Total RNA was extracted from B. glumae BGR1, BLONN (lon::Gm), and BLONC (lon::Gm/lon) grown in LB medium at 37°C for 10 h after subculture using RNeasy mini kits (Qiagen) following the manufacturer's protocols. Extracted total RNA was treated with RNase‐free DNase I (Ambion) to remove DNA. The quantity and quality of the total RNA were evaluated using RNA electropherograms (Agilent 2100 Bioanalyzer) and by assessing the RNA integrity number. From each sample with an RNA integrity number value greater than 8.0, 8 μg of total RNA was used as starting material and treated with the MICROBExpress mRNA Enrichment kit (Invitrogen). The resulting mRNA samples were processed for the sequencing libraries using the Illumina mRNASeq Sample Preparation kit (Illumina) following the manufacturer's protocols. One lane per sample was used for sequencing by the Illumina Genome Analyzer IIx (Illumina) to generate nondirectional, single‐ended, 36‐base‐pair reads. Quality‐filtered reads were mapped to reference genome sequences (NCBI BioProject accession: PRJNA59397 ID: 539397, http://www.ncbi.nlm.nih.gov/bioproject/59397 ) using the BWA package (Li & Durbin, 2009 (link)). The mRNA reads were normalized to reads per kilobase per million mapped reads (Mortazavi et al., 2008 (link)). The NCBI SRA accession number for the RNA sequencing data series of BGR1, BLONN, and BLONC is PRJNA727974.
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