| Salmonella enteritidis is an important foodborne pathogenic bacteria, has brought serious economic losses to the poultry business, At the same time, it can cause human intestinal disease through poultry related products and it is of great significance on veterinary public health. With different genetic background of different strains of chicken, the ability to resist the interference of salmonella enteritidis has significant difference.The methylation of genomic DNA is the most important part of the epigenetic modifications. the DNA methylation plays a vital role in the nervous system, cardiovascular immunodeficiency,and genetic disorder diseases such as sexual disorders study and so on. Recent studies suggest that methylation in resisting bacteria and virus infection is one of the key aspects.High throughput sequencing technology are becoming a more and more powerful tool to identify the various species of methylation. There were many reportes related to infection such as bacteria, viruses identification of methylation gene on the poultry. Whereas, the study of methylation regulatory mechanism with salmonella enteritidis infection is not clear.Transforming growth factor-beta is an important kind of bioactive functions of cytokines, it plays a very important role in that mediated especially by Smads family proteins signal transduction in maintaining the normal physiological function of cells.This study is based on artificial infection with salmonella enteritidis in shandong shouguang chicken and white leghorn SPF chicken for research object, euzymelinked immunosorbent assay, the next generation sequencing and PCR Array technology were used respectively to identificate the gene of significant difference in methylation with salmonella enteritidis infection. The result shows that:(1)By methylation euzymelinked immunosorbent assay analysis, we compared the shouguang chicken and SPF chicken inoculated days after 1,3,14 and 21 with salmonella enteritidis, the cecum groups treatment group and control group of genome-wide methylation level. Methylation of shouguang chicken treatment group were 78.06%,35.52%,31.21%, 34.55%, and the control group was 71.64%,57.33%,53.94%,38.38%. SPF chicken treatment group of methylation levels were 34.55%,42.79%,54.95%,52.42%, and the control group was 40.97%,50.3%,56.57%,57.98%. Different day age after inoculation and the different genetic background with salmonella enteritidis infection after detected by methylation level is different in the abstract.(2) 14 days after inoculation with SE, from constructing the two library of T and C,we got 39385230 and 46733556 High quality without repeated readings accounted for 98.33% and 98.39% of the total respectively, the results showed that the treatment group compared with control group, the distribution of DMRs number and length in different chromosomes is significantly different, and in different positions such as introns, gene promoter and intergenic regions, low methylation level of DMRs account for significantly more than the number of high methylation.we Found 751 different methylation geneS that respectively is 288 upregulated and463down regulated genes.(3) we did the functional analysis with the 751 different methylation genes with DAVID database and received significant enrichment GO classification annotations 142 (P< 0.05), including 103 biological processes comments,13 cell component annotation,26molecular functional annotation. Significant difference of all comments contained genes were count for the number of repeat statistics, TGFBR1, TGFBR2, APP, NOTCH1 gene duplication number is larger, respectively are 30,29,26,25. TGFBR1, TGFBR2, BMPR1B, IL6, MAP2K1 genes were part of transforming growth factor-beta signal pathway that play an important role in it.(4) The detection of the TGF-beta signaling pathways that infected with salmonella enteritidis after 7 days of age in spleen tissue, treatment group compared with the control group, there are 7 genes significantly lower in shouguang chicken and 52 genes in SPF chickens. In the control group, shouguang chicken had 4 up-regulated genes compared with SPF chickens,and in the treatment group,shouguang chicken has 62 genes were significantly lower and 2 gene were significantly raised compared with SPF chicken.To sum up, salmonella enteritidis has significant inhibitory effect on genome-wide methylation level, this effect had differences because of the genetic background and infection time; 751 methylation differential gene were found with SE Infection,288 of high methylation and 453 of low methylation. Comparing fold change, GO analysis, and the number of gene duplication, primarily determined the methylated candidate genes DPT, GPD1, ST5, MAP3K7, TGFBRland TGFBR2 associated with salmonella enteritidis infection. Methylation is not the only factor regulating the TGF-beta signaling pathway gene expression changes in chicken salmonella enteritidis infection. In this study, we confirmed the action of the methylation in the resistance of salmonella enteritidis preliminary, specific mechanisms need further study in various CpG sites. |
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