基于RNA-seq技术挖掘鹌鹑羽色自别雌雄相关基因  

Screening of genes related to auto-sexing on feather color based on RNA-seq technology

作　　者：王乾昆 张小辉[1,2] 庞有志 祁艳霞[1,2] 雷莹 白俊艳[1,2] 户运奇 赵毅威 苑志文[1] 王涛 WANG Qiankun;ZHANG Xiaohui;PANG Youzhi;QI Yanxia;LEI Ying;BAI Junyan;HU Yunqi;ZHAO Yiwei;YUAN Zhiwen;WANG Tao(College of Animal Science, Henan University of Science and Technology, Luoyang 471003, Henan, China;Luoyang Key Laboratory of Animal Genetic and Breeding, Luoyang 471003, Henan,China)

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003 [2]洛阳市动物遗传育种重点实验室,河南洛阳471003

出　　处：《浙江农业学报》2022年第3期498-506,共9页Acta Agriculturae Zhejiangensis

基　　金：河南省自然科学基金(202300410153);河南省科技攻关项目(202102110088)。

摘　　要：鹌鹑的羽色自别雌雄现象是由性染色体上的基因决定的,但具体的分子机制未知。本研究以北京白羽公鹌鹑和朝鲜栗羽母鹌鹑为研究对象,利用RNA-seq技术分析了F_(1)公鹑和F_(1)母鹑胚胎期第10天皮肤组织样品的转录组,以筛选调控鹌鹑羽色自别雌雄的关键基因,并利用qRT-PCR技术进行验证。结果表明:RNA-Seq共得到38.94 G的原始数据,平均每个样本获得了6.49 G的原始数据,6个库的Q30均在90%以上,GC含量平均值为49.7%,所测样品至少89%的reads比对到参考基因组上。通过数据库比对共得到16013个基因,其中,上调基因69个,下调基因22个。GO富集分析发现有13841个基因注释到GO数据库中,其中包含78个差异表达基因。KEGG富集结果显示,有22个差异基因富集到38条通路中。对全部差异基因进行筛选,得到了7个与羽色表型相关的基因,分别是DCT、MLANA、SLC45A2、TYRP1、TRPM1、FAM174A和KIT。qRT-PCR结果表明,候选基因均在栗羽鹌鹑中高表达,与RNA-seq结果一致,这7个候选基因可能与自别雌雄有关。The phenomenon of auto-sexingby feather color in quail is determined by genes on the sex chromosomes,but the specific molecular mechanism is unknown.In this study,Beijing male white feather quail and female maroon feather quail were selected as research animals.The transcriptome of F_(1) male and F_(1) female quails at the 10th day of embryo stage were analyzed by RNA-seq technique,and the key genes associated with feather color were screened and verified by qRT-PCR.The results showed that a total of 38.94 G of raw reads were obtained in RNA-seq,with an average of 6.49 G for each sample.And the Q30 of the six libraries was above 90%,average of GC content was 49.7%.At least 89%of the reads from the tested sample were aligned to the reference genome.A total of 16013 genes were obtained through database comparison,of which 69 were up-regulated genes and 22 were down-regulated genes.GO enrichment analysis revealed that 13841 genes were annotated into the GO database,including 78 differentially expressed genes.KEGG enrichment results showed that 22 differentially expressed genes were enriched in 38 pathways.All differentially expressed genes were screened and seven genes related to feather color phenotype were obtained:DCT,MLANA,SLC45A2,TYRP1,TRPM1,FAM174A and KIT.According to the results of qRT-PCR,the candidate genes were highly expressed in maroon feather quail,which was consistent with RNA-seq results.The seven candidate genes obtained in this study might be related to auto-sexing.

关 键 词：鹌鹑 羽色 自别雌雄 转录组 基因表达 

分 类 号：S839[农业科学—畜牧学]