Investigation of GDF9 (FecG1) Gene Polymorphism by PCR-RFLP Method in Of Sheep, a Local New Sheep Type in Turkey

Öz

Of sheep, registered as a local type in the 2020 year, are bred in the districts of Of, Çaykara, Dernekpazarı, Hayrat in Trabzon, Turkey. The number of sheep raised as pure is around 1800 and 2000 heads, and twin birth rates are up to 35-40 %. In this study, in a total of 24 head Of sheep, the GDF9 (FecG1) gene polymorphism, which is related to multiple births in sheep, was investigated by the PCR-RFLP method. After the digestion of PCR products with the Hhal restriction enzyme, two alleles (A and G) and two genotypes (GA and GG) were identified. The frequencies of alleles A and G were found to be 0.46 to 0.54. The frequencies of genotypes GA and GG were found to be 0.92 to 0.09. To conclude, many heterozygous individuals in terms of the GDF9 gene in Of sheep provide an important opportunity to conduct the studies on its relationships with multiple births in this genotype.

Anahtar Kelimeler

• Of sheep
• multiple births
• GDF9 (FecG1) gene
• PCR-RFLP
• Hhal enzyme

Türkiye Yöresel Yeni Koyun Tipi Of Koyunlarında GDF9 (FecG1) Gen Polimorfizmin PCR-RFLP Yöntemi ile Araştırılması

Öz

Of koyunu Trabzon ili Çaykara, Dernekpazarı, Hayrat ve Of ilçelerinde yetiştirilmekte olup 2020 yılında yöresel tip olarak tescil edilmiştir. Saf olarak yetiştiriciliği yapılan Of koyunu sayısı 1800 ile 2000 baş civarında olup bunlarda ikiz doğum oranları %35-40’lara kadar çıkmaktadır. Bu çalışmada toplam 24 baş saf Of koyununda çoklu doğum ile ilişkili GDF9 (FecG1) geni polimorfizmi, PCR-RFLP yöntemi ile araştırılmıştır. PCR ürünlerin Hhal restriksiyon emzimi ile kesimi sonucunda iki allel (G ve A) ve iki genotip (GA ve GG) tespit edilmiştir. A ve G allellerin frekansları; 0.46 ile 0.54, GA ve GG genotiplerin frekansları ise 0.92 ile 0.08 olarak bulunmuştur. Bu çalışma sonucunda Of koyununda GDF9 geni bakımından heterozigot birey sayısının fazla olması, bu genotipte çoklu doğum ile ilişkilendirme çalışmalarının yapılması için önemli bir fırsat sunmaktadır.

Anahtar Kelimeler

• Of koyunu
• çoklu doğum
• GDF9 (FecG1) geni
• PCR-RFLP
• Hhal enzimi

Kaynakça

1. Abdoli R, Zamani P, Deljou A, Rezvan H 2013. Association of BMPR-1B and GDF9genes polymorphisms and secondary protein structure changes with reproduction traits in Mehraban ewes. Gene, 524: 296–303.
2. Amini H, Ajaki A, Farahi M, Heidari M, Pirali A, Forouzanfar M, Eghbalsaied S 2018. The novel T755C mutation in BMP15 is associated with the litter size of Iranian Afshari, Ghezel, and Shal breeds. Archiv fuer Tierzucht, 61(1), 153.
3. Analla M, Munoz-Serrano A, Serradilla JM 1997. Analysis of the genetic relationship between litter size and weight traits in Segurena sheep. Canadian Journal of Animal Science, 77(1), 17-21.
4. Davis GH, McEwan JC, Fennessy PF, Dodds KG, Farquhar PA 1991. Evidence for the presence of a major gene influencing ovulation rate on the X chromosome of sheep. Biol Reprod 44:620–624.
5. Dincel D, Ardicli S, Soyudal B, Er M, Alpay F, Şamlı H, Balci F 2015. Analysis of FecB, BMP15 and CAST Gene Mutations in Sakiz Sheep. Kafkas Universitesi Veteriner Fakultesi Dergisi, 21(4).
6. Dinçel D, Ardıçlı S, Şamlı H, Balcı F 2018. Genotype frequency of FecXB (Belclare) mutation of BMP15 gene in Chios (Sakiz) sheep. Uludag Univ. J. Fac. Vet. Med, 37(2), 87-91.
7. Eghbalsaied S, Rashidi Khorasgani F, Amini HR, Farahi M, Davari M, Pi - rali A, Pourali S, Vatankhah M, Rostami M, Atashi H 2017. Variant GDF9 mRNA is likely not the main cause of larger litter size in Iranian Lori-Bakhtyari, Shal, Ghezel, and Afshari sheep breeds. Arch. Anim. Breed., 60: 119–129.
8. Escobar-Chaparro RA, Guillén G, Espejo-Galicia LU, Meza-Villalvazo VM, Peña-Castro JM, Abad-Zavaleta J 2017. qPCR and HRM-based diagnosis of SNPs on growth differentiation factor 9 (GDF9), a gene associated with sheep (Ovis aries) prolificacy. 3 Biotech, 7(3), 204.

