Turkish Journal of Biology Turk J Biol (2013) 37: 176-183 http://journals.tubitak.gov.tr/biology/ © TÜBİTAK Research Article doi:10.3906/biy-1203-64
Genetic variability among native dog breeds in Turkey
1, 2 3 1 Metin ERDOĞAN *, Cafer TEPELİ , Bertram BRENIG , Mine DOSAY AKBULUT , 1 4 5 Cevdet UĞUZ , Peter SAVOLAINEN , Ceyhan ÖZBEYAZ 1 Department of Medical Biology and Genetics, Faculty of Veterinary Medicine, Afyon Kocatepe University, Afyonkarahisar, Turkey 2 Department of Animal Science, Faculty of Veterinary Medicine, Selçuk University, Konya, Turkey 3 Institute of Veterinary Medicine, Göttingen University, Göttingen, Germany 4 Division of Gene Technology, KTH-Royal Institute of Technology, Science for Life Laboratory, Solna, Sweden 5 Department of Animal Science, Faculty of Veterinary Medicine, Ankara University, Ankara, Turkey
Received: 30.03.2012 Accepted: 07.09.2012 Published Online: 25.03.2013 Printed: 25.04.2013
Abstract: In this study, the genetic structures and relationships of native Turkish dog breeds were investigated using 20 polymorphic loci (17 microsatellites and 3 proteins). For this aim, a total of 141 blood samples were taken from Turkish shepherd dogs and Turkish
Greyhounds located in several geographical regions of Turkey. Multilocus FST values indicated that around 1.92% of the total genetic variation could be explained by breed differences and the remaining 98.08% by differences among individuals. The gene flow between populations within each generation varied between 8.4 (Akbash–White Kars Shepherd dog pairs) and 62.3 (Black–Grey Kars Shepherd dog pairs). Four different groups appeared in the 3-dimensional factorial correspondence analysis, and among these, dogs from the Akbash, Kangal, Kars Shepherd, and Turkish Greyhound breeds grouped in clearly separated clusters in distant parts of the 3-dimensional graph. These results clearly show that Akbash and Kangal Shepherd dogs are different populations with different genetic structures. Therefore, the generalised grouping of Turkish shepherd dogs into a single breed called Anatolian or Turkish shepherd dogs is incorrect.
Key words: Turkish dog breeds, microsatellite, F-statistics, genetic variability, factorial correspondence analysis, genetic distance
1. Introduction the origin of today’s dog breeds in Turkey. Dogs brought Archaeological findings show the presence of domestic by different cultures or nations conquering Anatolia also dogs in Germany, Israel, and North Iraq at 14,000–16,000 contributed to forming the modern dog breeds in Turkey years before the present (BP) (1,2). Still earlier dates for (11,12), e.g., the Turkish immigrants from Central Asia the presence of domestic dog have been suggested from who settled in Anatolia and brought dogs. However, Europe (3), but the evidence does not seem conclusive possible traces of the origins of Turkish dog breeds can be (4). Morphological, behavioural, and genetic data clearly found from across the Anatolian Plateau to Central Asia as indicate that domestic dogs (Canis familiaris) originated well as to the plateau of Afghanistan (13), and the history from domesticated wolves (5–7). It is not clear from of Turkish dog breeds is still obscure. Carvings of dog archaeological findings whether dogs originated from one pictures in caves (approximately 6000 BP) discovered in or more wolf population(s). Tibet resemble the modern shepherd dogs in Turkey, as Studies of mitochondrial DNA and Y-chromosomal well as mastiff breeds, Greyhound, and Saluki (11,12). DNA diversity world-wide indicate that dogs originated Kırmızı (11) claimed that the origins of Turkish in the southern part of East Asia (8–10). A study of shepherd dogs were in Central Asia and that these breeds genome-wide single-nucleotide polymorphism variation spread to Anatolia, the Middle East, and Europe through among domestic dogs and wolves showed dogs to share the emigration and immigration of Turks from Central more unique multilocus haplotypes with wolves from the Asia to Anatolia and Europe. These dogs were brought Middle East than with wolves from North China, Europe, to Europe by Turks during the Ottoman era or earlier, and America, but did not include samples from southern possibly serving as the origin of shepherd dog breeds East Asia (7). According to archaeological findings, dogs such as the Great Pyrenees, Chuvatch, Greek Shepherd similar to mastiff breeds lived in Anatolia at 9000 years dog, Kuvasz, Sharplaninatz, Komondor, and Maremma BP (11). It has been suggested that these dogs served as Shepherd dog in Europe (12,14). * Correspondence: [email protected] 176 ERDOĞAN et al. / Turk J Biol
