Selection of Reliable Reference Genes for Gene Expression Normalization in Sagittaria trifolia

Real-time quantitative PCR (RT-qPCR) is a method with high sensitivity and convenience that has been extensively used to analyze the expression level of target genes. A reference gene highly stable required ensure accuracy experimental results. However, report on appropriate genes in arrowheads (Sagittaria trifolia) still limited. In this study, eight candidate (ACT5, UBQ, GAPDH, CYP, NAC, IDH, SLEEPER PLA) were selected. The employed RT-qPCR assay different tissues at developmental stages same tissue (including corm, leaf leafstalk) arrowheads. Five statistical algorithms, GeNorm, NormFinder, BestKeeper, delta cycle threshold (ΔCt) RefFinder, evaluate stability these genes’ expressions order identify results showed UBQ was optimum leaf, leafstalk, root, stolon IDH exhibited most during expansion PLA respectively. Finally, reliability further confirmed by normalization PDS EXP1 under arrowhead This study constitutes important guidance for selection reliable analyzing tissue- developmental-stage-specific