsit was the emergence of a specific “Arctic” diet withvery high PUFA content, due to which maintaining the maximumfrequency of A-haplotypes was more preferable. It ispossible that these FADS variants in Eskimos possess evenlower desaturase activity .Meanwhile, a recent analysis of the FADS genes polymorphismin modern and ancient populations has shown that thewidespread distribution of haplotype A occurred under theinfluence of selection already in the Upper Paleolithic populationsof Eurasia, and therefore in the gene pool of theancient Beringians this haplotype could be fixed quite accidentallyunder the effects of genetic drift . Thus, the adaptive changes of FADSgenes in the Native American ancestors are questioned, andthe hypothesis that the FADS haplotypes inherited from theUpper Paleolithic Eurasians are preserved in the gene poolsof Native Americans looks more likelyAccording to the published data, another example of selectiveadvantages obtained by Native Americans from the ancientinhabitants of Beringia may be related to adaptive changesin the ACTN3 gene . This gene encodesthe protein α-actinin-3, which is expressed exclusively infast-twitch skeletal muscle fibers. Of the polymorphic loci ofthe ACTN3 gene, the most studied is rs1815739, a mutationin which leads to termination of protein synthesis at aminoacid position 577 of exon 16 (R577X substitution) . This causes a deficiency of α-actinin-3 in fasttwitchskeletal muscle fibers, which, in turn, can result in adecrease in the speed and power indicators of human physicalperformance . Genetic studies have shownthat the frequency of the 577X allele increases with distancefrom Africa to the north of Eurasia and reaches a maximumin the Native American populations .It is suggested that the advantages of the 577X allele, suchas increased endurance and improved protection against thecold, have contributed to the spread of this mutation .Recent physiological and metabolomic studies have shownthat carriers of the 577XX genotype tolerate cold much betterthan 577RR individuals , which maysupport the hypothesis of selection of the X allele at the initialstage of human expansion into Eurasia .According to this hypothesis, the elevated frequency of the577X allele in some human populations could be related toselection to improve metabolic efficiency and promote dynamicactivities .The high frequency of the 577X allele in Native Americanssuggests that this allele was also widespread in ancient Beringians,ancestral to Native Americans. However, the resultsof analysis of the distribution of the rs1815739 polymorphicvariants in the indigenous populations of northeastern Siberia have shown that thefrequencies of the 577X allele and 577XX genotype werenot the highest in Eurasia . Whilethe frequency of the 577X allele in Northeast Siberia is approximately36 % , the maximumfrequencies of this genetic variant are registered in South Asianand Native American populations – over 60 % according todbSNP database (www.ncbi.nlm.nih.gov/snp).A recent analysis of geographic distribution of rs1815739variants in modern and ancient populations has shown thatpopulation frequencies of the 577X allele correlate neitherwith geographic latitude nor with temperature . None of the statistical tests used in this study foundevidence of the effect of positive selection for the 577X alleleat high latitudes. Thus, the high frequency of this geneticvariant in Native Americans is most consistent with the effectof genetic drift, possibly occurring in Beringia at the stage oflow size of ancestral population. It is possible, however, thatsome advantages of this genetic variant, namely increasedendurance and resistance to cold, contributed to the increasedfrequency of the 577X allele in Native American ancestors.The results of genetic studies of modern and ancient populationsof Eurasia and America have demonstrated quite convincinglythat the role of Beringia in the settlement of theAmericas is very high. To date, sufficiently reliable estimatesof divergence times between the ancestral genetic lineages thatled to the formation of the various populations of NortheastAsia and the Americas have been obtained. The most probablescenario seems to be that during the last glacial maximum, thepopulation that gave rise to the Native Americans ancestors persistedin Beringia for several millennia . It is assumed that this was a relatively small ) group of people well adapted to high latitudes and cold environments, which is confirmed by the results of genomicstudies of modern Native American populations.Possible traces of adaptation to Arctic environments at thegenomic level are associated with various metabolic pathways– with melanin synthesis