Supplementary data Comparison of transcriptomes of chilling- and drought-tolerant and intolerant Nicotiana tabacum

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1 POJ 7(6): (214) ISSN: Supplementary data Comparison of transcriptomes of chilling- and drought-tolerant and intolerant Nicotiana tabacum varieties and identification of genes associated with stress tolerance Dahai Hao, Wenguang Ma, Yelong Sheng, Jianbo Zhang, Yunfeng Jin, Huiqin Yang, Zhongguang Li, Shasha Wang, Ming Gong * Supplementary Figures A

2 B Supplementary Figure 1 The length distribution of unigenes (A) and non-redundant unigenes (B) of Nicotiana. tabacum cv. Yunyan23 and MSK326.

3 Supplementary Figure 2. Unigenes of chilling and drought stressed N. tobacco associated with glutathione metabolic pathway for revealed unigenes by KEGG (from:

4 Supplementary Figure 3. Gene ontology (GO) enrichment analysis. The digital gene expressions (DGEs) of MSK326 (stress tolerance) and Yunyan23 (stress intolerance) cultured under control conditions. In this plot, the p values of all GO terms were obtained using Pearson chi-square test; the gene numbers between these two DGE libraries and were all significant.

5 Supplementary Figure 4. Gene ontology (GO) enrichment analysis of common down-regulated differentially expressed genes (DEGs) under chilling stress. Comparison of GO enrichment between the chilling stress-tolerant MSK326 and chilling stress-intolerant Yunyan23. The p values of all GO terms were obtained using Pearson chi-square test; the gene numbers of the two digital gene expression (DGE) libraries were all significant.

6 Supplementary Figure 5. Gene ontology (GO) enrichment analysis of common differentially expressed genes (DEGs) of drought-stressed samples. Comparison of GO enrichment analysis of drought-tolerant MSK326 and drought-intolerant Yunyan23. The p values of all GO terms were obtained using Pearson chi-square test; the gene numbers of the two digital gene expression (DGE) libraries were all significant.

7 Supplementary Figure 6. Differences between common differentially expressed genes (DEGs) of chilling and drought stress samples. The p values of all gene ontology (GO) terms were obtained using Pearson chi-square test; the gene numbers of the two digital gene expression (DGE) libraries and were all significant.

8 Multiple expressed genes in control CYP Msk326-drought-CYP Multiple expressed genes in control Yunyan23-chilling-CYP Yunyan23-drought-CYP 12 h 24 h 48 h 12 h 24 h 48 h 35 vs -MMT vs -MMT 3 Multiple expressed genes in control Multiple expressed genes in control Yunyan23-chilling-MMT Yunyan23-drought-MMT 12 h 24 h 48 h 12 h 24 h 48 h Supplementary Figure 7. Comparison of cytochrome P45 (CYP) and myricetin O-methyltransferase 2 (MMT) expression between control and stress-treated N. tabacum samples.

9 FPKM of Ø-6 fatty acid desaturase Yunyan23-chilling Yunyan23-drought FPKM of Ø-3 fatty acid desaturase Yunyan23-chilling Yunyan23-drought h 12h 24h 48h 2 h 12h 24h 48h Supplementary Figure 8. Expression pattern of fatty acid desaturase (FAD)6 and FAD3 of in chilling- and drought-treated N. tabacum samples MSK326-Chilling Yunyan23-Chilling MSK326-Drought Yunyan23-Drought FPKM of aquaporins h 12h 24h 48h Supplementary Figure 9. The sum of FPKM for aquaporins in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped.

10 1 8 Yunyan23-chilling Yunyan23-drought FPKM of LEA h 12 h 24 h 48 h Supplementary Figure 1. The sum of FPKM for late embryogenesis abundant (LEA) 5 proteins in 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped. 6 5 Yunyan23-chilling Yunyan23-drought FPKM of dehydrin h 12 h 24 h 48 h Supplementary Figure 11. The sum of FPKM for dehydrin (DHN) proteins in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped.

11 1 8 Yunyan23_chilling Yunyan23_drought FPKM of P5CS h 12h 24h 48h Supplementary Figure 12. The FPKM for 1 -pyrroline-5-carboxylate synthase (P5CS) in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped. FPKM of trehalose-6-phosphate synthase Yunyan23-chilling Yunyan23-drought h 12 h 24 h 48 h Supplementary Figure 13. The FPKM for trehalose-6-phosphate synthase in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped.

12 FPKM of Unigenes SOD -SOD -APX -APX h 12 h 24 h 48 h FPKM of Unigenes 22 Yunyan23-chilling-SOD Yunyan23-drought-SOD 2 Yunyan23-chilling-APX Yunyan23-drought-APX h 12 h 24 h 48 h 6 5 -CAT -CAT 6 5 Yunyan23-chilling-CAT Yunyan23-drought-CAT FPKM of Unigenes FPKM of Unigenes h 12 h 24 h 48 h h 12 h 24 h 48 h Supplementary Figure 14. The FPKM for superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped Yunyan23-chilling Yunyan23-drought FPKM of DREBs h 12 h 24 h 48 h Supplementary Figure 15. The FPKM for dehydration responsive element binding (DREB) proteins in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped.

13 NAC Yunyan23-chilling-NAC -NAC Yunyan23-drought-NAC FPKM of Unigenes h 12 h 24 h 48 h Supplementary Figure 16. The FPKM of NAC in the 14 digital gene expressions (DGEs) of N. tabacum. FPKM, fragments per kilobase of exon per million fragments mapped. Supplementary Figure 17. Some common reaction of N. tabaccum cv. Yunyan23 (intolerant) and MSK326 (tolerant) to drought/chilling stress.

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