Figure 4.

Expression profiles of wheat TaMET1 . A) Wheat RNA-seq. Data are expressed in Fragment Reads per Kilobase of Exon Model (FPKM) for each homoeologous group. FPKM were computed according to the following formula: FPKM = 10⁹ x (C/NL) where C is the number of mappable reads on a feature, N is the total number of reads in the experiment and L (length) is the sum of exonic sequences in base pairs. Genes were considered to be expressed only for FPKM values >0.15. Samples covering various tissues (root, leave, stem, spike and grain) and developmental stages indicated as Zadoks (Z) scales are indicated at the bottom. Expression levels from TaMET1 genes of a given homoeologous group were used to compute mean values. TaMET1 from group 2 are expressed at higher levels in comparison to the other MET1 copies and display two main peaks of expression in the stem (Z30) and spike (Z32). B) RT-PCR gel analysis. RT-PCR with (+) or without (−) reverse transcriptase was performed for all TaMET1 genes. Control PCR reactions were performed for two constitutively expressed genes corresponding to Ta4045 and Ta54227 selected according to Paolacci et al. 2009. C) Quantitative RT PCR. Reactions were performed for TaMET-2A1, 2B1 and 2D1. Expression is relative to Ta4045 (dark grey) and Ta54227 (light grey). D) Schematic representation of TaMET1 loci. Position of MET1 genes on chromosome maps of 2B, 5B and 7A (chromosome arms in grey and centromere as white ellipse, not to scale). Top and bottom positions on ITMI reference map are indicated in cM.