| Artificial insemination(Artificial insemination,AI)technology is an important link to improve the reproductive efficiency of livestock,so it is very important to screen and obtain high quality frozen semen from pigs.Testing the semen quality of breeding boars is an important link to identify whether the breeding boars have normal reproduction ability,and it is also the most basic measure index in the selection and breeding of pigs,which indirectly affects the production efficiency.Therefore,screening boars that can produce high quality semen is particularly important.The results were as follows:1.The semen with the best quality was divided into 7 groups on average,namely control group(no antioxidant and cryoprotectant),experimental group I(add 100 μmol/L CGA),experimental group II(add 40 μmol/L CAPE);experimental group III(0.5 g/L PVP),experimental group IV(add 100 μmol/L CGA and 0.5 g/L PVP),experimental group V(add40μmol/L CAPE and 0.5 g/L PVP),experimental group VI(add 100 μmol/L CGA、40 μmol/L CAPE and 0.5 g/L PVP).After 30 d of frozen storage in liquid nitrogen,semen was thawed,and the sperm motility,mitochondrial viability,DNA integrity,plasma membrane integrity,acrosome integrity,catalase(CAT)activity,superoxide dismutase(SOD)activity,glutathione peroxidase(GSH-Px)activity,reactive oxygen species(ROS)level,and malondialdehyde(MDA)level of frozen semen were measured.The results showed that the sperm motility,plasma membrane integrity,acrosome integrity,mitochondrial activity,CAT activity and GSH-Px activity of frozen semen in adding 100μmol/L CGA、40 μmol/L CAPE and 0.5 g/L PVP group were significantly improved compared with the control group(P<0.01);DNA integrity and SOD activity were significantly increased(P<0.05);the ROS level decreased significantly(P<0.01);and the MDA level decreased significantly(P<0.05).The results indicated that adding 100μmol/L CGA、40μmol/L CAPE and 0.5 g/L PVP to the frozen base solution of pig spermatozoa could improve the frozen semen quality of Xin Jilin Black Pig.2.After optimizing the formula,Freeze 88 boar semen,Extract the genome by frozen semen,Conduct DNA sequencing and HRM typing,Results showed: A SNP site C187 T,was detected on the pig AQP7 gene exon7 There are in all 3 genotypes,The amino acid sequence did not change;a SNP site T136 C,was detected on the pig LPAR1 gene exon2 There are 2genotypes,The amino acid sequence did not changed;A SNP site was detected each on the boar HSP90 gene exon3 and exon9,C129 A and C176 T,,respectively Each has 3 genotypes,The amino acid sequence did not change at the C129 A,Missense mutation occurs in the amino acid sequence at C176 T,Mutant from glutamate to alanine;A SNP site A159 G,was detected on the pig CRISP3 gene exon5 There are 2 genotypes,Missense mutations occur in the amino acid sequence,Mutant from threonine to alanine.The SNP sites of each candidate gene are evenly distributed in the detection pig population,each point in accordance with Hardy-Weinberg law,in Hardy-Weinberg genetic balance,and all in 0.25 <PIC< 0.5,in moderate polymorphisms.3.Frozen semen quality indicators such as sperm motility,plasma membrane integrity,acrosome integrity,mitochondrial activity and DNA integrity were detected.The results showed that the sperm motility,acrosome integrity rate,plasma membrane integrity rate,mitochondrial integrity rate and DNA integrity rate were the highest in the AQP7 gene C187 T locus,and the TT genotypes and CC genotypes had very significant differences in motility,acrosome integrity rate,plasma membrane integrity rate and mitochondrial integrity rate(P<0.01).There was no significant difference in the DNA integrity rate of each genotype(P>0.05).On the LPAR1 gene T136 C site,The sperm motility,acrosome integrity rate,plasma membrane integrity rate,mitochondrial integrity rate and DNA integrity rate the highest,The TT and TC genotypes showed significant differences in activity,acrosome integrity,plasma membrane integrity and mitochondrial integrity(P<0.01).There was no significant difference in the DNA integrity rate of each genotype(P>0.05).On the HSP90 gene C129 A site,The sperm motility,acrosome integrity rate,plasma membrane integrity rate,mitochondrial integrity rate and DNA integrity rate the highest,AA genotypes and CC genotypes showed significant differences in viability,acrosome integrity and plasma membrane integrity(P<0.01),There was significant difference between AA genotype and CA genotype and CC genotype in mitochondrial activity rate and DNA integrity rate(P<0.05).On the HSP90 gene C176 T site,CC genotypes had the highest sperm motility,acrosome integrity and plasma membrane integrity,TT genotypes and CC genotypes showed significant differences in viability,acrosome integrity and plasma membrane integrity(P<0.01),There was significant difference between TT genotype and CT genotype and CC genotype in mitochondrial activity rate and DNA integrity rate(P<0.05).On the CRISP3 gene A159 G site,The sperm motility,acrosome integrity rate,plasma membrane integrity rate,mitochondrial integrity rate and DNA integrity rate the highest,AA genotypes and AG genotypes showed significant differences in viability,acrosome integrity,plasma membrane integrity and mitochondrial integrity(P<0.01),The DNA integrity rate reached a significant difference(P>0.05). |
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