بررسی شناسایی فرم‏های مختلف آللی ژن میوستاتین و بررسی صفات بیومتری وزن بدن و لاشه اندازه‌گیری شده با اولتراسوند در گوسفند کردی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانش‌‌آموخته کارشناسی‌ارشد، گروه ژنتیک و اصلاح دام و طیور، دانشکده علوم دامی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان،

2 استاد، گروه ژنتیک و اصلاح دام و طیور، دانشکده علوم دامی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان،، ایران

3 استادیار، گروه ژنتیک و اصلاح دام و طیور ، دانشکده علوم دامی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، ایران

چکیده

چکیده
سابقه و هدف: ژن میوستاتین به عنوان عامل مهارکننده رشد عضله اسکلتی شناخته شده و اگر جهش در ناحیه کدکننده آن اتفاق افتد، باعث تغییر نقش مهاری آن و افزایش عضله می‏گردد. این پژوهش با هدف شناسایی چندشکلی موجود در اگزون 3 ژن میوستاتین و بررسی صفات بیومتری وزن بدن و لاشه اندازه‏گیری ‌‌‌‌شده با اولتراسونوگرافی در گوسفند کردی انجام شد.
مواد و روش: در این پژوهش برای اندازه‌گیری صفات ضخامت چربی پشت و مساحت ماهیچه چشمی از دستگاه اولتراسوند استفاده شد. خون‌گیری به طور تصادفی از تعداد 139 راس گوسفند کردی انجام و پس از استخراج DNA، قطعه 338 جفت بازی اگزون 3 ژن میوستاتین تکثیر شد. جهت تعیین ژنوتیپ‏ ژن میوستاتین، از تکنیک‏هایPCR-RFLP ، PCR-SSCP و تعیین توالی مستقیم استفاده شد. بررسی اثرات ثابت جنس، نوع تولد و سال تولد بر صفات مورد مطالعه با رویه GLM نرم‌افزار SAS و مقایسه میانگین‌های حداقل مربعات سطوح اثرات ثابت با آزمون توکی- کرامر در سطح معنی‌داری 5 درصد انجام شد. برای بررسی ارتباط بین صفات از همبستگی پیرسون استفاده شد.
یافته‏ها: در روش‏PCR-RFLP پس از هضم آنزیمی تمام نمونه‏ها، تنها یک ژنوتیپ مشاهده شد و این جایگاه مونومورف (تک شکل) می‏باشد. نتایج به‏دست آمده از روش PCR-SSCP، دو الگوی باندیA و B را نشان داد که شباهت زیادی بین دو الگوی مشاهده شده وجود داشت. لذا برای تعیین ژنوتیپ واقعی، برخی از نمونه‏ها با الگوی متفاوت با روش توالی‌یابی مستقیم مورد ارزیابی قرار گرفتند که نتایج نشان داد که در بین نمونه‌ها چندشکلی وجود ندارد. میانگین ضخامت چربی پشت و مساحت عضله راسته اندازه‏گیری شده با اولتراسونوگرافی در این مطالعه به‏ترتیب 46/0 سانتی‏متر و 78/7 سانتی‏متر مربع با میانگین وزن بدن 42/43 کیلوگرم بود. همبستگی بین دو صفت اولتراسوند (ضخامت چربی پشت و مساحت ماهیچه چشمی)، مثبت و 84/0 برآورد شد (01/0P<). کمترین ضریب همبستگی ضخامت چربی لاشه و مساحت ماهیچه چشمی با صفات مورد مطالعه به‏ترتیب 02/0 و 3/0 مربوط به صفات وزن تولد و از شیرگیری بود و بیشترین ضریب همبستگی صفات مورد مطالعه با ضخامت چربی لاشه و مساحت ماهیچه چشمی به‏ترتیب 82/0 و 87/0 مربوط به صفات وزن بدن در هنگام اولتراسونوگرافی و عرض دنبه میانی بود (001/0P<).
نتیجه‏گیری: در این پژوهش با استفاده از نشانگر ژنتیکیPCR-SSCP، PCR-RFLP و تعیین توالی مستقیم تنها یک ژنوتیپ در ژن میوستاتین تشخیص داده شد. با توجه به اینکه منابع مختلف، آلل جهش یافته ژن میوستاتین را به عنوان آلل موثر بر فنوتیپ عضله مضاعف و مطلوب جهت اصلاح نژاد و بهبود کیفیت و کمیت گوشت معرفی کرده اند، گله مورد بررسی فاقد این آلل بود. با توجه به تنوع نسبتا مناسب مشاهده شده در صفات مورد مطالعه بویژه صفات لاشه اندازه گیری شده با اولتراسوند، زمینه لازم برای اصلاح این صفات از طریق انتخاب وجود خواهد داشت.
کلمات کلیدی: چندشکلی، ژن میوستاتین، PCR-RFLP،PCR-SSCP ، گوسفند کردی

