Expression, Genetic Variation of IP3R3 Gene and Its Effect on Eggshell Quality in Sansui Duck (Anas platyrhyncha domestica)
TAN Guang-Hui1, 2, PING Li-Feng1, 2, XIONG Jian-Min1, 2, QIN Yuan-Yu1, 2, WU Lei1, 2, LI Jie-Zhang
1 College of Animal Science, Guizhou University, Guiyang 550025, China; 2 Key Laboratory of Genetics, Breeding and Reproduction of Plateau Mountain Animals, Ministry of education, Guiyang 550025, China
Abstract：Inositol triphosphate receptor type Ⅲ(IP3R3) is mediator of second messenger-induced intracellular calcium release. It can combine with inositol triphosphate (IP3) to form calcium channel and regulate intracellular calcium release. In order to explore the genetic effects of IP3R3 gene on duck (Anas platyrhyncha domestica) eggshell quality and enrich the molecular genetic basis of important economic traits related to laying ducks. In this study, qRT-PCR was used to detect the tissue expression of IP3R3 gene in Guizhou Sansui ducks. According to the sequence of IP3R3 gene in GenBank, PCR amplification primers were designed, and the amplified products were recovered, purified and sequenced directly.. The SNP loci of 120 Sansui ducks were screened and identified by DNA Star software MegAlign program and sequencing peak map, and SPSS18.0 general linear model (GLM) was used to analyze the relationship between SNPs. The results showed that IP3R3 gene expression was detected in 11 tissues of ducks. The results showed that the mRNA of IP3R3 gene was detected in 11 tissues of ducks, among which the expression abundance of eggshell gland was the highest, and the relative expression quantity of lung, chest muscle and muscle stomach was the lowest. The expression level of IP3R3 gene in different tissues of duck was as follow: Eggshell glands>heart>spleen>glandular stomach>pancreas>kidney>liver>small intestine>lung>pectorals>muscular stomach. SNPs identification results showed that 4 SNPs were found in the IP3R3 gene, including g.21663 C>T and g.21699 G>A in exon 20, g.21646 G>C in intron 19, and g.21825 C>A in intron 20. Further analysis showed that exons g.21663 C>T and g.21699 G>A were synonymous mutation sites, which did not cause changes in tyrosine (TAC) and lysine (AAG). The results of genetic analysis of 4 SNP loci showed that all four SNP loci were moderately polymorphic and deviated from Hardy-Weinberg equilibrium (P<0.05) and results of association analysis showed that the influence of g.21646 on the egg shape index reached a significant level (P<0.05), in which the GG egg shape index was significantly higher than the CC genotype, and the effect of g.21663 on eggshell strength reached a significant level (P<0.05), among which the CT genotype eggshell strength was significantly higher than the TT and CC genotypes, and the other 2 SNP sites did not reach a significant level. Comprehensive correlation analysis and expression results showed that IP3R3 gene expression in the uterus of laying ducks was higher than that in other tissues. It was preliminarily speculated that the high expression of IP3R3 in the eggshell glands might be related to the regulation of eggshell quality, and the mutations of g.21663 and g.21646 loci had significant effects on duck eggshell quality, which could be used as candidate markers assisted by molecular markers for duck eggshell quality traits and provide scientific data for improving duck eggshell quality and poultry breeding.
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