9. Gorlov IF, Kolosov YA, Shirokova NV, Getmantseva LV, Slozhenkina M., Mosolova NI, Zlobina EY 2018. GDF9 gene polymorphism and its association with litter size in two Russian sheep breeds. Rendiconti Lincei. Scienze Fisiche e Naturali, 29(1), 61-66.
10. Gursel E 2011. Determination of BMP-15, BMPR-1B and GDF-9 gene mutations of the indigenous sheep breeds in Turkey. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 17(5).
11. Janssens S, Vandepitte W, Bodin L 2004. Genetic parameters for litter size in sheep: natural versus hormone-induced oestrus.Genetics Selection Evolution, 36(5), 1-20.
12. Karslı T, Balcıoğlu MS 2010. Türkiye'de yetiştirilen altı yerli koyun ırkında BMPR-IB (Booroola) Geninde FecB allel varlığının PCR-RFLP yöntemiyle araştırılması. Kafkas Universitesi Veteriner Fakultesi Dergisi, 17(5).
13. Karsli T, Sahin E, Karsli BA, Alkan S, Balcioglu M S 2012. An investigation of mutations (FecXG, FecXI, FecXH, FecXB) on BMP-15 gene in some local sheep breeds raised in Turkey. Akdeniz Üniv. Ziraat Fak, 25(1), 29-33.
14. Kirikci K, Cam MA, Mercan L. Genetic diversity and relationship among indigenousTurkish Karayaka sheep subpopulations. Archives Animal Breeding 2020; 63 (2): 269-275.
15. Kolosov YA, LV Genmantseva, NV Shirockova, A Klimenko 2015. Polymorphism of the GDF9 Gene in Russian Sheep Brreds. Journal of Cytology & Histology, 6: 01-04.
16. Mohamed SEI, Ahmed RM, Jawasreh KI, Salih MAM, Abdelhalim DM, Abdelgadir AW, Ahmed MKA 2019. Genetic polymorphisms of fecundity genes in Watish Sudanese desert sheep. Veterinary world, 13(4), 614.
17. Moradband F, G Rahimi, M Gholizadeh 2011. Association of Polymorphisms in Fecundity Genes of GDF9, BMP15 and BMP15-1B with Litter Size in Iranian Baluchi Sheep. Asian-Australasian Journal of Animal Sciences, 24: 1179-1183.
18. Paz E, Quiñones J, Bravo S, Montaldo H, Sepúlveda N 2015. Genotyping of BMPR1B, BMP15 and GDF9 genes in Chilean sheep breeds and association with prolificacy. Animal Genetics; 46 (1): 98-99.
19. Yeh FC, Yang RC, Boyle TB J, Ye ZH, Mao JX 2000. POPGENE 32, Microsoft Windows-based software for population genetic analysis (version 1.32). Endmonton, AB: Molecular Biology and Biotechnology Centre, University of Alberta.
20. Safari E, Fogarty, NM, Gilmour, AR 2005. A review of genetic parameter estimates for wool, growth, meat and reproduction traits in sheep. Livestock Production Science, 92(3), 271-289.
21. Savas T, Röhe, R, Kalm, E 2000. Schätzung genetischer Parameter für die Fruchtbarkeitsleistung beim Schaf. Züchtungskunde, 72, 217-229.
22. Vage DI, Husdal M, Kent MP, Klemetsdal G, Boman IA 2013. A missense mutation in growth differentiation factor 9 (GDF9) is strongly associated with litter size in sheep. BMC Genetics 2013; (14): 1.
23. Vatankhah, M, Talebi, MA 2008. Heritability estimates and correlations between production and reproductive traits in Lori-Bakhtiari sheep in Iran. South African Journal of Animal Science, 38(2), 110-118.
24. Zamani P, Nadri S, Saffaripour R, Ahmadi A, Dashti F, Abdoli R 2015.A new mutation in exon 2 of the bone morphogenetic protein 15 gene is associated with increase in prolificacy of Mehraban and Lori sheep. Trop. Anim. Health. Prod., 47: 855–860.