Determination of CAST/MspI gene polymorphism in selected sheep breeds reared in Türkiye

Year 2025, Volume: 38 Issue: 2, 103 - 108, 20.08.2025

Göknur Bayrak , Yasemin Gedik

https://doi.org/10.29136/mediterranean.1712441

Abstract

Sheep breeding plays a crucial role in meeting the increasing demand for high-quality meat, milk, and wool. Traditional selection methods based on phenotypic traits have led to genetic improvement; however, progress remains slow for traits with low to moderate heritability. Marker-assisted selection (MAS) has emerged as a promising complementary approach, enabling faster genetic gains. Among candidate genes studied for MAS, the calpastatin (CAST) gene is notable for its association with meat yield and quality. This study aimed to identify CAST/MspI gene polymorphisms in four sheep breeds reared in Türkiye (Central Anatolian Merino-CAM, Pırlak-PRL, Romanov-RMV, and Suffolk-SFK) and assess their potential use in MAS. A total of 176 individuals were genotyped using the PCR-RFLP method. All breeds in this study were found to be polymorphic for the CAST gene. The frequency of the M allele ranged from 0.67 in CAM to 0.76 in SFK. Genotype frequencies for MM ranged from 0.40 (CAM) to 0.71 (SFK), for MN from 0.10 (RMV and SFK) to 0.54 (CAM), and for NN from 0.06 (CAM) to 0.20 (RMV). Significant deviations from Hardy-Weinberg equilibrium were observed in CAM, RMV, and SFK populations, but not in PRL. Observed heterozygosity ranged from 0.10 to 0.54 and expected heterozygosity from 0.37 to 0.44. The presence of all three genotypes and substantial genetic variation suggests that the CAST gene may be a valuable marker in MAS for these breeds. However, further association studies are required to confirm the relationship between CAST genotypes and economically significant traits related to meat production.

Keywords

Calpastatin gene , MspI , Polymorphism , Sheep

Ethical Statement

Ethical approval and permission for this study was obtained from Eskisehir Osmangazi University Animal Experiments Local Ethics Committee (Date: 09/10/2020; Decision No: 809).

