不同生理阶段边鸡卵巢转录谱的构建及卵泡发育相关基因的分析

doi:10.11843/j.issn.0366-6964.2022.02.010

摘要/Abstract

摘要： 本研究通过分析不同生理阶段边鸡卵巢转录表达谱，旨在找出与卵泡发育相关的基因并进行验证，以揭示边鸡就巢性强的遗传机制。构建代表不同生理期即70日龄（70 d，卵巢发育期）、165日龄（165 d，开产期）、220日龄（220 d，产蛋高峰期）和330日龄（330 d，就巢期）的边鸡卵巢基因表达谱，用转录组表达定量（RSEM）法对不同生理阶段边鸡卵巢基因表达进行相关性分析，通过基因差异表达、GO和KEGG分析筛选出与卵泡发育相关的候选基因，并应用实时定量PCR法对其表达进行验证。通过转录表达谱分析，共鉴定出18 891个基因，已知基因有18 063个，预测的新基因有828个；从转录表达谱中筛选出40个与卵泡发育相关的差异表达基因，其中，35个基因的表达量与发育卵泡数成正相关，FDX1L在70、220、165和330 d边鸡卵巢中表达量依次递减，NDRG4在70、165、220和330 d边鸡卵巢中表达量随日龄增加呈递减趋势，MC3R和PRLHR在70和330 d边鸡卵巢中表达量高于165和220 d的表达量；VIP仅在70、165和220 d边鸡卵巢中表达。随机取11个差异表达基因，经实时荧光定量PCR验证其相对表达量与转录表达谱一致；GO功能富集发现，差异表达基因主要集中在"生物过程"，其次是"细胞组分"，最后是"分子功能"；KEGG分析发现，差异基因主要集中在信号转导、免疫系统和信号分子与相互作用通路中。本研究通过分析不同生理阶段边鸡卵巢基因表达谱，挖掘与卵泡发育相关的基因，通过GO功能富集和KEGG分析，获得了差异表达基因的功能分类和信号通路，为揭示边鸡卵泡发育相关基因的分子调控及其就巢性提供依据。

关键词: 边鸡, 卵巢, 转录组, 差异表达基因, 卵泡发育, 就巢性

Abstract: To dig out the genes related to the follicular development and reveal the genetic resorce for strong nesting, the transcriptome profiles of ovaries of Bian chickens at different physiological stages were analyzed. The transcriptome profiles of ovaries of Bian chickens at 70 d (stage of ovary development), 165 d (beginning of egg laying), 220 d (eggs laying at peak) and 330 d (nesting) were constructed, RSEM method was used to analyze the correlations of ovarian gene expression at different ages, basing on the differentially expressed genes (DEGs), GO and KEGG analysis, the candidate genes related to follicular development were screened and verified by quantitative real-time PCR. The analysis of transcriptome profiles showed that in total, there were 18 891 genes were identified among which 18 063 were known genes and 828 were unknown genes. Among the differentially expressed genes, 40 genes correlated with follicles development were screened, of which 35 genes were positively related to the number of developed follicle. The expression level of FDX1L in Bian chicken ovary at 70, 220, 165 and 330 d gradually decreased, and the expression level of NDRG4 in Bian chicken ovary at 70, 165, 220 and 330 d showed a decreasing trend with the increase of age; The expression levels of MC3R and PRLHR were higher in ovaries of Bian chicken at 70 and 330 d than those at 165 and 220 d; VIP was only expressed in Bian chicken ovary on 70, 165 and 220 d. The 11 randomly selected DEGs were verified by real-time fluorescent quantitative PCR that their relative expression were consistent with the transcriptional expression profile. Furthermore, GO function enrichment found that the DEGs were mainly concentrated in "biological processes", followed by "cell components", and finally "molecular functions"; KEGG analysis found that the DEGs are mainly concentrated in signal transduction, immune system and signal molecules, and interaction pathways. By analyzing gene expression profiles of Bian chicken ovary at different physiological stages, mining genes related to follicular development, and using GO function enrichment analysis and KEGG analysis, the functional classification and signaling pathway of differentially expressed genes was obtained. It provides the basis for revealing the molecular regulation of the related genes and their nesting properties of the Bian chicken follicle development.

Key words: Bian chicken, ovary, transcriptome, differentially expressed gene, follicular development, nesting

中图分类号:

S831.2

张俊珍, 李彩娥, 刘博, 李建慧, 张蒙, 李亚莉, 常强强. 不同生理阶段边鸡卵巢转录谱的构建及卵泡发育相关基因的分析[J]. 畜牧兽医学报, 2022, 53(2): 423-435.

ZHANG Junzhen, LI Cai'e, LIU Bo, LI Jianhui, ZHANG Meng, LI Yali, CHANG Qiangqiang. Construction of Ovary Transcription Profile of Bian Chicken at Different Physiological Stages and Analysis of Genes Related to Follicular Development[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 423-435.

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