Figure 3.

Constitutive expression of GFP-REIL and RFP-REIL fusion proteins under the control of the UBQ10 promoter in reil1-1 reil2-1 restores single mutant morphology and development. Seedling development and vegetative rosette morphology of representative reil1-1 reil2-1 transformation events using the constructs UBQ10::GFP-REIL2 (b), UBQ10::RFP-REIL2 (c), UBQ10::GFP-REIL1 (e), and UBQ10::RFP-REIL1 (f) were compared with reil1-1 (a) and reil2-1 (d) plants. A, Seedlings after in vitro germination and cultivation at 10°C. Representative photographs of n ≥ 10 in vitro-grown plants per genotype were taken 30 d after imbibition (see C and D). B, Rosette plants after in vitro germination at 10°C and constant-temperature cultivation on soil at 10°C/8°C (day/night) according to cultivation scheme B in Figure 1. Representative photographs of n ≥ 10 plants per genotype were taken 6 to 7 weeks after transfer to soil. Racks of 6-cm-square pots were used. C, Appearance of the first juvenile rosette leaves after the introduction of REIL2 fusion proteins into reil1-1 reil2-1 (means ± sd of n = 4–5 plates per genotype; each plate had ∼80 seeds). D, Appearance of the first juvenile rosette leaves after the introduction of REIL1 fusion proteins into reil1-1 reil2-1 (means ± sd of n = 4–5 plates per genotype; each plate had ∼80 seeds). Leaf appearance was scored using the 10°C in vitro germination and cultivation assay described by Schmidt et al. (2013). The analysis included the reil1-1 reil2-1 mutant that is strongly delayed for the appearance and development of rosette leaves. Arrows indicate the absence of the first rosette leaf (compare with Fig. 1B), Arabidopsis Col-0 wild type (WT), and a nonfusion protein control, namely UBQ10::REIL2, transformed into reil1-1 reil2-1 (see C). The introduction of REIL1 restored reil2-1 morphology, and the introduction of REIL2 restored reil1-1 morphology. Note that reil1-1 morphology is virtually indistinguishable from that of the wild type (Schmidt et al., 2013).