Identification of Suitable Reference Genes for Barley Gene Expression Under Abiotic Stresses and Hormonal Treatments

Abstract

Barley (Hordeum vulgare L.) is an important cereal crop and also can be considered as a model plant for the Triticeae family. Quantitative real-time PCR (qRT-PCR) has emerged as a more practical technique to study barley gene expression, and the data normalization with reference genes is essential for accurate and reliable results. In this study, 17 candidate reference genes were evaluated from two distinct plant tissues (roots and leaves) at the barley seedling stage under different abiotic stresses (osmotic, salt, and heat) and hormonal treatments (gibberellin A3 (GA3) and methyl jasmonate (JA)). Our qRT-PCR data were analyzed with three commonly used software packages: geNorm, NormFider, and BestKeeper. In combination with the analysis of these three packages, four to five candidate genes were selected as suitable reference genes in each experimental set. These results confirmed that expression stability of reference genes depends on the experimental conditions. This study is the systematic analysis for the selection of superior reference genes for qRT-PCR in barley under different abiotic and hormonal treatments and will benefit further studies on gene expression in barley and other species of Triticeae family.