نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم دامی، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل، ایران. رایانامه: amirza@student.uma.ac.ir

2 نویسنده مسئول، گروه علوم دامی، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل، ایران. رایانامه: nhedayat@uma.ac.ir

3 گروه علوم دامی، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل، ایران. رایانامه: r.khalkhali@uma.ac.ir

4 گروه علوم دامی، دانشکده کشاورزی و منابع طبیعی، دانشگاه محقق اردبیلی، اردبیل، ایران. رایانامه: sharifi_r@uma.ac.ir

چکیده

به‌دلیل تنوع اقلیمی بالا در ایران، گوسفندان بومی کشور، از تنوع چشم‌گیری برخوردار هستند. به‌نظر می‌رسد تفاوت‌های اقلیمی، ردپایی در ژنوم نژادهای بومی مناطق مختلف جهان به جای گذاشته باشند. هدف از مطالعه حاضر، شناسایی نشانه‌های انتخاب مرتبط با گوسفندان ایرانی در مقایسه با نژاد غیرایرانی رومانف در سطح ژنوم می‌باشد. به این منظور، از داده‌های توالی‌یابی‌شده کل ژنوم گوسفندان ایرانی و غیرایرانی موجود در پایگاه داده‌ای NCBI استفاده گردید. این توالی‌ها، پس از سنجش و پالایش کیفی، به ژنوم مرجع گوسفند هم‌طراز شدند. پس از شناسایی تنوع‌های ژنومی، برای شناسایی نواحی تحت انتخاب مثبت در ژنوم گوسفندان ایرانی، از دو روش Fst و XP-EHH استفاده شد. پس از استخراج ژن‌های موجود در نواحی تحت انتخاب، با استفاده از نرم‌افزار BEDtools و فایل GTF مرتبط با ژنوم گوسفند، شرح‌نویسی عملکردی این ژن‌ها با استفاده از آنالیز هستی‌شناسی ژن صورت پذیرفت. براساس نتایج به‌دست‌آمده، به‌ترتیب 907 و 311 ژن کدکننده پروتئین توسط روش‌های Fst و XP-EHH شناسایی شدند که تعداد 29 ژن بین این دو روش مشترک بودند. ارزیابی‌های بیش‌تر نشان داد که تعدادی از این ژن‌ها، در صفات مرتبط با بهبود کیفیت چربی شیر (PCCB)، باروری (SPATA5، RAB35 و DICER1)، رشدونمو ماهیچه‌ای (NF1، AKAP6 و HDAC9)، وزن بدن (FBXL3، GRID2 و ADAMTS17)، سازگاری به نواحی سخت بیابانی و کوهستانی (BMPR2 و NF1) و شیر (EXOC6B) دخیل هستند. نتایج نشان داد که گوسفندان ایرانی احتمالاً برای سازگاری به مناطق خشک بیابانی و ارتقای کیفیت گوشت و شیر موردانتخاب قرار گرفته‌اند.

کلیدواژه‌ها

عنوان مقاله [English]

Identification of selection signatures associated with Iranian sheep compared to non-Iranian Romanov breed using whole genome sequencing data

نویسندگان [English]

  • Abbas Mirzapour-Abibagloo 1
  • Nemat Hedayat 2
  • Reza Khalkhali-Evrigh 3
  • Reza Seyedsharifi 4

1 Department of Animal Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran. E-mail: amirza@student.uma.ac.ir

2 Corresponding Author, Department of Animal Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran. E-mail: nhedayat@uma.ac.ir

3 Department of Animal Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran. E-mail: r.khalkhali@uma.ac.ir

4 Department of Animal Science, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran. E-mail: sharifi_r@uma.ac.ir

چکیده [English]

