| Congjiang Xiang pig is a rare miniature pig breed in China, however,the mechanism of slow growth trait is still remained to be unclear. The present paper focused on the genetic polymorphism investigation in Congjiang Xiang pig by using methylation-sensitive amplifying polymorphism technique(MSAP) and Illumina Porcine SNP60(60K) chips. The results were as following:1. Sixty piglets in 20 days were divided into two groups with high or low body weight. The methylation modification at cytosine in the genomic DNA from blood and liver was investigated by methylation-sensitive amplifying polymorphism technique(MSAP). A total of 5977 sites were counted based on results of ten pairs of primers chosen for amplification. DNA methylation levels in blood and liver genomic DNA in high-weight group were 32.43% and 36.36%, and 35.88% and 38.85% in the low-weight group, respectively. Five specific fragments from A1 to A5 were sequenced which were different between two groups. Both of A1 and A4 detected from high-weight group while the others got from low-weight group. Fragment A1 and A2 detected from blood genomic DNA and A3-A5 from liver samples. Fragment A1 located in the intron 1 of SPSB3 gene(SPRY domain-containing SOCS box protein). A2 was in the last exon of UDT2C1 gene(UDP-glucuronosyltransferase 2C1-like). A4 was in 5`-flank of CPEB4 gene(cytoplasmic polyadenylation element-binding protein 4). A5 was found from intron 3 of PIP4K2 A gene(phosphatidylinositol-5- phosphate 4-kinase type II alpha). A3 was apart far from two genes without coding region for RNA.2. The single nucleotide polymorphisms in genome wide of Xiang pig were detected by Illumina Porcine SNP60 chips. After strict quality control, 15 SNPs were deduced to be correlated with body length trait of Xiang pig by genome-wide association analysis(GWAS) approach. Three SNPs, which were INRA0011611, MARC0076145 and H3GA0051838 were verificated by AS-PCR. The result shows that AS-PCR had the same result with SNP chips.3. Using AS-PCR technique to test this three SNPs in seven population which were 12 mouth old Xiang Pigs, 18 mouth old Xiang pigs, 24 mouth old Xiang pigs, Dabai pig, Guanlin pig, Rongchang pig, Nuogu pig, Qianbei pig, Jiangkou pig and Zongdihua pig. The results show that: INRA0011611 had no predominant allele in each groups. The researchers also found a positive correlation between genotypes and 24 mouth old Xiang pigs` head length(0.287>0.200)。MARC0076145 was located in the seventh intro of ALDH5A1, the predominant allele in Xiang pigs is G. There were Negative correlations between genotypes and 18 mouth old and 24 mouth old Xiang pig group(-0.201>-0.200,-0.310>-0.200). H3GA0051838 was located in the 5`-flank of TCEAL4.The frequency of GG is the primary genotype in Congjiang and G is the predominant allele. Those SNPs were worth for further investigation about their effection and fuction, which might play an important role in Xiang pig`s growth. It suggested that the polymorphism bands or SNP locus found from the present paper would be useful for selection and breeding of new species to improve the growth trait of native pig breeds in future. |
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