Selection of Stable Reference Genes for Quantitative Real-Time PCR on Paeonia ostii T. Hong et J. X. Zhang Leaves Exposed to Different Drought Stress Conditions

Abstract

The definition of relatively stable expressed internal reference genes is essential in both traditional blotting quantification and as a modern data quantitative strategy. Appropriate internal reference genes can accurately standardize the expression abundance of target genes to avoid serious experimental errors. In this study, the expression profiles of ten candidate genes, ACT1, ACT2, GAPDH, eIF1, eIF2, α-TUB, β-TUB, TBP, RNA Pol II and RP II, were calculated for a suitable reference gene selection in Paeonia ostii T. Hong et J. X. Zhang leaves under various drought stress conditions. Data were processed by the four regularly used evaluation software. A comprehensive analysis revealed that RNA Pol II was the most stable gene and eIF2 was the least stable one. In addition, the geNorm program provided the optimal choice of two reference gene combination, RNA Pol II and β-TUB, for qRT-PCR normalization in P. ostii subjected to different drought stress levels. Our research provided convenience for gene expression analysis in P. ostii under drought stress and promoted research of effective methods to alleviate P. ostii drought stress in the future.

Selection of Stable Reference Genes for Quantitative Real-Time PCR on Herbaceous Peony (Paeonia lactiflora Pall.) in Response to Drought Stress

“…There were certain differences in each evaluation result in that the above 5 analysis methods adopt different algorithms. According to GeNorm Analysis, β-TUB and RNA Pol II were the optimal internal reference genes for P. ostii in response to different drought stresses [29]. It is different from P. ostii that even though P. ostii and P. lactiflora share 99% homology, the selections of reference genes are different in this research.…”

“…Many studies have emerged around the screening of reference genes in response to drought stress. In P. ostii, the most suitable reference gene was RNA polymerase II (RNA Pol II) under drought stress [29]. Eukaryotic initiation factor (eIF) and EF-1α are recommended as the steadiest ones in distinct tissues of grass pea [30] under drought stress.…”