Table 1.
Descriptions of the 12 candidate reference genes and two selected genes for validation of results.
Table 2.
Primers, amplicon sizes, and qPCR efficiency for the candidate internal control and results validation genes.
Fig 1.
Distribution of the average Cq values of all 12 reference genes evaluated in black locust samples: 18SrRNA, ACT, CAC, CYP, EF1a, GAPDH, Helicase, PP2A, SAMDC, SAND, βTUB, and UBQ.
The linear line shows median, dots for extremes, boxes for 25%-75% Cq value and whiskers for 5%-95% respectively.
Fig 2.
Average expression stability values (M) of 12 candidate reference genes under different conditions and tissues calculated by geNorm.
Fig 3.
Estimation of the required number of reference genes for normalization by pairwise variation (V) using geNorm.
Table 3.
Average expression stability values of the 12 reference genes in different organs, calculated by NormFinder.
Table 4.
Ranking of 10 reference genes in order of their expression stability, as calculated by BestKeeper.
Fig 4.
Validation of NAC2 expression was assessed using individual reference genes (ACT, GAPDH or UBQ) under diverse abiotic conditions, (A) 4°C for 0, 12, 24 and 48h, (B) 40°C for 0, 12, 24 and 48h, (C) NaCl (200 mM), (D) ABA (150 mM), (E) PEG (100%), (F) Tissues (Root, Stem, Leaf, Flower and Pod).
Fig 5.
Validation of BGL2 expression was assessed using individual reference genes (ACT, GAPDH or UBQ) under diverse abiotic conditions, (A) 4°C for 0, 12, 24 and 48h, (B) 40°C for 0, 12, 24 and 48h, (C) NaCl (200 mM), (D) ABA (150 mM), (E) PEG (100%), (F) Tissues (Root, Stem, Leaf, Flower and Pod).
Fig 6.
Expression profile of NAC2 gene in different tissues and in response to different stressors in Robinia pseudoacacia as investigated by qPCR with ACT and GAPDH used as dual reference genes.
Table 5.
Ranking of the candidate reference genes according to their stability values using geNorm, NormFinder, and BestKeeper.