Introduction Iranian sheep breeds, due to the climate diversity of country, show high diversity and have acquired high adaptability. Compromise with poor quality feed, tolerance of adverse weather and manageable body size are among the factors that probably caused sheep to adapt to different climates. Heretofore, several studies have been carried out in the field of identification of selection signatures in the different native breeds based on SNP-chip data. However, the use of whole genome data can provide researchers with more information about the differences between breeds and their genetic capacities. Identifying and evaluating the effects of climate on the genome of native breeds of Iranian sheep can be effective in designing breeding and conservation strategies. The aim of the present study was to identify the signs of selection related to Iranian sheep compared to the Romanov breed at the genomic level.
Materials and Methods For present study, we used the whole-genome sequencing data related to 43 Iranian and non-Iranian sheep available in the NCBI database. These reads, after performing quality control, were aligned to the sheep reference genome by BWA program. Here, RealignerTargetCreator and IndelRealigner commands available in the GATK program were used to realign around insertions and deletions. Then, the HaplotypeCaller algorithm was used to identify the variants of all samples in ERC GVCF mode. Further, using GenotypeGVCFs module, the variants of all samples were simultaneously identified and finally a VCF file containing raw variants was created. Using the SelectVariants command of the GATK program, all SNPs were separated from other variants. After applying multiple quality filters, high-quality SNPs were extracted and only bi-allelic SNPs present in autosomal chromosomes were used for downstream analysis. Putative selection signatures were identified by using two methods including Fst and XP-EHH. Genes located in positively selected genomic regions were extracted using BEDtools program and the GTF file related to the sheep genome. Gene ontology (GO) analysis was performed on selected genes by "g:Profiler" web-based tool.
Results and Discussion Here, Fst and XP-EHH methods were used to identify the signatures of selection related to Iranian sheep in comparison with Romanov sheep. After converting Fst values ​​to ZFst, 958 genomic windows containing 907 protein-coding genes were detected that had scores above the threshold (ZFst > 3.35). GO analysis on 907 genes identified by the ZFst method led to the identification of 157 significant GO terms in the field of biological processes. In addition, 26 significant terms related to molecular functions and 5 significant terms related to cellular components were also identified. The number of genomic windows identified by the XP-EHH method was 953, which contained a total of 311 protein-coding genes. Among identified genes for each method, 29 genes were detected by both methods as signatures of selection for Iranian sheep. From the GO analysis of 29 common genes, no significant term was obtained. However, these genes were involved on traits related to improving milk fat quality (PCCB), fertility (SPATA5, RAB35 and DICER1), muscle growth and development (NF1, AKAP6 and HDAC9), body weight (FBXL3, GRID2 and ADAMTS17), adaptability to harsh desert and mountain condition (BMPR2 and NF1) and also, milk related traits (EXOC6B).
Conclusion The results showed that Iranian sheep were probably selected to adapt to dry desert areas and improve the quality of meat and milk. The gradual accumulation of such information in different populations will improve the understanding and knowledge of researchers and breeders and will help them to implement efficient breeding programs.

کلیدواژه‌ها [English]

