Figure 1.
Characterization of dil1 mutant phenotype.
A. dil1 mutant plants (9-weeks-old) are characterized by their reduced plant height and smaller leaf angles as compared to the wild type plants. B. Stalks of wild type and dil1 mutant plants (9-weeks-old) with leaves and ears removed. C. Wild type (upper) and dil338 mutant (lower panel) seedlings six days after germination in dark. D. Abaxial surface of the first leaf below the ear in 9-weeks-old wild type and dil1 mutant plants. E and F. Close look of abaxial (E) and adaxial (F) surfaces of the first leaf below the ear in 9-weeks-old wild type and dil1 mutant plants.
Table 1.
Comparison of leaf angle, leaf length and width, internode length and plant height between wild-type and dil1 mutant plants.
Figure 2.
Cytological observations of dil1 mutant.
A and B: Multiphoton laser scanning micrograph (LSM) images of the adaxial surface of leaf epidermis from the 2nd leaf of V3 stage wild type (WT) (A) and dil338 (B) seedlings. C and D: Multiphoton LSM images of the adaxial surface of leaf epidermis at post-flowering stage in WT (C) and dil338 (D) plants. E and F: Multiphoton LSM images of stalk parenchyma cells from third internode (from base) at post-flowering stage in WT (E) and dil338 (F) plants. G and H: Multiphoton LSM images of stalk parenchyma cells from ninth internode (from base) at post-flowering stage in WT (G) and dil338 (H) plant. I and J: The adaxial surface of WT leaf with flat blade and regularly spaced macrohairs (I), and dil338 with an undulating leaf surface and clustered macrohairs (J, arrows), from fully expanded leaves. K and L: Stomata in the adaxial surface of WT leaves with paired subsidiary cells (K), whereas in dil338 the stomata sometimes had single subsidiary cells (arrows) (L).
Figure 3.
Fine mapping and cloning of dil1.
A. DIL1 was mapped on the long arm of chromosome 6, between Brugi030 and IDP8945. B. B73 BAC contig covering the DIL1 locus. C. Recombination events near DIL1 delimited DIL1 between c0137A18-A2 and c0427D16-D1. A: wild type (A632) genotype. B: mutant (PHN4611) genotype. H: heterozygous genotype.
Figure 4.
Single base pair mutations in dil1 mutants cause alternative splicing.
A. Structure of DIL1 gene and positions of single base pair mutations in dil338 and dil474 (in red and labeled with asterisk). B. Proportion of transcript variants in wild type and dil1 mutants.
Figure 5.
Expression analysis of DIL1 gene.
A. DIL1 mRNA levels in V3 leaves in wild type and mutant plants, as determined by RT-PCR with 30 cycles for DIL1 (Upper), with the actin gene as the control (Lower). B-D. RNA in situ hybridization on tissue sections from A619 wild type vegetative apices, showing expression of DIL1 in the leaf primordia and the P0 region of the meristem (B and C). Hybridization signal was not observed with control, sense probes (D).