processes, cardiovascular systemfunctioning, energy metabolism, and immune responsegenes .It has also been suggested that the relaxation of negative selectionin a number of protein-coding genes observed in NativeAmericans is also associated with the Beringian stage of populationadaptation . However, it isstill unclear to what extent such adaptive signals could havesurvived, given the approximately 15-thousand-year periodof American colonization following the Beringian Standstill.Obviously, answers to such questions can be obtained byusing paleogenomic data, but very few ancient human remainshave been found from the territory of ancient Beringia. Sofar, we can get some insight into the genetic features of theBeringians from only one genome of the Eastern Beringiandescendant  . Thus, investigationsinto the role of Beringia in the origins of the Native Americanancestors remain relevant and ongoing.The authors declare no conflict of interest.Achilli A., Perego U.A., Lancioni H., Olivieri A., Gandini F., HooshiarKashani B., Battaglia V., Grugni V., Angerhofer N., Rogers M.P.,Herrera R.J., Woodward S.R., Labuda D., Smith D.G., Cybulski J.S.,Semino O., Malhi R.S., Torroni A. 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Physalis angulata L.), an annual plant containing a variety of active ingredients, has great medicinal value. However, studies on the genetic diversity and population structure of P. angulata are limited. In this study, we developed chloroplast microsatellite (cpSSR) markers and applied them to evaluate the genetic diversity and population structure of P. angulata. A total of 57 cpSSRs were identified from the chloroplast genome of P. angulata. Among all cpSSR loci, mononucleotide markers were the most abundant (68.24%), followed by tetranucleotide (12.28%), dinucleotide (10.53%), and trinucleotide (8.77%) markers. In total, 30 newly developed cpSSR markers with rich polymorphism and good stability were selected for further genetic diversity and population structure analyses. These cpSSRs amplified a total of 156 alleles, 132 (84.62%) of which were polymorphic. The percentage of polymorphic alleles and the average polymorphic information content (PIC) value of the cpSSRs were 81.29% and 0.830, respectively. Population genetic diversity analysis indicated that the average observed number of alleles (Na), number of effective alleles (He), Nei’s gene diversity (h), and Shannon information indices (I) of 16 P. angulata populations were 1.3161, 1.1754, 0.1023, and 0.1538, respectively. Moreover, unweighted group arithmetic mean, neighbor-joining, principal coordinate, and STRUCTURE analyses indicated that 203 P. angulata individuals from 16 populations were grouped into four clusters. A molecular variance analysis (AMOVA) illustrated the considerable genetic variation among populations, while the gene flow (Nm) value (0.2324) indicated a low level of gene flow among populations. Our study not only provided a batch of efficient genetic markers for research on P. angulata but also laid an important foundation for the protection and genetic breeding of P. angulata resources.Cutleaf groundcherry ( Physalis angulata L.) is an important annual herbaceous plant from the Solanaceae family, mainly distributed in China, Japan, India, Australia, and the Americas [P. angulata has high potential medicinal value and a very long history of being used in traditional medicines around the world. Recent phytochemical and pharmacological studies have confirmed that P. angulata is rich in vitamins, minerals, antioxidants, and many important pharmacologically active constituents, including antibacterial, anti-inflammatory, and anticancer ingredients [P. angulata is often used to treat a variety of illnesses, including dermatitis, tracheitis, impaludism, rheumatism, hepatitis, and analogous conditions [P. angulata, and a variety of bioactive steroids with antitumor activities, including physagulins (A−Q), physangulidines (A−C), withangulatins (A−I), physalins , and withaminimin, have been isolated from the species [P. angulata plants have been widely cultivated in some regions of China for decades.Cutleaf groundcherry  were detected in the chloroplast genome of  A and T . Informa A and T . Of the  A and T . The ave A and T . Additio A and T .P. angulata chloroplast genome using Primer Premier 5 with manual correction  with good stability, high polymorphism, and clear electrophoresis bands were chosen for further genetic diversity analyses of the P. angulata populations  to 12 (KzcpSSR04), and an average of 5.2 bands per primer pair . The actP. angulata populations (n = 203) was evaluated, revealing high mean per population estimates of allele