WGS of Setaria viridis' beta-Carbonic anhydrase mutants
The M5 progeny of "Low CO2 responsive mutant 1" (lcr1) was crossed with wild-type (WT) plants, the latter as pollen donor, to obtain BC1F1 generation (following protocol of Jiang et al., 2013). BC1F1 lines were checked for a recovery of Fv/ Fm value in true F1, and then selfed to produce BC1F2 segregating population, which was used for whole genome sequencing. From a BC1F2 population of 300 plants, equal quantities of genomic DNA were pooled from 45 individuals exhibiting the lcr phenotype (homozygous mutant pool) and 54 individuals without the lcr phenotype (azygous or heterozygous pool). Total genomic DNA was extracted from young leaves of mid-tillering plant using CTAB protocol (Cetyl Trimethyl Ammonium Bromide protocol; Murray and Thompson, 1980). Genomic DNA was also pooled from 50 WT plants (WT pool). These pooled samples were sequenced using Illumina HiSeq2000 platforms at Beijing Genome Institute (BGI Tech Solutions (Hongkong) Co. Limited, Shenzhen, China). The reads were paired end (PE) of size 125 base pairs. Number of paired-end reads generated were ~108.1 millions for WT pool, ~91 millions for azygous pool and ~107.3 millions for mutant pool, yielding 23-33 Giga-bases of sequence data with a genome coverage of 45-65X for each.