کلیدواژه‌ها


عنوان مقاله [English]

Identification of different allelic forms of myostatin gene and investigation of body weight and carcass biometric traits measured by ultrasound in Kurdi sheep

نویسندگان [English]

  • Fatemeh Rayeji Yanesari 1
  • saeed Hassani 2
  • Mojtaba Najafi 3
1 Master's student, Animal and Poultry Genetics and Breeding Department, Faculty of Animal Sciences, Gorgan University of Agricultural Sciences and Natural Resources.
2 Professor of Animal Breeding and Genetics, College of Animal Science. Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
3 Assistant Professor of Animal Breeding and Genetics, College of Animal Science. Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
چکیده [English]

Abstrast
Background and Aim: Myostatin gene is an inhibitor of skeletal muscle developmentmutations in its coding regionpromotesmuscle growth in some breeds, mutation of the myostatin gene has a significant effect on the increase of the body weight and carcass traits. This study was aimed to investigate the polymorphisms in exon 3 of the myostatin gene and evaluate investigation of body weights and carcass traits measured by ultrasound in Kurdi sheep.
Materials and Methods: In this study, back fat thickness and loin muscel area traits were measured by ultrasound instrument. Blood samples were collected randomly from 139 Kurdi sheep and following DNA extraction, a 388 bp fragment from exon 3 of myostatin gene was amplified. For genotyping of myostatin gene, PCR-SSCP, PCR-RFLP, and direct sequencing techniques were used. To determine the fixed effects of gender, birth type and birth year on the studied traits, GLM procdure of SAS software was used. Least square means comparison of different fixed effects subclasses was carried out by Tukey-Kramer test at 5% probability level. To measure the relationships between tarits, pearson correlation was used.
Results: In PCR-RFLP technique, after enzymatic digestion of all samples, only one genotype was observed and this locus was monomorphic. PCR-SSCP analysis showed two band patterns A and B, which were very similar in shape. Therefore, to determine the actual genotype, some samples with different band patterns were evaluated by direct sequencing technique. The results showed that there is no polymorphism among samples. The mean ultrasonic fat thickness and loin muscle area in this study were 0.46 cm and 7.78 cm2, respectively, with an average body weight of 43.42 kg at the time of ultrasonography. The correlation between the two ultrasound carcass traits (UBF and UMA) was positive (r=0.84, P<0.0001). The lowest correlation coefficient of carcass fat thickness and ocular muscle area with the studied traits were 0.02 and 0.3, for birth weight and weaning traits, respectively. The highest correlation coefficient of the studied traits with carcass fat thickness and loin muscle area were 0.82 and 0.87 for body weight at the time of ultrasonography and middle tail width, respectively (P=0.001).

Conclusion: In this study, using PCR-SSCP, PCR-RFLP and direct sequencing, only one genotype was detected in the myostatin gene. Considering that various studies have introduced the mutated allele of the myostatin gene as an effective allele on the double muscling for breeding and improving the quality and quantity of meat, the studied herd did not have the allele. Considering the relatively suitable diversity observed in the studied traits, especially the carcass traits measured by ultrasound, there will be a possiblity to improve the traits through selection.
Keyword: Kurdi sheep, Myostatin Gene, PCR-RFLP, PCR-SSCP, Polymorphism

کلیدواژه‌ها [English]

