Genome wide association study between segregate wild-type and homozygous mutant

Purpose: We want to identify the perception components of cellooligomer based on cytoplasmic calcium elevation in Arabidopsis root. Aequorin-expressing wild-type Arabidopsis seedlings in pMAQ background were mutagenized with EMS. Seedlings from the F2 generation was screened based on cytoplasmic calcium elevation in root tissue upon cellobiose treatment. The non-responsive EMS mutant was back-crossed to the Aequorin wild-type. The subsquent F2 generation was separated into two groups: 1. segregated wild-type and heterogygotes (responsive) 2. homozygoutes (non-responsive). DNA from 50 seedlings per group was extracted and sequenced on Illumina sequencer by Novogene Co. UK (PE150). SIMPLE (v. 1.8.1) was used to compare SNPs found in the two groups. Candidate genes were selected for further confirmation.