DENİZ DİNÇEL, SENA ARDIÇLI, Bahadır SOYUDAL, Mehlika ER, FAZLI ALPAY, HALE ŞAMLI, Faruk BALCI

Analysis of FecB, BMP15 and CAST Gene Mutations in Sakiz Sheep [1]

Ovulasyon oranı ve bir batındaki yavru sayısı gibi dölverim özellikleri fekondite genleri tarafından regüle edilmektedir. Bu çalışma Sakız ırkı koyunlarda, Booroola (FecB), Galway (FecXG), Inverdale (FecXI ) ve Calpastatin (CAST) genlerindeki DNA polimorfizmlerini belirlemek amacıyla yapılmıştır. Araştırılan gen veya aleller açısından toplam 71 koyun PCR-RFLP metoduyla genotiplendirilmiştir. Çalışmadan çıkan sonuçlar, Sakız koyunların FecB, FecXG ya da FecXI ye ait mutasyonları taşımadığını göstermiştir. Ancak CAST geni açısından AA, AB ve BB genotiplerinin sırasıyla 0.59, 0.36 ve 0.05 frekans gösterdiği tespit edilmiştir. CAST geni açısından çalışılan sürünün Hardy-Weinberg dengesine uyduğu görülmüştür (P>0.05). Sonuç olarak yeni keşfedilen mutasyonların geliştirilmesi gibi daha yoğun tarama çalışmaları gerekmektedir. Ek olarak Sakız koyunlarında CAST genindeki polimorfizmin varlığı ilk defa bu çalışmada bildirilmektedir.

Sakız Irkı Koyunlarda FecB, BMP15 ve CAST Genlerindeki Mutasyonların Analizi

Fertility traits, such as the ovulation rate and the number of offspring at birth, are genetically regulated by fecundity genes. This study was performed to identify DNA polymorphisms in Booroola (FecB), Galway (FecXG), Inverdale (FecXI ) and Calpastatin (CAST) genes in Sakiz sheep. A total of 71 ewes were genotyped for gene or allelic polymorphisms in the genes listed above using the PCR-RFLP method. The results obtained from this study indicated that all of the Sakiz ewes sampled were non-carriers for FecB, FecXG or FecXI mutations. However, genotypic frequencies in the CAST gene were 0.59, 0.36 and 0.05 for AA, AB and BB, respectively. A significant deviation from Hardy-Weinberg equilibrium for the CAST gene was not observed in the investigated breed (P>0.05). As a result, more extensive screening is required as tests for newly discovered mutations are developed. Additionally, this study is the first to report a genetic polymorphism in the CAST gene in Sakiz sheep.

