Investigation of FecXI, FecXH and FecXB Polymorphism in BMP15 Gene in Lac Qashqai and Bahmaei Sheep Breeds at Kohkiloueh and Boyerahmad Province

Document Type : Genetics & breeding

Authors

• mostafa Muhaghegh-Dolatabady
• asma moradalipour
• farkhondeh Mansouri
• farangis Fattahy
• zienab Salehi

University of Yasouj

10.22067/ijasr.v1397i1.70188

Abstract

Introduction Litter size is an important economic trait in sheep breeding which is influenced by genetics and environment factors. Application of phenotypic data in traditional breeding programs is a time consuming process. Thus, marker assisted selection (MAS) play an important role for genetic improvement of reproduction efficiency. Since, there has been great interest in the identification of major genes that influence fecundity traits in sheep. Three of these fecundity identified genes are bone morphogenetic protein receptor type IB (BMPRIB) or Activin Like Kinase 6, known as FecB located on chromosome 6; growth differentiation factor (GDF9), known as FecG located on chromosome 5 and bone morphogenetic protein 15 (BMP15), known as FecX located on the X chromosome. The BMP15) is expressed in oocytes where encodes for a mature peptide of 125 amino acids that stimulate the proliferation rate of the granulose cells and thus it seems to be essential in folliculogenesis. Some mutations in BMP15 gene are reported to increase fertility rate. Six different point mutations i.e. FecXI and FecXH in the Inverdale and Hanna breeds, respectively, FecXL in the Lacaune, FecXG and FecXB in the Cam-bridge and Beclare breeds, respectively and the FecXR in Raza Aragoneza breed have been identified in BMP15 gene. In this study, the FecXB, FecXH and FecXI mutations at the BMP15 gene were investigated by PCR–RFLP in Lac Ghashghaei and Bahmaei sheep breeds. The FecXB (G to T nucleotide change) was detected using DdeI; the wild-type strand was cleaved. FecXH (C to T nucleotide change) was detected using SpeI, the mutation-type strand was cleaved and FecXI (T to A nucleotide change) was detected using XbaI; the mutation-type strand was cleaved.
Materials and Methods Blood samples were taken from 92 ewes according to data on litter size at the last lambing (Lac Ghashghaei: 24 single and 24 double lambing, Bahmaei: 22 single and 22 double lambing). The animals of the Lac Ghashghaei and Bahmaei breeds originate from two farms in Gachsaran and Dehdasht, respectively. Then, genomic DNA was isolated from whole blood using the DNA Extraction Kit according to the manufacturer’s instructions. Required parts of BMP15 gene were amplified using specific primers (fragment of 204 bp from FecXI, fragment of 235 bp of FecXH and fragment of 153 bp of FecXB) through Polymerase Chain Reaction (PCR). The PCR products of amplified fragments were digested by XbaI, SpeI and DdeI restriction enzymes and visualized by agarose gel electrophoresis, separately.
Results and Discussion Sheep breeding is the major activity in the Kohkiloueh and Boyerahmad province of Iran and is very important in the rural economy of this region. Although litter size is an important trait that affects the profitability of sheep production, but the litter size of almost all sheep breeds of this province is low. In this study, PCR-RFLP method was used to identify three mutations (FecXI, FecXH and FecXB) of BMP15 gene in Lac Ghashghaei and Bahmaei ewes. As expected, the size of PCR production for FecXI, FecXH and FecXB loci were 204, 235 and 153 bp, respectively. After digestion of amplified fragments with specific restriction enzyme, the three earlier mentioned mutation sites were not detected in Lac Ghashghaei and Bahmaei sheep and only the wild-type genotype (++) was observed in both breeds for each mutation of BMP15 gene. In more detail, restriction digestion of amplified fragments for FecXB locus resulted DNA fragments with 31 and 122 bp. The wild type of this locus has one restriction site and this means that FecXB mutation was not detected in two breeds of this study and animal are homozygous for wild type allele of this locus. The resulted PCR products were digested with SpeI and DdeI restriction enzymes for FecXH, FecXI loci revealed DNA fragments with 235 and 204 bp, respectively. The mutant allele of FecXH, FecXI loci has one restriction site and enzyme digestion will be resulted in 2 DNA fragments while the wild type allele with no restriction site, resulted in one DNA fragments with the size. It means all 92 Lac Ghashghaei and Bahmaei ewes sampled were homozygous wild allele (++) for FecXH, FecXI loci.
Conclusion The obtained results of this study indicated that there was an absence of three BMP15 mutations (FecXB, FecXH and FecXI) among the Lac Qashqai and Bahmaei sheep breeds sampled in the Kohkiloueh and Boyerahmad province, it seems genetic factor responsible for litter size is not related to reported mutated alleles of BMP15 gene and therefore other prolificacy related genes should be investigated in these breeds using a larger sample size.

