基于转录组测序筛选鸡蛋绿壳性状相关基因  

Screening of Candidate Genes for Green Shell Egg Shell Color Traits in Chishui Black Bone Chicken Based on Transcriptome Sequencing

作　　者：肖涛 李辉[1] 罗韦 叶涛 余欢 陈友波 石钰仕 赵德鹏 吴芸[2] XIAO Tao;LI Hui;LUO Wei;YE Tao;YU Huan;CHEN YouBo;SHI YuShi;ZHAO DePeng;WU Yun(Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountainous Region,Ministry of Education/Guizhou Provincial Key Laboratory of Animal Genetics,Breeding and Reproduction/College of Animal Science,Guizhou University,Guiyang 550025;College of Biology and Agriculture Technology,Zunyi Normal University,Zunyi 563006,Guizhou;Anshun City Animal Husbandary Technology Extension Station,Anshun 561000,Guizhou)

机构地区：[1]高原山地动物遗传育种与繁殖教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室/贵州大学动物科学学院,贵阳550025 [2]遵义师范学院生物与农业科技学院,贵州遵义563006 [3]安顺市畜牧技术推广站,贵州安顺561000

出　　处：《中国农业科学》2023年第8期1594-1605,共12页Scientia Agricultura Sinica

基　　金：贵州省地方家禽产业联合攻关项目(黔财农[2020]175号);贵州省科技支撑计划(黔科合支撑[2019]2285号);赤水河流域资源保护与开发研究院项目(CSH-[2019]-005)。

摘　　要：【背景】绿壳蛋深受消费者喜爱,蛋壳绿色的深浅在市场上是影响绿壳蛋定价销售的重要参考指标。绿壳蛋的形成受多基因共同调控,蛋壳绿色深浅不一,其分子机理尚不清楚。通过对赤水乌骨鸡蛋壳腺组织进行转录组测序,挖掘其调控绿壳蛋蛋壳颜色深浅的候选基因以及关键信号通路,探究影响蛋壳颜色遗传性,以期发展绿壳蛋的选种选育和提高经济效益。【目的】探讨赤水乌骨鸡的遗传基础,并通过SLCO1B3基因分型对其进行鉴定和筛选,以期通过分子标记在青壳鸡育种规划中提供新的见解,并在后期的选择策略中帮助控制和提高赤水乌骨鸡蛋壳品质的同质性。【方法】以纯系的280日龄赤水乌骨鸡为研究对象,屠宰产浅绿色蛋(QL)和深绿色蛋(SL)的母鸡各3只,采集蛋壳腺以RNA-seq技术检测分析,筛选与蛋壳颜色密切相关的差异表达基因(differentially expressed genes,DEGs),并对这些DEGs进行GO和KEGG富集分析。利用qRT-PCR技术检测与蛋壳颜色相关的6个候选基因的转录水平变化以验证转录组数据可靠性。【结果】在深绿组与浅绿组中共筛选到93个显著DEGs,有59个基因在深绿组中显著上调,34个显著下调。DEGs注释到GO数据库进行比对,主要显著性富集的是钠离子转运、负离子结合、肌质网膜等。KEGG分析富集程度最显著的是醛固酮调节钠的重吸收,以及富集的矿物质吸收、亚油酸的新陈代谢等信号通路。通过qRT-PCR验证,结果显示这些基因的表达趋势与转录组的测序结果一致。【结论】经功能分析,TF基因、SCNN1家族基因、CYP450家族基因、SLC家族基因和FAM家族基因,以及醛固酮调节钠的重吸收信号通路参与蛋壳色素合成、转运和沉积。上述基因和信号通路可能是影响赤水乌骨鸡蛋壳绿色深浅不一的候选基因和关键信号通路。【Background】Green shell eggs are loved by consumers,and the green color of eggshell is an important reference index affecting the pricing and sales of green shell eggs in the market.The formation of green shell eggs is regulated by multiple genes,and the color of green shell varies.However,the molecular mechanism of green shell eggs is still unclear.In this study,transcriptomic sequencing was conducted on the eggshell gland tissue of Chishui black bone chicken,and the candidate genes and key signal pathways that regulated the depth and depth of eggshell color of green shell eggs were excavated,so as to explore the heritability of eggshell color,and to develop the seed selection and breeding of green shell eggs and improve economic benefits.【Objective】The aim of this study was to investigate the genetic basis of Chishui black bone chicken and to identify and then screen them by SLCO1B3 genotyping,in order to provide new insights through molecular markers in the breeding planning of green-shelled hens,and to help control and improve the homogeneity of eggshell quality of Chishui black bone chicken in the later selection strategy.【Method】A pure 280-day-old Chishui black bone chicken was used as the research object.Three hens were slaughtered to produce light green eggs(QL)and dark green eggs(SL),and the eggshell glands were collected and analyzed by RNA-SEQ technology.Selected differentially expressed genes(DEGs)closely related to eggshell color,and analyzed for GO and KEGG enrichment.Qrt-pcr was used to detect the transcriptional level changes of six candidate genes related to eggshell color to verify the reliability of transcriptomic data.【Result】A total of 93 DEGs were screened in SL group and QL group,among which 59 genes were up-regulated and 34 genes were down-regulated in SL group.DEGs was annotated to GO database for comparison,and sodium ion transport,negative ion binding,and sarcoplasmic reticulum were mainly enriched significantly.KEGG analysis showed that aldosterone regulated sodium reab

关 键 词：转录组测序 赤水乌骨鸡 绿壳蛋 蛋壳颜色 QRT-PCR 

分 类 号：S831.2[农业科学—畜牧学]