There is no consensus on the origin of Akbash and PEZ12 labelled with JOE; PEZ6, PEZ8, FHC2079 labelled Kangal Shepherd dogs and their subvarieties (12). In a with NED), primer mix 2 (FH2247, FH2164 labelled study carried out by Erdoğan and Özbeyaz (15), Kangal with FAM; FH2001, FH2326 labelled with JOE; PEZ22, and Akbash Shepherd dogs were found to be located in FH2289 labelled with NED), or primer mix 3 (PEZ11, different clusters and to have different genetic structures FH2324 labelled with FAM; FH2161 labelled with NED). according to data from polymorphic loci. These findings The PCR buffer supplied by QIAGEN (Hilden, Germany) changed the perception about the idea that these 2 breeds contained 6 mM MgCl2. In a total reaction volume of 14 have a close relationship. µL with HotStar HiFidelity Taq polymerase (QIAGEN), The aim of the present study was to determine the 32 cycles were performed on an MJ Research Dyad Tetra microsatellite polymorphism in Turkish Greyhound, 4D thermocycler (MJ Research Inc., Waltham, MA, USA). Kangal, Akbash, and Kars Shepherd dog breeds, to estimate The DNA samples were denatured for 15 min at 95 °C and the genetic relatedness among these dog breeds, and to then subjected to cycles of 30 s at 94 °C, 1.5 min at 59 °C, address the following questions: What are the current and 1 min at 72 °C. Next, 1.5 µL of the PCR products was levels of gene flow between breeds? Has gene flow been the loaded on an ABI Prism 3100 Genetic Analyser (Applied main factor in the current genetic similarity between these Biosystems, Darmstadt, Germany). Profiles were analysed populations from different regions of Turkey? using Software 3100 GeneScan Analysis Module 2 and Genotyper V3.5 NT. 2. Materials and methods 2.2. Statistical analysis 2.1. Animal samples and microsatellite markers analysed The average heterozygosity in a population was calculated In this study, a total of 141 dogs from 6 breeds were used. according to the method of Nei (17) by using the values Kangal Shepherd dogs (n = 30), Akbash Shepherd dogs (n of heterozygosity calculated in each locus. The significant = 33), White Kars Shepherd dogs (KW, n = 15), Black Kars differences on an estimated average heterozygosity index Shepherd dogs (KB, n = 23), Grey Kars Shepherd dogs (KG, between populations were calculated with the t-test. n = 9), and Turkish Greyhounds (TG, n = 31) were chosen F-statistics were estimated in the form of F, θ, and ƒ as nonrelative to each other and as best representing their as described by Weir and Cockerham (18) according to breed characteristics (Figure 1). In the phenotyping of the FIT, FST, and FIS, respectively. The gene flow (Nem) between transferrin, postalbumin-1 (Poa-1) and postalbumin-3 populations was calculated with Nem = (1 ‒ FST) / 4 FST
(Poa-3), with the methods described by Erdoğan and based on the FST value of one locus and all loci (17,19). Özbeyaz (15), were used. DNA from blood samples was These were calculated using the GENETIX 4.05 computer extracted according to standard phenol–chloroform programme (20). methods (16). DNA concentrations were determined Classification of dogs according to their neighbourhood using a NanoDrop ND-1000 spectrophotometer (Peqlab, in the factorial space was drawn using the GENETIX 4.05 Erlangen, Germany) and adjusted to 25 ng/µL. One computer packet programmes (20). Further assessments microlitre of each DNA was mixed with primer mix 1 of genetic structures of populations were performed with (PEZ1, FHC2054, FHC2010 labelled with FAM; PEZ5, STRUCTURE version 2.3.2 (21).
