Article Figures & Data
Figures
- Figure 1.
Transcript levels of RGL1, RGL2, RGL3, RGA, and GAI throughout development. The level of gene expression in each tissue sample, as determined by qPCR, is shown as the number of cDNA copies per 103 copies of UBQ11 cDNA. The tissues and developmental stages tested were, from left to right in the graph, 12-h- and 24-h-imbibed seeds, 2-d-old and 5-d-old seedlings, 10-d-old shoots, roots of 10-d-old seedlings, 14-d-old rosettes, and the following tissues from 33-d-old plants: rosette leaves, cauline leaves, stems, flower clusters, and siliques. The means of three experiments ±se are shown. Although UBQ11 mRNA levels are constant in most tissues, they are approximately 3- to 4-fold higher in 33-d-old rosettes, flowers, stems, and cauline leaves (Supplemental Table I). Therefore, the expression levels of the DELLA protein genes in this figure are slightly underestimated in these tissues.
- Figure 2.
Locations of the T-DNA insertion sites in the new alleles of rga and rgls. The schematic diagram of each gene shows the encoded functional domains and motifs as labeled: DELLA, DELLA motif; S/T, polymeric Ser and/or Thr; LR, Leu heptad repeat; NL, nuclear localization signal. The numbers refer to the nucleotide length of the coding region of each gene and the insertion site of each T-DNA. n.t., nucleotide.
- Figure 3.
Germination of rgl2 seeds is resistant to PAC. Seeds of wild-type plants (WT) and of homozygous single (A) and double (B) rgl mutants were treated with 120 μm PAC. The germination percentages in A and B are the means of three separate experiments. Error bars indicate the se of the mean. For each experiment, approximately 100 to 160 seeds were scored per genotype.
- Figure 4.
Phenotypic effects of loss-of-function rga and rgl mutations in a GA-deficient background. A, ga1-3 and rgl mutant combinations without rga-28. B, ga1-3, rgl, and rga-28 mutant combinations and a wild-type control. C, Primary inflorescence stems of mutants and wild type, as labeled. D, Flowers of mutant plants and wild type, as labeled. The front-most floral organs (sepal, petal, and in some cases stamen) were removed to expose the interior of the flower. Wild-type and homozygous mutant plants were grown on soil under long-day conditions. The plants shown in panels (A) and (B) are 73 d old, except for wild type, which is 44 d old.
- Figure 5.
Effect of the rga, rgl1, and rgl2 mutations on fertility in a ga1-3 background. A, Percent of fertile siliques on the primary inflorescence. B, Number of seeds per fertile silique on the primary inflorescence. Seeds were counted for a minimum of 17 fertile siliques for each wild-type plant and for all possible fertile siliques for each mutant plant. Both A and B show the means ± se for 14 to 16 plants per genotype.
- Figure 6.
SLY1 regulates GA-induced RGL2 degradation. A and B, protein or RNA was isolated from imbibed seeds of various homozygous mutants as labeled. Before harvesting the tissues, the seeds were imbibed in water for 2 d, and then treated for 5, 12, or 24 h with 10 μm GA4 (+) or water (–). A, The protein blot contains 60 μg total proteins from imbibed seeds of homozygous mutants (±GA for 5 h) and was probed with polyclonal anti-RGA antibodies from rat. B, Relative RGL2 mRNA levels determined by qRT-PCR. At least three reactions were performed for each sample, and the RGL2 mRNA level was normalized using 18S rRNA. The means ± se are shown. The value of RGL2 mRNA in 5-h water-treated ga1-3/rga/rgl1 is arbitrarily set to 1.0.
Tables
Genotype Days to Flower Maximum Rosette Radius Final Height mm cm ga1-3 42.4 ± 1.5 16.1 ± 0.4 1.50 ± 0.16a ga1-3/rgl1 38.6 ± 1.1 16.4 ± 0.5 2.31 ± 0.23a ga1-3/rgl2 43.8 ± 1.9 16.0 ± 0.2 1.92 ± 0.25a ga1-3/rgl1/rgl2 38.8 ± 1.3 15.5 ± 0.5 2.32 ± 0.26a ga1-3/rga-28 27.0 ± 0.5 21.9 ± 0.5 18.21 ± 0.31 ga1-3/rga-28/rgl1 29.1 ± 0.4 24.6 ± 0.9 20.18 ± 0.30 ga1-3/rga-28/rgl2 29.8 ± 0.4 19.7 ± 0.6 17.08 ± 0.32 ga1-3/rga-28/rgl1/rgl2 29.5 ± 0.5 23.8 ± 1.4 17.13 ± 0.36 Wild type 20.2 ± 0.5 27.8 ± 0.8 32.32 ± 0.59 The values shown are means ± se for 11–18 plants per genotype.
↵a Height 105 d after planting, rather than the final height. Unlike the mutants homozygous for rga-28, which all began bolting immediately after flowering, these plants did not begin bolting until day 65 or later. Only a fraction of the plants (35%, 75%, 38%, and 76% for ga1-3, ga1-3/rgl1, ga1-3/rgl2, and ga1-3/rgl1/rgl2, respectively) had bolted by the end of the experiment.
LexA DB Fusion Gal4 AD Fusion His− Media + 3-ATa β-gal Unitsb mm SLY1 RGL1 2 1.5 ± 0.2 SLY1 RGL2 0 0.1 ± 0.1 SLY1 RGL3 5 1.4 ± 0.2 SLY1 RGA 2 0.7 ± 0.1 sly1-d RGL1 5 7.1 ± 1.3 sly1-d RGL2 30 43.0 ± 5.9 sly1-d RGL3 30 34.0 ± 1.5 sly1-d RGA 60 118.7 ± 16.1 LexA RGL1 1 1.0 ± 0.0 LexA RGL2 0 0.1 ± 0.1 LexA RGL3 0 0.1 ± 0.0 LexA RGA 0 0.1 ± 0.1 SLY1 Gal4 −c Ndd sly1-d Gal4 − Nd ↵a The relative growth on His− plates containing 3-aminotriazole (3-AT) at 0, 1, 2, 5, 10, 30, and 60 mm. 3-AT is a competitive inhibitor of the His3 enzyme. The experiment was repeated twice.
↵b β-gal activity (units). For each sample, at least three independent enzyme assays were performed and the means ± se are shown.
↵c No growth on His− plates at 0 mm 3-AT.
↵d Nd, Not determined.
Supplemental Data
Supplemental Tables
Files in this Data Supplement:
- Supplemental Data - Table 3
- Supplemental Data - Table 1
- Supplemental Data - Table 2
