| Summary: | (1) Background: the species of <i>Corylus</i> 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 “<i>Dawei</i>” 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 <i>ChaActin</i>, <i>VvActin,</i><i>ChaUBQ14,</i> and <i>ChaEF1-α</i> 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, <i>ChaActin</i> and <i>ChaEF1-α</i> 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, <i>CavPrx</i>, which the expression profiles in <i>Corylus</i> have been reported, was quantified by using <i>ChaActin</i> and <i>ChaEF1-α</i> as reference genes. (4) Conclusions: our study of reference genes selection in hazelnut shows that the two reference genes, <i>ChaActin</i> and <i>ChaEF1-α</i>, are suitable for the evaluation of gene expression, and can be used for the analysis of pollen-pistil interaction in <i>Corylus</i>. The results supply a reliable foundation for accurate gene quantifications in <i>Corylus</i> species, which will facilitate the studies related to the reproductive biology in <i>Corylus.</i>
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