Figure 1. Geographical locations and illustrations of the 6 Turkish dog breeds’ samples collected.
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3. Results breeds, FST, was calculated to be 1.92% (P < 0.001), which
The estimated values of the heterozygosity index (He was relatively low. The gene flow value, assigned for the and Ho) of the investigated dog breeds based on each number of individuals migrating between populations for locus and over all loci are given in Table 1. The average each generation and calculated from all loci, was 12.8. heterozygosity values were between 0.664 (KW) and 0.778 The gene flow between the population pairs and the FST (KB). The mean heterozygosity values calculated for each estimations are shown in Table 3. After 1000 permutations breed were not statistically significant. between breed pairs, all FST values were found to be The population differentiation was tested with a fixation significantly different from 0 (P < 0.001). The gene flow index, with FIT, FST, and FIS values for each locus and over occurring between populations in each generation varied all loci. The results of the F-statistical analysis of the 20 between 8.4 (Akbash–KW pairs) and 62.3 (KB and KG loci for all investigated breeds are shown in Table 2. The pairs). lack of heterozygosity level was around 3.27% (P < 0.001) The Reynolds genetic distance matrix is given in Table for each of the analysed breeds and 5.12% (P < 0.001) for 4. The genetic distance values vary between 0.0011 and the whole population. The genetic differentiation between 0.0358. The smallest genetic distance value (0.0011) was
Table 1. The heterozygosity index, calculated according to locus and all loci of dog breeds (He and Ho), average heterozygosity, and allele numbers.
Akbash Kangal KW KB KG TG Locus (n = 33) (n = 30) (n = 15) (n = 23) (n = 9) (n = 31)
Allele He Ho Allele He Ho Allele He Ho Allele He Ho Allele He Ho Allele He Ho FH2001 6 0.840 0.684 6 0.808 0.833 5 0.683 0.615 8 0.854 0.923 5 0.824 0.889 7 0.815 0.750
FH2161 5 0.789 0.818 8 0.823 0.733 7 0.708 0.539 11 0.836 0.913 5 0.758 0.667 9 0.809 0.625
FH2164 14 0.886 0.833 12 0.838 0.867 8 0.831 0.846 9 0.851 0.769 11 0.948 0.889 13 0.902 0.969
FH2247 24 0.950 0.976 26 0.957 0.967 15 0.954 0.923 21 0.953 1.000 9 0.909 0.889 23 0.950 0.969
FH2289 16 0.918 0.854 17 0.893 0.867 10 0.779 0.461 15 0.919 0.923 10 0.915 0.864 14 0.891 0.844
FH2324 17 0.917 0.758 19 0.918 0.867 11 0.877 0.692 12 0.863 0.826 12 0.954 0.901 11 0.880 0.813
FH2326 15 0.915 0.878 16 0.912 0.833 13 0.939 0.846 16 0.911 0.808 9 0.928 0.876 18 0.940 0.781
FHC2010 7 0.740 0.737 7 0.580 0.548 5 0.742 0.692 4 0.655 0.692 4 0.765 0.444 5 0.675 0.625
