بررسی ساختار ژنتیکی اسب نژاد عرب با استفاده از نشانگرهای ریزماهواره

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشیار، بخش تحقیقات بیوتکنولوژی، موسسه تحقیقات علوم دامی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

2 استادیار، بخش تحقیقات علوم دامی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان ایلام، سازمان تحقیقات، آموزش و ترویج کشاورزی، ایلام، ایران

3 استادیار، بخش تحقیقات ژنتیک و اصلاح نژاد دام، موسسه تحقیقات علوم دامی کشور، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

هدف از این تحقیق، بررسی تنوع ژنتیکی اسب­های عرب ایران با استفاده از نشانگرهای ریزماهواره بود. به این منظور از تعداد 10 نشانگر ریزماهواره شامل AHT04، AHT05، ASB02، ASB17، ASB23، HMS03، HMS06، HMS07 و VHL20 مورد تایید انجمن بین­المللی ژنتیک دام استفاده شد. تعداد 8673 نمونه موی اسب از استان­های خوزستان، یزد، کرمان، اصفهان، لرستان و البرز جمع آوری شد و سپس استخراج DNA از نمونه­های اخذ شده انجام گرفت. نشانگرهای ریزماهواره با روش Multiplex PCR تکثیر شدند. سپس محصولات تکثیر شده به وسیله سیستم Genetic analyzer و با روش الکتروفورز مویینه تعیین ژنوتیپ شدند. بیشترین و کمترین تعداد آلل مشاهده شده به ازای هر نشانگر، به ترتیب مربوط به نشانگرهای ASB17 (۱۷ آلل)، HMS06 (هشت آلل) و HMS07 (هشت آلل) بودند. مجموع تعداد کل آلل­های مشاهده شده در همه جایگاه­ها و میانگین تعداد آلل مشاهده شده به ترتیب برابر با 113 و 3/11 بودند. میانگین تعداد آلل موثر، شاخص شانون، میزان هتروزیگوسیتی مشاهده شده و مورد انتظار و شاخص تثبت به ترتیب 97/3، 58/1، 721/0، 736/0 و 021/0 به­دست آمد. میانگین تنوع ژنتیکی در جمعیت اسب عرب مورد بررسی برابر با 736/۰ محاسبه شد. بیشترین تنوع ژنتیکی در نشانگر VHL20 با فراوانی برابر با 803/۰ و کمترین میزان تنوع ژنتیکی مربوط به نشانگر ASB2 با مقدار ۶۲0/۰ بود. فراسنجه­های جمعیتی در اسب عرب نشان داد که چندشکلی و تنوع ژنتیکی در این نژاد نسبتاٌ بالا است. وجود تنوع ژنتیکی در این نژاد جهت اجرای برنامه­های اصلاحی و حفاظت نژادی از اهمیت ویژه­ای برخوردار است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Investigation of the genetic structure of Arabian horses using microsatellite markers

نویسندگان [English]

  • H. R. Seyedabadi 1
  • J. Ahmadpanah 2
  • A. Javanrouh 3
  • H. Baneh 3
1 Associate Professor, Department of Biotechnology, Animal Science Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran
2 Assistant Professor, Department of Animal Science, Ilam Agricultural and Natural Resources Research and Education Center, Agricultural Research, Education and Extension Organization, Ilam, Iran
3 Assistant Professor, Department of Animal Breeding and Genetics, Animal Science Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran
چکیده [English]

