Supplementary table 1

Genes related to thermotolerance and their primers for qRT-PCR in Arabidopsis"

Fig. 1

Amino acid sequences of TaHsfA2e from wheat and structure domains DBD: conserved DNA binding domain of Hsf family; OD: oligomerization; NLS: nuclear localization signal; AHA: aromatic, large hydrophobic and acidic amino residues; NES: nuclear export signal."

Fig. 2

Subcellular localization of TaHsfA2e in onion epidermal cells under normal growth conditions A: bright field; B: green fluorescence of GFP; C: red fluorescence of DAPI; D: merged image."

Supplementary fig. 1

Expression levels of TaHsfA2e in different tissues and organs at seedling and anthesis stages under normal growth conditions There are three replicates for each sample and the data are mean ± standard error. The relative expression level of TaHsfA2e by qRT-PCR in young roots was set to 1 as controls."

Fig. 3

Changes of TaHsfA2e relative expression levels in leaves of wheat seedlings subject to heat stress at 37 °C (A), 0.8 mmol L-1 SA (B), and 10 mmol L-1 H2O2 (C) Each bar value represents ±SD of triplicate experiments. The relative expression level of TaHsfA2e by qRT-PCR of 0 h was set to 1 as controls."

Fig. 4

Thermotolerances of yeast harboring pYES2-TaHsfA2e and pYES2 after heat shock at 50°C A: culture at 30 °C; B: HS at 50 °C for 45 min, then culture at 30 °C for 3 days; C: OD600 of transformed yeast cells at 30 °C; D: OD600 of transformed yeast cells at 50 °C for 45 min and then culture at 30 °C. The error bar represents ±SD of triplicate experiments. * means significant difference between transgenic yeast cells and the controls (harboring pYES2) (P<0.05)"

Supplementary fig. 2

TaHsfA2e relative expression in different Arabidopsis TaHsfA2e transgenic lines by semi-RT-PCR The wild type (WT) was used as negative control."

Fig. 5

Assays of basal and acquired thermotolerance in three TaHsfA2e-transgenic Arabidopsis and its wild type (WT) The seeds of TaHsfA2e transgenic Arabidopsis lines (8_1, 12_2, and 14_19) and wild type were planted in plate, incubated for 3 d, and grew for 5 d. Then all seedlings were subjected to heat stress. A: plants under normal growth conditions; B: plants subjected to HS treatment of BT (45°C 40 min, recovered growth for 8 d at 22°C); C: plants subjected to HS treatment of AT (37°C 1 h, recovered growth for 2 d at 22°C, and 46°C 50 min, then recovered growth for 8 d at 22°C); D: survival rates of transgenic and wild type plants at normal conditions and subjected to BT and AT treatments. The represented values are the means of at least 15 individual plants of each line, and the experiment was repeated three times."

Fig. 6

Chlorophyll content (A) and REC (B) of transgenic Arabidopsis lines overexpressing TaHsfA2e and wild type subject to different heat shock regimes CK: normal condition; BT: heat shock regime for basal thermotolerance; AT: heat shock regime for acquired thermotolerance. REC: electrical relative conductivity. The represented values are the means of at least 15 individual plants of each line, and the experiment was repeated three times."

Fig. 7

Relative expression levels of eight Hsps in transgenic Arabidopsis line overexpressing TaHsfA2e subject to heat shock regimes for basal (BT) and acquired thermotolerance (AT) All values represent that at 0 h after heat stress in BT and at 2 h after heat stress in AT. Wild type subjected to same heat stress was used as controls. The represented values are the means of at least fifteen individual plants of each line, and the experiment was repeated three times."