Table 1.
Candidate reference genes, primers and amplicons.
Table 2.
Primer efficiencies and correlation of reference gene candidates.
Fig 1.
RT-qPCR Cq values of candidate reference genes in all treatments.
The box includes all data points between the 25% quantile and the 75% quantile. The whiskers span all values below and above, excluding outliers which are displayed as dots. The median, which marks the 50% quantile, is displayed as a line.
Table 3.
Ranking of gene expression stability under stress conditions and hormone stimuli.
Genes were ranked using the three commonly used statistical algorithms NormFinder, BestKeeper and geNorm. The stability value describes the variance (NormFinder and geNorm) or standard deviation (BestKeeper).
Fig 2.
Optimal number of reference genes for various conditions.
The geNorm algorithm was used to determine the pairwise variation (V) between the reference genes for treatments with cold, drought, heat, salt and gibberellic acid. The threshold for adequate normalisation is V≤0.15, indicated by the green dashed line.
Table 4.
Abiotic stress and hormone responsive genes, primers and amplicons.
Fig 3.
Comparison of specific stress response genes normalised with different reference genes.
Normalisation of the stress response genes RD29A (RESPONSIVE TO DESICCATION 29A, cold and drought responsive), HSP81.2/90 (heat responsive), TSPO (OUTER MEMBRANE TRYPTOPHAN-RICH SENSORY PROTEIN-RELATED, salt responsive) and GA3ox1 (GIBBERELLIN 3-OXIDASE 1, GA responsive) was carried out with one or two reference genes (RG1 and RG2): cold–RAN3 and HCF, drought–UBQ10 and TUA5, GA and salt–PSB33 and TUA5, heat–RAN3 and PSB33.