Supporting Institution

ESOGU BAP

Project Number

1562-2021230A5

References

• Afanasyeva A, Sarychev V, Goncharenko G (2019) Phenotypic effects of polymorphism of the calpastatin gene (CAST), associated with growth and development indicators, in West Siberian mutton breed. International Scientific and Practical Conference “Digital Agriculture-Development Strategy” 167: 116-120.
• Asadi N, Nanekarani S, Khederzadeh S (2014) Genotypic frequency of calpastatin gene in lori sheep by polmerase chain reaction-restriction fragment lengt polymorphism (PCR-RFLP) method. African Journal of Biotechnology 13: 1952-1954.
• Avanus K (2015) Genetic variability of CAST gene in native sheep breeds of Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 21(6): 789-794.
• Balcıoğlu MS, Karslı T, Şahin E, Ulutaş Z, Aksoy Y (2014) Türkiye’de yetiştirilen bazı yerli koyun ırklarında kalpastatin (CAST) geni polimorfizminin PCR-RFLP yöntemiyle belirlenmesi. Tarım Bilimleri Dergisi 20: 427-433.
• Bayraktar M, Shoshin O (2022) Association between CAST and MSTN gene polymorphisms with growth traits in Awassi sheep. Kuwait Journal Science 49(2): 1-15.
• Bayram D, Akyüz B, Arslan K, Özdemi̇r F, Aksel EG, Çınar MU, (2019) Akkaraman kuzularında DGAT1, CAST ve IGF-I gen polimorfizmleri ve sütten kesim yaşına kadarki canlı ağırlık artışı üzerine etkileri. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 25(1): 9-15.
• Bozhilova-Sakova M, Dimitrova I, Tzonev TI, Petrov N (2020) Genotype frequencies in calpastatin (CAST) and callipyge (CLPG) genes in Northeast Bulgarian Merino sheep breed using PCR-RFLP method. Bulgarian Journal of Agricultural Science 26(2): 475-479.
• Chung H, Davis M (2012) PCR-RFLP of the ovine calpastatin gene and its association with growth. Asian Journal of Animal and Veterinary Advances 7(8): 641-652.
• Ciobanu DC, Bastiaansen JWM, Lonergan SM, Thomsen H, Dekkers JCM, Plastow GS, Rothschild MF (2004) New alleles in calpastatin gene are associated with meat quality traits in pigs. Journal of Animal Science 82: 2829-2839.
• Dehnavi E, Ahani Azari M, Hasani S, Nassiry MR, Mohajer M, Khan Ahmadi AR, Shahmohamadi L, Yousefi S (2012) Association between yearling weight and calpastatin and calpain loci polymorphism in Iranian Zel sheep. Iranian Journal of Applied Animal Science 2(2): 131-135.
• Demir E, Ceccobelli S, Bilginer U, Pasquini M, Attard G, Karsli T (2022) Conservation and selection of genes related to environmental adaptation in native small ruminant breeds: a review. Ruminants 2(2): 255-270.
• Esteves C, do Livramento KG, Paiva LV, Peconick AP, Garcia IF, Garbossa CAP, Cardoso TB, Faria PB (2020) Polymorphism of CAST and GDF-8 genes and association with physical-chemical parameters of sheep meat. Genetics and Molecular Research 19(3): 1-12.
• Gabor M, Trakovicka A, Miluchova M (2009) Analysis of polymorphism of cast gene and clpg gene in sheep by pcr-rflp method. Scientific Papers Animal Science and Biotechnologies 42: 470-476.
• Georgieva S, Hristova D, Dimitrova I, Stancheva N, Bozhilova-Sakova M (2015) Molecular analysis of ovine calpastatin (CAST) and myostatin (MSTN) genes in synthetic population Bulgarian Milk sheep using PCR-RFLP. Journal of BioScience Biotechnology 4(1): 95-99.
• Gharahveysi S, Abbasi HA, Irani M, Abdullahpour R, Mirhabibi S (2012) Polymorphism investigation of calpastatin gene in Zel sheep population of Iran by PCR-RFLP method. African Journal Biotechnol 11: 3211-3214.
• Goll DE, Thompson VF, Taylor RG, Ouali A (1998) The calpain system and skeletal muscle growth. Canadian Journal of Animal Science 78(4): 503-512.
