Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, The Jiangsu Provincial Platform for Conservation and Utilization of Agricultural Germplasm, Qian Hu Hou Cun No. 1, Nanjing 210014, China.
Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, 159 Longpan Road, Nanjing 210037, China.
Int J Mol Sci. 2021 Sep 29;22(19):10533. doi: 10.3390/ijms221910533.
Due to the lack of effective and stable reference genes, studies on functional genes in , a genus of economically important small berry crops, have been greatly limited. To select the best internal reference genes of different types, we selected four representative cultivars of blackberry and raspberry (red raspberry, yellow raspberry, and black raspberry) as the research material and used RT-qPCR technology combined with three internal stability analysis software programs (geNorm, NormFinder, and BestKeeper) to analyze 12 candidate reference genes for the stability of their expression. The number of most suitable internal reference genes for different cultivars, tissues, and fruit developmental stages of was calculated by geNorm software to be two. Based on the results obtained with the three software programs, the most stable genes in the different cultivars were and . Finally, to validate the reliability of selected reference genes, the expression pattern of the gene was analyzed, and the results highlighted the importance of appropriate reference gene selection. and were screened as reference genes for their relatively stable expression, providing a reference for the further study of key functional genes in blackberry and raspberry and an effective tool for the analysis of differential gene expression.
由于缺乏有效和稳定的参照基因，对经济上重要的小浆果作物属中的功能基因的研究受到了极大的限制。为了选择不同类型的最佳内参基因，我们选择了黑莓和覆盆子的四个代表性品种（红树莓、黄树莓和黑树莓）作为研究材料，利用 RT-qPCR 技术结合三个内参稳定性分析软件程序（geNorm、NormFinder 和 BestKeeper）分析了 12 个候选内参基因的表达稳定性。geNorm 软件计算出不同品种、组织和果实发育阶段的最适内参基因数量为两个。基于这三个软件程序的结果，在不同品种中最稳定的基因是 和 。最后，为了验证所选参考基因的可靠性，分析了 基因的表达模式，结果突出了选择合适参考基因的重要性。 和 被筛选为表达相对稳定的参考基因，为黑莓和覆盆子中关键功能基因的进一步研究提供了参考，也为差异基因表达的分析提供了有效工具。
