RNA-Seq of Lens lamottei
To obtain leaves for RNA extraction, 2-3 seeds of each accession were germinated for 2-3 weeks in glasshouse at 22  2 C under a 16/8h (light/dark) photoperiod, in individual pots filled with standard potting mix at the premises of Agriculture Victoria, AgriBio Centre, Bundoora, Victoria, Australia. Fresh leaf tissues were sampled from single seedlings and immediately frozen in liquid nitrogen before storage at -80 C. Total RNA extraction was performed using the RNeasy 96 Plant Kit (QIAGEN, Hilden, Germany) following the manufacturers instructions. Using a NanoDropTM UV-Visible spectrophotometer (Thermo-Scientific, Wilmington, DE, USA), the concentration and quality of RNA was confirmed at the wavelength ratios of A260/230 and A260/280 nm. The integrity of extracted RNA samples was evaluated using TapeStation 2200 platform with RNA ScreenTape System (Agilent Technologies, Santa Clara, CA, USA), following manufacturers guidelines. RNA-Seq libraries were prepared using the Agilent SureSelect Strand Specific RNA Library Preparation Kit and evaluated on TapeStation 2200 platform with D1000 ScreenTape System (Agilent Technologies, Santa Clara, CA, USA). Each RNA-seq library was paired-end (2x151 bp) sequenced using Illumina Hiseq 3000 Sequencing platforms (Illumina Inc., San Diego, USA).