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The present study has analyzed the allelic-specific expression in Purebred Sistani (Bos Indicus) and their crossbreed with Holstein, Simmental, and Montbeliarde breeds (Bos Taurus). The blood samples were taken from the caudal vein of purebred Sistani cows and crossbreed Sistani's with Holstein, Simental, and Montbeliarde (4 treatments). We discovered 152,496 (Purebred Sistani), 134,285 (Sistani × Simmental), 163,362 (Sistani × Montbeliarde), and 177,042 (Sistani × Holstein) SNPs on the assembled transcriptomes. In the Purebred Sistani, 8295 (5%), Sistani × Holstein crossbreed 11,900 (7%), Sistani × Simmental crossbreed 13,187 (10%), and Sistani × Montbeliarde crossbreed 16,666 (10%) number of SNPs were identified as ASE-SNPs. In the present study, 12 SNPs types identify, of which four were transition and eight were transversion. The most common SNPs were transition types. These SNPs were present in purebred Sistani 71.84%, Sistani × Holstein crossbreed 72.65%, Sistani × Simmental crossbreed 72.60%, and Sistani × Montbeliarde crossbreed 71.94%. Ontology analysis of the expressed genes in these cows revealed the involvement of these genes in different Biological classifications. Conducting such studies in parts of the world, such as the Sistan region, where it is not possible to record accurate records of cows, is a suitable and economical method for identifying genes with different expressions.
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Hibridização Genética , Feminino , Bovinos/genética , Animais , AlelosRESUMO
The coat color of dromedary is usually uniform and varies from black to white, although dark- to light-brown colors are the most common phenotypes. This project was designed to gain knowledge on novel color-related variants using genotyping-by-sequencing (GBS). The association between the SNPs and coat color was tested using MLM (mixed linear models) with kinship matrix. Three GWAS models including white color vs. non-white color, black vs. non-black color, and light-brown vs. dark-brown color were performed. There were no distinct genetic clusters detected based on the color phenotypes. However, admixture occurred among all individuals of the four different coat color groups. We identified nine significant SNPs associated with white color after Bonferroni correction, located close to ANKRD26, GNB1, TSPYL4, TEKT5, DEXI, CIITA, TVP23B, CLEC16A, TMPRSS13, FXYD6, MPZL3, ANKRD26, HFM1, CDC7, TGFBR3, and HACE1 genes in neighboring flanking regions. The 13 significant SNPs associated with black color and the candidate genes were: CAPN7, CHRM4, CIITA, CLEC16A, COL4A4, COL6A6, CREB3L1, DEXI, DGKZ, DGKZ, EAF1, HDLBP, INPP5F, MCMBP, MDK, SEC23IP, SNAI1, TBX15, TEKT5, TMEM189, trpS, TSPYL4, TVP23B, and UBE2V1. The SNAI1 gene interacted with MCIR, ASIP and KIT genes. These genes play a key role in the melanin biosynthetic and pigmentation biological process and melanogenesis biological pathway. Further research using a larger sample size and pedigree data will allow confirmation of associated SNPs and the identified candidate genes.
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Growth is an important heritable economic trait for dromedaries and necessary for planning a successful breeding program. Until now, genome-wide association studies (GWAS) and QTL-mapping have identified significant single nucleotide polymorphisms (SNPs) associated with growth in domestic animals, but in dromedaries, the number of studies is very low. This project aimed to find biological themes affecting growth in dromedaries. In the first step, 99 candidate SNPs were chosen from a previously established set of SNPs associated with body weight, gain, and birth weight in Iranian dromedaries. Next, 0.5 kb upstream and downstream of each candidate SNP were selected from NCBI (assembly accession: GCA_000803125.3). The annotation of fragments with candidate SNPs regarding the reference genome was retrieved using the Blast2GO tool. Candidate SNPs associated with growth were mapped to 22 genes, and 25 significant biological themes were identified to be related to growth in dromedaries. The main biological functions included calcium ion binding, protein binding, DNA-binding transcription factor activity, protein kinase activity, tropomyosin binding, myosin complex, actin-binding, ATP binding, receptor signaling pathway via JAK-STAT, and cytokine activity. EFCAB5, MTIF2, MYO3A, TBX15, IFNL3, PREX1, and TMOD3 genes are candidates for improving growth in camel breeding programs.