Introduction: Horse has been of great interest to humans due to its speed, strength, and endurance. Based on its role in human history and civilization, it is considered the most important domesticated animal. Iranian horse breeds are bred in the north, south, and west regions of the country. Arabian horse is one of the international breeds in the world, which is bred in more than 60 countries. Preliminary evidence shows that the Arabian horse breed was established about 3000 years ago and was bred in small populations in many countries of the Middle East, including Egypt, Iran, Saudi Arabia, and Syria. These separate populations have led to the development of different maternal strains of the Arabian horse. Genetic diversity is necessary for genetic conservation and survival of the breeds. Molecular markers are used to assess the structure and genetic diversity of different populations. Therefore, one of the suitable markers is a microsatellite. Regarding the importance of genetic diversity in the survival of the breed, and designing the genetic conservation and breeding programs, the purpose of this study was to investigate the population structure and genetic diversity of Arabian horse breeds using microsatellite markers.
Materials and Methods: In this study, hair root samples were obtained from 8673 (5602 mares and 3071 stallions) Arabian horses from Khuzestan, Yazd, Kerman, Isfahan, Lorestan, and Alborz provinces. DNA samples were extracted from hair roots using the Direct PCR Kit (Thermo Fisher Scientific, Vilnius, Lithuania). The quantity and quality of the extracted DNA were controlled by spectrophotometry and agarose gel methods. The number of 10 microsatellite markers including AHT04, AHT05, ASB02, ASB17, ASB23, HMS03, HMS06, HMS07, HTG10, and VHL20 were used based on ISAG recommendation. Then, the amplification of genomic fragments and the genotype of samples was determined by COrDIS Horse Reagent Kit (Moscow, Russia). Forward primers were labeled with fluorescent dye at 5'-end. These microsatellite loci were amplified by the multiplex PCR method. Then, the PCR products were genotyped by the Genetic Analyzer system and the capillary electrophoresis method. Obtained data were used to estimate demographic parameters. The number of effective alleles, the number of observed alleles, the observed heterozygosity, the expected heterozygosity, and the Shannon index were calculated by GenAlex 6.5 software. Genpop 4.7.5 software was used to calculate the FIS statistic or Fixation index in the population.
Results and discussion: The results showed that the lowest and highest number of observed alleles were related to ASB17 (17 alleles), HMS06 (eight alleles), and HMS07 (eight alleles) markers, respectively. The total number of alleles observed at all loci was 113 alleles. The average number of observed alleles was estimated to be 11.3, which indicates high allelic diversity in the Persian Arabian horse. The mean number of effective alleles, Shannon index, observed and expected heterozygosity and Fixation index were 3.97, 1.58, 0.721, 0.736, and 0.021, respectively. The mean genetic diversity in the Arabian horse’s population was calculated to be 0.736. The highest and lowest genetic diversity was observed in VHL20 (0.803) and ASB02 (0.620) markers, respectively. The average number of effective alleles per locus in studies conducted in different countries on Arabian horses, including Egypt, Syria, and Western countries, with Persian Arabian horses in this research, showed that number of effective alleles is higher in Persian Arabian horses. The average Shannon's index has been reported to be slightly lower in previous studies of Persian Arabian horses, which is probably due to the small number of samples in these studies compared to the present study.  The highest and lowest observed heterozygosities in this study were related to AHT4 (0.788) and ASB02 (0.613) markers, which were consistent with the results of the research conducted on Arabian horses of Iran, Syria, and Egypt. The positive FIS value in Arabian horses made a decrease in heterozygosity in the population, an increase in homozygosity and consequently inbreeding in Arabian horse populations. In this research, all studied loci showed a significant deviation from the Hardy-Weinberg equilibrium. Except for the AHT04 marker, which deviated from the Hardy-Weinberg equilibrium at P<0.01, the others have deviated at P<0.001. Deviation from the Hardy-Weinberg equilibrium can indicate the presence of some factors disturbing Hardy-Weinberg equilibrium, such as migration and selection, which in Persian Arabian horses, the entry of stallions from outside the herd and the gene flow between different maternal strains and also the existence of a selection program among breeders are the main factors of deviation from Hari-Weinberg equilibrium.
Conclusions: Evaluation of the genetic structure of Arabian horses of Iran and comparison with other Arabian horses in the world, including those in the Middle East region, showed that Arabian horses of Iran have a higher genetic diversity, which is probably due to the presence of different maternal strains in Iran, high gene flow among the different strains, a large import of Arabian horses and crossbreeding with Arabian horses of Iran, as well as a high number of samples and so identification of new alleles in this research. On the other hand, the rate of inbreeding was positive according to the FIS in Iran Arabian horses, indicating a risk of genetic diversity loss and increasing inbreeding in these horses. Therefore, management of genetic diversity and prevention of mating among related animals should be considered.

کلیدواژه‌ها [English]

  • Arabian horse
  • Genetic diversity
  • Population parameters
  • Microsatellite marker

