农业生物技术学报2013，Vol.21Issue(3)：328-337,10.DOI:10.3969/j.issn.1674-7968.2013.03.010
磷脂酶C-zeta基因(PLCz)多态性与中国荷斯坦公牛精液性状相关性研究
Association between Polymorphism of Phospholipase C-zeta Gene (PLCz)and Sperm Quality Traits in Chinese Holstein Bulls
摘要
Abstract
The testis-specific phospholipase C isoform, PLC-zeta (PLCz), is widely considered to be a quite essential role in the oocyte activation and embryo development owing to its ability to trigger intracellular a series of Ca2+ oscillations in all mammals. Recently, while PLCz as the molecular trigger for development of a fertilised egg into an embryo has been reported, the exact molecular mechanism for generating Ca2+ oscillations and the PLCz gene expression and regulation has not been established. Accordingly, the objectives of this study were to clone and characterize the expression and regulation of bovine (Bos taurus) PLCz in sperm through the analysis of single nucleotide polymorphisms (SNPs) using DNA sequencing methods and PCR-restriction fragment length polymorphism (PCR-RFLP) to study their association with sperm quality and fertility of 424 normal Chinese Holstein Bulls from three bull stations. Molecular cloning of the bovine PLCz yielded a 53 160 bp sequence which transcripted into a 2 176 bp codons and encoded a number of 634 amino acid. We detected eight novel SNPs, g.27529 T>A and g.27597 T>C located in the seventh intron, g.47969 G>A, g.48020 T>C, g.48079 G>A, g.48127 G>T and g.48384 G>T in the twelveth intron, thus the SNP g.27768 G>C in the eighth exon was identified as a non-synonymous mutation [GCC(Ala)>CCC (Pro)] at 337th amino acid. Significantly, linkage disequilibrium analysis showed that g.27529 T>A, g.27597 T>C and g.27768 G>C (SNP1); g.47969 G>A, g.48020 T>C, g.48079 G>A, g.48127 G>T and g.48384 G>T (SNP2) were completely linked. Correlation analysis with the sperm quality traits showed that the SNP2 marker had a respective effect on the percentages of viable sperm (P<0.05). Whereas, the SNP1 marker had no significantly relationship with the sperm production traits (P>0.05). In addition, The association analysis between nine combined haplotypes and bull reproductive traits showed that the bulls with the combined haplotype TTAB were significantly higher with ejaculate volume (P<0.05), sperm density(P<0.01) and frozen semen motility (P<0.05) than that with the other haplotype combinations; as well as the TTAB haplotype with initial sperm motility was higher than that with the TTBB haplotype combination (P<0.05); but the TTAB haplotype with deformity rate was lower than that with the CTAB haplotype combination (P<0.05), The result indicated that the TTAB combined haplotype may be regard as a new molecular marker associated with sperm quality traits which in marker-assisted selection of Chinese Holstein bulls breeding programs.关键词
PLCz基因/中国荷斯坦公牛/单核苷酸多态性/完全连锁/单倍型组合Key words
PLCz gene/ Chinese holstein bulls/ Single nucleotide polymorphisms/ Completely linked/ Haplotype combination引用本文复制引用
潘晴,鞠志花,张燕,黄金明,李建斌,王玲玲,齐超,侯明海,仲跻峰..磷脂酶C-zeta基因(PLCz)多态性与中国荷斯坦公牛精液性状相关性研究[J].农业生物技术学报,2013,21(3):328-337,10.基金项目
国家"科技支撑计划"项目(No.2011BAD19BO4)、现代农业产业技术体系(No.CARS-37)和山东省良种工程项目(No.2010LZ015-06) （No.2011BAD19BO4）