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Semen cryopreservation is an effective strategy for distributing spermatozoa for artificial insemination, but this process reduces the fertility potential of post-thawed spermatozoa. The current study was conducted to assess effects of the novel mitochondria-targeted antioxidant "Mito-TEMPO" on ram sperm quality and fertility potential during the freeze-thawing process in a plant-based extender. Semen samples were diluted in extenders supplemented with 0, 0.5, 5, 50 and 500 µM Mito-TEMPO and then frozen using a standard protocol. Motility, abnormal morphology, acrosome integrity, membrane integrity, mitochondria membrane potential, viability, apoptotic-like changes, lipid peroxidation, DNA fragmentation, H2O2 concentration and fertility potential were assessed after thawing. Results indicated that with the 5 and 50 µM Mito-TEMPO there was a greater (P ≤ 0.05) percentage of sperm total motility, progressive motility, acrosome integrity and viability as well as less (P ≤ 0.05) lipid peroxidation and late apoptotic-like changes. Membrane integrity and mitochondria membrane potential were greater (P ≤ 0.05) with the 50 µM Mito-TEMPO extender supplementation. Furthermore, with 0.5, 5 and 50 µM Mito-TEMPO supplementations there was a greater (P ≤ 0.05) average path velocity and lesser (P ≤ 0.05) percentages of spermatozoa with DNA fragmentation and relatively greater H2O2 concentration. Results from the fertility experiment indicated the 5 and 50 µM Mito-TEMPO treatments resulted in greater pregnancy, parturition and lambing rates. It, therefore, is concluded that Mito-TEMPO may enhance quality and fertility potential of post-thawed semen of rams.
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For thousands of years, camels have produced meat, milk, and fiber in harsh desert conditions. For a sustainable development to provide protein resources from desert areas, it is necessary to pay attention to genetic improvement in camel breeding. By using genotyping-by-sequencing (GBS) method we produced over 14,500 genome wide markers to conduct a genome- wide association study (GWAS) for investigating the birth weight, daily gain, and body weight of 96 dromedaries in the Iranian central desert. A total of 99 SNPs were associated with birth weight, daily gain, and body weight (p-value < 0.002). Genomic breeding values (GEBVs) were estimated with the BGLR package using (i) all 14,522 SNPs and (ii) the 99 SNPs by GWAS. Twenty-eight SNPs were associated with birth weight, daily gain, and body weight (p-value < 0.001). Annotation of the genomic region (s) within ± 100 kb of the associated SNPs facilitated prediction of 36 candidate genes. The accuracy of GEBVs was more than 0.65 based on all 14,522 SNPs, but the regression coefficients for birth weight, daily gain, and body weight were 0.39, 0.20, and 0.23, respectively. Because of low sample size, the GEBVs were predicted using the associated SNPs from GWAS. The accuracy of GEBVs based on the 99 associated SNPs was 0.62, 0.82, and 0.57 for birth weight, daily gain, and body weight. This report is the first GWAS using GBS on dromedary camels and identifies markers associated with growth traits that could help to plan breeding program to genetic improvement. Further researches using larger sample size and collaboration of the camel farmers and more profound understanding will permit verification of the associated SNPs identified in this project. The preliminary results of study show that genomic selection could be the appropriate way to genetic improvement of body weight in dromedary camels, which is challenging due to a long generation interval, seasonal reproduction, and lack of records and pedigrees.
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Peso Corporal/genética , Camelus/crescimento & desenvolvimento , Camelus/genética , Animais , Feminino , Estudo de Associação Genômica Ampla , Masculino , Fenótipo , Polimorfismo de Nucleotídeo Único , GravidezRESUMO
The development of camel husbandry for good production in a desert climate is very important, thus we need to understand the genetic basis of camels and give attention to genomic analysis. We assessed genome-wide diversity, linkage disequilibrium (LD), effective population size (Ne) and relatedness in 96 dromedaries originating from five different regions of the central desert of Iran using genotyping-by-sequencing (GBS). A total of 14,522 Single Nucleotide Polymorphisms (SNPs) with an average minor allele frequency (MAF) of 0.19 passed quality control and filtering steps. The average observed heterozygosity in the population was estimated at 0.25 ± 0.03. The mean of LD at distances shorter than 40 kb was low (r2 = 0.089 ± 0.234). The camels sampled from the central desert of Iran exhibited higher relatedness than Sudanese and lower than Arabian Peninsula dromedaries. Recent Ne of Iran's camels was estimated to be 89. Predicted Tajima's D (1.28) suggested a bottleneck or balancing selection in dromedary camels in the central desert of Iran. A general decrease in effective and census population size poses a threat for Iran's dromedaries. This report is the first SNP calling report on nearly the chromosome level and a first step towards understanding genomic diversity, population structure and demography in Iranian dromedaries.