Abstract:The selection of high stablely expressed reference genes is an important prerequisite for improving the accuracy of Real-time quantitative PCR (qRT-PCR). We investigated the expression of 6 candidate reference genes including eukaryotic initiation factor 4 alpha (eIF4A), Ti-binding peptide-1 (TBP-1), ubiquitin-conjugating enzyme (E2), ubiquitin (UBQ), zeitlupe (ZTL) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) under various abiotic stresses in perennial ryegrass (Lolium perenne L.). The analysis with geNorm (ver. 3.5), Normfinder (ver. 0.953), BestKeeper (ver. 1.0), Delta Ct and RefFinder revealed that under drought, salt, heat and ABA stresses, eIF4A gene was the most stable reference gene; under waterlogging stress, UBQ gene was the most stable reference gene. As a result, eIF4A and UBQ genes were selected as reference genes for perennial ryegrass under different abiotic stresses, which provides a foundation for gene expression investigation of perennial ryegrass.