半番鸭卵巢发育不全关键基因的筛选与差异基因表达分析  

Screening of Key Genes for Ovarian Hypoplasia and Analysis of Differential Gene Expression in Mulard Ducks(Cairina moschata)

作　　者：李丽[1,2] 章琳俐 缪中纬[1] 朱志明[1] 邱君志[2] 王娟[3] 辛清武 郑嫩珠[1] LI Li;ZHANG Lin-Li;MIAO Zhong-Wei;ZHU Zhi-Ming;QIU Jun-Zhi;WANG Juan;XIN Qing-Wu;ZHENG Nen-Zhu(Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Ganzhou Agriculture College,Ganzhou 341100,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福州350013 [2]福建农林大学生命科学学院,福州350002 [3]赣州农业学校,赣州341100

出　　处：《农业生物技术学报》2021年第9期1761-1773,共13页Journal of Agricultural Biotechnology

基　　金：福建省自然科学基金(2018J01041);福建省省属公益类项目(2020R10260016);福建省科技计划项目(2019R1026-6);福建省家禽产业技术体系(闽农综[2019]144号);福建省农业科学院科技创新平台专项(CXPT0022019007);福建省农业科学院自由探索科技创新项目(ZYTS2019026)。

摘　　要：半番鸭(Cairina moschata)为番鸭和家鸭(Anas plalyrhynchas var.domestica)的属间杂交后代,具有很强的杂种优势,但无实际种用价值。在实际生产中需要饲养半番鸭父母本,并借助于人工授精技术进行繁殖,过程繁琐且饲养管理成本高,严重限制了半番鸭生产的发展。由于半番鸭不育性状的特殊性,其与母本北京鸭卵巢组织差异表达基因的分析尚未有报道,GenBank序列数据库中相关基因的信息也相对匮乏,其不育遗传基础需进一步深入研究。转录组测序(RNA-Seq)技术不仅可用于病毒感染后组织变化分析,还可用于基因挖掘及功能注释。为了探究半番鸭卵巢发育不全相关基因与不育的分子机制,本研究利用转录组高通量测序技术筛选半番鸭和北京鸭卵巢组织分化发育相关的差异表达基因,并进行基因本体(Gene Ontology,GO)和KEGG功能注释,再利用qRT-PCR技术对差异表达基因进行验证。本次测序共获得43.84 Gb高质量测序标签(clean data)和193535条Unigene。两种类型鸭间共发现89个包括与卵巢发育、性激素合成及生殖细胞分化发育相关的差异表达基因,其中72个基因在半番鸭中上调表达,17个基因下调表达,GO功能注释差异基因富集到591个GO条目(term)中,其中显著富集的钙介导的信号传导与分化发育相关。KEGG通路(pathway)分析结果显示,差异表达基因共富集到15条信号通路中,包括与性激素合成、分泌相关的糖苷脂类生物合成-神经节系列、类固醇激素生物合成通路和酪氨酸代谢。qRT-PCR结果显示半番鸭中上调、下调基因的表达趋势与转录组测序结果一致,表明转录组测序数据较为可靠。本研究为探究半番鸭特殊的生殖系统分化机理提供理论参考依据。Mulard duck is an inter-genus hybrid product of Muscovy duck(Cairina moschata)and domestic duck(Anas plalyrhynchas var.domestica),which has strong heterosis but no reproductive ability.In actual production,it is necessary to raise the parents of Mulard ducks,and with the help of artificial insemination technology,the process is cumbersome and the feeding management cost is high,which severely restricts the development of Mulard duck production.Due to the particularity of the sterile traits of Mulard ducks,the analysis of differentially expressed genes in the ovarian tissues of Mulard ducks has not been reported.The information on related genes in the GenBank sequence database is relatively scarce.The genetic basis of infertility needs further research.Transcriptome sequencing(RNA-seq)technology could not only be used for analysis of tissue changes after virus infection,but also for gene mining and functional annotation.In order to explore the molecular mechanism of genes related to ovarian hypoplasia and infertility in Mulard duck,transcriptome sequencing was performed on the ovarian tissues of Mulard ducks and Pekin ducks,the differentially expressed genes related to ovarian differentiation and development were screened,and the genes were annotated with Gene Ontology(GO)and KEGG,and then the qRT-PCR was used to verify the sequencing results.A total of 43.84 Gb clean data and 193535 Unigenes were obtained in the sequencing.A total of 89 differentially expressed genes related to ovarian development,sex hormone synthesis and germ cell differentiation and development were found between two types of ducks,among them 17 genes were downregulated,72 genes were up-regulated in Mulard ducks,Go functional annotation differential genes were enriched in 591 GO terms,in which the significantly enriched calcium-mediated signal transduction was related to germ cell differentiation and development.KEGG analysis showed that differentially expressed genes are enriched in 15 signal pathways,Including glycosides biosynthesis-ganglio

关 键 词：半番鸭 卵巢发育不全 差异表达基因 转录组 

分 类 号：S813.22[农业科学—畜牧学]