Whole Genome Bisulfite Sequencing in TaMET1-1 mutants
Mutants in TaMET1-1 were obtained from the Cadenza TILLING population. The parental lines were Cadenza0465 and C0884. We collected leaf samples from the fourth leaf at Zadok's growth stage Z15.6/23 for wild type segregant, single and double mutants and Cadenza WT, or from the sixth leaf at Z16 for the Aabbdd mutant. DNA was extracted using the DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) from three biological replicates per genotype, excluding the Aabbdd genotype, of which only one plant was recovered. DNA concentration was estimated for each sample using an Invitrogen Qubit 4 fluorometer (Thermo Fisher Scientific, Massachusetts, USA) and the three replicates per genotype were pooled in equimolar ratios. Library preparation, bisulfite conversion, and sequencing by Illumina NovaSeq 6000 was performed by BMKgene (Münster, Germany), to obtain 150 bp paired-end reads.