Hou Sihao, Zhao Tiantian, Yang Dan, Li Qing, Liang Lisong, Wang Guixi, Ma Qinghua
Key Laboratory of Tree Breeding and Cultivation of the State Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China.
Hazelnut Engineering and Technical Research Center of the State Forestry and Grassland Administration, Beijing 100091, China.
Plants (Basel). 2021 Jan 15;10(1):159. doi: 10.3390/plants10010159.
(1) Background: the species of 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 "" 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 , and 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, and 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, , which the expression profiles in have been reported, was quantified by using and as reference genes. (4) Conclusions: our study of reference genes selection in hazelnut shows that the two reference genes, and , are suitable for the evaluation of gene expression, and can be used for the analysis of pollen-pistil interaction in . The results supply a reliable foundation for accurate gene quantifications in species, which will facilitate the studies related to the reproductive biology in
(1) 背景：该物种具有孢子体型自交不亲和性。在榛子中已鉴定出几个与花粉和柱头识别反应相关的基因。为了更好地理解自交不亲和（SI）反应，我们首次在榛子中通过定量实时逆转录PCR（qRT-PCR）分析筛选了合适的内参基因。(2) 方法：以主要品种“”为材料。共鉴定出12个候选基因，并通过RT-qPCR比较了它们在不同组织以及对各种处理（自花授粉和异花授粉后不同时间）的表达谱。使用geNorm、NormFinder、BestKeeper、Delta Ct和RefFinder程序评估这12个候选内参基因的表达稳定性。(3) 结果：RefFinder的综合排名表明，、和是最合适的内参基因。根据基于四个软件包对每个样本组的12个候选内参基因的稳定性分析，和分别在自花授粉后不同时间以及自花授粉和异花授粉后4小时最稳定。为了进一步验证本研究中鉴定的内参基因的适用性，使用和作为内参基因对已报道其在中的表达谱的进行了定量分析。(4) 结论：我们对榛子内参基因选择的研究表明，两个内参基因和适用于基因表达评估，可用于分析中的花粉 - 雌蕊相互作用。研究结果为物种中准确的基因定量提供了可靠基础，将促进与生殖生物学相关的研究。
