College of Plant Protection, Henan Agricultural University, Zhengzhou 450046, China.
Henan Key Laboratory of Creation and Application of New Pesticide, Henan Agricultural University, No. 218, Ping'an Avenue, Zhengzhou 450046, China.
Int J Mol Sci. 2023 Oct 15;24(20):15198. doi: 10.3390/ijms242015198.
Reference genes are important for the accuracy of gene expression profiles using reverse-transcription quantitative PCR (RT-qPCR). However, there are no available reference genes reported for ; it actually has a pretty diverse and wide host range. In this study, seven candidate reference genes (, , , , , and ) were validated for their expression stability in under conditions of different developmental stages, populations, fungicide treatments, photoperiods and pHs. Four algorithm programs (geNorm, Normfinder, Bestkeeper and ΔCt) were used to evaluate the gene expression stability, and RefFinder was used to integrate the ranking results of four programs. Two reference genes were recommended by RefFinder for RT-qPCR normalization in . The suitable reference genes were and across developmental stages, and across populations, and across fungicide treatments, and across photoperiods, and across pHs and and across all samples. Four target genes (, , and ) were selected for the validation of the suitability of selected reference genes. However, using one or two reference genes in combination to normalize the expression of target genes showed no significant difference in . In short, this study provided reliable reference genes for studying the expression and function of genes in .
参考基因对于使用反转录定量 PCR (RT-qPCR) 进行基因表达谱的准确性非常重要。然而，目前还没有报道适用于 的参考基因；实际上， 具有非常多样化和广泛的宿主范围。在这项研究中，验证了 7 个候选参考基因（ 、 、 、 、 、 和 ）在不同发育阶段、种群、杀菌剂处理、光周期和 pH 值条件下的表达稳定性。使用了 4 种算法程序（geNorm、Normfinder、BestKeeper 和 ΔCt）来评估基因表达稳定性，并使用 RefFinder 整合了 4 种程序的排名结果。RefFinder 推荐了 2 个参考基因用于 RT-qPCR 在 中的归一化。适用于 RT-qPCR 归一化的参考基因在发育阶段是 和 ，在种群中是 和 ，在杀菌剂处理中是 和 ，在光周期中是 和 ，在 pH 值中是 和 ，在所有样本中是 和 。选择了 4 个靶基因（ 、 、 、 ）来验证所选参考基因的适用性。然而，使用一个或两个参考基因组合来归一化靶基因的表达，在 中没有显示出显著差异。总之，这项研究为研究 在 中的基因表达和功能提供了可靠的参考基因。
