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| Effect of g.46547645T>G Locus of GDF9 Gene on Promoter Activity and Litter Size of Mongolia Sheep (Ovis aries) |
| CHENG Zi-Xuan1,*, LU Da-Tong1,*, LIU Jia-Seng2, LI Yun-Hua2, ZHANG Xiu-Ying3, CHAO Lu-Meng3, CANG Ming1, WANG Jian-Guo1, YU Hai-Quan1, LI Guang-Peng1,**, TONG Bin1,** |
1 State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock/School of Life Sciences, Inner Mongolia University, Hohhot 010071, China;
2 Animal Husbandry Institute, Inner Mongolia Academy of Agricultural & Animal Husbandry, Hohhot 010031, China;
3 Xilinhot Animal Husbandry Station, Xilinhot 026000, China |
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Abstract Increasing litter size is the primary aim of breeding in Mongolia sheep (Ovis aries). Growth and differentiation factor 9 (GDF9) is an oocyte-derived growth factors that play an important role in ovarian folliculogenesis in mammals. Many studies have shown that the mutations in GDF9 were associated with litter size and ovulation rate in several sheep breeds. The aims of this study were to detect the association between the g.46547645T>G SNP of the GDF9 gene and litter size in Mongolia sheep, and detect the genetic diversity in Mongolia, Ujimqin, Hulunbuir (big and short tail types), Small-tailed Han and Hu sheep breeds, as well as to validate the effect of the g.46547645T>G SNP on the promoter activity of GDF9. A total of 416 sheep were used in this study including 260 Mongolia sheep, 36 Ujimqin sheep, 30 Hulunbuir sheep (big tail type), 30 Hulunbuir sheep (short tail type), 30 Small-tailed Han sheep and 30 Hu sheep. The g.46547645T>G in GDF9 were genotyped using the PCR-restriction fragment length polymorphism (PCR-RFLP) method, and then performed the association and genetic diversity analyses. The effect of g.46547645T>G on the promoter activity of the GDF9 was detected by the luciferase reporter assay. The χ2 test indicated that the g.46547645T>G SNP is under Hardy-Weinberg equilibrium in the 6 sheep breeds. There were 3 genotypes TT, TG and GG of the g.46547645T>G SNP in Mongolia sheep, 2 genotypes TT and TG in Ujimqin sheep, Hulunbuir sheep (big tail type), Hulunbuir sheep (short tail type), Small-tailed Han sheep and Hu sheep populations. The values of polymorphism information content (PIC) of the g.46547645T>G SNP presented with related low polymorphism in the Mongolia sheep, Ujimqin sheep, Small-tailed Han sheep and Hu sheep breeds, and presented with moderate polymorphism in the Hulunbuir sheep (big tail type), Hulunbuir sheep (short tail type) breeds. The result of association analysis showed that the individuals with TG genotype of g.46547645T>G SNP had extremely significantly more lambs than the individuals with TT genotype in Mongolia sheep (P<0.01). In addition, GDF9 promoter activity analysis showed that the G allele at g.46547645T>G could significantly decrease luciferase activity compared with that of the control (P<0.05), the T allele at g.46547645T>G could significantly increase luciferase activity compared with that of the control (P<0.05), and the G allele at g.46547645T>G could extremely significantly decrease luciferase activity compared with that of the T allele (P<0.01). This study may facilitate effective marker-assisted selection to increase litter size in Mongolia sheep populations, as well as bring new insights into GDF9 expression.
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Received: 11 May 2020
Published: 01 March 2021
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Corresponding Authors:
**gpengli@imu.edu.cn; tongbin87@163.com
*The authors who contribute equally
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