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Stipa grandis  P. Smirn. is a dominant plant species in the typical steppe of the Xilingole Plateau of Inner Mongolia. Selection of suitable reference genes for the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) is important for gene expression analysis and research into the molecular mechanisms underlying the stress responses of  S .  grandis . In the present study, 15 candidate reference genes ( EF1 beta ,  ACT ,  GAPDH ,  SamDC ,  CUL4 ,  CAP ,  SNF2 ,  SKIP1 ,  SKIP5 ,  SKIP11 ,  UBC2 ,  UBC15 ,  UBC17 ,  UCH , and  HERC2 ) were evaluated for their stability as potential reference genes for qRT-PCR under different stresses. Four algorithms were used: GeNorm, NormFinder, BestKeeper, and RefFinder. The results showed that the most stable reference genes were different under different stress conditions:  EF1beta  and  UBC15  during drought and salt stresses;  ACT  and  GAPDH  under heat stress;  SKIP5  and  UBC17  under cold stress;  UBC15  and  HERC2  under high pH stress;  UBC2  and  UBC15  under wounding stress;  EF1beta  and  UBC17  under jasmonic acid treatment;  UBC15  and  CUL4  under abscisic acid treatment; and  HERC2  and  UBC17  under salicylic acid treatment.  EF1beta  and  HERC2  were the most suitable genes for the global analysis of all samples. Furthermore, six target genes,  SgPOD ,  SgPAL ,  SgLEA ,  SgLOX ,  SgHSP90  and  SgPR1 , were selected to validate the most and least stable reference genes under different treatments. Our results provide guidelines for reference gene selection for more accurate qRT-PCR quantification and will promote studies of gene expression in  S .  grandis  subjected to environmental stress.

Selection of Reference Genes for qRT-PCR Analysis of Gene Expression in Stipa grandis during Environmental Stresses