The main purpose of this study was to assess the presence of the previously reported single nucleotide polymorphisms (SNPs) in the sheep growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) genes and their association with litter size at lambing in Iran-Black sheep breed. Blood samples were taken from 120 Iran-Black ewes. DNA extraction was conducted using a modified salting-out method. DNA fragments with sizes of 462bp and 141bp for the GDF9 and BMP15 genes were amplified using PCR with specific primers, respectively. The PCR-RFLP approach was adopted for detecting the genotypes. The results indicated that the SNP in the exon 2 of BMP15 is a monomorphic locus in Iran-Black sheep. However, the substitution of G to A nucleotide was determined in the GDF9 locus. Digestion of the 462bp PCR product from exon 1 of theGDF9 using the HhaI restriction enzyme produced fragments of 52, 156, and 254bp. However, DNA fragments containing the A nucleotide yielded only two fragments (52 and 410bp). The heterozygous animals for this mutation in GDF9 locus had fragments of all four sizes (52, 156, 254, and 410bp). The frequency (0.75) of the wild type allele (+) in GDF9 locus was higher than the frequency (0.25) of mutant allele (G). The observed frequencies for the GG, G+ and ++ genotypes were 0.05, 0.40 and 0.55, respectively. The association results indicated that the mutation of GDF9 gene has a substantial impact on lambing rate and the Iran-Black ewes with the GG and G+ genotypes had higher lambing rate than those with the ++ genotype. Thus, a gene assisted selection program to improve lambing rate in this breed can be designed based on the GDF9 gene mutation.
Barzegari, A., Atashpaz. S., Ghabili, K., Nemati, Z., Rustaei, M. Azarbaijani, R., 2010. Polymorphisms in GDF9 and BMP15 associated with fertility and ovulation rate in Moghani and Ghezel sheep in Iran. Reproduction in Domestic Animals 45, 666–669.
Bindon, B.M., Piper, L.R., Hilliard, M.A., 1996. Reproductive physiology and endocrinology of prolific sheep. In: Fahmy, M.H. (Ed.), Prolific Sheep. CAB International, Wallingford, UK pp. 453–470.
Bodensteiner, K.J., Clay, C.M., Moeller, C.L., Sawyer H.R., 1999. Molecular cloning of the ovine growth/differentiation factor-9 gene and expression of growth/differentiation factor-9 in ovine and bovine ovaries. Biology of Reproduction 60, 381-386.
Bodin, L., Di Pasquale, E., Fabre, S., Bontoux, M., Monget, P., Persani, L., Mulsant, P., 2007. A novel mutation in the bone morphogenetic protein 15 gene causing defective protein secretion is associated with both increased ovulation rate and sterility in Lacaune sheep. Endocrinology 148, 393-400.
Bodin, L., Lecerf, F. Pisselet, C., SanCristobal, M., Bibe, B., Mulsant, P., 2003. How many mutations are associated with increased ovulation rate and litter size in progeny of Lacaune meat sheep? Proceedings of the 3rd International Workshop on Major Genes and QTL in Sheep and Goats. INRA, Toulouse, France.
Caballero, A., Toro, M.A., 2000. Interrelations between effective population size and other pedigree tools for the management of conserved populations. Genetics Research 75, 331–343.
Davis, G.H., 2005. Major genes affecting ovulation rate in sheep. Genetics Selection Evolution 37, S11-S23.
Davis, G.H., Galloway, S. M., Ross, I.K., Gregan, S.M., Ward, J., Nimbkar, B.V., Ghalsasi, P.M., Nimbkar, C., Gray, G.D., Subandriyo, Inounu, I., Tiesnamurti, B., Martyniuk, E., Eythorsdottir, E., Mulsant, P., Lecerf, F., Hanrahan, J.P., Bradford, G.E. and Wilson, T., 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.
Davis, G.H., Montgomery, G.W., Allison, A.J., Kelly, R.W., Bray, A.R., 1982. Segregation of a major gene influencing fecundity in progeny of Booroola sheep. New Zealand Journal of Agricultural Research 25, 525–529.
