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Selection of reference genes for RT‐qPCR normalization in blueberry (Vaccinium corymbosum?×?angustifolium) under various abiotic stresses

机译:在各种非生物应激下,蓝莓(醋杆菌瘤β×β×angustifolium)中RT-QPCR标准化参考基因的选择

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As a small fruit rich in anthocyanins, blueberry ( Vaccinium corymbosum × angustifolium ) has become a focus of research in recent years for identifying genes related to anthocyanin transport and stress resistance mechanisms based on transcriptome sequencing. However, the lack of validated, stably expressed reference genes greatly limits the functional study of blueberry genes. Therefore, in this study, we selected 14 candidate reference genes from a blueberry transcriptome database and used three algorithms (geNorm, NormFinder and BestKeeper) to evaluate the expression stability of these genes in various organs at different fruit developmental stages under five abiotic stress conditions. EF1α , EIF and TBP were observed to be the most stable and were thus chosen as reference genes for quantitative real‐time PCR. Measurement of the relative expression of VcMATE1 (European Nucleotide Archive accession number KF875433) in blueberry further verified the reliability of these reference genes, which may have great utility for determining the accuracy of gene expression analyses in future research on blueberry.
机译:作为富含花青素的小果实,蓝莓(VacciniumBoSum×Angustifolium)已成为近年来识别与基于转录体测序有关的基因与花青素输送和胁迫阻力机制相关的基因。然而,缺乏验证的稳定表达的参考基因大大限制了蓝莓基因的功能研究。因此,在本研究中,我们从蓝莓转录组数据库中选择了14个候选参考基因,并使用了三种算法(Genorm,Normfinder和BestKeeper)来评估在五种非生物胁迫条件下不同果实发育阶段的各种器官中这些基因的表达稳定性。观察到EF1α,EIF和TBP是最稳定的,因此选择作为参考基因进行定量实时PCR。在蓝莓中测量VCMATE1(欧洲核苷酸档案登录号KF875433)的相对表达进一步验证了这些参考基因的可靠性,这可能具有很大的实用性,用于确定蓝莓未来研究中基因表达分析的准确性。

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