and genetic diversity  to 85 (YN and XS), with an average of 49, while the PPL ranged from 6.41% (DQ) to 54.49% (YN and XS), with an average of 31.61%. The Na per population ranged from 1.0641 (DQ) to 1.5449 (XS and YN), with an average of 1.3161. The Ne across the 16 populations ranged from 1.0371 (DQ) to 1.2984 (XS), with a mean of 1.1754. The I per population ranged from 0.0325 (DQ) to 0.2640 (XS), with an average of 0.1538. At the population level, the PPL, Na, Ne, and I values were the highest in the XS population, indicating that this population had the highest level of genetic diversity. The 16 populations were ranked by genetic diversity as follows: XS > YN > WZ > XJ > PJ > HG > JJ > NJZ > YW > TZ > LA > NH > LH > NJX > HZ > DQ.The genetic diversity of the 16 P. angulata revealed no significant correlation .Nei’s genetic distance, calculated on the basis of a pairwise comparison, ranged from 0.0329 (between DQ and YW) to 0.8268 (between NH and NJX), with an average of 0.3401. The majority of Nei’s genetic identity ranged from 0.4374 (between NH and NJX) to 0.9677 (between TZ and LH), with an average of 0.7386 . A ManteHt of the P. angulata population was 0.3224, the Hs was 0.1023, and the Gst was 0.6827 , which was consistent with the results from the analysis of Nei’s genetic diversity, indicating that the genetic differentiation between populations was higher than that within populations.At the species level, the s 0.6827 . The resulations . The resP. angulata populations was constructed on the basis of Nei’s genetic distance, which is an accurate reflection of the genetic relationships among populations. The UPGMA tree showed that the 16 populations could be divided into four major clusters  (The UPGMA dendrogram of  and IV) . ClusterP. angulata individuals were grouped into four distinct clusters, consistent with the results of the UPGMA analysis. The findings indicated that most of the individuals from the same population clustered together, but there were also a few individuals from the same population who grouped separately into different clusters. For example, almost all individuals in the XJ population were grouped into Cluster I, and the remaining individuals were grouped into Cluster II and Cluster III  were the most common SSR loci detected in our study, followed by tetranucleotides . These proportions are similar to those of cpSSR marker types found in many plants ,34,35. PP. angulata populations, accounting for 55.56% of the newly developed markers. The polymorphic ratio of the cpSSR markers ranged from 25% to 100%, with an average of 81.29%, which was higher than the polymorphic ratios detected by SSR of 73.5% among Dendrocalamus hamiltonii [P. angulata populations.A total of 30 out of 54 cpSSR markers amplified electrophoretic bands with high levels of polymorphism, good stability, and high resolution in miltonii , and 53.miltonii . The PICP. angulate populations . The proportion of polymorphism among P. angulata populations ranged from 6.41% to 54.49%, indicating that there were significant differences in polymorphic loci among populations. Furthermore, the XS population showed greater genetic diversity than the other 15 populations. Correspondingly, the complexity of genetic diversity was as follows: XS > YN > WZ > XJ > PJ > HG > JJ > NJZ > YW > TZ > LA > NH > LH > NJX > HZ > DQ. Compared to other populations, the DQ population from Deqing, Zhejiang Province had the lowest level of genetic diversity, with the smallest number of polymorphic loci, i.e., the lowest Na and Ne values. The reason for the lowest genetic diversity in the DQ population is probably because it contains a limited number of individuals adapted to specific habitats due to their isolation from other populations. Therefore, to more accurately evaluate the genetic diversity of this population in the future, more samples will be required. The average h and I values among the P. angulata populations were 0.1023 and 0.1538, respectively (<0.5), similar to those of P. philadelphica and P. peruviana [Genetic diversity is the sum of genetic information in a population, which is necessary for population persistence, adaptation, and evolution ,39. The eruviana ,28,41,42P. angulata populations was low (Nm = 0.2324), similar to that found in P. philadelphica, P. peruviana, and M. savatieri [P. angulata populations may be related to the genetic mode and the dispersal distance of the seeds and pollen. The Ht value of P. angulata (0.3224) was higher than that of P. philadelphica (Ht = 0.292) [Gst value of 0.6827 for P. angulata indicates that 68.27% of the genetic variation occurred among populations, while 31.73% of the genetic variation occurred within