Figure 4.

Disruption of TIE Genes Led to Epinastic Leaves.

(A) to (C) Phenotypes of 35S-TIE3-RNAi plants in the TIE1 null mutant tie1-455 background: epinastic true leaves (A), single cotyledon (B), or cup-shaped cotyledon (C). Bar = 1 mm.

(D) to (F) Expression of TIE1mEAR in wild-type Arabidopsis led to epinastic leaves.

(D) Top, schematic representation of TIE1mEAR protein. The three conserved Leu residues of EAR motif were mutated into Ser residues in TIE1mEAR. Bottom, left, 28-d-old wild-type plant. Bar = 1 cm. Right, 28-d-old TIE1mEAR-7 transgenic line. Bar = 2 mm.

(E) The close-up views of the 6th and 7th leaves from 28-d-old wild-type (left) and TIE1mEAR-7 plants (right). Bar = 1 mm.

(F) Transverse sections of leaves from 28-d-old wild-type (left) and TIE1mEAR-7 plants (right). Bar = 1 mm.

(G) Scanning electron micrographs of the leaf epidermal cells at the base of abaxial sides of the 5th leaves from 27-d-old wild-type (top) and TIE1mEAR-7 plants (bottom). Bars = 50 µm.

(H) Distribution of cell size of the leaf epidermal cells at the base of abaxial sides of the 5th leaves from 27-d-old wild-type and TIE1mEAR-7 plants.

(I) to (M) Expression of TIE1 chimeric protein of TIE1mEARVP16 caused severe cotyledon and leaf phenotypes.

(I) Top, schematic representation of TIE1mEARVP16 fusion protein. Bottom, 9-d-old seedlings from wild-type and TIE1mEARVP16 plants.

(J) and (K) Twenty-one-day-old plants from wild-type (J) and TIE1mEARVP16 (K) plants. Bar = 2 mm.

(L) Close-up views of leaves from 28-d-old wild-type and TIE1mEARVP16 plants. From left to right, the 5th leaf from the wild type and nine 5th leaves from nine independent TIE1mEARVP16 transgenic lines. Bar = 2 mm.

(M) The 8-d-old seedlings from wild-type and TIE1mEARVP16 plants. From left to right, two wild-type seedlings and three seedlings from three independent TIE1mEARVP16 transgenic lines. The first one displayed one cotyledon, the second one had two cotyledons fused, and the third one had three cotyledons fused. Bar = 2 mm.