Abstract: 【Objective】 The objective of this study was to analyze the transcriptome sequencing of pituitary glands of Jinghai Yellow Chickens under different light color conditions,to understand the expression profile of the pituitary glands of Jinghai Yellow Chickens under different light color conditions, to screen out the key genes of red light affecting the synthesis of reproductive hormones in Jinghai Yellow Chickens,so as to provide theoretical basis for improving the laying performance of breeding chickens.【Method】 The egg production of Jinghai Yellow Chickens in the red light group(90.61 eggs)was significantly higher than that of the white light group(87.44 eggs) (P<0.05). Three Jinghai Yellow Chickens each were selected under red light and white light conditions.The pituitary glands of chickens were sequenced through high-throughput sequencing. The differentially expressed genes(DEGs)were obtained by HISAT and DESeq methods, and GO functional annotation and KEGG signaling pathway enrichment analysis were performed.【Result】A total of 4892794884-4980565639 raw reads were obtained by high-throughput sequencing,with an alignment rate of over 92.12% and a GC content of 48.54%-49.11%. A total of 155 DEGs were obtained by comparing the pituitary glands of Jinghai Yellow Chickens under red and white lights, of which 90 gene expressions were up-regulated and 65 gene expressions were downregulated in the red light group compared with the white light group(RP vs WP). GO annotation analysis of DEGs was performed,mainly in important biological pathways such as biological regulation,cell process,cell,cell component, binding and catalytic activity. KEGG signaling pathway analysis suggested that DEGs were mainly enriched in the pathways of cytokine-cytokine receptor interaction,steroid hormone biosynthesis pathway,retinol metabolism,arachidonic acid metabolism,neuroactive ligand-receptor interaction signaling pathways. The expressions of gonadotropin releasing hormone(GNRH1),secreted phosphoprotein 1(SPP1),drosophila mothers against decapentaplegic protein(SMAD1), gremlin 2(GREM2)and Epiregulin(EREG)in these pathways may be affected by the light colors,which may affect the laying performance of breeding chickens. Furthermore,the results of real-time fluorescence quantitative PCR(qRT-PCR) confirmed that the transcrtiptome results were accurate and reliable.【Conclusion】Compared with white light,red light can significantly increase the egg production of Jinghai Yellow Chickens. The genes obtained from the screening,such as GNRH1,SPP1,SMAD1,GREM2 and EREG,may play important roles in the light color regulation of the laying performance of Jinghai Yellow Chickens,and can be used as key genes for the study of the molecular mechanism of pituitary gland tissue of breeding chickens to regulate their laying performance.