مراجع

Abdoli, M., Zandi, M. B., Harkinezhad, M. T., & Khalili, M. (2021). Genetic structure survey of Iranian native horse breeds by microsatellite markers. Journal of Animal Production, 23(2), 155-163. [In Persian]
Ala-Amjadi, M., Mehrabani Yeganeh, H., & Sadeghi, M. (2017). Study of genetic variation in Iranian Kurdish horse using microsatellite markers. Iranian Journal of Animal Science Research, 11(4), 481-487. [In Persian]
Almarzook, S., Abdel-Shafy, H., Ahmed, A. S., Reissmann, M., & Brockmann, G. A. (2022). Genetic diversity of Arabian horses using microsatellite markers. Egyptian Journal of Animal Production, 59(1),19-27.
Badbarin, S., Seyedsharifi, R., & Falahi, H. (2022). Investigation the genetic diversity of Arabian horses and their different strains using SSR markers. Research on Animal Production, 13(37), 158-165. [In Persian]
Benhamadi, M. E. A., Naima Berber, N., Benyarou, M., Ameur, A. A., Haddam, H. Y., Piro, M., & Gaouar, S. B. S. (2020). Molecular characterization of eight horse breeds in Algeria using microsatellite markers. Biodiversitas, 21(9), 4107-4115.
Binn, M. M., Uolmes, N. G., Holliman, A., & Scott, A. M. (1995). The identification of polymorphic microsatellite loci in the horse and their use in Troroughbred parentage testing. British Veterinary Journal, 151, 9-15.
Breen, M., Lindgren, G., Binns, M. M., Norman, J., & Irvin, Z. (1997). Genetical and physical assignments of equine microsatellite first integration of anchored markers on horse genome mapping. Mammalian Genome, 8, 267-273.
Dorji, J., Tamang, S., Tshewang, T., Dorji, T., & Dorji, T. Y. (2018). Genetic diversity and population structure of three traditional horse breeds of Bhutan based on 29 DNA microsatellite markers. PLoS ONE, 13(6), e0199376.
FAO. (2021). Commission on genetic resources for food and agriculture. CGRFA/WG-AnGR-11/21/Inf.6.
Guerin, G., Bertaud, M., & Amigues, Y. (1994). Cgaracterization of seven new horse microsatellites: HMS1, HMS2, HMS3, HMS5, HMS6, HMS7 and HMS8. Animal Genetics, 25, 62.
Hedayat-Evrigh, N., Azadmard, E., SeyedSharifi, R., Nikbin, S., Shakouri, M. D., & Khalkhali-Evrigh, R. (2019). Investigation of genetic diversity of Iran northwest horses using microsatellite markers. Agricultural Biotechnology Journal, 11(4), 35-50. [In Persian]
Hedrick, P. W. (2000). Genetic of populations. Second edition. Jones and Bartlett Publishers, Sudbury, MA, USA. 553 p.
Irvin, Z., Giffard, J., Brandon, R., Breen, M., & Bell, K. (1998). Equine dinucleotide repeat polymorphisms at loci 21, 23, 25 and 37-43. Animal Genetics, 29(1), 67.
Jabbari, S., Mashayekhi, M. R., Hasanpour, A., & Shirmohammadly, B. (2020). Evaluation of the genetic diversity of Iranian Arabian horses. Iranian Journal of Animal Sceince Research, 11(4), 481-487. [In Persian]
Javanrouh, A., & Khodamoradi, S. (2022). Study of the genetic structure of the Shin Bash sheep population by molecular markers. Animal Production Research, 11(3), 27-40. [In Persian]
Jemmali, B., Haddad, M. M., & Barhoumi, N. (2017). Genetic diversity in Tunisian horse breeds. Archives Animal Breeding, 60, 153-160.
Khalili, M. (2009). Horses and my expertise. (ed. by M. Khalili), Nashr-e Zare Publication, Iran. 694 p. [In Persian]
Khalili, M. (2015). Asil stud book of the Islamic Republic of Iran, Volume III, Published by Equestrian Federation, Tehran, Iran. [In Persian]
Khanshour, A., Conant, E., & Juras, R. (2013). Microsatellite analysis of genetic diversity and population structure of Arabian horse populations. Journal of Heredity, 104, 386-398.
Khanshour, A., Juras, R., Blackburn, R., & Cothran, E. G. (2014). The Legend of the Canadian Horse: Genetic Diversity and Breed Origin. Journal of Heredity, 106(1), 37-44.
Leroy, L., Callède, L., Verrier, E., Mériaux, J. C., Ricard, R., Danchin-Burge, C., & Rognon, X. (2009). Genetic diversity of a large set of horse breeds raised in France assessed by microsatellite polymorphism. Genetics Selection Evolution, 41, 1-12.