  • Polymorphism
  • Myostatin Gene
  • PCR-RFLP
  • PCR- SSCP
  • Kurdi sheep
Ansary, M., Tahmoorespur, M., Nassiry, M.R., Taheri, A. and Vafayevaleh, M. 2011. Polymorphism in Intron-1 of myostatin gene and its association with estimated breeding values of growth traits in Baluchi sheep (Ovis aries). Indian Journal of Animal Science, 81(8): 75-100.
Askari, N., Baghizadeh, A. and Mohammadabadi, M.R. 2008. Analysis of the genetic structure of Iranian indigenous Raeni Cashmere goat populations using microsatellite markers. Biotechnology, 2(3): 1-4.
Clop, A., Marcq, F., Takeda, H., Pirottin, D., Tordoir, X., Bibe, B., Bouix, J., Caiment, F., Elsen, JM., Eychenne, F., Larzul, C., Laville, E., Meish, F., Milenkovic, D., Tobin, J., Charlier, C. and Georges, M. 2006. A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep. Nature Genetic, 38(7): 813-818.
Dehnavi, A., Ahani Azeri, M., Hassani, S., Nasiri, M.R., Mohajer, M. and Khan Ahmadi, A.R. 2004. Investigation of myostatin gene polymorphism (GDF-8) in Zell sheep using the method. PCR-RFLP 4th Congress of Animal Sciences, September 20-29, Agricultural Campus, University of Tehran, Karaj. (In Persian).
Hadjipavlou, G., Matika, O., Clop, A. and Bishop, S.C. 2008. Two single nucleotide polymorphisms in the myostatin (GDF8) gene have significant association with muscle depth of commercial Charollais sheep. Animal Genetics, 39(4): 346-353.
Han, J., Zhou, H., Forres, R., Sedcole, J., Frampton, C. and Hickford, J. 2010. Effect of myostatin (MSTN) g+ 6223G> A on production and carcass traits in New Zealand Romney Sheep. Asian-Australasian Journal of Animal Sciences, 3(7):863-866.
Hosseini Vardanjani, S.M., Miraei Ashtiani, S.R., Pakdel, A. and Moradi Shahrebabak, H. 2014. Accuracy of real-time ultrasonography in assessing carcass traits in Torki-Ghashghaii sheep. Journal of Agricultural Science and Technology, 16(4): 791-800.
Jahnson, P.L., McEwant, J.C., Dodds, K.G., Purchas, R.W. and Blari, H.T. 2005. A directed search in the region of GDF8 for quantitative trait loci affecting carcass trait in Texel sheep. Journal of Animal Science, 83(9):1988-2000.
Julianna, K. and Gocza, E. 2002. The role of the myostatin protein in meat quality. Archiv Tierzucht, 45:159-170.
Khorramtaei, R., Nezam Abadi, M., Herki Nejad, T., Eskandari Nasab, M., Shahir, M.H. and Salimi, D. 2012. Evaluation of the relationship between direct measurement and ultrasound of muscle and back fat to estimate livestock carcass traits in Afshari sheep. Journal of Animal Science Research, 22)2):161-171. (In Persian).
Kiyanzad, M.R. 2004. Predicting carcass physical and chemical composition of Moghani and Makui sheep in breeding flocks. (Body measurements and carcass characteristics). Pajouhesh Va Sazandgi, 64: 2-11. (In Persian).
Leeds, T., Mousel, M., Notter, D., Zerby, H., Moffet, C. and Lewis, G. 2008. B-mode, real-time ultrasound for estimating carcass measures in live sheep: Accuracy of ultrasound measures and their relationships with carcass yield and value. Journal of Animal Science, 86(11): 3203-3214.
Masri, A.Y., Lambe, N.R., Macfarlane, J.M., Brotherstone, S., Haresign, W. and Bünger, L. 2011. Evaluating the effects of a single copy of a mutation in the myostatin gene (c.*1232 G>A) on carcass traits in crossbred lambs. Meat Science, 87(4):412-418.
Masoudi, A., Imrani, J., Abbasi, A., Nejati Jorami, A., Konida, T., Farhang, K.h., Ismail Khanian, S. and Ziaee, F. 2005. Evaluation of myostatin gene polymorphism and its relationship with growth traits in Baluchi sheep by PCR-SSCP method, 4th Iranian National Biotechnology Conference, Kerman. (In Persian).
Masumi, N., Akihiro, Y., Shinichirou, N., Tsutomu, N., Takashi, Y., Mitsuo, I., Keiji, M., Hideyo, O. and Sumihare, N. 2002. A missense mutant myostatin causes hyperplasia without hypertrophy in the mouse muscle. Biochemical and Biophysical Recearch Commonications, 293(1): 247-251.
Maxa, J., Norberg, E., Berg, P. and Milerski, M. 2007. Genetic parameters for body weight, longissimus muscle depth and fat depth for Suffolk sheep in the Czech Republic. Small Ruminant Research, 72(2-3): 87-91.