Dekkers, J.C.M., 2012. Application of genomics tools to animal breeding. Current Genomics13, 207–212.
Duguma, G., Schoeman, S.J., Cloete, S.W.P., Jordaan, G.F., 2002. Genetic and environmental parameters for ewe productivity in Merinos. South African Journal of Animal Science 32, 154-159.
Esmailizadeh, A., 2010. A partial genome scan to identify quantitative trait loci affecting birthweight in Kermani sheep. Small Ruminant Research 94, 73–78.
Esmailizadeh, A., 2014. Detection of chromosomal segments underlying scrotal circumference in ram lambs and age at onset of puberty in ewe lambs. Animal Production Science55, 1018-1024.
Esmailizadeh, A., Miraei-Ashtiani, S.R., Mokhtari, M.S., Asadi Fozi, M., 2011. Growth performance of crossbred lambs and productivity of Kurdi ewes as affected by the sire breed under extensive production system. Journal of Agricultural Science and Technology 13, 701–708.
Eusebi, P.G., Martinez, A., Cortes, O., 2020. Genomic tools for effective conservation of livestock breed diversity. Diversity12, 8.
Farid, A., Makarechian, M., Sefidbakht, N., 1977. Crossbreeding of Iranian fat-tailed sheep: lamb performance of Karakul, Mehraban and Naeini breeds. Journal of Animal Science 44, 542-548.
Galloway, S.M., McNatty, K.P., Cambridge, L.M., Laitinen, M.P., Jennife, J.L., Jokiranta, S., 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.
Hanrahan, J.P., Gregan, S.M., Mulsant, P., Mullen, M., Davis, G.H., Powell, R., Galloway, S.M., 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 Reproduction70,900–909.
Juengel, J.L., Hudson, N.L., Whiting, L., McNatty, K.P., 2004. Effects of immunization against bone morpHogenetic protein 15 and growth differentiation factor 9 on ovulation rate, fertilization, and pregnancy in ewes. Biology of Reproduction 7, 557-561.
Liao, W.X., Moore, R.K., Shimasaki, S., 2004. Functional and molecular characterization of naturally occurring mutations in the oocytesecreted factors bone morphogenetic protein-15 and growth and differentiation factor-9. Journal of Biological Chemistry 17, 17391- 17396.
Matos, C.A.P., Thomas, D.L., Gianola, D., Perez-Enciso, M., Young, L.D., 1997. Genetic analysis of discrete reproductive traits in sheep using linear and nonlinear models: II. Goodness of fit and predictive ability. Journal of Animal Science 75, 88-94.
Michailidis, G., Avdi, M., Pappa V., 2008. Reproductive performance and investigation of BMPR-IB and BMP-15 gene mutations in Greek Chios and Florina sheep breeds.Archiva Zootechnica 11, 24-31.
Miller, S.A, Dykes, D.D, Polesky, H.F. 1988. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research 16, 1215.
Mishra, A.K., Arora, A.L., Kumar, S., Prince., L.L, 2009. Studies on effect of Booroola (FecB) genotype on lifetime ewes’ productivity efficiency, litter size and number of weaned lambs in Garole×Malpura sheep.Animal Reproduction Science 113, 293–298.
Montaldo, H.H., Meza-Herrera, C.A., 1998. Use of molecular markers and major genes in the genetic improvement of livestock. Electronic Journal of Biotechnology 1, 1-7.
Monteagudo, L.V., Ponz, R., Tejedor, M.T., Laviña, A., Sierra, I., 2009. A 17bp 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.
Moore, R.K., Erickson, G.F., Shimasaki, S., 2004. Are BMP-15 and GDF-9 primary determinants of ovulation quota in mammals? Trends in Endocrinology and Metabolism 15, 356–361.
Moradband, F., Rahimi, G., Gholizadeh, M., 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.
Nosrati, M., Asadollahpour Nanaei, H., Amiri Ghanatsaman, Z., Esmailizadeh, A., 2019. Whole genome sequence analysis to detect signatures of positive selection for high fecundity in sheep. Reproduction in Domestic Animals 54, 358–364.
