Association Between Polymorphisms in B4GALNT2 Gene and Litter Size in Mongolia Sheep and Ujimqin Sheep (Ovis aries)
GAO Yi-Jie1,*, WANG Jin2,*, CHAO Lu-Meng3, CANG Ming1, YU Hai-Quan1, WANG Jian-Guo1, BAO Si-Qin1, LIU Yong-Bin4, ZHANG Wen-Guang5, MA Qing2,**, TONG Bin1,**
1 State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock/School of Life Sciences, Inner Mongolia University, Hohhot 010071, China;
2 Institute of Animal Sciences, Ningxia Academy of Agricultural and Forestry Sciences, Yinchuan 750001, China;
3 Agricultural and Animal Husbandry Technology Extension Center of Xilinhot City, Xilinhot 026000, China;
4 Animal Husbandry Institute, Inner Mongolia Academy of Agricultural & Animal Husbandry, Hohhot 010031, China;
5 College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010071, China
Abstract：Litter size is an important economic trait in sheep (Ovis aries) breeding, which determines the production efficiency of mutton sheep industry. Studies have shown that the g.36938224A>T SNP in β-1,4-N-acetyl-galactosaminyl transferase 2 gene (B4GALNT2) was a major candidate marker associated with the ovulation rate of French Lacaune sheep breed, therefore, the B4GALNT2 gene was considered as a new candidate gene for fecundity in sheep. The aims of this study were to confirm the association between the known SNPs in the B4GALNT2 and litter size of Mongolia and Ujimqin sheep breeds, and identify novel breed-specific variants of Mongolia sheep as well as perform the association study with litter size of Mongolia and Ujimqin sheep breeds. A total of 480 sheep were used in this study including 250 Mongolia sheep, 110 Ujimqin sheep, 30 Hulunbuir sheep, 30 Tan sheep, 30 Small-tailed Han sheep, and 30 Hu sheep. Then, direct sequencing was used to identify novel polymorphism and detect the g.25929884A>T SNP of B4GALNT2 gene (GenBank No. NC_040262.1), the MassARRAY technology was used to genotype the g.25921552C>T and g.25935026C>T SNPs of B4GALNT2 gene and c.*803A>G SNP of distal-less homeobox 3 (DLX3) gene in 480 samples of Mongolia sheep, Ujimqin sheep, Hulunbuir sheep, Tan sheep, Small-tailed Han sheep and Hu sheep. Therefore, association studies were performed between the novel variants and litter size in Mongolia and Ujimqin sheep breeds. The results showed that the g.25929884A>T SNP of B4GALNT2 and the c.*803A>G SNP of DLX3 were absent in any sheep breed. The g.25921552C>T and g.25935026C>T SNPs of B4GALNT2 gene were not associated with litter size of Mongolia and Ujimqin sheep. Furthermore, 8 novel SNPs were found in B4GALNT2 gene, among them, the g.25929637G>A (P<0.05), g.25929679T>C (P<0.01), g.25929819A>G (P<0.01), and g.25929965A>T (P<0.05) SNPs were associated with litter size of Mongolia sheep. Except for g.25921552C>T site in Tan sheep, the other 9 polymorphic loci were under Hardy-Weinberg equilibrium. According to PIC standard, all SNPs presented low and moderate polymorphism in 6 sheep breeds. The presnet study might facilitate effective marker-assisted selection to increase litter size in Mongolia sheep, and provide scientific basis for genetic improvement of sheep.
[1] 程子轩, 路大同, 刘佳森, 等. 2021. GDF9基因g.46547645T>G位点对启动子活性和蒙古羊产羔数的影响[J]. 农业生物技术学报, 29(3): 540-549.
(Cheng Z X, Lu D T, Liu J S, et al.2021. Effect of g.46547645T>G locus of GDF9 gene on promoter activity and litter size of Mongolia sheep (Ovis aries)[J]. Journal of Agricultural Biotechnology, 29(3): 540-549.)
[2] 国家畜禽遗传资源委员会. 2011. 中国畜禽遗传资源志: 羊志[M]. 北京: 中国农业出版社, pp. 20-23.
(China National Commission of Animal Genetic Resources. 2011. Animal Genetic Resources in China: Sheep[M]. China Agriculture Press, Beijing, China, pp. 20-23.)
[3] 梁富武. 2010. 乌珠穆沁羊的选育攻关[J]. 中国畜牧业, (17): 38-39.
(Liang F W. 2010. Breeding of Ujimqin sheep[J]. China Animal Industry, (17): 38-39.)
[4] 李亚星, 成功, 赵云博, 等. 2020. 秦川牛ANGPTL3基因g.-38T>C和g.509A>G的遗传多样性及其与生长和胴体性状的关联性分析[J]. 农业生物技术学报, 28(9): 1543-1553.
(Li Y X, Cheng G, Zhao Y B, et al.2020. Genetic diversity of g.-38T>C and g.509A>G in ANGPTL3 and their association analysis with growth and carcass traits in Qinchuan Cattle (Bos taurus)[J]. Journal of Agricultural Biotechnology, 28(9): 1543-1553.)
[5] Abdoli R, Zamani P, Mirhoseini S Z, et al.2016. A review on prolificacy genes in sheep[J]. Reproduction in Domestic Animals, 51(5): 631-637.
[6] Ben Jemass S, Ruesche J, Sarry J, et al.2019. The high prolificacy of D'man sheep is associated with the segregation of the FecLL mutation in the B4GALNT2 gene[J]. Reproduction in Domestic Animals, 54(3): 531-537.