Keywords

• BMP15 gene
• PCR-RFLP
• Polymorphism
• Sheep
• Twin

20.1001.1.20083106.1398.11.2.10.2

---

References

1- Abulyazid, I., H. M. Sharada, M. S. Abdalla, H. M. Saleh, and W. F. Hassanin. 2011. FecX1 gene as a monitor for the production of twins in the Egyptian sheep. Research Opinions in Animal and Veterinary Sciences, 1(4): 217-219.

2- Alinaghizadeh, H., M. R. MohammadAbadi, and S. Zakizadeh. 2010. Exon 2 of BMP15 gene polymorphism in Jabal Barez Red Goat. Iranian Journal of Agricultural Biotechnology, 2(1): 69-80. (In Persian).

3- Amiri, A., G. H. Rahimi Miyanji, and M. Vatankhah. 2008. Absence of allelic polymorphisms in Broula (FecB) and FecX1 gene in Lori-Bakhtiari breed sheep. Journal of Genetics-Novin, 3(4): 57-63. (In Persian).

4- Bodin, L., E. Di Pasquale, S. Fabre, M. Bontoux, P. Monget, L. Persani, and P. Mulsant. 2007. A novel mutation in the BMP15 gene causing defective protein secretion is associated with both increased ovulation rate and sterility in Lacunae sheep. Endocrinology, 148: 393-400.

5- Borni, J., B. Sonia, and D. M. Naouer. 2011. Study for identification FecXI and FecXH mutations in Tunisian Barbarine sheep. Research Opinions in Animal & Veterinary Sciences, 1(2): 112-115.

6- Chu, M. X., Z. H. Liu, C. L. Jiao, Y. Q. He, L. Fang, S. C. Ye, G. H. Chen, and J. Y. Wang. 2006. Mutations in BMPR-IB and BMP15 genes are associated with litter size in small tailed Han sheep (Ovis aries). Animal Science, 85: 598-603.

7- Chu, M., R. Cheng, G. Chen, L. Fang, and S. Ye. 2004. Study on bone morphogenetic protein 15 as a candidate gene for prolificacy of small tail Han sheep and Hu sheep. Journal of Anhui Agricultural University, 32(3): 278-282.‏

8- Davis, G. H. 2005. Major genes affecting ovulation rate in sheep. Genetics Selection Evolution, 37: 11–23.

9- Davis, G. H., S. M. Galloway, I. K. Ross, S. M. Gregan, J. Ward, B. V. Nimbkar, P. M. Ghalsasi, C. Nimbkar, G. D. Gray, Subandriyo, I. Inounu, B. Tiesnamurti, E. Martyniuk, E. Eythorsdottir, P. Mulsant, F. Lecerf, J. P. Hanrahan, G. E. Bradford, and T. Wilson. 2002. DNA tests in prolific sheep from eight countries provide new evidence on origin of the Booroola (FecB) mutation. Biology of Reproduction, 66: 1869-1874.

10- Davis, G.H., L. Balakrishnan, I. K. Ross, T. Wilson, S. M. Galloway, B. M. Lumsden, et al. 2006. Investigation of the Booroola (FecB) and Inverdale (FecXI) mutations in 21 prolific breeds and strains of sheep sampled in 13 countries. Animal Reproduction Science, 92: 87–96.

11- Dinçel, D., S. Ardiçli, B. Soyudal, M. Er., F. Alpay, H. Șamli, and F. Balci. 2015. Analysis of FecB, BMP15 and CAST gene mutations in Sakiz sheep. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 21(4): 483-488.‏

12- Eghbalsaied, S., H. Amini, S. Shahmoradi, and M. Farahi. 2014. Simultaneous presence of G1 and G4 mutations in growth differentiation factor 9 gene of Iranian sheep. Iranian Journal of Applied Animal Research, 4: 781-785. (In Persian).

13- Eghbalsaied, S., K. Ghaedi, S. Shahmoradi, A. Pirestani, H. Amini, T. Saiedi, L. Nicol, and A. McNeilly. 2012. Presence of SNPs in GDF9 mRNA of Iranian Afshari sheep. Iranian International Journal of Fertility and Sterility, 5: 225–230 (In Persian).