PDF

___

1. Davis GH: Major genes affecting ovulation rate in sheep. Genet Sel Evol, 37 (Suppl. 1): S11-S23, 2005. DOI: 10.1051/gse:2004026
2. Işık S: Bafra koyununun (Sakiz × Karayaka G1) Kazım Karabekir Tarım İşletmesi şartlarında dölverimi, yaşama gücü ve büyüme özellikleri. Doktora Tezi, Kafkas Univ. Sağlık Bil. Enst, 2010.
3. Ulker H, Bas S: Koyunlarda major etkili fekondite genleri. 2- Fekonditeye etki eden mutasyonlarin henüz belirlenemediği genler. Hayvansal Üretim, 50 (1): 68-73, 2009.
4. Davis GH: Fecundity genes in sheep. Anim Reprod Sci, 82-83, 247-253, 2004. DOI: 10.1016/j.anireprosci.2004.04.001
5. Davis GH, Balakrishnan L, Ross IK, Wilson T, Galloway SM, Lumsdenb BM, Hanrahan JP, Mullen M, Maoe XZ, Wang GL, Zhaoe ZS, Zenge YQ, Robinson JJ, Mavrogenis AP, Papachristoforou C, Peter C, Baumungi R, Cardyn P, Boujenane I, Cockett NE, Eythorsdottirm E, Arranz JJ, Notter DR: Investigation of the Booroola (FecB) and Inverdale (FecXI) mutations in 21 prolific breeds and strains of sheep sampled in 13 countries. Anim Reprod Sci, 92, 87-96, 2006. DOI: 10.1016/j. anireprosci.2005.06.001
6. Mishra C: Genetic basis of prolificacy in sheep. Int J Livest Res, 4, 46-57, 2014. DOI: 10.5455/ijlr.20131227083421
7. Piper LR, Bindon BM, Davis GH: The single gene inheritance of the high litter size of the Booroola Merino. In, Land RB, Robinson DW (Eds): Genetics of Reproduction in Sheep. 115-125, Butterworths, London, 1985.
8. Montgomery GW, Lord EA, Penty JM, Dodds KG, Broad TE, Cambridge L, Sunden ALF, Stone RT, Crawford AM: The Booroola fecundity (FecB) gene maps to sheep chromosome 6. Genomics, 22, 148- 153, 1994. DOI: 10.1006/geno.1994.1355.
9. Mulsant P, Lecerf F, Fabre S, Schibler L, Monget P, Lanneluc I, Pisselet C, Riquet J, Monniaux D, Callebaut I, Cribiu E, Thimonieri J, Teyssieri J, Bodin L, Cognie Y, Chitour N, Elsen J: Mutation in bone morphogenetic protein receptor-IB is associated with increased ovulation rate in Booroola Merino ewes. Proc Natl Acad Sci U S A, 98, 5104-5109, 2001. DOI: 10.1073/pnas.091577598
10. Karsli T, Balcioğlu MS: Türkiyede yetiştirilen altı yerli koyun ırkında BMPR-IB (Booroola) geninde FecB allel varlığının PCR-RFLP yöntemiyle araştırılması. Kafkas Univ Vet Fak Derg, 16, 1033-1036, 2010. DOI: 10.9775/ kvfd.2010.2333
11. Wilson T, Wu XY, Juengel JL, Ross, IK, Lumsden JM, Lord, EA, Dodds KG, Walling GA, Mcewan JC, Oconnel AR, Mcnatty KP, Montgomery GW: Highly prolific Booroola sheep have a mutation in the intracellular kinase domain of Bone Morphogenetic Protein IB Receptor (ALK-6) that is expressed in both oocytes and granulosa cells. Biol Reprod, 64, 1225- 1235, 2001.
12. Kumar S, Mishra AK, Kolte AP, Dash SK, Karim SA: Screening for Booroola (FecB) and Galway (FecXG) mutations in Indian sheep. Small Rumin Res, 80, 57-60, 2008. DOI: 10.1016/j.smallrumres.2008.09.007
13. Davis GH, Galloway SM, Ross IK, Gregan SM, Ward J, Nimbkar BV, Ghalsasi PM, Nimbkar C, Gray DG, Inounu I, Tiesnamurti B, Martyniuk E, Eythorsdottir E, Mulsant P, Lecerf F, Hanrahan JP, Bradford GH, Wilson T: DNA tests in prolific sheep from eight countries provide new evidence on origin of the Booroola (FecB) mutation. Biol Reprod, 66, 1869- 1874, 2002.
14. Kasiriyan MM, Hafezeyan H, Sayahzadeh H, Jamshidi R, Asgrahi SR, Irajeyan GH, Buesagh H: Genetic polymorphism FecB and BMP15 genes and association with litter size in Sangsari sheep breed of Iran. J Anim Vet Adv, 8 (5): 1025-1031, 2009.
15. Galloway, SM, Mcnatty KP, Cambridge LM, Laitinen MPE, Juengel JL, Jokiranta TS, Mclaren RJ, Luiro K, Dodds KG, Montgomery GW, Beattie AE, Davis GH, Ritvos O: Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner. Nat Genet, 25, 279-283, 2000. DOI: 10.1038/77033
16. Hanrahan JP, Gregan SM, Mulsant P, Mullen M, Davis GH, Powell R, Galloway SM: Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both ıncreased ovulation rate and sterility in Cambridge and Belclare sheep (Ovis aries). Biol Reprod, 70, 900-909, 2004. DOI: 10.1095/biolreprod.103.023093
