基于转录组测序筛选半番鸭性腺分化候选基因  

Screening of Candidate Genes for Gonadal Differentiation Mule Duck Using RNA-seq

作　　者：杨宇 叶胜强 翟明丽 李雪莲 童新红 李中心 徐小娟[3] 龚萍 YANG Yu;YE Shengqiang;ZHAI Mingli;LI Xuelian;TONG Xinhong;LI Zhongxin;XU Xiaojuan;GONG Ping(Institute of Animal Husbandry and Veterinary Medicine,Wuhan Academy of Agricultural Sciences,Wuhan,Hubei 430208;Huazhong Agricultural University,Wuhan,Hubei 430070;Fruit and Tea Research Institute,Hubei Provincial Academy of Agricultural Sciences,Wuhan,Hubei 430000)

机构地区：[1]武汉市农业科学院畜牧兽医研究所,湖北武汉430208 [2]华中农业大学,湖北武汉430070 [3]湖北省农业科学院果树茶叶研究所,湖北武汉430000

出　　处：《中国家禽》2025年第5期17-24,共8页China Poultry

基　　金：国家水禽产业技术体系(CARS-42-28);“动物胚胎工程及分子育种”湖北省重点实验室开放课题(KLAEMB-2022-03)。

摘　　要：为筛选到更多与禽类性腺分化相关的候选基因,研究收集半番鸭、金定鸭和番鸭在1/2胚胎期雌性和雄性性腺组织样品进行RNA-seq测序,通过转录组测序分析半番鸭和亲本性腺分化过程中SNP、Indel位点以及可变剪接事件,对获取的SNP和InDel位点在外显子(Exonic)、UTR5以及内含子(Intronic)等8种功能元件上的分布进行统计。结果显示:共获得798216036条cleanreads,9471个差异基因(Differentially expressed genes,DEGs)和691个蛋白质网络互作对(Protein-protein interaction,PPI)。突变位点在外显子上发生的比率占整体的21.26%。共筛选出非同义单核苷酸突变(Non-synonymous SNP,nsSNPs)引起的ATP2C2、GTF2A1L和HHIPL2等8个雌性特异表达基因和STK10、GDF7和SLC5A8等6个雄性特异表达基因,Indel位点引起的NTN4、RAB5A、SHPRH等12个雄性特异表达基因。DEGs可变剪接分析显示,半番鸭雌性和雄性比较组发生的差异可变剪接事件最少(17793次)。分析各比较组外显子使用(Exonusage)差异,发现雌雄性腺之间的外显子使用差异数量总体要少于不同鸭种同种性别之间的数量。研究表明,ATP2C2、GTF2A1L、HHIPL2、STK10、GDF7和SLC5A8等基因外显子SNP位点非同义突变以及NTN4、RAB5A、SHPRH等基因外显子Indel位点移码突变在雌雄性腺分化中发挥着重要作用;性腺分化和发育以及半番鸭的不育与DEGs中发生的可变剪接是密不可分的;位于Z染色体上的STARD6、STARD4、SPEF2、HSD17B3和25号染色体HSD17B1基因发生了可变剪接并参与了性腺性别分化等生物学过程。To identify more candidate genes associated with poultry gonadal differentiation,the study collected male and female gonadal tissue samples from mule duck,Cairina moschata and Jinding duck at the 1/2 embryonic stage for RNA-seq sequencing.Transcriptome sequencing was conducted to analyze SNPs,Indels,and alternative splicing during the gonadal differentiation process in mule duck and its parental species.The distribution of SNPs and Indels on eight functional elements including exonic,UTR5,and intronic regions was statistically analyzed.The results showed that a total of 798216036 clean reads,9471 differentially expressed genes(DEGs),and 691 protein-protein interaction(PPI)pairs were obtained.The mutation rate on exons accounted for 21.26%of the total mutations.A total of eight female-specific expressed genes such as ATP2C2,GTF2A1L and HHIPL2 and six male-specific expressed genes such as STK10,GDF7 and SLC5A8 caused by non-synonymousc single-nucleotide polymorphisms(nsSNPs),and twelve shared genes specifically expressed in the male gonads of three species such as NTN4,RAB5A and SHPRH,which caused by the Indel locus were screened.Variable splicing analysis of DEGs showed that the number of differential splicing events(17793)in the male groups and female muleduck groups was the lowest.Differential exon usage analysis revealed that the overall number of differential exon usage events between male and female gonads was less than that between conspecific genders of different duck species.The results showed that important mutations at the exonic SNP sites of ATP2C2,GTF2A1L,HHIPL2,STK10,GDF7 and SLC5A8,and code-shifting mutations at the exonic Indel sites of NTN4,RAB5A,and SHPRH played an irreplaceable role in male and female gonadal differentiation;poultry differentiation and development,as well as sterility in mule ducks,were inextricably linked to the variable splicing that occurred in DEGs;Genes located on the Z chromosome such as STARD6,STARD4,SPEF2,HSD17B3,and on chromosome 25 like HSD17B1 underwent alternative spl

关 键 词：转录组 番鸭 性别分化 性腺 可变剪接 SNP/Indel 

分 类 号：S831.2[农业科学—畜牧学]