【摘要】 黑番鸭(Cairna moschata)是优良的瘦肉型肉鸭，耐旱、耐粗饲，在养禽业中具有重要价值和特殊地位；但是就巢性强、繁殖力低，严重制约其产业发展。本研究分别采集就巢期和产蛋期各3只黑番鸭的卵巢组织，以TRIzol法提取卵巢组织总RNA，构建文库并分析2组miRNA的差异表达；随后对miRNA进行靶基因预测，并对靶基因进行GO和KEGG分析；最后通过qPCR验证高通量测序结果的可靠性。结果显示，测序产出的原始数据(raw reads)均超过11 099 443条，过滤后序列所占比例均高于96.15%，可用于后续分析；经比对鉴定获得344个miRNAs，包括275个已知miRNAs和69个新发现的miRNAs；与产蛋期比较，就巢期黑番鸭卵巢组织中筛选到16个差异miRNA，其中5个上调、11个下调，并预测到440个靶基因，其中生长激素促分泌素受体(growth hormone secretagogue receptor, GHSR)、卵泡抑素(follistatin, FST)等靶基因与繁殖及卵巢发育相关，推测其对应的gga-miR-16c-5p、gga-miR-146a-3p、novel＿247、novel＿325等miRNAs可能与黑番鸭就巢性状相关；GO富集分析表明，与繁殖发育相关的过程有染色体组织(chromosome organization)、核染色体(nuclear chromosome)、纺锤体微管(spindle microtubule)、核酸结合(DNA binding)和核苷酸激酶活性(nucleotide kinase activity)等；KEGG通路注释结果显示，234个靶基因注释到101个信号通路上，包括Wnt和胰岛素信号通路等可能与生殖细胞增殖分化及发育相关的通路。qPCR验证结果显示，上调和下调miRNAs的相对表达量变化趋势均与高通量测序结果一致。本研究筛选了黑番鸭就巢性状关键miRNAs，为从转录或转录后调控层面分析黑番鸭就巢机制、加快高产新品系的选育提供基础资料。更多还原

【Abstract】 The black Muscovy duck(Cairna moschata) is excellent lean meat ducks that is drought-tolerant and rough-fed, which has important value and special status in the poultry industry. However, the black Muscovy duck has strong broodiness and low fecundity, which restricts rapid development of the industry. The ovarian tissues of 3 black Muscovy ducks at the laying and broody stages were collected, respectively. The TRIzol method was used to extract total RNA from the ovarian tissues, the library was constructed and highthroughput sequencing technology was used to analyze the differential expression of miRNA between the 2groups, then the known, novel miRNAs were predicted for target genes, and the functions of the target genes were enriched and analyzed by GO and KEGG, at last the high-throughput sequencing results were verified by qPCR. The results showed that the raw reads produced by sequencing exceeded 11 099 443 pieces in both groups, and the proportion of clean reads after filtering was all higher than 96.15%, which suggested the data could be used for subsequent analysis. A total of 344 miRNAs were identified, including 275 known miRNAs and 69 newly discovered miRNAs. Sixteen significantly differently expressed miRNAs were screened,including 5 down-regulated and 11 down-regulated in broody ducks with 440 predicted target genes, in which growth hormone secretagogue receptor(GHSR), follistatin(FST) and other target genes were related to reproduction and ovarian development. Thus, it was speculated that the corresponding gga-mir-16c-5p, ggamir-146a-3p and novel＿ 247、novel＿ 325 and other miRNAs might be related to broodiness of black Muscovy duck. GO enrichment analysis showed that chromosome organization, nuclear chromosome, spindle microtubule, DNA binding, and nucleotide kinase activity were related to reproduction and development.KEGG pathway annotation placed 234 target genes into 101 signaling pathways, including Wnt and insulin signaling pathways which might be related to germ cell differentiation and development. qPCR confirmed that relative expression levels of up-regulated and down-regulated miRNAs were consistent with the highthroughput sequencing results. This study screened key miRNAs related to broodiness in Muscovy ducks,providing basic material for analyzing broodiness mechanism of Muscovy duck from transcriptional or post transcriptional regulatory level and accelerating the breeding of new high-yield strains.更多还原