背景:B染色体是一些真核生物中存在的额外的非必要元件。与一个物种的所有个体中必不可少的染色体不同,B染色体对于生物体的正常功能不是必需的。以前被认为是基因不活跃的,已经发现B染色体不仅表达自己的基因,而且对A染色体的基因表达也有影响。最近的研究表明,在一些Psalidodon(Characiformes,Characidae)物种,B染色体可能与表型效应有关,例如生殖周期和基因表达的变化。
结果:在这项研究中,我们的目的是建立稳定的内参基因,用于对Psalidodon属中三种鱼类的性腺进行RT-qPCR实验,在存在和不存在B染色体的情况下。使用NormFinder评估了五个选定的参考基因的稳定性,geNorm,BestKeeper,和RefFinder算法。我们确定ppiaa和pgk1是P.fasciatus中最稳定的基因,而ppiaa和hmbsa在P.bockmanni中显示出最高的稳定性。对于P.paranae,tbp和hprt1是雌性中最稳定的基因,男性中ppiaa和hprt1最稳定。
结论:考虑到B染色体的存在,我们确定了三种Psalidodon物种性腺中最稳定的参考基因。这是该属中参考基因稳定性的第一份报告,并提供了有价值的工具来更好地了解B染色体在基因表达水平上的作用。
BACKGROUND: B chromosomes are extra non-essential elements present in several eukaryotes. Unlike A chromosomes which are essential and present in all individuals of a species, B chromosomes are not necessary for normal functioning of an organism. Formerly regarded as genetically inactive, B chromosomes have been discovered to not only express their own genes, but also to exert influence on gene expression in A chromosomes. Recent studies have shown that, in some Psalidodon (Characiformes, Characidae) species, B chromosomes might be associated with phenotypic effects, such as changes in the reproductive cycle and gene expression.
RESULTS: In this study, we aimed to establish stable reference genes for RT-qPCR experiments conducted on gonads of three fish species within Psalidodon genus, both in the presence and absence of B chromosomes. The stability of five selected reference genes was assessed using NormFinder, geNorm, BestKeeper, and RefFinder algorithms. We determined ppiaa and pgk1 as the most stable genes in P. fasciatus, whereas ppiaa and hmbsa showed the highest stability in P. bockmanni. For P. paranae, tbp and hprt1 were the most stable genes in females, and ppiaa and hprt1 were the most stable in males.
CONCLUSIONS: We determined the most stable reference genes in gonads of three Psalidodon species considering the presence of B chromosomes. This is the first report of reference gene stability in the genus and provides valuable tools to better understand the effects of B chromosomes at gene expression level.