Differentially Expressed Genes in Wild-type Tobacco Leaves and NtCBL5A-OE Transgenic Tobacco Leaves Under Salt Stress

Purpose: To further identify the genes and pathways involved in the necrotic phenotype of NtCBL5A-OE lines, the leaf transcriptome profiling of WT and OE-2 lines grown under control conditions and salt stress (100 mM NaCl) at 4 DAT were sequenced and compared. Methods: Two datasets of differentially expressed genes (DEGs) were made in which we identified the genes that were differentially expressed as a result of the overexpression of NtCBL5A: Control-WT vs Control-OE2 (C-WT/C-OE2), Salt-WT vs Salt-OE2 (S-WT/S-OE2). Another two datasets were also used to identify the transcripts that were responsive to the salt treatments: Control-WT vs Salt-WT (C-WT/S-WT) and Control-OE2 vs Salt-OE2 (C-OE2/S-OE2). DEGs from C-WT/C-OE2 and S-WT/S-OE2 were compared to select the transcripts affected by NtCBL5A overexpression only under salt stress. We also compared DEGs from C-WT/S-WT and C-OE2/S-OE2 to identify the specific transcripts affected by salt stress and only in NtCBL5A-OE lines. This procedure was done for two independent experiments, and only DEGs that were identified in both experiments were considered. Results: The OE-affected DEGs and salt-affected DEGs together resulted in 2079 up-regulated DEGs and 1154 down-regulated DEGs, strongly affected by the combination of NtCBL5A overexpression and salt stress. Overall design: Tobacco leaf mRNA profiles of 4 days after salt treatment (100 mM NaCl in hydropoic system) in wild type (WT) and NtCBL5A-OE lines