• Gorlov IF, Shirokova NV, Randelin AV, Voronkova VN, Mosolova NI, Zlobina EY, Kolosov YA, Bakoev NF, Leonova MA, Bakoev SY, Leonova MA, Bakoev SY, Kolosov AY, Getmantseva LV (2016) CAST/MspI gene polymorphism and its impact on growth traits of Soviet Merino and Salsk sheep breeds in the South European part of Russia. Turkish Journal of Veterinary and Animal Sciences 40(4):399-405.
• Ibrahim AHM, Ismail IM, Shehata MF, El-Beltagy AR (2015) Calpastatin polymorphism in Barki lambs and their effects on growth and carcass traits. Journal of American Science 11(3): 106-112.
• Javanmard A, Panandam JM, Sugnaseelan S, Yusoff K (2010) Allele frequencies at six candidate genes associated with growth and carcass quality traits in the Boer goats. African Journal of Biotechnology 9(43): 7236-7238.
• Jawasreh KI, Jadallah R, Al-Amareen AH, Abdullah AY, Al-Qaisi A, Alrawashdeh IM, Al-Zghoul MBF, Ahamed M, Obeidat B (2017) Association between MspI calpastatin gene polymorphisms, growth performance, and meat characteristics of Awassi sheep. Indian Journal of Animal Science 87(5): 635-639.
• Jawasreh KI, Al-Amareen AH, Aad PY (2019) Relationships between Hha1 calpastatin gene polymorphism, growth performance, and meat characteristics of Awassi sheep. Animals 9(9): 667.
• Jawasreh KI, Ismail ZB (2019) Effects of calpastatin gene polymorphism on hematology and selected serum biochemical parameters in Awassi lambs. Journal of Advanced Veterinary and Animal Research 6(2): 193-196.
• Kania GM, Gruszecki TM, Junkuszew A, Juszczuk-Kubiak E, Florek M (2019) Association of CAST gene polymorphism with carcass value and meat quality in two synthetic lines of sheep. Meat Science 154: 69-74.
• Karslı T, Balcıoğlu MS, Demir E, Fidan HG, Aslan M, Aktan S, Şahin, E (2017) Determination of polymorphisms in IGF-I and NPY candidate genes associated with egg yield in pure layers chicken lines reared in the Ankara Poultry Research Institute. Turkish Journal of Agriculture-Food Science and Technology 5(9): 1051-1056.
• Kawasaki H, Kawashima S (1996) Regulation of the calpain-calpastatin system by membranes. Molecular Membrane Biology 134: 217-224.
• Khan SH, Riaz MN, Ghaffar A, Ullah MF (2012) Calpastatin (CAST) gene polymorphism and its association with average daily weight gain in Balkhi and Kajli sheep and Beetal goat breeds. Pakistan Journal of Zoology 44: 377-382.
• Knight MI, Daetwyler HD, Hayes BJ, Hayden MJ, Ball AJ, Pethick DW, McDonagh MB (2012) Discovery and trait association of single nucleotide polymorphisms from gene regions of influence on meat tenderness and long-chain omega-3 fatty acid content in Australian lamb. Animal Production Science 52(7): 591.
• Kolosov YA, Gorlov IF, Kolosov AY, Shirokova NV, Kulikova AY, Kolosova MA, Slozhenkina MI, Vorontsova ES, Kolosova NN (2021) Determination of CAST gene polymorphism in sheep of the Volgograd breed. IOP Conference Series: Earth and Environmental Science 677(5): 052112.
• Kumar SN, Jayashankar MR, Ruban W, Sreesujatha RM, Devi YSR (2018) Association of calpastatin polymorphism with carcass and meat quality traits of Bandur sheep. International Journal of Livestock Research 8(5): 232-243.
• Nanekarani S, Khederzadeh S, Kaftarkari AM (2011) Genotypic frequency of calpastatin gene in Atabi sheep by PBR method. International Conference on Food Engineering and Biotechnology 9: 189-192.
• Nassiry MR, Tahmoures PM, Javad MA, Soltani M, Foroutani FS (2006) Calpastatin polymorphism and its association with daily gain in Kurdi sheep. Iranian Journal of Biotechnology 4(3): 188-192.
• NCBI (2021) National Center for Biotechnology Information, CAST calpastatin [Ovis aries (sheep)], https://www.ncbi.nlm.nih.gov/gene/?term=443364, Accessed 28 December, 2021.
• Nikmard M, Molaee V, Eskandarinasab MP, Dinparast Djadid N, Vajhi AR (2012) Calpastatin polymorphism in Afshari sheep and its possible correlation with growth and carcass traits. Journal of Applied Animal Research 40(4): 346-350.