14- Galloway, S. M., K. P. McNatty, L. M. Cambridge, M. P. E. Laitinen, J. L. Juengel, T. S. Jokiranta, et al. 2000. Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner. Nature Genetics, 25: 279–283.

15- Galloway, S. M., S. M. Gregan, T. Wilson., K. P. McNatty, J. L. Jungel, O. Ritvos, and G. H. Daivis. 2002. BMP15 mutations and ovarian function. Molecular and Cellular Endocrinology, 191: 15-18.

16- Ghaffari, M., A. Nejati-Javaremi, and G. Rahimi-Mianji. 2009. Lack of polymorphism in the oocyte derived growth factor (GDF9) gene in the Shal breed of sheep. South African Journal of Animal Science, 39: 355-360.

17- GhangAhi. H., Gh. L. Mohamade, H. Ghale dare, and M. R. Afzal Zadeh. 2015. The study of mutations FECXB and FECXG in Kamvry goats in Khuzestan province. Iranian Journal of Genetics in the Third Millennium, 2: 3528-3535. (In Persian).

18- Guan, F., S. R. Liu, G. Q. Shi, and L. G. Yang. 2007. Polymorphismof FecB gene in nine sheep breeds or strains and its effects on litter size, lamb growth and development. Animal Reproduction Science, 99: 44-52.

19- Gursel, F. E., I. Akis, H. Durak, A. Mengi, and K. Oztabak. 2011. Determination of BMP15, BMPR-1B and GDF-9 gene mutations of the indigenous sheep breeds in Turkey. Kafkas Universitesi Veteriner Fakultesi Dergisi, 17(5): 725-729.

20- Hanrahan, J. P., S. M. Gregan, P. Mulsant, M. Mullen, G. H. Davis, R. Powell, and S. M. Galloway. 2004. Mutations in the genes for oocyte-derived growth factors GDF9 and BMP15 are associated with both increased ovulation rate and sterility in Cambridge and Belclare sheep (Ovis aries). Biology of Reproduction, 70: 900-909.

21- Hedayat Iveregh, N., R. Meraeiashtiani, and A. Nejati Javaremi. 2011. Investigate the presence of polymorphisms in genes FecXI and FecXH associnted with twin formation in Shal sheep. Journal of Genetic Novin, 6(4): 29-34. (In Persian).

22- Hua, G. H., S. L. Chen, J. T. Ai, and L. Yang. 2008. None of polymorphism of ovine fecundity major genes FecB and FecX was tested in goat. Animal Reproduction Science, 108: 279-286.

23- Khanahmadi, A., S. H. Gharehveysi, R. Khataminejad, and J. Arab. 2014. Polymorphic variants of G1 and B4 from GDF9 of Dalagh sheep. Iranian Journal of Research on Animal Production, 10: 148-155. (In Persian).

24- Kumar, S., A. K. Mishra, A. P. Kolte, S. K. Dash, and S. A. Karim. 2008. Screening for Booroola (FecB) and Galway (FecXG) mutations in Indian sheep. Small Ruminant Research, 80 (1-3): 57-61.

25- Lin, J., Z. Du, C. Qin, J. f. Wang, and X. Ran. 2007. Polymorphism of BMP15 gene in Guizhou white goats. Animal Husbandry and Veterinary Medicine, 39(12): 21-24.

26- Lio, S. F., Y. L. Jiang, and L. X. Du. 2003. Study of BMPR-IB and BMP15 as candidate genes for fecundity in little tailed Han sheep. Acta Genetica Sinica, 8: 755–760.

27- Luis, V., P. Ricardo, M. T. Tejedor, L. Adolfo, and S. Isidro. 2009. A 17 bp deletion in the Bone Morphogenetic Protein 15 (BMP15) gene is associated to increased prolificacy in the Rasa Aragonesa sheep breed. Animal Reproduction Science, 110: 139–146.

28- Martinez-Royo, A., J. J. Jurado, J. P. Smulders., et al. 2008. A deletion in the bone morphogenetic protein 15 gene causes sterility and increased prolificacy in Rasa Aragonesa sheep. Animal Genetics, 39 (3): 294–297.

29- Michaidilis, G., M. Avdi, and V. Pappa. 2008. Reproductive performance and investigation of BMPR-IB and BMP15 gene mutations in Greek Chios and Florina sheep breeds. Arch Zootech, 11(1): 24-31.

30- Moghadaszadeh, M., M. R. Mohammadabadi, and A. K. Esmailizadeh. 2015. Association of Exon 2 of BMP15 Gene with the litter Size in the Raini Cashmere goat. Iranian Journal of Genetics in the Third Millennium, 13: 4062-4067. (In Persian).