populations. This is a similar result to that observed in Rhodiola alsia [Gene flow, negatively correlated with the genetic differentiation coefficient, is a basic microevolutionary phenomenon that often hinders genetic differentiation between populations and affects the maintenance of genetic diversity ,43,44. Iavatieri ,41,45. Tla alsia . The AMOP. angulata. Many studies have reported a correlation between genetic distance and geographical location in the populations of certain plants [P. angulata populations from these provinces and those from Zhejiang Province was relatively large, and the level of gene flow was very low. Interestingly, some populations did not show a clear correlation with geographical location. For example, the NJZ population from Jiangsu Province and four populations  from Zhejiang Province were clustered together, although they were geographically distant. In our previous investigation of morphological phenotypic traits, we found that the plant height in these five populations was higher than that in the other populations. This might explain why they clustered together. Our study showed that the genetic diversity of P. angulata was not only related to geographical location but also likely associated with the geographical environment, human selection, and other factors. Furthermore, the PCoA and Structure analysis results were identical, and both supported the results of the UPGMA and NJ trees analyses. In a future study, more P. angulata population samples and more genome-wide molecular markers  will be used to more comprehensively evaluate the genetic diversity and structure of P. angulata populations.The population genetic structure reveals the distribution pattern of genetic diversity within and among populations ,39,47. Tn plants ,39,45,47P. angulata representing 16 populations were randomly collected from their main areas of distribution in China . The collected samples were planted during the regular crop growing season (early May) in 2018 at the field site of the Zhejiang Provincial Key Laboratory for Genetic Improvement and Quality Control of Medicinal Plants, Hangzhou Normal University, China. The field site was located at a geographic position of 30°32′18′′ N, 120°40′07′′ E and an altitude of 10 m. The phenotypes of some of the P. angulata materials are shown in In total, 203 samples of wild in China . The ideFresh, young leaf tissues from three individuals of each sample were randomly collected for genomic DNA isolation. Genomic DNA was isolated from these samples as described in our previous studies ,48. The P. angulata was sequenced, annotated, and submitted to the National Center for Biotechnology Information (NCBI) GenBank database (GenBank accession no. MH045574) [P. angulata chloroplast genome were screened using MISA software  [In our earlier work, the complete chloroplast genome sequence of H045574) . The SSRay 2022) . The cpSay 2022) . The par4)2SO4, 20 mM MgCl2, and 1% TritonX-100), 0.8 μL of dNTPs (10 mmol/L), 1 µL each of forward and reverse primers (10 µM), 1 µL of Taq DNA polymerase (2 U/µL) , 50 ng of genomic DNA template, and 4.7 μL of ddH2O. The amplification program for the cpSSR-PCR was as follows: pre-denaturation at 94 °C for 5 min, followed by 32 cycles of denaturation at 94 °C for 50 s, annealing at 55–61 °C  for 50 s, 72 °C for 1.5 min, and a final extension at 72 °C for 10 min. PCR products were separated on 8% nondenaturing-polyacrylamide gel (acrylamide: bisacrylamide 19:1) using Trans2K DNA markers  as size standards before being stained with silver.In total, 54 cpSSR primer pairs were selected and synthesized by Shanghai Sangon Biological Engineering Technology and Service Co. Ltd. . After a trial run of the 54 newly developed cpSSR primer pairs, 30 cpSSRs showing high definition, good stability, and abundant polymorphism were chosen for further analysis. cpSSR amplification was performed in 10 μL volumes including 1 μL of 10× buffer (200 mM Tris–HCl (pH 8.8), 100 mM KCl, 100 mM , the number of observed alleles (Na), the number of effective loci (Ne), Shannon’s information diversity index (I), Nei’s genetic diversity index (h), and the total genetic variation of the population (Ht) were calculated using PopGene32 Version 1.32 . The genetic variation within the population (Hs), population genetic differentiation coefficient (Gst), and gene flow (Nm) were also calculated using PopGene32 Version 1.32. On the basis of the genetic consistency of the populations, an unweighted group arithmetic mean (UPGMA) cluster diagram and three-dimensional principal coordinate analysis (PCoA), which enable the visualization of genetic variation distribution across populations, were