RAPID DUPLICATION AND LOSS OF NBS-ENCODING GENES IN EUROSIDS II

摘要

Hydrogen peroxide (H2O2) acts as a signaling molecule modulating the expression of various genes in plants. However,the reference gene(s) used for gene expression analysis of H2O2 signaling is still arbitrary. A reliable result obtained byquantitative real-time RT-PCR (RT-qPCR) highly depends on accurate transcript normalization using stably expressedreference genes, whereas the inaccurate normalization could easily lead to the false conclusions. In this report, by usinggeNorm and NormFinder algorithms, 12 candidate reference genes were evaluated and compared in root and shoot tissues ofArabidopsis upon different doses of H2O2. The results revealed that, in our experimental conditions, three novel referencegenes (TIP41-like, UKN, and UBC21) were identified and validated as suitable reference genes for RT-qPCR normalizationin both root and shoot tissues under oxidative stress. This conclusion was further confirmed by publicly available microarraydata of methyl viologen and drought stress. In comparison with a single reference gene (EF-1a), the expression pattern ofZAT12 modulated by H2O2, when using TIP41-like, UKN, and UBC21 as multiple reference gene(s), was similar with theprevious reports by using northern blotting. Thus, we proposed that these three reference genes might be good candidates forother researchers to include in their reference gene validation in gene expression studies under H2O2-related oxidative stress.