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. 2020 Sep 12;11(9):1073. doi: 10.3390/genes11091073

Figure 2.

Figure 2

Figure 2

Expression heat map of 25 selected TaNAC genes under stress due to stripe rust or powdery mildew. (a) Pst treatment; (b) Bgt treatment. (left) Expression determined by RNA-seq, and values represent fold change of average reads per kilobase of transcript per million mapped (RPKM) across three replicate samples; (right) expression of corresponding TaNAC genes according to qRT-PCR, and value is relative expression calculated using 2−ΔΔCt. S: stripe-rust stress; P: powdery-mildew stress; 0, 24 h, etc.: time interval between sampling and inoculation. Group I contained genes that were differentially expressed under both Pst and Bgt stress. (a) Group II contained genes that were non-DE under Pst stress, but DE under Bgt stress; (b) Group II contained genes that were non-DE under Bgt stress, but DE under Pst stress; genes in Group II were determined to be DE under all fungal stress. Group III contained genes that were non-DE under both Pst and Bgt stress. Notes: TaNAC015_3A and TaNAC020_2A, marked in green in Figure 2a, were upregulated both in RNA-seq and qRT-PCR but with inconsistent expression patterns; TaNAC001_7A, marked in yellow in Figure 2a, was upregulated in RNA-seq and downregulated in qRT-PCR, but with consistent expression patterns; TaNAC047_7D, marked in red in Figure 2b, was upregulated in RNA-seq and downregulated in qRT-PCR, and with inconsistent expression patterns. The four sets of comparison result above showed no change in DE classification attribute. Six other sets of comparison results, marked in blue—namely TaNAC018_7B and TaNAC024_3A in Figure 2a, and TaNAC006_3A, TaNAC018_7B, TaNAC020_2A, and TaNAC001_7A in Figure 2b—were non-DE in RNA-seq but DE in qRT-PCR, or the opposite. They all showed inconsistent expression patterns. However, upon combining the two sets of RNA-seq and qRT-PCR comparison results between Pst stress and Bgt stress, respectively, there was no change in DE classification attribute under fungal stress for any gene. In addition, only TaNAC013_3A.1, marked with a red star, showed a changed classification attribute from non-DE in RNA-seq to DE in qRT-PCR, with inconsistency of specific expression patterns under fungal stress.