Ling, Y. H., Ma, Y. H., Guan, W- J., Cheng, Y. J., Wang, Y. P., Han, J. L., Mang, L., Zhao, Q. J., He, X. H., Pu, Y. B., & Fu, B. L. (2011). Evaluation of the genetic diversity and population structure of Chinese indigenous horse breeds using 27 microsatellite markers. Animal Genetics, 42(1), 56-65.
Machmoum, M., Boujenane, I., Azelhak, R., Badaoui, B., Petit, D., & Piro, M. (2020). Genetic diversity and population structure of Arabian horse populations using Microsatellite markers. Journal of Equine Veterinary Science, 93, 103200.
Mahrous, K. F., Hassanane, M., Abdel Mordy, M., Shafey, H. I., & Hassan, N. (2011). Genetic variations in horse using microsatellite markers. Journal of Genetic Engineering and Biotechnology, 9, 103-109.
Marklund, S., Ellegren, H., Eriksson, S., Sandberg, K., & Anderson, L. (1994). Parentage testing and linkage analysis in the horse using a set of highly polymorphic microsatellites. Animal Genetics, 25, 19-23.
Moravcikova, N., Kasarda, R., Kukuckova, V., Vostry, L., & Kadleclk, O. (2016). Genetic diversity of old Kladruber and Nonius horse populations through microsatellite variation analysis. Acta Agriculturae Slovenica, 5, 46.
Moshkelani, S., Rabiee, S., & Javaheri-Koupaei, M. (2011). DNA fingerprinting of Iranian Arab horse using fourteen microsatellites marker. Research Journal of Biological Sciences, 6, 402-405.
Mostafavi, A., Asadi Fozi, M., Esmailizadeh Koshkooieh, A., Mohammadabadi, M. R., Ivanivna Babenko, O., & Ihorivna Klopenko, N. (2019). Effect of LCORL gene polymorphism on body size traits in horse populations. Acta Scientiarum. Animal Sciences, 42, e47483.
Ott, J. (1992). Strategies for characterizing highly polymorphic markers in human gene mapping. American Journal of Human Genetics, 51, 283-290.
Peakall, R., & Smouse, P. E. (2012). GenALEx 6.5: Genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics, 28, 2537-2539.
Sadeghi, R., Moradi-Shahrbabak, M., MiraeiAshtiani, S. R., Schlamp, F., Cosgrove, E. J., & Antczak, D. F. (2019). Genetic diversity of Persian Arabian horses and their relationship to other native Iranian horse breeds. Journal of Heredity, 110(2), 173-182.
Sargious, M. A., Ahmed, H. A., El-Shawarby, R. M., Bakery, H. H., Ramadan, S., Cothran, E. G., & Farid, A. S. (2022). Genetic diversity and population assignment of Arabian Horses. Pakistan Journal of Zoology, 54, 825-833.
Seo, J. H., Park, K. D., Lee, H. K., & Kong, H. S. (2016). Genetic diversity of Halla horses using microsatellite markers. Journal of Animal Science and Technology, 58, 40.
Seyedsharifi, R., Badbarin, S., Hedayat-Evrigh, N., Savarsofla, S., Seifdavati, J., & Khamisabadi, H. (2019). Investigation of the genetic structure and phylogenic relationships of Caspian, Arabic and Taleshi horses. Iranian Journal of Animal Science Research, 11(2), 223-232. [In Persian]
Shamsalddini, S., Mohammadabadi, M. R., & Esmailizadeh, A. K. (2016). Polymorphism of the prolactingene and its effect on fiber traits in goat. Russian Journal of Genetics, 52, 405-408.
Rousset, F. (2008). genepop’007: A complete re‐implementation of the genepop software for Windows and Linux. Molecular Ecology Resources, 8(1), 103-106. 
Van Haeringen, H., Bowling, A. T., Stott, M. L., Lenstra, J. A., & Zwaagstra, K. A. (1994). A highly polymorphic horse microsatellite locus: VHL20. Animal Genetics, 25(3), 207.
Van Lent, R., & Upton, P. (1999). Arabians (ed. H. Amirsadeghi). Chronicle Books.
Vázquez-Armijo, J., Parra-Bracamonte, M., Velazquez, M., Sifuentes Rincon, A., Tinoco-Jaramillo, J., Ambriz-Morales, P., Arellano-Vera, W., & Moreno-Medina, V. (2017). Diversity and effective population size of four horse breeds from microsatellite DNA markers in south-central Mexico. Archives Animal Breeding, 60, 137-143.
تحقیقات تولیدات دامی
دوره 12، شماره 1
خرداد 1402
صفحه 53-64

فایل ها

هم رسانی

ارجاع به این مقاله

آمار