| Salmonella enteritidis(SE)is a common zoonotic pathogen,seriously affecting the healthy development of poultry industry,but also endangering human health.It can infect the digestive tract of poultry,cause acute inflammation of digestive system and even death,even infect people causing food poisoning.Transcriptome is the basis and key point for the study of gene function and structure.Through a new generation of high-throughput sequencing,almost all transcripts sequence information of a specific tissue or organ in a certain state can be obtained quickly and comprehensively.It has been widely used in basic research,clinical diagnosis and drug research and development.Quantitative proteomics can be used to select for differentially expressed proteins between samples caused by any factor,and reveal the regulation mechanism of protein expression in combination with bioinformatics.Therefore,the study of proteome can directly explain the mechanism of gene regulation of biological functions.But the process from transcriptome to protein is very complicated.To date,there is little known about the regulation of chicken in response to SE infection at both transcriptome and proteome levels.In this study,Jining Bairi chickens were inoculated with Salmonella enteritidis.The effects of Salmonella enteritidis infection on genome methylation were detected by transcriptome sequencing and isobaric tags for relative and absolute Quantification(iTRAQ).The results showed that:A total of 49.27 G raw data were obtained by transcriptome sequencing.Comparing the clean data with the reference genome,16190 genes were obtained,and 365 differentially expressed genes(Fold Change > 2,P < 0.05)were identified,including 200 up-regulated genes and 165 down-regulated genes.The differentially expressed genes were significantly enriched in 28 biological processes(P < 0.05),mainly focused on immune related like inflammatory response,regulation of inflammatory response,immune response,oxidative-reduction process,toll-like receptor 6 signaling pathway,positive regulation of heterotypic,positive regulation of NF-kappa B import into nuclear and some metabolicrelated terms like glutathione metabolic process,mitochondrial electron transport and so on.Differentially expressed genes were significantly enriched in six signaling pathways,like JAK-STAT signaling pathway,CAMs,drug metabolism-cytochrome P450(P < 0.05)and so on.A total of 3528 proteins were identified using iTRAQ,including 563 differentially expressed proteins,of which 225 were up-regulated and 338 were down-regulated.The differentially expressed proteins were significantly enriched in 35 biological processes(P <0.05),mainly focused on metabolic related process like fatty acid metabolism process,malate metabolic process,mitochondrial electron transport NADH to ubiquinone and other mitochondrial related processes.KEGG pathway analysis showed that 563 differentially expressed proteins were significantly enriched in 27 pathways like carbon metabolism and metabolic pathway(P < 0.05).225 up-regulated proteins were significantly enriched in three signaling pathways like RNA transport,Ribosome,protein processing in endoplasmic reticulum(P < 0.05).338down-regulated proteins were significantly enriched in metablic related pathways like biosynthesis of antibiotics,metabolic pathway,carbon metabolism(P < 0.05)and so on.57 differentially expressed proteins(10.1% of differentially expressed proteins)were correlated with 74 differentially expressed genes(20.2% of differentially expressed mRNA)with integrated analysis of transcriptome and proteomic.These genes were mainly enriched in oxidation-reduction process and cell adhesions(P < 0.05).The results of qPCR showed that the expression level of TLR1 LA,TLR1LB and CCL4 in the treat group was significantly higher than that in the control group with a fold change of2.56,1.94 and 3.4,respectively(P < 0.05).The expression level of PRDX6 in the treat group was significantly lower than that in the control group with a fold change of 1.85(P < 0.05).The results of Western Blot showed that the expression level of PRDX6 in the treat group was significantly lower than that in the control group with a fold change of 3.3(P < 0.05).These valided information was in accordance with the results of RNA-seq and iTRAQ,which indicated the information was incredible.Jining Bairi Chicken would response to Salmonella enteritidis infection in both transcriptome and proteome with different ways.At the transcriptome level,the regulation mainly focused on immune-related terms,while,at the protein level,the regulation mainly focused on metabolism and mitochondria-related process.Our research provided theoretical and scientific basis for further understanding of the regulation mechanism to Salmonella enteritidis infection in chicken and molecular diseaseresistance genetic breeding of chicken. |
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