17. Davis GH, McEwan JC, Fennessy PF, Dodds KG, Farquhar PA: Evidence for the presence of a major gene influencing ovulation rate on the x-chromosome of sheep. Biol Reprod, 44, 620-624, 1991. DOI: 10.1095/ biolreprod44.4.620
18. Monteagudo LV, Ponz R, Tejedor MT, Lavina A, Sierra I: A 17 bp deletion in the Bone Morphogenetic Protein 15 (BMP15) gene is associated to increased prolificacy in the Rasa Aragonesa sheep breed. Anim Reprod Sci, 110, 139-146, 2009. DOI: 10.1016/j.anireprosci.2008.01.005
19. Chu M, Liu ZH, Jiao CL, He YQ, Fang L, Ye SC, Chen GH, Wang JY: Mutations in BMPR-IB and BMP15 genes are associated with litter size in Small Tailed Han sheep (Ovis aries). J Anim Sci, 85, 598-603, 2006. DOI: 10.2527/jas.2006-324
20. Esen Gursel F, Akis I, Durak H, Mengi A, Oztabak K: Determination of BMP15, BMPR-1B and GDF-9 gene mutations of the indigenous sheep breeds in Turkey. Kafkas Univ Vet Fak Derg, 17, 725-729, 2011. DOI: 10.9775/kvfd.2011.4256
21. Mohammadi M, Beigi Nasiri MT, Alami-Saeid KH, Fayazi J, Mamoee M, Sadr AS: Polymorphism of calpastatin gene in Arabic sheep using PCR-RFLP. Afr J Biotechnol, 7 (15): 2682-2684, 2008.
22. Chung H, Davis M: PCR-RFLP of the ovine Calpastatin and its association with growth. Asian J Anim Vet Adv, 7, 641-652, 2012. DOI: 10.3923/ajava.2012.641.652
23. Dehnavi E, Azari MA, Hasani S, Nassiry MR, Mohajer M, Ahmadi ARK: Genetic variability of calpastatin and calpain genes in Iranian Zel sheep using PCR-RFLP and PCR-SSCP methods. Iranian J Biotech, 10 (2): 136-139, 2012.
24. Ranjbari M, Hashemi A, Mardani K, Darvishzadeh R: Allelic polymorphism of Makoei sheep Calpastatin gene identified by polymerase chain reaction and single strand conformation polymorphism. J Agr Sci Tech, 14, 533-538, 2012.
25. Szkudlarek-Kowalczyk M, Wiśniewska E, Mroczkowsi S: Polymorphisms of calpastatin gene in sheep. JCEA, 12, 425-432, 2011. DOI: 10.5513/JCEA01/12.3.934
26. Garcia MD, Mıchal JJ, Gaskins CT, Reeves JJ, Ott TL, Liu Y, Jiang Z: Significant association of the calpastatin gene with fertility and longevity in dairy cattle. Anim Genet, 37, 304-305, 2006. DOI: 10.1111/j.1365- 2052.2006.01443.x
27. Michaidilis G, Avdi M, Pappa V: Reproductive performance and investigation of BMPR-IB and BMP15 gene mutations in Greek Chios and Florina sheep breeds. Arch Zootech, 11 (1): 24-31, 2008.
28. Kurar E, Ozsensoy Y, Bulut Z, Guzeloglu A, Nizamlioglu M: Investigaiton of BMP15 and BMRP-1B gene mutations in profilic Sakiz sheep. Reprod Domest Anim, 46 (3): 121, 2011.
29. Doyle JJ, Doyle JL: A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 19, 11-15, 1987.
30. Killefer J, Koohmaraie M: Bovine skeletal muscle calpastatin: Cloning, sequence analysis, and steady-state mRNA expression. J Anim Sci, 72, 606- 614, 1994.
31. Yeh FC, Yang RC, Boyle TBJ, Ye ZH, Mao JX: Popgene (v.1.32) the user-friendly shareware for population genetic analysis, 2000. http:// www.ualberta.ca/~fyeh/, Accessed: 01.01.2015.
32. Moradband F, Rahimi G, Gholizadeh M: Association of polymorphisms in fecundity genes of GDF9, BMP15 and BMP15-1B with litter size in Iranian Baluchi sheep, Asian-Aust J Anim Sci, 24, 1179-1183, 2011. DOI: 10.5713/ajas.2011.10453
33. Roy J, Polley S, De S, Mukherjee A, Batabyal S, Pan S, Brahma B, Datta TK, Goswami SL: Polymorphism of fecundity genes (FecB, FecX, and FecG) in the Indian Bonpala sheep. Anim Biotechnol, 22, 151-162, 2011. DOI: 10.1080/10495398.2011.589239
34. Abulyazid I, Sharada HM, Abdalla MS, Saleh HM, Hassanin WF: FecXI gene as a monitor for the production of twins in the Egyptian sheep. Roavs, 1 (4): 217-219, 2011.
35.Byun SO, Zhou H, Frampton CM, Hickford JGH: No association between variation in the ovine calpastatin gene and either longevity or fertility in sheep. Anim Sci, 41, 222-224, 2009. DOI: 10.1111/j.1365- 2052.2009.01982.x