constructed and carried out using NTSYS-PC 2.10e software [http://taylor0.biology.ucla.edu/structureHarvester; accessed on 22 August 2022), the results of the structure calculation were analyzed to find the best group K value [To ensure the accuracy of the results, each pair of primers was used for PCR amplification and electrophoresis detection twice, and only the cpSSR fragments with high definition and good stability were scored. Genetic variation at each locus was characterized in terms of the number of alleles. The PIC was calculated using the following formula: h allele . The persoftware . On the software . Analysisoftware . The popsoftware . The est K value ,58.P. angulata populations. Our study revealed that most of the newly developed cpSSR markers had a high level of stability and polymorphism. The cpSSR analysis confirmed that P. angulata populations contain considerable genetic diversity, and there are high levels of genetic differentiation among populations. The 16 populations of P. angulata were clustered into four groups with some significant geography-related population structure and extensive admixture. Our study demonstrated that cpSSR markers can be used as a powerful tool for evaluating genetic diversity and population structure in P. angulata.In conclusion, this is the first study to develop a novel set of cpSSR markers and apply them to the investigation of genetic diversity and genetic structure in


The Arabian horse is probably one of the most historic horse breeds worldwide. It has been selectively bred by the Bedouin tribes in the steppes of Arabia for over 2000 years. Bedouin tribes have created their own classification of strains based on maternal lines. The aim of our study was to investigate genetic differentiation at the level of (i) the current populations  and (ii) the traditional Bedouin strains. Two hundred and eleven Arabian individuals, representing 12 strains, were sampled. Mitochondrial DNA D-loop analysis showed higher genetic diversity in the Egyptian and Polish Arabian populations than in the Desert-Bred. No genetic markers distinguishing the traditional strains were highlighted in this study, a fact probably due to the lack of interest in this component in modern breeding programs. Some Polish Arabian individuals, who could not be traced back to the Bedouin tribes, appeared to be genetically distinct from the other studied horses, requiring more in-depth study. This research, complemented by future SNP and Y-chromosome analyses, may provide a more accurate evaluation of the relationships between Arabian horse populations′ genetic and traditional classification.Genetic diversity and phylogenetic relationships within the Arabian show horse populations are of particular interest to breeders worldwide. Using the complete mitochondrial DNA D-loop sequence (916 pb), this study aimed (i) to understand the genetic relationship between three populations, the Desert-Bred (DB), a subset of the Kingdom of Saudi Arabia (KSA), United Arab Emirates (UAE) and Bahrain (BAH), the Straight Egyptian (EG) and the Polish bloodline (PL), and (ii) to assess the accuracy of the traditional strain classification system based on maternal lines, as stated by the Bedouin culture. To that end, we collected 211 hair samples from stud farms renowned for breeding Arabian show horses from Nejd KSA, Bahrain, Egypt, Qatar, Morocco, UAE, and Poland. The phylogenetic and network analyses of the whole mitochondrial DNA D-loop sequence highlighted a great genetic diversity among the Arabian horse populations, in which about 75% of variance was assigned to populations and 25% to strains. The discriminant analysis of principal components illustrated a relative distinction between those populations. A clear subdivision between traditional strains was found in PL, in contrast to the situation of DB and EG populations. However, several Polish horse individuals could not be traced back to the Bedouin tribes by historical documentation and were shown to differ genetically from other studied Bedouin strains, hence motivating extended investigations. Assil/Atiq). Firstly, its pedigree (Al-Nasab) must be solely based on breeding from Arabia and free of any evidence of morphological or genealogical sign of impurity ; therefore, it must be related to a recognized strain name (Rasan) and a sub-strain (Marbat). Secondly, its morphological features must match those of the breed (Al-Muwasafat). The strain names came from the original mares (or group of mares) that founded different families of Asil/Atiq horses centuries ago within foundational Marbat [Arabian horses are a worldwide breed appreciated for their unparalleled beauty, human character, and