Piper, L.R, Bindon, B.M., 1982. Genetic segregation for fecundity in Booroola Merino sheep. In: Barton, R.A., Smith, W.C. (Eds.), Proceedings of the World Congress on Sheep and Beef Cattle Breeding, Palmerston North, New Zealand, pp. 395–400.
Rashidi, A., 2013. Genetic parameter estimates of body weight traits in Iran-Black sheep. Journal of Livestock Science and Technologies 1, 50-56.
SAS, 2002. SAS User’s Guide: Statistics. Version 9.1. SAS Institute Inc., Cary, North Carolina, USA.
Silva, B.D., Castro, E.A., Souza, C.J., Paiva, S.R., Sartori, R., Franco, M.M., Azevedo, H.C., Silva, T.A., Vieira, A.M., Neves, J.P. , Melo, E.O., 2011. A new polymorphism in the growth and differentiation factor 9 (GDF9) gene is associated with increased ovulation rate and prolificacy in homozygous sheep. Animal Genetics 42, 89–92.
Smith, C., Simpson, S.P., 1986. The use of genetic polymorphisms in livestock improvement. Journal of Animal Breeding and Genetics 103, 205–217.
Snowder, G.D., 2008. Genetic improvement of overall reproductive success in sheep: A review. Asociacion Latinoamericana de Produccion Animal 16, 32–40.
Snyman, M.A., Olivier, J.J., Erasmus, G.J., Van Wyk, J.B., 1997. Genetic parameter estimates for total weight of lamb weaned in Afrino and Merino sheep. Livestock Production Science 48, 111-116.
Souza, C.J.H., MacDougall, C., Campbell, B.K., McNeilly, A.S., Baird, D.T., 2001. The Booroola (FecB) phenotype is associated with a mutation in the bone morphogenetic receptor type 1 B (BMPR1B) gene. Journal of Endocrinology 169, R1-R6.
Woolliams, J.A., Oldenbroek, J.K., 2018. Genetic diversity issues in animal populations in the genomic era. In: Oldenbroek, J.K. (Ed.), Genomic Management of Animal Genetic Resources, 1st ed., Wageningen Academic Publisher, Gelderland, The Netherlands, pp. 13–47.
Rezaei, V., Esmailizadeh, A., Ayatollahi Mehrgardi, A., & Dehghani Qanatqestani, M. (2020). Allelic polymorphism in exon 1 of GDF9 and exon 2 of BMP15 genes and its impact on litter size at lambing in Iran-Black sheep. Journal of Livestock Science and Technologies, 8(2), 57-65. doi: 10.22103/jlst.2020.15007.1299
MLA
Vahideh Rezaei; Ali Esmailizadeh; Ahmad Ayatollahi Mehrgardi; Mostafa Dehghani Qanatqestani. "Allelic polymorphism in exon 1 of GDF9 and exon 2 of BMP15 genes and its impact on litter size at lambing in Iran-Black sheep", Journal of Livestock Science and Technologies, 8, 2, 2020, 57-65. doi: 10.22103/jlst.2020.15007.1299
HARVARD
Rezaei, V., Esmailizadeh, A., Ayatollahi Mehrgardi, A., Dehghani Qanatqestani, M. (2020). 'Allelic polymorphism in exon 1 of GDF9 and exon 2 of BMP15 genes and its impact on litter size at lambing in Iran-Black sheep', Journal of Livestock Science and Technologies, 8(2), pp. 57-65. doi: 10.22103/jlst.2020.15007.1299
VANCOUVER
Rezaei, V., Esmailizadeh, A., Ayatollahi Mehrgardi, A., Dehghani Qanatqestani, M. Allelic polymorphism in exon 1 of GDF9 and exon 2 of BMP15 genes and its impact on litter size at lambing in Iran-Black sheep. Journal of Livestock Science and Technologies, 2020; 8(2): 57-65. doi: 10.22103/jlst.2020.15007.1299