Native cattle breeds can carry specific signatures of selection reflecting their adaptation to the local environmental conditions and response to the breeding strategy used. In this study, we comprehensively analysed high-density single nucleotide polymorphism (SNP) genotypes to characterise the population structure and detect the selection signatures in Russian native Yaroslavl and Kholmogor dairy cattle breeds, which have been little influenced by introgression with transboundary breeds. Fifty-six samples of pedigree-recorded purebred animals, originating from different breeding farms and representing different sire lines, of the two studied breeds were genotyped using a genome-wide bovine genotyping array (Bovine HD BeadChip). Three statistical analyses—calculation of fixation index (FST) for each SNP for the comparison of the pairs of breeds, hapFLK analysis, and estimation of the runs of homozygosity (ROH) islands shared in more than 50% of animals—were combined for detecting the selection signatures in the genome of the studied cattle breeds. We confirmed nine and six known regions under putative selection in the genomes of Yaroslavl and Kholmogor cattle, respectively; the flanking positions of most of these regions were elucidated. Only two of the selected regions (localised on BTA 14 at 24.4–25.1 Mbp and on BTA 16 at 42.5–43.5 Mb) overlapped in Yaroslavl, Kholmogor and Holstein breeds. In addition, we detected three novel selection sweeps in the genome of Yaroslavl (BTA 4 at 4.74–5.36 Mbp, BTA 15 at 17.80–18.77 Mbp, and BTA 17 at 45.59–45.61 Mbp) and Kholmogor breeds (BTA 12 at 82.40–81.69 Mbp, BTA 15 at 16.04–16.62 Mbp, and BTA 18 at 0.19–1.46 Mbp) by using at least two of the above-mentioned methods. We expanded the list of candidate genes associated with the selected genomic regions and performed their functional annotation. We discussed the possible involvement of the identified candidate genes in artificial selection in connection with the origin and development of the breeds. Our findings on the Yaroslavl and Kholmogor breeds obtained using high-density SNP genotyping and three different statistical methods allowed the detection of novel putative genomic regions and candidate genes that might be under selection. These results might be useful for the sustainable development and conservation of these two oldest Russian native cattle breeds.
Analysis of ancient and historical DNA has great potential to trace the genetic diversity of local cattle populations during their centuries-long development. Forty-nine specimens representing five cattle breeds (Kholmogor, Yaroslavl, Great Russian, Novgorod, and Holland), dated from the end of the 19th century to the first half of the 20th century, were genotyped for nine polymorphic microsatellite loci. Using a multiple-tube approach, we determined the consensus genotypes of all samples/loci analysed. Amplification errors, including allelic drop-out (ADO) and false alleles (FA), occurred with an average frequency of 2.35% and 0.79%, respectively. A significant effect of allelic length on ADO rate (r2 = 0.620, p = 0.05) was shown. We did not observe significant differences in genetic diversity among historical samples and modern representatives of Kholmogor and Yaroslavl breeds. The unbiased expected heterozygosity values were 0.726–0.774 and 0.708–0.739; the allelic richness values were 2.716–2.893 and 2.661–2.758 for the historical and modern samples, respectively. Analyses of FST and Jost’s D genetic distances, and the results of STRUCTURE clustering, showed the maintenance of a part of historical components in the modern populations of Kholmogor and Yaroslavl cattle. Our study contributes to the conservation of biodiversity in the local Russian genetic resources of cattle.
