Identification of Key Candidate Genes Affecting Translucent Egg Production of Laying Hens Based on Transcriptome Sequencing Technology

Du Yuanjun; Han Haixia; Liu Wei; Li Dapeng; Lei Qiuxia; Zhou Yan; Liu Jie; Wang Jie; Li Fuwei; Chen Fu

doi:10.14083/j.issn.1001-4942.2023.12.019

Abstract：The aim of this study was to screen the functional genes and signaling pathways associated with translucent egg production in chicken duodenum by transcriptome sequencing(RNA-seq).Ten Jining Bari hens with low continuous translucent egg laying rate were selected as the control group,and ten ones with high continuous translucent egg laying rate were as the eggshell translucence group.Duodenum samples was collected,and the mRNA expression level of calcium metabolism related genes(DEGs)were measured.The differentially expressed genes(DEGs)were detected by RNA-seq,and the GO(gene ontology)functional an-notation and KEGG(kyoto encyclopedia of genes and genomes)signaling pathway enrichment were performed.The results showed that the total data obtained from the transcriptome was 108.75 Gb,and Q30 was＞97%in all samples.A total of 471 DEGs were screened out,of which,327 ones were up-regulated and 144 ones were down-regulated.Seven DEGs were randomly selected for real-time quantitative fluorescence PCR(qRT-PCR)verification,and it was found that their expression trend was basically consistent with that of the transcripome sequencing results.GO functional annotation analysis showed that SLC8A3,SLC24A4 and SLC9A2 of the solute carrier family were mainly enriched into the mitochondrial calcium homeostasis,sodium ion transmembrane transport and calcium ion binding processes.KEGG pathway enrichment results showed that the DEGs such as GSTT1,GSTO2,GSTA3,CLDN2 and CLDN10 were mainly enriched into the cytochrome P450-mediated for-eign substance metabolism pathway,drug metabolity-cytochrome P450,calcium signaling pathway,etc.Com-pared with the control group,the mRNA expression of duodenal CALB1 in the eggshell translucence group was significantly decreased(P＜0.05).In conclusion,the up-regulation of GSTT1,GSTO2 and GSTA3 genes and the down-regulation of SLC8A3 and SLC24A4 genes in the duodenum of laying hens were the key factors affect-ing the formation of translucent eggs,suggesting that calcium signaling pathway might play an important role in the formation of translucent eggs.The results of this study could provide basic materials for the mechanism re-search of translucent eggs.