Fig. 5

eGFP is strongly induced only at or above 38 °C. a–t One-week old barley seedings were subjected to a heat shock of 30 min, at one of five different temperatures: 20 °C (a–d), 26 °C (e–h), 32 °C (i–l), 38 °C (m–p), and 44 °C (q–t). The mCHERRY (i, magenta) and eGFP (ii, green) expression is shown for each tissue sample. Images of leaf 1 (a, e, i, m, q), leaf 2 (b, f, j, n, r), leaf 3 (c, g, k, o, s), and the meristem with attached leaf primordia (d, h, l, p, t) are shown for each temperature treatment. All plants were imaged 3 days following treatment. The scale bar is 100 µm. In 1-week old wheat seedlings, NAM-B1 is induced most strongly at 38 °C, though also to a lesser extent at 32 °C in the single copy line 2020-20-02 (u). Statistical comparisons were carried out using the Wilcox test due to the non-normality of the data, with N = 4 for all conditions; *p < 0.05, ns, p > 0.05. Gene expression in u was determined using qRT-PCR and is reported relative to the housekeeping gene TaActin (RE_Actin)