FHC2054 9 0.870 0.868 10 0.852 1.000 6 0.803 0.769 8 0.847 0.889 7 0.863 0.889 7 0.844 0.831
FHC2079 6 0.679 0.658 3 0.579 0.710 2 0.409 0.231 6 0.661 0.741 6 0.758 0.778 3 0.514 0.500
PEZ1 9 0.803 0.806 12 0.786 0.807 6 0.825 0.846 4 0.750 0.889 4 0.712 0.667 10 0.816 0.844
PEZ5 7 0.767 0.686 8 0.749 0.742 4 0.754 0.846 6 0.741 0.667 4 0.726 0.778 5 0.719 0.719
PEZ6 15 0.882 0.842 19 0.918 0.839 10 0.902 0.846 16 0.895 0.815 7 0.869 0.889 11 0.865 0.656
PEZ8 11 0.820 0.632 11 0.841 0.839 10 0.905 0.846 12 0.873 0.692 8 0.889 0.667 10 0.843 0.844
PEZ11 13 0.893 0.727 11 0.847 0.800 10 0.892 0.769 12 0.893 0.870 9 0.928 0.889 9 0.836 0.625
PEZ12 15 0.861 0.895 15 0.810 0.741 6 0.628 0.692 10 0.809 0.778 8 0.876 0.667 11 0.836 0.813
PEZ22 13 0.875 0.838 11 0.816 0.900 7 0.794 0.692 9 0.895 1.000 7 0.863 0.667 10 0.827 0.813
Tf 2 0.504 0.559 2 0.509 0.400 2 0.443 0.308 2 0.485 0.409 2 0.400 0.250 2 0.482 0.516
Poa-1 2 0.395 0.529 2 0.325 0.400 3 0.489 0.500 2 0.426 0.500 2 0.458 0.625 2 0.252 0.290
Poa-3 2 0.479 0.706 2 0.503 0.633 2 0.271 0.308 2 0.495 0.455 2 0.533 0.500 2 0.389 0.452
Mean 10.4 0.789 0.764 10.85 0.763 0.766 7.1 0.731 0.664 9.25 0.781 0.778 6.55 0.794 0.735 9.1 0.754 0.715 estimates
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Table 2. F-statistic values and number of individuals migrating between populations in each generation.
Locus FIS = ƒ FIT = F FST = θ Nem FH2001 0.0485 0.0533 0.0050 FH2161 0.0781 0.0896 0.0124 FH2164 0.0129 0.0212 0.0084 FH2247 –0.0186 –0.0035 0.0148 FH2289 0.0693 0.0887 0.0209 FH2324 0.1033 0.1282 0.0278 FH2326 0.0966 0.1038 0.0080 FHC2010 0.0522 0.0590 0.0072 FHC2054 –0.0434 –0.0285 0.0144 FHC2079 –0.0320 0.0068 0.0376 PEZ1 –0.0455 –0.0444 0.0011 PEZ5 0.0322 0.0672 0.0362 PEZ6 0.1019 0.1037 0.0021 PEZ8 0.1177 0.1490 0.0355 PEZ11 0.1343 0.1519 0.0203 PEZ12 0.0292 0.0463 0.0176 PEZ22 –0.0027 0.0089 0.0116 Tf 0.0763 0.1058 0.0320 Poa-1 –0.2302 –0.2189 0.0091 Poa-3 –0.2124 –0.0937 0.0979 Mean estimates 0.0327 (0.016)*** 0.0512 (0.016)*** 0.0192 (0.003)*** 12.8
ƒ, estimation of pure breeding within the population; F, estimation of total pure breeding; θ, measure of population differentiation. The standard deviation is given in parentheses. ***P < 0.001, from permutation tests in the TFPGA programme.
Table 3. TheF ST statistics (vertical triangle) and gene migration Nem (inverted vertical triangle) among dog breeds in Turkey.
Breeds Akbash Kangal KW KG KB TG Akbash *** 17.6 8.4 35.5 35.5 10.2 Kangal 0.014 *** 9.0 14.5 15.4 9.8 KW 0.029 0.027 *** 41.4 12.9 10.2 KG 0.007 0.017 0.006 *** 62.3 11.1 KB 0.007 0.016 0.019 0.004 *** 11.7 TG 0.024 0.025 0.024 0.022 0.021 ***
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Table 4. The Reynolds genetic distance matrix among the 6 Turkish dog breeds.