Mohammadabadi, M.R. 2017. Inter-simple sequence repeat loci associations with predicted breeding values of body weight in Kermani sheep. Genetics 3rd Millennium, 14(4): 4383-4390.
Mohammadifar, A., Faghih Imani, S.A., Mohammadabadi, M.R. and Soflaei, M. 2014. The effect of TGFb3 gene on phenotypic and breeding values of body weight traits in Fars native fowls. Agricultural Biotechnology Journal, 5(4): 125-136. (In Persian).
Mohammadifar, A. and Mohammadabadi, M. 2018. Melanocortin-3 receptor (MC3R) gene association with growth and egg production traits in Fars indigenous chicken.  Malaysian Applied Biology, 47(3): 85-90.
Mohammadifar, A. and Mohammadabadi, M.R. 2011. Application of microsatellite markers for a study of Kermani sheep genome. Iranian Journal of Animal Science (IJAS), 42(4): 337-344. (In Persian).
Mohammadi, A., Hassani, S., Zerehdaran, S., Bagheri, M. and Mirshahi, A. 2018. Genetic evaluation of some carcass characteristics assessed by in vivo real time ultrasonography in Baluchi sheep. Iranian Journal of Applied Animal Science. 3(8): 457-468.
Mokhaber, M., Moradi Shahrbabak, H. and Khalat Abadi Farahani, A.M. 2013. Application of principal component analysis method to predict tail weight and carcass weight in Makui lambs. Iranian Journal of Animal Sciences, 44(4): 354-347. (In Persian).
Moradi Shahrbabak, H. 2009. Association of polypathin, myostatin, leptin and potassium genes with important economic traits, blood metabolites and carcass traits in Makui and Zel sheep. PhD Thesis, Faculty of Agricultural Sciences, University of Tehran. (In Persian).
Moradi Shahr Babak, H., Moradi Shahr Babak, M., Rahimi, Q. and Nik Khah, A. 2004. Estimation of variance-covariance components and genetic and environmental parameters of growth traits in Kerman sheep. Proceedings of the First Congress of Animal and Aquatic Sciences of the country. Faculty of Agriculture, University of Tehran, 797-795. (In Persian).
Najafi, M. and Rahimi, G.h. 2010. Cloning, sequencing and comparative analysis of gene structure in promoter site of alpha-s1casein gene in Naeinian goat and sheep breed (thesis), Sari Agricultural Sciences and Natural Resources University, Iran, P 150. (In Persian).
Rychlik, W. 2007.  OLIGO 7 primer analysis software.  PCR primer design, 35-59.
Sahin, E., Yardimci, M., Cetingul, I., Bayram, I. and Sengor, E. 2008. The use of ultrasound to predict the carcass composition of live Akkaraman lambs. Meat Science, 79(4): 716-721.
Salimi, D., Eskandari Nasab, M.P. And Herki Nezhad, D. 2012. The effect of time delay from weaning on compensatory growth of Merino × Afshari lambs. The First National Congress of New Agricultural Sciences and Technologies. University of Zanjan. (In Persian).
Sanjari, A., Zia al-Dini, M., Asadi Fawzi, M. and Ismailizadeh Kashkuyeh, A. 2017. Myostatin gene polymorphism and its relationship with growth traits in Fluffy goats. Livestock Products, 20(3):365-373. (In Persian).
Sufi, B., Shojaeian, K., Mohammadabadi, M., Baghizadeh, A. and Frosti, S. 2009. Myostatin gene polymorphism in Squirrel sheep using PCR-RFLF marker, Journal of Animal Science Research, 9(1): 89-81. (In Persian).
Theriault, M., Pomar, C. and Castonguay, F. 2009. Accuracy of real-time ultrasound measurements of total tissue, fat, and muscle depths at different measuring sites in lamb. Journal of Animal Science, 87(5):1801-1813.
Vatankhah, M., Moradi-Shahrbabak, M., Nejatijavaremi, A., Miraei-Ashtiani, S.R. and Vaez Torshizi, R. 2004. The relationships between body and fat-tail measurments with weights of live, hot carcass and hot carcass without fat-tail in Lori-Bakhtiari sheep. Animal Science Journal (Pajouhesh and Sazandgi), 64, 66-74. (In Persian).
Wiener, P., Woolliams, J., Frank-Lawale, A., Ryan, M., Richardson, R., Nute, G., Wood, J., Homer, D. and Williams, J. 2009. The effects of a mutation in the myostatin gene on meat and carcass quality. Meat Science, 83(1):127-134.