Article Figures & Data
Figures
- Figure 1.
Atxyn3::YFP marker gene expression. Wild type (A–F) and ate3 mutant (G and H) are shown. YFP signals were examined by fluorescent microscope (A, D, F, and H) and by a confocal laser scanning microscope (B and C). E and G are daylight images. In the wild-type seedlings, YFP fluorescence signals appeared along a line of the vasculatures of the root (A) and cotyledon (D and F). The YFP signal is detected in the nucleus of a developing TE (B), but it is lost during maturation of a TE (C). In the ate mutant, ectopic YFP signals appear independent of vasculatures (H). White arrows in F show YFP signals in the developing TEs. White arrowheads in H show ectopic YFP signals. Scale bars, 100 μm (A), 10 μm (B and C), and 1 mm (D–H).
- Figure 2.
Expression pattern of the Atxyn3::YFP marker gene and phenotypes of seedlings. Wild type (A, E, I, and O), ate1 (B, F, J, L, and P), ate2 (C, G, K, M, and Q), and ate3 (D, H, N, and R) are shown. Seedlings were grown for 1 d (A–D), 3 d (E–H), 10 d (I–N), 14 d (Q), and 21 d (O, P, and R). Arrows in H, L, M, and N show ectopic Atxyn3::YFP marker expression. Arrowheads indicate the ectopic Atxyn3::YFP marker gene expression (P, inset), and dying cell region (P). Scale bars, 100 μm (A–H) and 1 mm (I–R).
- Figure 3.
Observation of the lignin deposition in rosette leaves of 3-week-old plants. Wild type (A and B), ate3 (C–G), ate1 (H, I, K, and L), and ate2 (J and M) are shown. Lignin was visualized by phloroglucinol-HCl staining (B, D, G, and H–J) or UV illumination (A, C, and K–M). A region of the daylight image of ate3 leaf (E, inset) is magnified in F (YFP fluorescent) and G (phloroglucinol staining). The inset of G shows ectopically formed lignified cells with spiral secondary wall thickenings, which indicates TE formation. White arrows in D also indicate ectopically formed TEs with spiral lignin deposition. Scale bars, 25 μm (A–D) and 100 μm (F–M).
- Figure 4.
In vitro TE induction in the ate mutants. Hypocotyls of the wild type (A) and the ate1 mutant (B) at 4 d after treatment are shown. The frequency of TE transdifferentiation in the ate mutants is shown (C). Twenty-five hypocotyls were examined in each sampling point, and the sd was calculated from data of six independent experiments. Arrows indicate ectopic transdifferentiated TE cell. Scale bars, 50 μm (A–F).
- Figure 5.
pAt3g62160::YFP marker gene expression. Wild type (A and C) and the ate1 mutant (B and D) are shown. Note the ectopic distribution of fluorescence signals in the hypocotyl (B) and cotyledon (D) of the ate1 mutant. Scale bars, 500 μm (A–D).
- Figure 6.
Expression analysis of TE formation-related genes in the ate mutants. Two pairs of primers in the same reaction mixture were used in the quantitative RT-PCR experiment. One pair of primers was used to amplify the internal control gene, TUA4, while other gene-specific primers were used to amplify Atxyn3, XCP1, laccase (At2g38080), IRX3, or AGP (At1g03820). Only the TUA4 control for the AGP amplification is shown.
- Figure 7.
Genetic analyses of the ate1 mutant. Histochemical localization of GUS activity in the cotyledon of the 10-d-old wild type (A) and ate1 mutant (B) carrying the pDR5::GUS gene is shown. Cotyledon vein patterns of the 10-d-old pin1 (C), pin1 ate1 (D), van3 (E), and van3 ate1 (F) mutants are shown. The ate1 mutation was confirmed by ectopic YFP signals (data not shown). Light field (A–D) and dark field microscopy was used to clarify the discontinuous vein pattern (E and F). Scale bars, 500 μm (A–F).