Breeds Akbash Kangal KW KG KB TG Akbash ------Kangal 0.0141 - - - - - KW 0.0358 0.0320 - - - - KG 0.0020 0.0145 0.0150 - - - KB 0.0076 0.0147 0.0289 0.0011 - - TG 0.0244 0.0269 0.0307 0.0214 0.0232 -
found between KB and KG. In terms of genetic distance a structure test analysis using K = 6 to determine which analysis, the breeds Akbash and KW were genetically far breed or breeds an individual belongs to, and to group the away from each other. The genetic distance value between individuals (Figure 3). these 2 breeds was 0.0358. To investigate and show the relationship between 4. Discussion individuals, 3-dimensional factorial correspondence The observed heterozygosity values from all loci in all analysis (3D-FCA) and the GENETIX 4.05 computer breeds were found to be between 0.664 (KW) and 0.778 packet programme (20) were employed (Figure 2). (Kangal) (Table 1). These estimates of heterozygosity The 3D-FCA grouped all populations into 4 clusters. values show that the genetic variations are high in all The Akbash breed, Turkish Greyhound breed, and Kangal animals investigated in this study, and that there is no breed formed 3 of these groups, which were clearly separated statistically significant difference between the breeds for and located in different parts of the 3-dimensional graph. the heterozygosity values. This means that all the dogs The dogs from the Kars region (KB, KG, KW) constituted have the same level of genetic variations, and it is difficult a single fourth group placed in the middle of all the other to differentiate the dog breeds from each other in terms of groups (Figure 2). Within this fourth group, it could be mean heterozygosity levels. seen that there were 3 breeds: KB, KG, and KW. However, It has been reported that the Ho and He values for the breeds KW and KB were grouped into the same cluster Akbash, Kangal, and Turkish Greyhound breeds are but settled in different positions (Figure 2). 0.715, 0.701, and 0.710, and 0.620, 0.701, and 0.705 (22), It can be seen that the Turkish Greyhound is more respectively, and that the average heterozygosity values for pure or a closer relative compared to the other breeds in Akbash and Kangal are 0.367 and 0.410, respectively (15). Axis 2 (22.51%) Axis
Axis 3 (21.22%)
Axis 1 (26.63%)
Figure 2. Three-dimensional factorial correspondence analysis depiction of the genetic relationships among indigenous dog breeds in Turkey.
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1.00 0.80 0.60 0.40 0.20 0.00 AKBASHKANGALKARS KARS KARS TURKISH WHITE GREY BLACK GREYHOUND Figure 3. Structure analysis of dog breeds of Turkey.
The estimated oH and He values for Akbash, Kangal, and For population mating at random, genes are equally
Turkish Greyhound are 0.764, 0.766, and 0.715, and 0.789, related within or between individuals. In this case, FIT =
0.763, and 0.754, respectively (Table 1). The heterozygosity FST or FIS = 0. Therefore, the significant difference in the levels for Akbash, Kangal, and Turkish Greyhound are estimates of FIT and FST indicates a departure from random higher than those reported by Erdoğan and Özbeyaz mating. Avoidance of mating between relatives will cause
(15) and Altunok et al. (22); for dog breeds in Japan (23), positive FST values that exceed the negative values of FIT
Bangladesh (24), Germany (25), and Finland (26); in and FIS. Generally, if FIS is positive (FIT > FST), it could be
Golden Retriever, Labrador Retriever, and Rottweiler (27), interpreted as evidence of inbreeding (34). FIS was positive and Beagle and Labrador Retriever (28); and in 16 street (0.0327) while FIT was 0.0512, which was greater than the dogs (29), 11 East Asian domestic dog breeds (30), 28 dog 0.0192 value of FST (Table 1). The estimate of the FIT and breeds (31), Bali street dogs (32), and Hannover hunting FIS values was positive for some loci. This means that the dogs (33). The reason for attaining higher heterozygosity frequencies of heterozygote genotypes are in accordance values in comparison to other researchers could be with the Hardy–Weinberg balance, but they are lower the selection of dogs from different regions that were than those of expected heterozygote values in these unrelated to each other, and the fact that the loci searched loci in all breeds. It is