Fig 1.
Annotation of the scaffolds containing (a) TPS and (b) TRE genes in Adineta vaga.
Color-filled rectangles represent genes, numbers refer to the list at the bottom of the figure. Genes were colored in green or red if AI<45 or AI≥45, respectively. The genes annotated automatically in the A. vaga genome are surrounded with continuous lines, whereas discontinued lines surround additional genes detected during manual curation. The gene orientation is indicated by the position above (forward direction) or below (reverse direction) the line.
Fig 2.
(a.) 3D alignment of the OtsA chain B crystal structure obtained for E. coli (white) with the two protein structures of TPS1 modeled using SWISS-MODEL (light blue) and PHYRE2 (orange).
UDP (green) and G6P (red) are localized as observed in the OtsA crystal structure. (b.) Close-up on the conserved sites involved in the binding of G6P (red). The residues of the E. coli OtsA are shown in white and those of the AvTpsA predicted using SWISS-MODEL in light blue. SwissModel TPS1 A. thaliana (light green) and SwissModel predicted TPS1 P. brassicacea (gold) are also displayed in order to compare their topologies. (c.) Close-up on the conserved sites related to the binding of UPD (green). The residues are labeled based on the 3D structure of AvTpsA predicted using SWISS-MODEL.
Table 1.
Summary of top BLASTP hits for the trehalose-6-phosphate synthase (TPS) and trehalase (TRE) genes of Adineta vaga against GenBank.
Fig 3.
Maximum-likelihood phylogenetic tree based on the amino acid sequences available for trehalose-6 phosphate synthase (TPS) domains.
The tree displayed was generated using RaxML with 1000 rapid bootstrap according to the model WAG+I+G+. Numbers above the branches are bootstrap support percentages and posterior probability of 1 obtained with MrBayes (See M&M). When the node was not recovered in the Bayesian tree, the bootstrap value was replaced by “-“. Bootstrap support values above 70% for MetaPIGA and NJ are indicated by lines respectively drawn above and below the ML bootstrap values and posterior probabilities*. Color code: green (plant), red (metazoans), black (bacteria and archeae), cyan (protists) and brown (fungi).
Fig 4.
Maximum likelihood phylogenetic tree based on the amino acid sequences available for trehalase (TRE) domains.
The tree displayed was generated using RaxML with 1000 rapid bootstrap according to the LG+I+G evolutionary model. Numbers above the branches are bootstrap support percentages and posterior probability of 1 obtained with MrBayes (See M&M). When the node was not found in a Bayesian tree, the bootstrap value was replaced by “-“. Bootstrap support values above 70% for MetaPIGA and NJ are indicated by lines respectively drawn above and below the ML bootstrap values and posterior probabilities. Color code: green (plant), red (metazoans), black (bacteria and archeae), cyan (protists) and brown (fungi).
Fig 5.
Relative expression of the trehalose-6-phosphate synthase (TPS) and trehalase (TRE) genes in Adineta vaga submitted to drying or rehydration: (a.) AvTpsA&A’; (b.) AvTreA&A”; (c.) AvTreC&C’ (d.) AvTreB&B’.
The five time points are as follows: (1) hydrated; (2) 28h drying/humidity effect; (3) 37h drying/early desiccated state; (4) 5min rehydration after 14 days of desiccation; (5) 1h30 rehydration after 14 days of desiccation; (6) 3h rehydration after 14 days of desiccation. The expression level of each gene was normalized by reference to the housekeeping gene for the ribosomal protein L40; relative level of expression changes were calculated by reference to that that of the hydrated time point (value = 1). Error bars represent mean value 95% confidence intervals based on three replicates (one-way analysis of variance RM, *P<0.05 ** P<0.01 *** P<0.001).