31- Monteagudo, L. V., R. Ponz, M. T. Tejedor, A. Lavina, and I. Sierra. 2009. A 17 bp deletion in the bone morphogenetic protein 15 gene is associated to increased prolificacy in the Rasa Aragonesa sheep breed. Animal Reproduction Science, 110: 139-146.

32- Moradband, F., G. Rahimi, and 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, 9: 1179-1183.

33- Gemmell, N. J. and J. Slate. 2006. Heterozygote advantage for fecundity. PLoS One, 1(1):125.

34- Pardeshi, V. C., M. N. Sainani, J. F. Maddox, P. M. Ghalsasi, C, Nimbkar, and V. S. Gupta. 2005. Assessing the role of FecB mutation in productivity of Indian sheep. Current Science, 89 (5): 887–890.

35- Paz, E., J. Quinones, S. Bravo, H. H Montaldo, and N. Sepulveda. 2015. Genotyping of BMPR1B, BMP15 and GDF9 genes in Chilean sheep breeds and association with prolificacy. Animal Genetics, 46: 98-99.

36- Pirkhezranian, Z., A. Mohammad Hashemi, M. Tahmorthpour, and N. Pirani. 2015. Polymorphhism in second exon of BMP 15 gene in Sangsari sheep of Iran. Iranian Journal of Animal Science Research, 6 (4): 381-387. (In Persian).

37- Polley, S., S. De, B. Brahma, A. Mukherjee, P. Vinesh, S. Batabyal, J. S. Arora, S. Pan, A. K. Samanta, T. K. Datta, and S. L. Goswami. 2010. Polymorphism of BMPR1B, BMP15 and GDF9 fecundity genes in prolific Garole sheep. Tropical Animal Health and Production, 42(5): 985-993.

38- Polley, S., S. De, S. Batabyal, R. Kaushik, P. Yadav, and J.S. Arora. 2009. Polymorphism of fecundity genes (BMPR1B, BMP15 and GDF9) in the Indian prolific Black Bengal goat. Small Ruminant Research, 85: 122–129.

39- Shahmohammadi, L. 2010. Identify polymorphisms GDF9, BMP15 and FecB genes effective twining by PCR-RFLP and PCR-SSCP techniques in Zel sheep. M.Sc Thesis, University of Gorgan. (In Persian).

40- Solimani, B., M. G. Rahimi, and B. Chaharaein. 2011. The segregation of exon 2 BMP15 gene on twining and traits of weight in Sanjabi sheep. Iranian Journal of Biology, 24(4): 487-493. (In Persian).

41- Tajangookeh, H. D., A. R. Shahneh, M. J. Zamiri, M. Daliri, H. Kohram, and A. Nejati-Javaremi. 2009. Study of BMP15 gene polymorphism in Iranian goats. African Journal of Biotechnology, 8(13): 2929-2932.

42- Vacca, G. M., A. Dhaouadi, M. Rekik, V. Carcangiu, M. Pazzola, and M. L. Dettori. 2010. Prolificacy genotypes atBMPR 1B, BMP15 and GDF9 genes in North African sheep breeds. Small Ruminant Research, 88(1): 67-71.

43- Wang, Q. G., F. G. Zhong, H. Li, X. H. Wamg, S. R. Liu, X. J. Chen, and S. Q. Gan. 2005. Detection of major gene on litter size in sheep. Yi. Chuan xue bao, 27(1):8.

44- Wang, Q. G., F. G. Zhong, H. Li, X. H. Wang, S. R. Liu, and X. J. Chen. 2003. The polymorphism of BMPR-IB gene associated with litter size in sheep. Grass-feeding Livestock, 2: 20-23.‏

45- Wang, Y., L. Yuanxiao, Zh. Nana, W. Zhanbin, and B. Junyan. 2011. Polymorphism of Exon 2 of BMP15 Gene and Its Relationship with litter size of Two Chinese goats. Asian-Australian Journal of Animal Sciences, 24(7): 905-911.

46- Zamani, P., R. Abdoli, A. Deljou, and H. Rezvan. 2015a. Polymorphism and bioinformatics analysis of growth differentiation factor 9 gene in Lori sheep. Annals of Animal Science, 15(2):337-348.

47- Zamani, P., S. Nadri, R. Saffaripour, A. Ahmadi, F. Dashti, and R. Abdoli. 2015b. A new mutation in exon 2 of the bone morphogenetic protein 15 gene is associated with increase in prolificacy of Mehraban and Lori sheep. Tropical Animal Health and Production, 47(5): 855-860.