Departamento de Bioquímica y Biología Molecular e Instituto Andaluz de Biotecnología, Campus Universitario de Rabanales y Campus de Excelencia Internacional Agroalimentario-CEIA3, Universidad de Córdoba, Córdoba, Andalucía, Spain.
PLoS One. 2013 Aug 5;8(8):e70603. doi: 10.1371/journal.pone.0070603. Print 2013.
Strawberry (Fragaria spp) is an emerging model for the development of basic genomics and recombinant DNA studies among rosaceous crops. Functional genomic and molecular studies involve relative quantification of gene expression under experimental conditions of interest. Accuracy and reliability are dependent upon the choice of an optimal reference control transcript. There is no information available on validated endogenous reference genes for use in studies testing strawberry-pathogen interactions. Thirteen potential pre-selected strawberry reference genes were tested against different tissues, strawberry cultivars, biotic stresses, ripening and senescent conditions, and SA/JA treatments. Evaluation of reference candidate's suitability was analyzed by five different methodologies, and information was merged to identify best reference transcripts. A combination of all five methods was used for selective classification of reference genes. The resulting superior reference genes, FaRIB413, FaACTIN, FaEF1α and FaGAPDH2 are strongly recommended as control genes for relative quantification of gene expression in strawberry. This report constitutes the first systematic study to identify and validate optimal reference genes for accurate normalization of gene expression in strawberry plant defense response studies.
草莓（Fragaria spp）是蔷薇科作物中发展基础基因组学和重组 DNA 研究的新兴模式。功能基因组学和分子研究涉及在感兴趣的实验条件下相对定量基因表达。准确性和可靠性取决于选择最佳的参考控制转录本。目前尚无关于在用于测试草莓-病原体相互作用的研究中使用的经过验证的内源性参考基因的信息。对 13 种潜在预选的草莓参考基因进行了不同组织、草莓品种、生物胁迫、成熟和衰老条件以及 SA/JA 处理的测试。通过五种不同的方法评估参考候选物的适用性，并合并信息以确定最佳的参考转录本。使用所有五种方法的组合进行参考基因的选择性分类。最终推荐 FaRIB413、FaACTIN、FaEF1α 和 FaGAPDH2 作为草莓中基因表达相对定量的控制基因。本报告首次对草莓植物防御反应研究中基因表达的准确归一化进行了最佳参考基因的系统鉴定和验证。