Comparison of genomic footprints in chicken breeds with different selection history is a powerful tool in elucidating genomic regions that have been targeted by recent and more ancient selection. In the present work, we aimed at examining and comparing the trajectories of artificial selection in the genomes of the native egg-type Russian White (RW) and meat-type White Cornish (WC) breeds. Combining three different statistics (top 0.1% SNP by FST value at pairwise breed comparison, hapFLK analysis, and identification of ROH island shared by more than 50% of individuals), we detected 45 genomic regions under putative selection including 11 selective sweep regions, which were detected by at least two different methods. Four of such regions were breed-specific for each of RW breed (on GGA1, GGA5, GGA8, and GGA9) and WC breed (on GGA1, GGA5, GGA8, and GGA28), while three remaining regions on GGA2 (two sweeps) and GGA3 were common for both breeds. Most of identified genomic regions overlapped with known QTLs and/or candidate genes including those for body temperatures, egg productivity, and feed intake in RW chickens and those for growth, meat and carcass traits, and feed efficiency in WC chickens. These findings were concordant with the breed origin and history of their artificial selection. We determined a set of 188 prioritized candidate genes retrieved from the 11 overlapped regions of putative selection and reviewed their functions relative to phenotypic traits of interest in the two breeds. One of the RW-specific sweep regions harbored the known domestication gene, TSHR. Gene ontology and functional annotation analysis provided additional insight into a functional coherence of genes in the sweep regions. We also showed a greater candidate gene richness on microchromosomes relative to macrochromosomes in these genomic areas. Our results on the selection history of RW and WC chickens and their key candidate genes under selection serve as a profound information for further conservation of their genomic diversity and efficient breeding.
Summary Local breeds can serve as an important source of genetic variability in domestic animal species. This study aimed to assess the genetic diversity and population structure of Belarusian Red cattle and their differentiation from other European cattle populations based on genome‐wide SNP genotypes. Twenty pedigree‐recorded non‐closely related cows of Belarusian Red cattle were genotyped using the Illumina BovineHD BeadChip. Genotypes of 22 other European cattle breeds were included in the study for comparison. A total of 28 562 SNPs passed through the quality control checks and were selected for analysis. The Belarusian Red cattle displayed a moderate level of genetic variability (UHE = 0.341, HO = 0.368), and the highest heterozygote excess (UFIS = −0.066), among the studied breeds; this reflects the contribution of multiple breeds to their formation. The principal component analysis, FST‐based Neighbor‐Net tree and Admixture clustering, clearly distinguished the Belarusian Red cattle from the other European cattle breeds. Moreover, the presence of ancestral genomic components of Danish Red and Brown Swiss breeds were clearly visible, which agrees with the breed's history and its recent development. Our study highlights the importance of maintaining the specific genomic components, which makes a significant contribution to the global genetic diversity in the modern population of Belarusian Red cattle, allowing us to consider them a valuable national genetic resource. Our research results will be useful for the development of conservation programs for this local cattle breed.
Poaching is one of the major types of wildlife crime in Russia. Remnants of goats (presumably the wild endemic species, the Caucasian tur) were found in an area of the Caucasian mountains. The case study involves a suspected poacher whose vehicle was found to have two duffel bags containing pieces of a carcass, which he claimed was that of a goat from his flock. The aim of the forensic genetic analysis for this case was to (i) establish individual identity and (ii) perform species identification. DNA typing based on fourteen microsatellites revealed that STR-genotypes generated from pieces of evidence found at crime scene fully matched those obtained from the evidence seized from the suspect. The results of genome-wide SNP-genotyping, using Illumina Goat SNP50 BeadChip, provided evidence that the poached animal was a wild Caucasian tur (Capra caucasica). Thus, based on comprehensive molecular genetic analysis, evidence of poaching was obtained and sent to local authorities. To our knowledge, this case study is the first to attempt to use DNA chips in wildlife forensics of ungulates.
The comparative molecular genetic study of museum and modern representatives of cattle breeds can help to elucidate the origin and maintenance of historical genetic components in modern populations. We generated the consensus genotypes for 11 microsatellite loci for 24 museum samples of Kalmyk, Kyrgyz, and Kazakh cattle, dated from the first quarter of the 20th century, and compared them with those of modern Kalmyk, Kyrgyz, and Kazakh white-headed breeds. The level of genetic diversity of the modern Kalmyk and Kyrgyz cattle (uHe = 0.771–0.778) was similar to those observed in the museum samples (uHe = 0.772–0.776), while a visible decrease in genetic variability in the modern Kazakh white-headed breed compared to museum Kazakh cattle was detected (uHe = 0.726 and 0.767, respectively). The PCA plot, FST- and Jost’s D-based networks, and STRUCTURE clustering provided strong evidence of the maintenance of the historical genetic background in modern populations of Kalmyk and Kyrgyz cattle. In spite of the allele pool of Kazakh white-headed cattle having undergone great changes compared to the museum Kazakh cattle, several animals still carry the visible aspect of the historical genetic components. Our results can be used for the selection of individuals for the creation of gene banks and may significantly improve the efficiency of conservation programs aimed at preserving genetic diversity in the national genetic resources of cattle.