[7] Bodin L, SanCristobal M, Lecerf F, et al.2002. Segregation of a major gene influencing ovulation in progeny of Lacaune meat sheep[J]. Genetics Selection Evolution, 34(4): 447-464.
[8] Chang T S.1979. Livestock production in China with particular reference to sheep[J]. Wool Technology & Sheep Breeding, 27(2): 19-28.
[9] Chantepie L, Bodin L, Sarry J, et al.2018. Presence of causative mutations affecting prolificacy in the Noire du Velay and Mouton Vendéen sheep breeds[J]. Livestock Science, 216: 44-50.
[10] Chu M X, Liu Z H, Jiao C L, et al.2007. Mutations in BMPR-IB and BMP-15 genes are associated with litter size in Small Tailed Han sheep (Ovis aries)[J]. Journal of Animal Science, 85(3): 598-603.
[11] Drouilhet L, Lecerf F, Bodin L, et al.2009. Fine mapping of the FecL locus influencing prolificacy in Lacaune sheep[J]. Animal Genetics, 40(6): 804-812.
[12] Drouilhet L, Mansanet C, Sarry J, et al.2013. The highly prolific phenotype of lacaune sheep is associated with an ectopic expression of the B4GALNT2 gene within the ovary[J]. PLOS Genetics, 9(9): 809-1003.
[13] Gabriel S, Ziaugra L, Tabbaa, D.2009. SNP genotyping using the Sequenom MassARRAY iPLEX platform[J]. Current Protocols in Human Genetics, 60(1): 2-12.
[14] Galloway S M, McNatty K P, Cambridge L M, et al.2000. Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner[J]. Nature Genetics, 25(3): 279-283.
[15] Gao Y Y, Hao Q, Cang M, et al.2021. Association between novel variants in BMPR1B gene and litter size in Mongolia and Ujimqin sheep breeds[J]. Reproduction in Domestic Animals, 56(12):1562-1571
[16] Guo X, Wang X, Liang B, et al.2018. Molecular cloning of the B4GALNT2 gene and its single nucleotide polymorphisms association with litter size in Small Tail Han sheep[J]. Animals, 8(10): 160.
[17] Hanrahan P J, Gregan S M, Mulsant P, et al.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)[J]. Biology of Reproduction, 70(4): 900-909.
[18] Hedrick P W.1987. Gametic disequilibrium measures: Proceed with caution[J]. Genetics, 117(2): 331-341.
[19] Hua G H, Yang L G.2009. A review of research progress of FecB gene in Chinese breeds of sheep[J]. Reproduction in Domestic Animals, 116(1-2): 1-9.
[20] Mulsant P, Lecerf F, Fabre S, et al.2001. Mutation in bone morphogenetic protein receptor-IB is associated with increased ovulation rate in Booroola Mérino ewes[J]. Proceedings of the National Academy of Sciences of the USA, 98(9): 5104-5109.
[21] Neigenfind J, Gyetvai G, Basekow R, et al.2008. Haplotype inference from unphased SNP data in heterozygous polyploids based on SAT[J]. BMC Genomics, 30(9): 356.
[22] Nei M, Li W H.1979. Mathematical model for studying genetic variation in terms of restriction endonuclease[J]. Proceedings of the National Academy of Sciences of the USA, 76(10): 5269-5273.
[23] Olson D M, Dinerstein E, Wikramanayake E D, et al.2001. Terrestrial ecoregions of the world: A new map of life on earth[J]. Bioscience, 51(11): 933-938.
[24] Purcell S, Neale B, Todd-Brown K, et al.2007. PLINK: A tool set for whole-genome association and population-based linkage analyses[J]. American Journal of Human Genetics, 81(3): 559-75.
[25] Rochus C M, Tortereau F, Plisson-Petit F, et al.2017. Revealing the selection history of adaptive loci using genome-wide scans for selection: An example from domestic sheep[J]. BMC Genomics, 19(1): 71.
[26] Sham P, Bader J S, Craig I, et al.2002. DNA Pooling: A tool for large-scale association studies[J]. Nature Reviews. Genetics, 3(11): 862-871.
[27] Souza C J H, MacDougall C, Campbell B K, et al.2001. The Booroola (FecB) phenotype is associated with a mutation in the bone morphogenetic receptor type 1 B (BMPR1B) gene[J]. Journal of Endocrinology, 169(2): 1-6.
[28] Stwora-Wojczyk M M, Kissinger J C, Spitalnik S L, et al.2004. O-glycosylation in Toxoplasma gondii: Identification and analysis of a family of UDP-GalNAc: Polypeptide N-acetylgalactosaminyltransferases[J]. Glycobiology, 34(3): 309-322.
[29] Ten Hagen K G, Fritz T A, Tabak L A, et al.2003. The UDP-GalNAc: Polypeptide N-acetylgalactosaminyltransferases[J]. Glycobiology, 13(1): 1R-16R.
[30] Tong B, Wang J P, Cheng Z X, et al.2020. Novel variants in GDF9 gene affect promoter activity and litter size in Mongolia sheep[J]. Genes, 11(4): 375.
[31] Vinet A, Drouilhet L, Bodin L, et al.2012. Genetic control of multiple births in low ovulating mammalian species[J]. Mammalian Genome, 23(11-12): 727-740.
[32] Wilson T, Wu X Y, Juengel J L, et al.2001. 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[J]. Biology of Reproduction, 64(4): 1225-1235.