possible that the selection factors have multiple alleles. have a positive effect on the frequencies of homozygote Population differentiation was tested with a fixation genotypes based on selected loci at individual and index of FIT, FST, and FIS for each locus and all loci. The population levels. Neglecting the effects of migration, and F-statistics results, calculated from all 20 loci for all 6 assuming a low contribution of mutations to the genetic dog breeds, are given in Table 2. The observed mean diversity between these breeds, the differences in allele heterozygote deficiency was 3.27% for each investigated frequencies may be interpreted as primarily the result of breed and 5.12% in all populations, and these values random genetic drift. The genetic differentiation (1.92%) are statistically significant (P < 0.001). The frequency in may be seen as the result of an increased mean inbreeding homozygote genotypes in all populations is higher than coefficient FIT over a rather short period of time. We that expected in the Hardy–Weinberg balance. This means therefore consider the relatively low mean FIS value that there is not random mating in populations or that (0.0327) to be the result of a reduction of heterozygosity there is inbreeding. The genetic differentiation among within the breeds studied and the relatively low mean FIT breeds or the calculated FST was 1.92% (P < 0.001). This value (0.0512) as indicative of ineffective barriers to gene value of FST shows that the populations have different flow between populations. genetic structures. It has been reported that the genetic distance among
The FIS, FIT, and FST values calculated from all loci were Spanish dog breeds ranges from 0.000 to 0.051 (35). 0.085, 0.083, and 0.160, respectively, for 7 dog breeds in Although there was not a dramatic difference among dog
Turkey (15), whereas the FIS, FIT, and FST values were 0.072, breeds, the genetic distance varied between 0.013 and 0.214, and 0.154, respectively, in 11 East Asian domestic 0.242 for 7 dog breeds (15). The genetic distance between dog breeds (30), and the values of FIS and FIT vary between Akbash and Kangal Shepherd dogs has been estimated
0.01 and 0.13 and the FST value varies between 0.602 and to be 0.093 and the Nem value to be 1.3 (15). The genetic 0.975 in 5 dog breeds of Finland (26). However, Altunok distance between dog breeds ranged from 0.0836 to 0.3235 et al. (22) determined that the pairwise FST values were and the Nem value ranged between 0.43 and 10.83 (30), 0.167 and 0.121 in Kangal–Akbash and Kangal–Turkish while Koskinen and Bredbacka (26) demonstrated that the Greyhound, respectively. The calculated F-statistic values genetic distance between dog breeds ranged between 0.182 in this study are much lower than those of other reports and 0.291. In this study, the estimated genetic distance (15,22,26,30). value ranged from 0.0011 (KB–KG) to 0.0358 (Akbash–
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KW), and the Nem value ranged between 8.4 (Akbash– However, KW and KB were grouped into the same cluster
KW) and 62.3 (Akbash–KW). The expected Nem value but in different positions. KG was also situated in the calculated from all loci was 12.8. The calculated genetic middle of other Kars dog breeds. These findings indicate distance among dog breeds in this study is in accordance that KW and KB are a variety of Kars Shepherd dogs with the findings in Spanish dog breeds (35), but lower separately, and KG is also a crossbreed of these 2 Kars than those in other reports. The Nem value was higher Shepherd dog breeds. than other researchers’ findings, as well. In conclusion, our analysis indicates that native
