Fferentially expressed genes in four sorts of comparisons in WT and VaNAC26-OE plants under standard situations and drought tension. (A) and (B) show the numbers of overlapping upregulated and downregulated genes, respectively. The numbers in brackets represent the total numbers of differentially expressed genes in distinct comparisons.target of VaNAC26. To confirm the NACRS-binding capability of VaNAC26, the coding region of VaNAC26 was ligated towards the yeast expression vector pGADT7 to make a recombinant plasmid pGADT7-VaNAC26, along with a 4 tandem repeated NACRS motif (CACGCATGTG) and its mutant sequence (CAttttTGTG), which was substituted for four bases (lower letters) compared with NACRS, had been ligated to pAbAi (Fig. 9A). AbA is usually a cyclic depsipeptide antifungal agent with activity against yeast cells (Takesako et al., 1991). The AbA resistant gene URA-3 was integrated into Y1HGold yeast by the pAbAi vector, and it was made use of as a reporter gene to screen for putative binding activity of protein NA interactions. The result (Fig. 9B) showed successfully transformed Y1Hgold grew on SD-LEU-URA medium, and only the good control and those cotransformed with VaNAC26 and NACRS could develop on AbA-containing medium (Fig. 9C), indicating that VaNAC26 could bind to NACRS but not its mutant sequence.Endogenous JA content elevated in VaNAC26-OE lines and drought-treated V. amurensisJA is an significant signaling molecule within a plant’s defense against biotic and abiotic stresses (Sasaki-Sekimoto et al.,2840 | Fang et al.Table 1. Pathway enrichment analysis of 4 types of comparisons from WT and OE microarrays beneath standard and drought strain conditions.OE0d vs WT0d NF five.33 4.63 three.85 three.69 3.6 three.54 three.38 three.16 two.43 p-value 0.017 4.952E-10 0.201 0.039 0.01 0.005553 9.141E-05 0.231 0.025 OE5d vs WT5d NF 1.02 2.61 1.11 1.42 1.03 two.9 two.18 1.82 1.99 six.59 p-value 0.273 three.06E-09 0.369 0.158 0.177 7.05E-05 four.93E-05 0.202 three.34E-03 0.033 OE5d vs OE0d NF 1.43 1.58 1.33 1.98 1.66 2.25 2.01 1.82 two.24 3.95 5.27 2.84 two.84 two.three two.27 1.42 10.32 4.05 3.04 two.33 1.56 0.74 0.52 0.46 0.38 0.088 0.194 0.112 0.064 0.114 0.353 0.044 0.126 0.192 0.43 three.03 0.36 four.3 0.93 0.67 1.55 1.92 0.74 0.56 0.19 0.0002963 0.0000714 0.78 0.68 0.44 0.17 0.233 two.56E-05 eight.98E-03 four.94E-08 0.07 0.051 0.031 0.312 0.352 three.46E-04 two.47E-03 0.239 1.20E-09 0.93 two.81 1.34 1.12 1.03 1.05 1.86 0.97 0.76 3.27 0.73 0.93 0.17 0.07 0.73 0.46 0.273 1.47E-04 0.039 0.06 0.116 0.04 2.90E-07 0.086 0.358 four.25E-04 3.92E-10 0.029 0.02 8.55E-31 0.192 0.251 p-value 0.1 six.18E-05 0.203 6.76E-03 9.11E-03 9.31E-06 three.07E-08 0.083 1.21E-07 0.033 0.048 0.253 0.253 0.287 0.046 0.355 WT5d vs WT0d NF 1.8 1.59 two.44 1.4 1.77 two.17 two.21 three.2 1.61 three.61 5.78 three.11 1.55 1.26 1.75 1.55 0.65 0.76 3.08 0.92 1.72 1.18 0.82 2.27 0.95 1.26 2.61 0.71 0.94 0.58 0.06 0.93 0.51 p-value 8. 91E-03 7.64E-08 five.15E-03 0.036 1.98E-04 1.77E-08 three.08E-18 2.61E-05 5.64E-05 8.97E-03 three.58E-03 0.109 0.35 0.37 0.059 0.235 0.335 0.203 1.06E-08 0.075 two.74E-07 0.058 2.26E-03 7.37E-22 0.063 0.213 2.78E-04 1.32E-19 0.019 0.057 six.64E-57 0.151 0.Groups IPathways Nucleotide metabolism Misc Metal handling Amino acid metabolism Secondary metabolism Hormone metabolism Strain Main CHO metabolism Improvement Biodegradation of Xenobiotics Fermentation Gluconeogenesis glyoxylate cycle S-assimilation Polyamine metabolism Co-factor and vitamine metabolism N-metabolismIIOPP TCA org transformation Redox Cell wall Transport Lipid metabolism RNA Signalling Cell Tetrapyrrole TCID manufacturer synthesis M.