Fig 1.
Phylogenetic tree of common wheat, Brachypodium and rice TIFY proteins.
Phylogenetic tree was obtained using MEGA6.06 with the Neighbor-Joining method based on TIFY protein sequences. Wheat, Brachypodium and rice gene identifiers are indicated such as in Table 1. Wheat proteins are indicated by red dots, rice proteins by dark blue squares and Brachypodium proteins by green triangles. The red open circle represent durum wheat TdTIFY11a. Scale bar indicates evolutionary distances inferred using the Neighbor-Joining method calculated by the number of amino acid substitutions per site as conducted by MEGA6.06.
Table 1.
List of the common wheat (Triticum aestivum) TIFY genes and proteins identified.
Fig 2.
Conserved domains in wheat TIFY proteins.
Schematic representation of 49 wheat TIFY proteins and conserved domains drawn with GPS tool (A). Blue boxes represent the TIFY domain, yellow highlight the Jas domain, green stand for divergent CCT motifs and red represent C2C2-GATA-Zinc-finger DNA binding domain. EAR motif is shown by black box. Grey bars represent non-conserved sections. The scale at the bottom (indicating the number of amino acid) corresponds to the proteins length. Consensus of sequences conservation of TIFY/ZIM domain (B), Jas motifs (C) and GATA domain (D) using MEME.
Fig 3.
Expression pattern of 6 durum wheat TIFY genes in response to stresses.
Seven-day-old durum wheat seedlings were treated with NaCl (150 mM), JA (50 μM) or ABA (100 μM) for 1 or 6 hours. Relative expression of TIFY genes was analyzed by quantitative real-time qPCR using wheat actin as control. Log2 transformed values were used to generate the color-coded heatmap. The color-coded scale bar is indicated below the heatmap.
Fig 4.
Over-expression of TdTIFY11a variants confers salt tolerance to Arabidopsis seedlings.
(A) 7-day-old seedlings (N = 20 to 50) of the different over-expressing lines germinated on control media or in presence of 100 or 150 mM NaCl. Percentage of radicle emergence of 2 day-old seedlings (B), percentage of seedlings with green cotyledons at day 5 after germination (C) and on control media or supplemented with NaCl (100 or 150 mM). Data presented as box-plots; horizontal lines are medians, boxes show the interquartile range and error bars show the full data range. The experiments were repeated at least 4 times with similar results. Asterisks (B-C) indicate statistical significance (One-way ANOVA with post-hoc Tukey HSD, * p<0.05, ** p<0.01).