The mean estimation of genetic differentiation (FST) Turkish dog breeds have different genetic structures on the among breeds was 1.92% (P < 0.001). This value is lower basis of the analysed loci, since they are located in clearly than that of other breeds, and the effective number of separated clusters in 3D-FCA. These findings disprove individuals exchanged between populations per generation the beliefs that Kangal, Akbash, and Kars Shepherd dogs was 12.8 (Table 3). The gene flow ranges from 8.4 to 62.3 are close relatives of each other. The results clearly show between populations. If gene flow is higher than 1 (in an that Akbash, Kangal, and Kars Shepherd dogs are different endless island model), this means that Nem is large enough populations. Therefore, the generalised grouping of to decrease the genetic differentiation between populations Turkish shepherd dogs into a single breed called Anatolian (36). Genetic drift could be a factor supporting genetic or Turkish shepherd dogs is not correct. It is proper differentiation between breeds. If gene exchange occurs to differentiate between Kangal Shepherd and Akbash between breeds at a high frequency, the FST value gets lower, Shepherd dogs as separate breeds. which means that genetic differentiation occurs through To determine the genetic structure of the dogs, gene flow. These results could be interpreted to indicate polymorphic biochemical systems and microsatellite loci that the investigated shepherd dog breeds originated from can be employed to compare the dog populations and a common ancestor a long time ago and lived together the average heterozygosity values, and F-statistics, the in the same geographical area. Gene flow plays a very individual numbers migrating in each generation, and the important role in populations living within the same or FCA method can be used for successful classification of close geographical areas. The reason for high gene flow the breeds. occurring among the investigated dog breeds could be that Since Turkey is located in both Asia and Europe as a the dogs (except the Turkish Greyhound) live in the same junction for migration roads, and since it is the cradle of geographical areas. Although the Turkish Greyhound is many different cultures, further determination of genetic not considered among shepherd dogs, this breed has a structure in Turkish dog breeds is important for better high rate of gene flow with the other dog breeds (Table 3). understanding of the genetic origin of dogs in Asia and The high gene flow between Turkish Greyhound and other Europe. The determination of genetic structure and genetic dog breeds could be due to uncontrolled mating between relatedness in Turkish dogs will also help to determine the Turkish Greyhound and other dog breeds. migration paths not only for dogs but also for people or Four different clusters were obtained as a result of civilisations in history. 3D-FCA applied to determine and show the relationship between individuals (Figure 2). The 3-dimensional graph Acknowledgements shows that Akbash, Turkish Greyhound, and Kangal dogs This research was supported by the Scientific and are distinctly separated and positioned in different sectors. Technological Research Council of Turkey (TÜBİTAK In between these 3 groups, the 3 Kars breeds constitute TOVAG 103V024), and additional support was provided a fourth group. Populations are genetically separated by Afyon Kocatepe University (AKÜ BAPK 041 VF 06). We into distinct clusters. Direct evidence supporting this are grateful to S. Rose and S. Loos for excellent technical conclusion derives from 3D-FCA analyses (Figure 2). assistance. Peter Savolainen is a Royal Swedish Academy Interestingly, despite their different exterior morphology, of Sciences Research Fellow supported by a grant from the the Kars dog breeds do not show a clear genetic divergence. Knut and Alice Wallenberg Foundation.
References
1. Clutton-Brock J. Evolution of domesticated animals. In: Mason 3. Sablin MV, Khlopachev GA. The earliest ice age dogs: evidence IL. ed. Dog. Longman; 1984: pp. 198–211. from Eliseevichi I. Curr Anthropol 43: 795–799, 2002. 2. Zeder MA. Domestication and early agriculture in the 4. Napierala H, Uerpmann HP. A ‘new’ palaeolithic dog from Mediterranean basin: origins, diffusion, and impact. P Natl Central Europe. Int J Osteoarchaeol 22: 127–137, 2010. Acad Sci-Biol 105: 11597–11604, 2008.
182 ERDOĞAN et al. / Turk J Biol
