Selection and validation of reference genes for gene expression analysis in apomictic and sexual Cenchrus ciliaris

Table 3 Tissue specific reference gene stability analyses of B-2S (sex) and B-12-9 (apo) using GeNorm and NormFinder

Genotype_Tissue type		GeNorm - M values						NormFinder_Gene ranking (SD values)
Samples		GeNorm - M values						NormFinder_Gene ranking (SD values)
		ACT2	EF1alpha	EIF4A	GAPDH	TUBA	UBCE	ACT2	EF1alpha	EIF4A	GAPDH	TUBA	UBCE
Ovary (early/tetrad/DOP)	Sex	0.23	0.30	0.19	0.19	0.47	0.37	1 (0.09)	4 (0.34)	3 (0.25)	2 (0.13)	6 (0.68)	5 (0.50)
	Apo	0.15	0.15	0.37	0.30	Oc	0.24	1 (0.07)	1 (0.07)	4 (0.48)	2 (0.22)	5 (0.87)	3 (0.38)
Ovary (early/tetrad/DOP/DOP + 5)	Sex	0.44	Oc	0.16	0.16	Oc	0.39	4 (0.53)	6 (1.06)	1 (0.07)	2 (0.08)	5 (0.64)	3 (0.39)
	Apo	0.20	0.20	0.32	0.27	Oc	0.38	2 (0.14)	1 (0.10)	4 (0.40)	3 (0.18)	6 (0.74)	5 (0.48)
Anther (early/tetrad/DOP)	Sex	Oc¹	Oc	0.47	0.47	Oc	Oc	5 (1.20)	3 (0.53)	2 (0.45)	1 (0.10)	5 (1.20)	4 (0.85)
	Apo	Oc	0.43	0.27	0.36	Oc	0.27	6 (0.82)	4 (0.64)	2 (0.19)	3 (0.63)	5 (0.72)	1 (0.13)
All tissues	Sex	Oc	Oc	0.38	0.38	Oc	Oc	4 (0.80)	5 (0.97)	1 (0.20)	2 (0.40)	6 (1.10)	3 (0.60)
	Apo	Oc	Oc	0.46	Oc	Oc	Oc	3 (0.84)	4 (0.86)	1 (0.50)	2 (0.60)	6 (0.95)	5 (0.90)

GeNorm stability value is represented by M value (the selected cut off for M value is ≤0.5). NormFinder stability value is represented by SD (standard deviation). There is no cut-off for SD, hence the ranking of genes based on the SD value is used as a measure to evaluate stability (1 stands for most stable, 6 is least stable). The ovary and anther stages are as follows: early stands for early premeiosis, tetrad, DOP for day of pollination, and DOP + 5 is 5 days after day of pollination.
¹Oc refers to value outside the cut-off range.

ISSN: 1756-0500