Barley Gene Expression Microarray Analysis

The synthesis, labeling, and hybridization of cDNA and cRNA were performed by the Genomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany. The microarray data were analyzed using GeneSpring GX 13.0 software (Agilent Technologies) as described earlier (Daszkowska-Golec et al., 2017 (link)). A gene was considered to be differentially expressed when the level of its expression differed between the analyzed conditions by at least two times (fold change (FC) ≥ 2; P ≤ 0.05 after FDR correction). The annotation of the Agilent Barley Gene Expression Microarray (Agilent Technologies) was performed against IBSC_v2 of barley genome deposited in Ensembl Plants (v. 45). Functional annotation of differentially regulated genes was carried out using Ensembl Plants tools and the IPK Barley BLAST Server as references (https://plants.ensembl.org/index.html, https://webblast.ipk-gatersleben.de/barley_ibsc/). Three biological replications were used for microarray expression analysis (each biological replicate represented leaf of one seedling).