versatility under saddle. All of these attributes are legacies of the Arab Bedouin tribes who lived across the steppes of Arabia for several centuries. The Bedouin tribes, who developed the breed, established specific requirements before the horse could be considered purebred , including Syrian, Bahraini and Saudi populations, were the sole ancestors of the Straight Egyptian (EG) Arabian and that they contributed largely to the creation of the Polish Arabian (PL) ,4. MoreoAs for the EG horses, the background sources confirm that they were based on premium Bedouin horses from Arabia ,3. The mMicrosatellite ,11 and mThrough the sequence analysis of DB, EG and PL Arabian populations, we aimed to first challenge the level of maternal genetic diversity in the Arabian horse populations as highlighted by previous studies and the significant differentiation between them ,13. It wHair samples were collected from 211 Arabian horses composed of three populations DB (n = 54), PL (n = 51) and EG (n = 106), all coming from stud farms localized in KSA, UAE, Qatar, BHR, Poland, Egypt and Morocco. All samples were unrelated from the mother’s side at least for three generations and had the phenotype of an Arabian horse breed, and were registered in a WAHO-recognized studbook. The geographical location of these stud farms is described in Rasan) using their pedigrees and origins. Thus, 193 horses out of 211 were assigned to 12 strains: Dahman, Hadban, Hamdani, Kuhailan, Obeyan, Saqlawi, Suwaiti, Subeyli, Al-Maanaqy, Al-Musannah, Al-Radba and Al-Tuwaisah. The individual Polish horses derived from Szamrajówka, Ukrainka and Wołoszka dam lines, whose founders’ mothers could not be traced to the Bedouin tribes’ horses, were included as an unknown origin (PLS)   Table S1® DNA purification kit  following the manufacturer’s instructions. DNA quality was assessed with the NanoDrop 8000 spectrophotometer . The whole mitochondrial D-loop region was amplified using the pairs of specific primers tested previously by Khanshour and Cothran [Total genomic DNA was extracted from hair follicles using the PUREGENE Cothran  in the u Cothran .http://www.ncbi.nlm.nih.gov/the accession numbers NCBI: MZ735748–MZ735958 (accessed on 6 August 2021). The genetic variations within populations were evaluated by using the DNAsp 5.10.1 software [Haplotype sequences included in this study were registered into the National Center for Biotechnology Information (NCBI) GenBank database available at software . The folsoftware , Fu Fs [software  and D ofsoftware  using DNTo estimate the genetic differentiation between studied populations, the pairwise FST was calculated using the Kimura 2-parameter model with 1000 permutations and was carried out with Arlequin 3 . For theThe phylogenetic analysis of the haplotype using the whole D-loop sequence was conducted with MEGA software version 10.0 . The phyp-value was deduced from a set of 9999 permutations. At the level of strains, the chi-square (χ2) test was applied for each combination of studied lines, and the p-value was calculated by the Monte Carlo test.To verify if there was a significant variation between haplogroup compositions in maternal lines, two separate approaches were applied using PAST 2.17c software . At the To visualize the proximities between the studied strains, a Cluster analysis was conducted with the Chord distance index implemented in PAST 2.17c, and the significance of the branches was assessed by 1000 bootstrap iterations.Discriminant analysis of principal components (DAPCs), implemented in the adegenet package for R , was appUsing the gl.manova function from the DartR package, a multivariate analysis of variance (MANOVA) was conducted on genetic distance data, with a focus on horse populations and horse strains. The Phi-statistic was determined to estimate the proportion of genetic differentiation among populations relative to overall genetic differentiation.p < 0.05) according to the Chi-2 test  using DNAsp . Sixty-ni-2 test .Twelve haplogroups: A, B, C, D, G, I, L, M, N, O/P and Q according to Achilli’s classification , were idKuhailan, Dahman, Saqlawi, Hamdani, and Obeyan with 36, 27, 19, 12 and 10 haplotypes, respectively. The Al-Musannah and Subeyli strains were represented uniquely by one haplotype each, H1 and H6, respectively. The MJ network based on 916 bp of the D-loop for strains is shown in p < 0.05) according to the Chi-2 test  and Al-Musannah strain were only present in haplogroup L, and they were close to the Dahman, Hadban, Hamdani, Kuhailan, Saqlawi and Suwaiti.Eight strains shared two or more haplogroups, and individuals from the same strain were found in separate haplogroups. The The polymorphism