Tamechika Masami M, Yusa Yoichi, Akita Shingo, Sekino Masashi
Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1 Minato- cho, Hakodate, Hokkaido, 041-8611, Japan.
Faculty of Science, Nara Women's University, Nara, 630-8506, Japan.
Mol Biol Rep. 2025 Aug 28;52(1):853. doi: 10.1007/s11033-025-10957-7.
Simultaneously hermaphroditic animals allocate resources to both male and female reproductive functions (sex allocation). Although male reproductive success is a key factor in determining sex allocation, it is seldom evaluated in marine hermaphrodites, as it requires the development of suitable DNA markers to trace paternity. The sessile, simultaneously hermaphroditic barnacle Chthamalus challengeri Hoek, 1883 is widely found along rocky intertidal shores in the northwestern Pacific. In this report, we describe microsatellite DNA markers for C. challengeri and multiplex panels of the markers, which can be used to reveal reproductive success via paternity identification.
We obtained 632 microsatellite sequences by using next-generation sequencing. We examined 189 primer sets, of which 11 passed our filtering criteria. Through simulation analysis, we demonstrated that these primers can be used as microsatellite markers for accurate paternity determination. Additionally, we established five multiplex panels by using all the developed markers.
We developed 11 microsatellite DNA markers for paternity analysis. These markers, which exhibited a moderate to high degree of polymorphism in a population sample, should be useful in paternity inference. We also established multiplex panels for the markers, thus enabling low-cost and efficient analysis. These advancements will enable us to evaluate male reproductive success and contribute to a better understanding of sex allocation in C. challengeri and in hermaphroditic barnacles more broadly.
同步雌雄同体动物会将资源分配给雄性和雌性生殖功能（性别分配）。虽然雄性生殖成功率是决定性别分配的关键因素，但在海洋雌雄同体动物中很少进行评估，因为这需要开发合适的DNA标记来追踪父系。固着性的同步雌雄同体藤壶Chthamalus challengeri Hoek，1883在西北太平洋的岩石潮间带广泛分布。在本报告中，我们描述了用于C. challengeri的微卫星DNA标记及其多重引物组，可用于通过父系鉴定揭示生殖成功率。
我们通过下一代测序获得了632个微卫星序列。我们检测了189对引物，其中11对通过了我们的筛选标准。通过模拟分析，我们证明这些引物可作为微卫星标记用于准确的父系鉴定。此外，我们使用所有开发的标记建立了五个多重引物组。
我们开发了11个用于父系分析的微卫星DNA标记。这些标记在群体样本中表现出中度到高度的多态性，应有助于父系推断。我们还为这些标记建立了多重引物组，从而实现低成本和高效分析。这些进展将使我们能够评估雄性生殖成功率，并有助于更广泛地了解C. challengeri和雌雄同体藤壶的性别分配。