Развитие молекулярно-генетических методов дает возможность уточнить происхождение и демографическую историю пород сельскохозяйственных животных. Сохранившиеся в краниологических коллекциях образцы костей и зубов служат источником ДНК для подобных исследований. Работа с историческими образцами осложняется наличием очень малых количеств ДНК, высокой степенью ее деградации и загрязнением образцов ингибиторами ПЦР. Целью настоящей работы было сравнение результативности различных методов экстракции ДНК из исторических черепов крупного рогатого скота, пригодной для проведения молекулярно-генетических исследований. Материалом служили зубы, извлеченные из исторических черепов крупного рогатого скота ярославской и холмогорской пород, хранящихся в краниологической коллекции Музея животноводства им. Е.Ф. Лискуна РГАУ-МСХА им. К.А. Тимирязева. На первом этапе сравнивали различные методы выделения ДНК согласно протоколам к соответствующим коммерческим наборам, модифицируя используемое количества костного материала и условия лизиса: Prep Filer™ BTA Forensic DNA Extraction Kit («Thermo Fisher Scientific, Inc.», США), COrDIS «Экстракт» декальцин (ООО «ГОРДИЗ», Россия), М-сорб-кость (ООО «Синтол», Россия), QIAamp DNA Investigator Kit («Qiagen», США). По результатам предварительных исследований для более детального анализа отобрали QIAamp DNA Investigator Kit («Qiagen», США), в котором реализована технология выделения на колонках с силикагелевой мембраной, и Prep Filer™ BTA Forensic DNA Extraction Kit («Thermo Fisher Scientific, Inc.», США), основанный на использовании магнитных частиц. Количественные и качественные характеристики полученной ДНК оценивали измерением концентрации двухцепочечной ДНК на флуориметре Qubit™ («Invitrogen, Life Technologies», США) и по соотношению поглощения при = 260 нм и = 280 нм на приборе NanoDrop 8000 («Thermo Fisher Scientific, Inc.», США). Пригодность полученных препаратов ДНК для молекулярно-генетических исследований оценивали на основании мультиплексного анализа 11 микросателлитных локусов (TGLA227,
Native coarse wool fat-tailed sheep breeds are the main producers of mutton in mountainous, steppe and semi-desert regions of Southern Russia. An assessment of homozygosity is required to prevent loss of genetic diversity and inbreeding depression onset. The aim of our work is estimation of genomic inbreeding and distribution of the runs of homozygosity (ROH) segments in Russian sheep breeds. Sample collection included Andean (n = 20), Buryat (n = 18), Buubei (n = 11), Edilbai (n = 20), Karachaev (n = 20), Kalmyk (n = 22), Karakul (n = 20), and Lezgin (n = 15) breeds. All sheep were genotyped using Ovine Infinium® HD SNP BeadChip (Illumina, USA). Quality control and raw data processing were performed in PLINK 1.9. A consecutive runs method implemented in the R package “detectRUNS” was used for estimation of ROH segments. We detected the ROH in all breeds with mean length ranging from 0.35 in Lezgin to 11.18 Mb in Karachaev. Andean had the greatest number of the ROH segments (total of 8496), while 3094 ROH segments were found in Buubei. Short ROH segments (< 2 Mb) were predominant in all breeds and varied from 96.83% in Andean to 99.87% in Buubei. The frequencies of the ROH segments of 8–16 Mb ranged from 0.03% in Karakul to 0.18% in Andean. ROH segments of 8–16 Mb did not occur in the Buubei and Lezgin breeds. The ROH segments were found on all autosomes. The largest genome coverage in the ROH was identified on OAR1, OAR 12 and OAR 20 and the lowest was accounted for OAR 17, OAR 18 and OAR 19. The values FROH ranged from 0.042 in Buubei to 0.084 in Andean. Except of Andean, no traces of long-term inbreeding were detected in local breeds. The obtained data are relevant for future sustainable management of native fat-tailed sheep breeds in Russia. The study was funded by RSF No. 19-16-00070.
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