analysis using the whole D-loop sequence  showed tRasans . The Kuhailan was the most variable strain with NHap:36, followed by Dahman (NHap:23) and Saqlawi (NHap:21). Suwaity and Hamdani showed the lowest number of haplotypes . As the number of haplotypes by strain is strictly proportional to the size of the strains , the use of this data is limited and will not be considered in the following.The mtDNA polymorphism analysis results concerning the strains are shown in a priori assumptions of population membership (K = 3) and strain membership (K = 8). The number of retained principal components was defined using the α-score optimization proposed by Jombart et al. (2010) [In the first step, discriminant analysis of principal components was undertaken to investigate the relationships between the populations. The clusters in DAPC were defined by . (2010) , resultiThe population structure deduced from DAPC revealed a clear genetic differentiation between the three tested populations with a narrow overlap among them a. The ovPairwise FST used to explore the genetic relationships among the populations confirmed that the Polish bloodline was closer to the Straight Egyptian than to the Desert-Bred .A DACP analysis was conducted on the strains in parallel with the population approach. To complete this analysis, the matrix with the number of individuals of each haplogroup per strain was submitted to a Cluster analysis b. The stTo investigate how the strains are distributed within the populations, a new DAPC was conducted. Given the strain sizes within each population, only two strains were retained in the DB, four in the EG, and three in the PL. It showed that the strain envelopes in DB and EG overlapped or merged, while the PL population strains were separated. In other words, it seems not possible to distinguish the strains within the EG and DB populations, while certain strains in the PL population are clearly distinct see  below. p-values were less than 10−4, indicating a significant difference from what would be expected by chance. These Phi-statistic values suggest a moderate level of differentiation among the horse populations and strains under investigation. Moreover, the variance coefficients provided additional evidence of the impact of these factors on the overall variability in the data. The variance coefficient for horse populations was calculated as 65.8, while the coefficient for horse strains was 18.3. These relatively high coefficients indicated that both horse populations and strains contributed significantly to the observed variability in the genetic distance data. Overall, the Phi-statistics and variance coefficients indicated that there is moderate but significant differentiation among horse populations and strains, which is supported by the gl.manova analysis results. These results imply that both variables significantly contribute to the explanation of the genetic variation found in the data.Following the MANOVA, the Phi-statistics associated with the proportion of genetic differentiation among populations relative to the total genetic differentiation was found to be 0.07 for horse populations and 0.17 for horse strains. The corresponding This study used the mtDNA control region to highlight the genetic variability of DB, EG and PL Arabian horse populations, including the strain classification system. It has been proved that the whole mtDNA D-loop for genetic diversity analysis is more robust and powerful than using the hyper-variable region 1 (HVR1) alone for this purpose ,28,29. TAs mentioned in the introduction, it has been established that the DB horse, developed by Bedouins under harsh climatic conditions in the steppes of Arabia, is the origin of all modern Arabian horse populations, including the EG and most PL lines ,37,38.Based on the pedigree analysis from the stud books data, several founding broodmares of the PL and EG populations were brought from the Arab region  ,36. So, The study of haplotype distribution, based on the consensus NJ tree, showed that 18 haplotypes were only found in the DB individuals, while 38 and 31 haplotypes were found in the EG and PL individuals, respectively. The same trend was retrieved for the average number of nucleotide differences in DB, EG and PL populations. On the other hand, the number of haplotypes and haplotype diversity showed lower values in the DB population than in the EG and even more so in the PL. The number of polymorphic sites was intermediate in the PL population compared to the EG with the highest value and the DB with the lowest. The MJ network confirmed this result, where DB individuals were present in only eight haplogroups, while both PL and EG populations were