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利用 qPCR 研究南瓜(Cucurbita pepo)基因表达的参考基因选择。

Selection of reference genes for gene expression studies in zucchini (Cucurbita pepo) using qPCR.

机构信息

Área de Mejora y Biotecnología, IFAPA, Centro Alameda del Obispo, Córdoba, Spain.

出版信息

J Agric Food Chem. 2011 May 25;59(10):5402-11. doi: 10.1021/jf200689r. Epub 2011 Apr 22.

Abstract

The zucchini (Cucurbita pepo) is an important food crop, the transcriptomics of which are a fundamental tool to accelerate the development of new varieties by breeders. However, the suitability of reference genes for data normalization in zucchini has not yet been studied. The aim of this study was to assess the suitability of 13 genes for their potential use as reference genes in quantitative real-time PCR. Assays were performed on 34 cDNA samples representing plants under different stresses and at different developmental stages. The application of geNorm and NormFinder software revealed that the use of a combination of UFP, EF-1A, RPL36aA, PP2A, and CAC genes for the different experimental sets was the best strategy for reliable normalization. In contrast, 18S rRNA and TUA were less stable and unsuitable for use as internal controls. These results provide the possibility to allow more accurate use of qPCR in this horticultural crop.

摘要

南瓜(Cucurbita pepo)是一种重要的粮食作物,其转录组学是加速育种者培育新品种的重要工具。然而,南瓜中数据归一化的参考基因的适用性尚未得到研究。本研究旨在评估 13 个基因作为定量实时 PCR 中参考基因的适用性。在代表不同胁迫和不同发育阶段的 34 个 cDNA 样本上进行了测定。geNorm 和 NormFinder 软件的应用表明,对于不同的实验集,使用 UFP、EF-1A、RPL36aA、PP2A 和 CAC 基因的组合是可靠归一化的最佳策略。相比之下,18S rRNA 和 TUA 不太稳定,不适合用作内参。这些结果为在这种园艺作物中更准确地使用 qPCR 提供了可能性。

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