Selection of Suitable Reference Genes for RT-qPCR Gene Expression Analysis in Siberian Wild Rye () under Different Experimental Conditions.

, which is a perennial and self-pollinated grass, is the typical species of the genus , which plays an important role in forage production and ecological restoration. No reports have, so far, systematically described the selection of optimal reference genes for reverse transcriptase quantitative real-time polymerase chain reaction (RT-qPCR) analysis in . The goals of this study were to evaluate the expression stability of 13 candidate reference genes in different experimental conditions, and to determine the appropriate reference genes for gene expression analysis in . Five methods including Delta Ct (ΔCt), BestKeeper, NormFinder, geNorm, and RefFinder were used to assess the expression stability of 13 potential reference genes. The results of the RefFinder analysis showed that and were the most stable reference genes in different genotypes. and had the most stable expression in different developmental stages. and were suitable reference genes in different tissues. Under salt stress, and were identified as the most stable reference genes. and showed the most stability under heat stress. For cold stress, and presented the highest degree of expression stability. and were considered as the most stable reference genes under osmotic stress. The optimal reference genes were selected to investigate the expression pattern of target gene in different conditions. This study provides suitable reference genes for further gene expression analysis using RT-qPCR in .