Validation of Novel Reference Genes for Reverse Transcription Quantitative Real-Time PCR in Drought-Stressed Sugarcane

Table 4

Relative expression rates of target genes (PFP𝛼1, AS, PIP1-1, and ACC oxidase) based on RT-qPCR with roots, cDNAs of sugarcane accessions under abiotic stress, and respective unitag regulation by SuperSAGE analysis covering drought^a stress (24 h of continuous dehydration) or salt^b stress (100 mM NaCl).

Unitag	Annotation	SuperSAGE [FC/Regulation*]		RT-qPCR^*&
		Tolerant	Sensitive	Tolerant	Sensitive
SD282748^a	AS	1.92^#/UR	−1.10^#/ns	1.473^#/UR	1.038^#/ns
D179780^a	PFP𝛼1	1.99^#/ns	−1.07^#/ns	0.756^#/ns	1.403^#/ns
ASS122537^b	ACC oxidase	1.95/UR	—	2.174/UR (30′); 1.830/ns (90′)	—
ASS140030^b	PIP1-1	−1.31/ns	—	0.805/ns (30′); 1.057/ns (90′)	—

AS: glutamine-dependent asparagine synthetase (EC 6.3.5.4); PFP𝛼1: pyrophosphate fructose-6-phosphate 1-phosphotransferase alpha subunit (EC 2.7.1.90); ACC oxidase: 1-aminocyclopropane-1-carboxylate oxidase (EC 1.14.17.4); PIP1-1: plasma membrane intrinsic protein. ^#Bulk with four accessions by each library; FC: fold change [ratio of the frequencies (normalized to 1,000,000) observed in the stressed library in relation to the control library]; ^&relative expression level by REST software (v.2.0.13) after the ΔΔCq method, *𝑃<0.05 [27]; UR: upregulated; ns: not significant at 𝑃<0.05. The time in the parentesis represents the salt stress exposition (min).