Development of an Accurate Polymerase Chain Reaction (PCR) Assay for Genetic Sex Identification in Lumpfish (Cyclopterus lumpus) Based on Male-Specific Anti-Mullerian Hormone (amh) Gene
Development of an Accurate Polymerase Chain Reaction (PCR) Assay for Genetic Sex Identification in Lumpfish (Cyclopterus lumpus) Based on Male-Specific Anti-Mullerian Hormone (amh) Gene
复制标题
DOI：
10.3390/fishes8060327
复制
发表时间：
2023-06
期刊：
影响因子：
2.3
通讯作者：
Kasidis Chaiyasut;Paul Merviel;P. Palma;Johanna H. Perschthaler;E. Jiménez-Fernández;A. Davie;Alejandro P. Gutiérrez
中科院分区：
文献类型：
--
作者：
Kasidis Chaiyasut;Paul Merviel;P. Palma;Johanna H. Perschthaler;E. Jiménez-Fernández;A. Davie;Alejandro P. Gutiérrez
在鲑鱼水产养殖中，团头鱼(Cyclopterus Lumpus)的生产已成为控制海虱水平的关键。为了改善它们的繁殖，有必要及早进行性别鉴定。含有抗苗勒氏激素(AMH)基因的基因组区域被认为是团头鱼性别决定的候选主控基因。然而，团头鱼的基因组包含三个性别特异性不明确的AMH拷贝，分别命名为amh1、amh2和amh3。本研究旨在分析这些AMH同源基因之间的男性特异性区域，以便将其应用于性别标记。在这项研究中，我们利用基于聚合酶链式反应(PCR)的分析方法在团头鱼中鉴定雄性特异的AMH标记，并估计雄性特异区域在团头鱼基因组中的长度。我们的结果表明，包含amh1和amh2基因的约27kb的特定基因组区域表现出男性特异性，而amh3在两性中都存在。建立的针对amh1和amh2的基因性别鉴定方法与表型记录的符合率达97%以上。在环翅虫科的其他成员中的进一步实验发现，只有A ventricosus ventricosus、Eummicrotreus taranetzi和E.asperrimus有雄性特有的AMH基因组区域。利用现有的环翅科AMH序列进行的系统发育分析表明，雄性特有的AMH早期出现在环翅科的谱系中。随着聚合酶链式反应检测技术的发展，我们的研究结果为硬骨鱼水产养殖领域带来了巨大的希望，也将对未来旨在加强我们对硬骨鱼性别决定系统和性染色体进化的了解的研究做出重大贡献。
The production of lumpfish (Cyclopterus lumpus) has become crucial in controlling sea lice levels in salmonid aquaculture. To improve their breeding, there is a need for early sex identification. The genomic region containing the anti-Müllerian hormone (amh) gene was suggested as the candidate master sex-determining gene in lumpfish. However, the genome of lumpfish contains three copies of amh with ambiguous sex specificity, designated amh1, amh2, and amh3. The study aims to analyse the male-specific region between these amh paralogues for its application as a sex marker. In this study, we utilised polymerase chain reaction (PCR)-based assays to identify the male-specific amh markers in lumpfish and estimate the length of the male-specific region in the lumpfish genome. Our results indicate that a specific genomic region of approximately 27 kilobases (kb), encompassing amh1 and amh2 genes, exhibits male specificity, whereas amh3 is present in both sexes. The developed PCR-based genetic sex identification assays targeting amh1 and amh2 exhibited over 97% concordance with phenotypic records. Further experiments in other members of the Cyclopteridae: Aptocyclus ventricosus, Eumicrotremus taranetzi, and E. asperrimus revealed male-specific amh genome region only in A. ventricosus. Phylogenetic analyses using the available Cyclopteridae amh sequences suggest that male-specific amh arose early in the Cyclopteridae lineage. Our findings, along with the development of the PCR test, hold great promise for the field of lumpfish aquaculture and will also contribute significantly to future investigations aiming to enhance our understanding of the sex-determination system and the evolution of sex chromosomes in teleostean fish.