(1) Background: the species of Corylus have sporophytic type of self-incompatibility. Several genes related to recognition reaction between pollen and stigma have been identified in hazelnuts. To better understand the self-incompatibility (SI) response, we screened the suitable reference genes by using quantitative real-time reverse transcription PCR (qRT-PCR) analysis in hazelnut for the first time. (2) Methods: the major cultivar “Dawei” was used as material. A total of 12 candidate genes were identified and their expression profiles were compared among different tissues and in response to various treatments (different times after self- and cross-pollination) by RT-qPCR. The expression stability of these 12 candidate reference genes was evaluated using geNorm, NormFinder, BestKeeper, Delta Ct, and RefFinder programs. (3) Results: the comprehensive ranking of RefFinder indicated that ChaActin, VvActin,ChaUBQ14, and ChaEF1-α were the most suitable reference genes. According to the stability analysis of 12 candidate reference genes for each sample group based on four software packages, ChaActin and ChaEF1-α were most stable in different times after self-pollination and 4 h after self- and cross-pollination, respectively. To further validate the suitability of the reference genes identified in this study, CavPrx, which the expression profiles in Corylus have been reported, was quantified by using ChaActin and ChaEF1-α as reference genes. (4) Conclusions: our study of reference genes selection in hazelnut shows that the two reference genes, ChaActin and ChaEF1-α, are suitable for the evaluation of gene expression, and can be used for the analysis of pollen-pistil interaction in Corylus. The results supply a reliable foundation for accurate gene quantifications in Corylus species, which will facilitate the studies related to the reproductive biology in Corylus.

中文翻译：

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太子参定量RT-PCR分析参考基因的选择和验证。×榛子

（1）背景：榛属种具有自交不亲和的孢子体类型。在榛子中已鉴定出几种与花粉和柱头之间的识别反应有关的基因。为了更好地了解自我不相容性（SI）响应，我们首次通过在榛子中使用定量实时逆转录PCR（qRT-PCR）分析筛选了合适的参考基因。（2）方法：主要品种“大围””用作材料。总共鉴定了12个候选基因，并通过RT-qPCR比较了它们在不同组织之间以及对各种处理（自花授粉和交叉授粉后的不同时间）的表达情况。使用geNorm，NormFinder，BestKeeper，Delta Ct和RefFinder程序评估了这12个候选参考基因的表达稳定性。（3）结果：RefFinder的综合排名表明，ChaActin，VvActin，ChaUBQ14和ChaEF1-α是最合适的参考基因。根据基于ChaActin和ChaEF1-α四个软件包的每个样品组的12个候选参考基因的稳定性分析自花授粉后的不同时间和自花授粉后和交叉花粉后4 h的稳定性最强。为了进一步验证本研究中鉴定的参考基因的适宜性，CavPrx，其在表达谱榛已经报道了，通过使用定量ChaActin和ChaEF1-α作为参考基因。（4）结论：我们对榛子中参考基因选择的研究表明，ChaActin和ChaEF1-α这两个参考基因适合评估基因表达，并可用于分析榛属中的花粉雌蕊相互作用。。该结果为正确确定Corylus物种的基因定量提供了可靠的基础，这将有助于与Corylus生殖生物学相关的研究。