Department of Genetics, Wrocław University of Environmental and Life Sciences, Kożuchowska 7, 51-631 Wrocław, Poland.
Genes (Basel). 2022 Mar 12;13(3):507. doi: 10.3390/genes13030507.
In palaeognathous birds, several PCR-based methods and a range of genes and unknown genomic regions have been studied for the determination of sex. Many of these methods have proven to be unreliable, complex, expensive, and time-consuming. Even the most widely used PCR markers for sex typing in birds, the selected introns of the highly conserved gene (primers P2/P8, 1237L/1272H, and 2550F/2718R), have rarely been effective in palaeognathous birds. In this study we used eight species of Palaeognathae to test three PCR markers: CHD1i9 ( gene intron 9) and NIPBLi16 ( gene intron 16) that performed properly as Psittaciformes sex differentiation markers, but have not yet been tested in Palaeognathae, as well as the CHD1iA intron ( gene intron 16), which so far has not been used effectively to sex palaeognathous birds. The results of our research indicate that the CHD1i9 marker effectively differentiates sex in four of the eight species we studied. In , , and , the electrophoretic patterns of the amplicons obtained clearly indicate the sex of tested individuals, whereas in , sexing is possible based on poorly visible female specific bands. Additionally, we present and discuss the results of our in silico investigation on the applicability of CHD1i9 to sex other Palaeognathae that were not tested in this study.
在古颌鸟类中，已经研究了几种基于 PCR 的方法和一系列基因和未知基因组区域，以确定性别。其中许多方法已被证明不可靠、复杂、昂贵且耗时。即使是鸟类性别分型最广泛使用的 PCR 标记物，即高度保守基因的选定内含子（引物 P2/P8、1237L/1272H 和 2550F/2718R），在古颌鸟类中也很少有效。在这项研究中，我们使用了八种古颌鸟类来测试三个 PCR 标记物：CHD1i9（基因内含子 9）和 NIPBLi16（基因内含子 16），它们作为凤头鹦鹉性别分化标记物表现良好，但尚未在古颌鸟类中进行测试，以及 CHD1iA 内含子（基因内含子 16），迄今为止，它尚未有效地用于鉴定古颌鸟类的性别。我们的研究结果表明，CHD1i9 标记物在我们研究的八种物种中的四种中有效地区分了性别。在凤头鹦鹉科、鹦鹉科和鹤鸵科中，获得的扩增子的电泳图谱清楚地表明了测试个体的性别，而在鸵鸟科中，根据女性特异性带的可见度差，可以进行性别鉴定。此外，我们提出并讨论了我们对 CHD1i9 应用于本研究中未测试的其他古颌鸟类性别的计算机模拟研究结果。