  • Iranian Sheep
  • Romanov
  • Signature of Selection
  • Whole-Genome Sequencing
منابع
Cheng, J., Zhao, H., Chen, N., Cao, X., Hanif, Q., Pi, L., Hu, L., Chaogetu, B., Huang, Y., Lan, X., & Lei, C. (2020). Population structure, genetic diversity, and selective signature of Chaka sheep revealed by whole genome sequencing. BMC genomics, 21, 1-10.
Guðmundsdóttir, Ó. Ó. (2015). Genome-wide association study of muscle traits in Icelandic sheep (Doctoral dissertation).
Guo, J., Zhong, J., Liu, G. E., Yang, L., Li, L., Chen, G., Song, T., & Zhang, H. (2020). Identification and population genetic analyses of copy number variations in six domestic goat breeds and Bezoar ibexes using next-generation sequencing. BMC Genomics, 21(1), 1-13.
 Haberland, M., Arnold, M. A., McAnally, J., Phan, D., Kim, Y., & Olson, E. N. (2007). Regulation of HDAC9 gene expression by MEF2 establishes a negative-feedback loop in the transcriptional circuitry of muscle differentiation. Molecular and Cellular Biology, 27(2), 518-525.
Hong, X., Luense, L. J., McGinnis, L. K., Nothnick, W. B., & Christenson, L. K. (2008). Dicer1 is essential for female fertility and normal development of the female reproductive system. Endocrinology, 149(12), 6207-6212.
Hu, L., Zhang, L., Li, Q., Liu, H., Xu, T., Zhao, N., Han, X., Xu, S., Zhao, X., & Zhang, C. (2022). Genome-wide analysis of CNVs in three populations of Tibetan sheep using whole-genome resequencing. Frontiers in Genetics, 13, 971464.
Khalkhali-Evrigh, R., Hedayat, N., Ming, L., & Jirimutu. (2022). Identification of selection signatures in Iranian dromedary and Bactrian camels using whole genome sequencing data. Scientific Reports, 12(1), 9653.
Kijas, J.W., Townley, D., Dalrymple, B.P., Heaton, M.P., Maddox, J.F., McGrath, A., Wilson, P., Ingersoll, R.G., McCulloch, R., McWilliam, S., & Tang, D. (2009). A genome wide survey of SNP variation reveals the genetic structure of sheep breeds. PloS one, 4(3), p.e4668.
Lee, Y. L., Bosse, M., Mullaart, E., Groenen, M. A., Veerkamp, R. F., & Bouwman, A. C. (2020). Functional and population genetic features of copy number variations in two dairy cattle populations. BMC Genomics, 21(1), 1-15.
Li, G., Tang, J., Huang, J., Jiang, Y., Fan, Y., Wang, X., & Ren, J. (2022). Genome-Wide Estimates of Runs of Homozygosity, Heterozygosity, and Genetic Load in Two Chinese Indigenous Goat Breeds. Frontiers in Genetics, 13.
Li, R., Zhao, Y., Liang, B., Pu, Y., Jiang, L., & Ma, Y. (2023). Genome-Wide Signal Selection Analysis Revealing Genes Potentially Related to Sheep-Milk-Production Traits. Animals, 13(10), 1654.
Liu, J., Shi, L., Li, Y., Chen, L., Garrick, D., Wang, L., & Zhao, F. (2021). Estimates of genomic inbreeding and identification of candidate regions that differ between Chinese indigenous sheep breeds. Journal of Animal Science and Biotechnology, 12, 1-14.
Mohammadi, H., Rafat, S. A., Moradi Shahrbabak, H., Shodja, J., & Moradi, M. H. (2020). Genome-wide association study and gene ontology for growth and wool characteristics in Zandi sheep. Journal of Livestock Science and Technologies, 8(2), 45-55.
Pausch, H., Jung, S., Edel, C., Emmerling, R., Krogmeier, D., Götz, K. U., & Fries, R. (2012). Genome‐wide association study uncovers four QTL predisposing to supernumerary teats in cattle. Animal Genetics, 43(6), 689-695.
Remsburg, C., Testa, M., & Song, J. L. (2021). Rab35 regulates skeletogenesis and gastrulation by facilitating actin remodeling and vesicular trafficking. Cells & Development, 165, 203660.
Serranito, B., Cavalazzi, M., Vidal, P., Taurisson-Mouret, D., Ciani, E., Bal, M., Rouvellac, E., Servin, B., Moreno-Romieux, C., Tosser-Klopp, G., & Hall, S. J. (2021). Local adaptations of Mediterranean sheep and goats through an integrative approach. Scientific Reports, 11(1), p.21363.
Shi, H., Li, T., Su, M., Wang, H., Li, Q., Lang, X., & Ma, Y. (2023). Whole genome sequencing revealed genetic diversity, population structure, and selective signature of Panou Tibetan sheep. BMC Genomics, 24(1), 1-15.
Sujit, K. M., Singh, V., Trivedi, S., Singh, K., Gupta, G., & Rajender, S. (2020). Increased DNA methylation in the spermatogenesis‐associated (SPATA) genes correlates with infertility. Andrology, 8(3), 602-609.
Van Poucke, M., Sjoberg, A., Mattheeuws, M., Van Zeveren, A., Bouquet, Y., Chowdhary, B. P., & Peelman, L. J. (1997). Mapping of the ATP2B2 and PCCB genes on porcine chromosome 13. Mammalian Genome, 8(11), 852.
Wei, C., Wang, H., Liu, G., Zhao, F., Kijas, J. W., Ma, Y., Lu, J., Zhang, L., Cao, J., Wu, M., & Wang, G. (2016). Genome-wide analysis reveals adaptation to high altitudes in Tibetan sheep. Scientific Reports, 6(1), 26770.
Wiener, P., Robert, C., Ahbara, A., Salavati, M., Abebe, A., Kebede, A., Wragg, D., Friedrich, J., Vasoya, D., Hume, D.A., & Djikeng, A. (2021). Whole-genome sequence data suggest environmental adaptation of Ethiopian sheep populations. Genome Biology and Evolution, 13(3), evab014.
Xu, S. S., Gao, L., Xie, X. L., Ren, Y.L., Shen, Z. Q., Wang, F., Shen, M., Eyϸórsdóttir, E., Hallsson, J. H., Kiseleva, T., & Kantanen, J. (2018). Genome-wide association analyses highlight the potential for different genetic mechanisms for litter size among sheep breeds. Frontiers in Genetics, 9, 118.
Yang, J. I., Li, W. R., Lv, F. H., He, S. G., Tian, S. L., Peng, W. F., Sun, Y. W., Zhao, Y. X., Tu, X. L., Zhang, M., & Xie, X. L. (2016). Whole-genome sequencing of native sheep provides insights into rapid adaptations to extreme environments. Molecular Biology and Evolution, 33(10), 2576-2592.
Zhang, L., Wang, F., Gao, G., Yan, X., Liu, H., Liu, Z., Wang, Z., He, L., Lv, Q., Wang, Z., & Wang, R. (2021). Genome-wide association study of body weight traits in Inner Mongolia cashmere goats. Frontiers in Veterinary Science, 8, 752746.
Zhao, H., Guo, T., Lu, Z., Liu, J., Zhu, S., Qiao, G., Han, M., Yuan, C., Wang, T., Li, F., & Zhang, Y. (2021). Genome-wide association studies detects candidate genes for wool traits by re-sequencing in Chinese fine-wool sheep. BMC Genomics, 22, 1-13.