represented in nine and 10 haplogroups. All of these data confirmed the expected high genetic diversity of the DB population, supporting the previous finding by microsatellite analyses [Rasan”, there is no guarantee that all the diversity was captured, which may explain the relatively low genetic diversity of the DB. In addition, the current estimated size of the EG and DB populations between 3–10% of the global Arabian horse population would be far less than that of the PL population. Despite the exact size of the respective populations, this effect remains a possible cause of the low DB genetic diversity due to allele loss and genetic erosion. In addition, it is possible that the DB population has fallen below the critical minimum for survival, resulting in a bottleneck effect, explaining the low level of genetic diversity.The interpretation of these data is an open question. Despite the sampling within the DB population being performed based on the “Regarding the origin of the PL population, our analysis showed that PL shared one and five exclusive haplotypes with DB and EG, respectively. We cannot exclude that some haplotypes have disappeared or were not sampled in our study, but the most likely hypothesis is that the majority of the maternal contribution for the PL population was Egyptian, although the possibility of the contribution of DB stallions is not excluded. However, the microsatellite data indicated a closer genetic relationship between PL and DB; this suggests that the DB stallions participating in the breeding program of the PL had a major impact on the genetic pool of the PL population . An SNP Rasan), which are transmitted through the maternal lines, and sub-strain (Marbat), both constituting the identity of pure Arabian (Assil/Atiq) individual horses [Marbat) of our samples, we limited our study to the strains.According to history, the traditional Bedouins tribe’s Arabian classification was based on the notions of strain  when the population level was not considered. It should be noted that only Suwaiti and Saqlawi strain individuals were found to belong to haplogroups B and M, respectively, which may support the genetic reality of these strains. In contrast, all seven strains were represented in haplogroup A. The possibility of some discrimination between strains in the Syrian Arabian horse population was previously mentioned [Szamrajówka, Ukrainka, and Wołoszka have had their origins traced back not far away from the stocks of Sławuta stud [Saqlawi and Kuhailan strains observed in the PL population, the explanation still needs to be determined. For a more detailed and objective interpretation of the origin of these Polish Arabian horses using Y sequences and SNP data, it is necessary to integrate the human population history’s results and the European/Asian migrations of peoples.To highlight the genetic structuring between strains, the low sample size of the strains entioned . Howeveruta stud . As for Rasan-Marbat) both constitute the identity of pureblood Arabian horses (Assil/Atiq) [Hence, when we consider the reality of strains between populations, a low number of strain markers appears since the population effect dominates the strain effect. However, our phylogenetic results and DAPC did not reveal a clear distribution that fits strain subdivisions. This finding is consistent with some previous works ,28,33. Iil/Atiq) ,39. TherRasan-Marbat) data as well as SNP data may shed additional light on the relationship between genetics and the traditional Bedouin classification system.Based on the whole mtDNA D-loop sequence, the present study confirmed the large genetic diversity of the Arabian horse populations and illustrated the genetic differences between DB, EG, and PL. Even though the DB is at the origin of EG and PL, it has been observed that it has the lowest genetic diversity, which may be due to genetic erosion related to the low population size. The maternal lineages of the PL population would be mostly of EG origin, and we hypothesize that a major paternal part could come from Desert-Bred ancestors, which would fit the results of microsatellite markers. Once again, this point would need to be investigated by Y chromosome analysis. A special mention must be stated about the maternal lineage of the PLS group, which needs to be investigated more deeply, as no connection was found with the Desert-Bred individuals of our sample. Concerning the traditional strain system of the Bedouins, there are only some elements in the mtDNA, but it is evident that the differentiation of the populations has erased more or less the genetic markers of the strains. Further, detailed studies including both strain and sub-strain (