5. Fiennes R, Fiennes A. The Natural History of the Dog. 22. Altunok V, Koban E, Chikhi L et al. Genetic evidence for the Wedenfeld & Nicolson Press. London; 1968. distinctness of Kangal dogs. B Vet I Pulawy 49: 249–254, 2005. 6. Savolainen P, Zhang Y, Luo J et al. Genetic evidence for an East 23. Tanabe Y, Ôta K, Ito S et al. Biochemical-genetic relationships Asian origin of domestic dogs. Science 298: 1610–1613, 2002. among Asian and European dogs and the ancestry of the Japanese native dogs. J Anim Breed Genet 108: 455–478, 1991. 7. vonHoldt BM, Pollinger JP, Lohmueller KE et al. Genome-wide SNP and haplotype analyses reveal a rich history underlying 24. Kobayashi R, Miyakawa H, Tanabe Y et al. Blood protein dog domestication. Nature 464: 898–902, 2010. polymorphism in Bangladesh native dogs. Report of the Society for Researches on Native Livestock 12: 269–289, 1987. 8. Pang JF, Kluetsch C, Zou XJ et al. mtDNA data indicate a single origin for dogs south of Yangtze River, less than 16,300 years 25. Lachmann C. Verteilung genetischer Polymorphismen bei ago, from numerous wolves. Mol Biol Evol 26: 2849–2864, einigen deutschen Hunderassen. PhD, University of Veterinary 2009. Medicine, Hannover, 1993. 9. Ding ZL, Oskarsson M, Ardalan A et al. Origins of domestic 26. Koskinen MT, Bredbacka P. Assessment of the population dog in Southern East Asia is supported by analysis of structure of five Finnish dog breeds with microsatellites. Anim Y-chromosome DNA. Heredity 108: 507–514, 2012. Genet 31: 310–317, 2000. 10. Ardalan A, Kluetsch CFC, Zhang A et al. Comprehensive 27. Altet L, Francino O, Sánchez A. Microsatellite polymorphism study of mtDNA among Southwest Asian dogs contradicts in closely related dogs. J Hered 92: 276–279, 2001. independent domestication of wolf, but implies dog–wolf 28. Ichikawa Y, Takagi K, Tsumagari S et al. Canine parentage hybridization. Ecology and Evolution 3: 373–385, 2011. testing based on microsatellite polymorphisms. J Vet Med Sci 11. Kırmızı E. Türk çoban köpeklerinin tarihçesi. Türk Veteriner 63: 1209–1213, 2001. Hekimliği Dergisi 6: 39–41, 1994. 29. Jouquand S, Priat C, Hitte C et al. Identification and 12. Nelson DD. A general classification of the native dogs of characterization of a set of 100 tri- and dinucleotide Turkey. In: The International Symposium on Turkish Shepherd microsatellites in the canine genome. Anim Genet 31: 266– Dogs, Selçuk University, Konya, Turkey; 1996: pp. 19–94. 272, 2000. 13. Özbeyaz C. Kangal köpeklerinde bazı morfolojik özellikler. 30. Kim KS, Tanabe Y, Park CK et al. Genetic variability in East Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 34: 38–46, Asian dogs using microsatellite loci analysis. J Hered 92: 398– 1994. 403, 2001. 14. Reed S. The history of Turkish shepherd dogs. In: The 31. Irion DN, Schaffer AL, Famula TR et al. Analysis of genetic International Symposium on Turkish Shepherd Dogs, Selçuk variation in 28 dog breed populations with 100 microsatellite University, Konya, Turkey; 1996: pp. 97–109. markers. J Hered 94: 81–87, 2003. 15. Erdoğan M, Özbeyaz C. Investigation of blood protein 32. Irion DN, Schaffer AL, Grant S et al. Genetic variation analysis polymorphism and estimation of genetic distances in some of the Bali street dog using microsatellites. BMC Genet 6: 1–13, dog breeds in Turkey. Turk J Vet Anim Sci 28: 583–590, 2004. 2005. 16. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A 33. Lupke L, Distl O. Microsatellite marker analysis of the genetic Laboratory Manual. Cold Spring Harbor Press. Cold Spring variability in Hanoverian hounds. J Anim Breed Genet 122: Harbor, NY, USA; 1989. 131–139, 2005. 17. Nei M. Molecular Evolutionary Genetics. Columbia University 34. Weir BS. Genetic Data Analysis. Sinauer Associates Press. Press. New York; 1987. Sunderland, MA, USA; 1996. 18. Weir BS, Cockerham CC. Estimating F-statistics for the analysis 35. Jordana J, Piedrafita J, Sánchez A. Genetic relationships of population structure. Evolution 38: 1358–1370, 1984. in Spanish dog breeds, II. The analysis of biochemical polymorphism. Genet Sel Evol 24: 245–263, 1992. 19. Hartl DL, Clark AG. Principles of Population Genetics. Sinauer Associates Press. Sunderland, MA, USA; 1989. 36. Trexler JC. Hierarchical organization of genetic variation in the Sailfin Molly, Poecilia latipinna (Pisces: Poeciliidae). Evolution 20. Belkhir K, Borsa P, Chikhi L et al. GENETIX 4.00 WindowsTM 42: 995–1005, 1988. Software for Population Genetics. University of Montpellier Press. Montpellier, France; 1996. 21. Pritchard JK, Stephens M, Donnelly P. Inference of population structure using multilocus genotype data. Genetics 155: 945– 959, 2000.
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