Nodule NGR234 rhcN- L1
Phaseolus vulgaris BAT93 seeds surface-sterilized, seeds were placed on B and D medium agar for germination in 25C for 2 days. Seeds were transferred to Magenta jars with vermiculite and B and D medium. Plants were inoculated 3 days after with Sinorhizobium fredii NGR234 strains and harvested 10 days after inoculation to collect nodule primordia or 21 days to collect mature nodules. Total RNA was purified from primordia and nodules using RNAqueous Total RNA Isolation Kit (Thermo Fisher Scientific). RNA integrity was evaluated in an Agilent 2100 Bioanalyzer System (Agilent Technologies) and RNA concentration was determined by NanoDrop ND-8000 spectrophotometer (Thermo Fisher Scientific). Enrichment of messenger RNA was obtained by depleting ribosomal RNA using RiboMinus Plant Kit for RNA-Seq (Thermo Fisher Scientific). Library construction and cDNA sequencing on a Life Technologies SOLiD 4 platform were performed according to the manufacturers instructions.