• Palmer BR, Roberts N, Kent MP, Ilian MA, Le Couteur C, Morton J, Hickford JGH, Bickerstaffe R (1997) Calpastatin as a candidate gene for sheep meat tenderness. Annual International Congress of Meat Science and Technology 43: 590-591.
• Palmer BR, Roberts N, Hickford JG, Bickerstaffe R (1998) Rapid communication: PCR-RFLP for MspI and NcoI in the ovine calpastatin gene. Journal of Animal Science 76: 1499-1500.
• Palmer BR, Morton JD, Roberts N, Ilian MA, Bickerstaffe R (1999) Marker–assisted selection for meat quality and the ovine calpastatin gene. Proceedings of the New Zealand Society of Animal Production 59: 266-268.
• Peirvisi N, Noshary A, Hemati B (2020) The calpastatin gene polymorphism study and its effect on early body weight traits in Zandi sheep. Iranian Journal of Applied Animal Science 10(2): 297-302.
• Pomitun IA, Rossokha VI, Boyko YA, Guzevatyi OE, Shpilka MV, Kulibaba RO (2019) Analysis of calpastatin and сallipyge genes polymorphism in Prydniprovska meat sheep. Agricultural Science and Practice 6(2): 58-65.
• Ramadevi B, Kumari BP, Sudhakar K, Gangaraju G, Vinod U (2020) Polymorphism of the ovine calpastatin (CAST) gene and its association with productive traits in Nellore sheep. Journal of Animal Research 10(6): 881-887.
• Ropka-Molik K, Bereta A, Tyra M, Różycki M, Piórkowska K, Szyndler-Nędza M, Szmatoła T (2014) Association of calpastatin gene polymorphisms and meat quality traits in pig. Meat Science 97(2): 143-150.
• Selionova MI, Chizhova LN, Surzhikova ES, Podkorytov NA, Podkorytov AT, Voblikova TV (2020) Meat productivity of sheep of the Altai Mountain breed of different genotypes according to the CAST and GDF9 genes. IOP Conference Series: Earth and Environmental Science 613(1): 012130.
• Shahroudi FE, Nassiry MR, Valizadh R, Moussavi A H, Pour MT, Ghiasi H (2006) Genetic polymorphism at MTNR1A, CAST and CAPN loci in Iranian Karakul sheep. Iranian Journal Biotechnology 4:117-122.
• Suleman M, Khan SU, Riaz MN, Yousaf M, Shah A, Ishaq R, Ghafoor A (2012) Calpastatin (CAST) gene polymorphism in Kajli, Lohi and Thalli sheep breeds. African Journal of Biotechnology 11(47): 10655-10660.
• Sunilkumar MC, Nagaraja M, Jayashankar N, Fairoze B, Veeregowda B (2014) Molecular studies on meat quality gene in Bandur sheep. Journal of Cell and Tissue Research 141: 40-49.
• Sutikno M, Yamin M, Sumantri C (2011) Association of polymorphisms calpastatin gene with body weight of Local sheep in Jonggol, Indonesia. Media Peternakan 34(1): 1-6.
• Szkudlarek-Kowalczyk M, Wiśniewsk E, Mroczkowsk S (2011) Polymorphisms of calpastatin gene in sheep. Journal of Central European Agriculture 12: 425-432.
• Tohidi R (2013) Molecular analysis of ovine calpastatin gene in six Iranian sheep breeds using PCR-RFLP. Journal of Animal Production Advances 3: 271-277.
• Valencia CPL, Franco LÁÁ, Herrera DH (2022) Association of single nucleotide polymorphisms in the CAPN, CAST, LEP, GH, and IGF-1 genes with growth parameters and ultrasound characteristics of the Longissimus dorsi muscle in Colombian hair sheep. Tropical Animal Health and Production 54(1): 82.
• Yeh FC, Yang RC, Boyle TBJ, Ye ZH, Mao JX (1997) Popgene, The user-friendly shareware for population genetic analysis. Molecular Biology and Biotechnology Centre, University of Alberta, Canada.
• Yılmaz O, Sezenler T, Ata N, Yaman Y, Cemal İ, Karaca O (2014a) Polymorphism of the ovine calpastatin gene in some Turkish sheep breeds. Turkish Journal of Veterinary and Animal Sciences 38: 354-357.
• Yılmaz O, Cemal İ, Karaca O, Ata N (2014b) Association of calpastatin (CAST) gene polymorphism with weaning weight and ultrasonic measurements of loin eye muscle in Kıvırcık lambs. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 20: 675-680.