Article Figures & Data
Figures
- Fig. 1.
In vivo complementation of the albino phenotype in the cla1-1 seedlings by DX. A, Phenotypic analysis of 10-d-old seedlings of wild type grown on GM medium (left);cla1-1 grown on GM medium supplemented with 0.02% (w/v) of DX (center); and cla1-1 grown on GM medium (right). The arrow indicates the first pair of leaves on the DX supplemented mutant plant, where a green phenotype is clearly visible. An upper view is shown for a wild-type plant grown on GM medium (B), a 15-d-oldcla1-1 seedling grown on GM medium supplemented with 0.02% (w/v) DX (C) in which the two pairs of true leaves can be seen, and a 15-d-old cla1-1 seedling grown on GM medium (D).
- Fig. 2.
Analysis of CLA1 gene transcript and protein accumulation in Arabidopsis plants. A, RNA-blot analysis of theCLA1 transcript. Five micrograms of total RNA was purified from 15-d-old wild-type seedlings (lane 1), cla1-1 plants (lane 2), and different tissues of wild-type plants including mature leaves (lane 3), cauline leaves (lane 4), buds (lane 5), roots (lane 6), and 5-d-old seedlings (lane 7). The probes used wereCLA1 and RBCS, as well as rRNA as an RNA-loading control as indicated in the left side of the panel. B, CLA1 protein accumulation in different tissues. Western-blot analyses were performed using total protein extracts obtained from 15-d-old cla1-1seedlings (lane 1), 15-d-old wild-type seedlings (lane 2), young rosette leaves (lane 3), mature rosette leaves (lane 4), roots (lane 5), 24-h-imbibed seeds (lane 6), flowers (lane 7), and immature siliques (lane 8). Fifteen micrograms of total protein extracts was loaded in each lane except for roots and seeds, where 30 and 45 μg was used, respectively. C, CLA1 protein developmental expression. Western-blot analysis shows CLA1 protein accumulation in 15-d-oldcla1-1 seedlings (lane 1), 5-d-old (lane 2), 8-d-old (lane 3), 15-d-old (lane 4), 20-d-old (lane 5), and 25-d-old (lane 6) wild-type seedlings. In each lane, 15 μg of total protein was loaded.
- Fig. 3.
Histochemical analyses of GUS activity in Arabidopsis plants expressing the GUS gene under the control of theCLA1 gene promoter. A, Water-imbibed (48-h) germinating seeds; B, 3-d-old seedlings; C, 5-d-old seedlings; D, 15-d-old seedlings; E, immature seeds and siliques from Arabidopsis; F, transverse section of the inflorescence; G, fully developed Arabidopsis flower; and H, individual stigma and anthers from a fully developed Arabidopsis flower.
- Fig. 4.
Microscopic analysis of the plastids and mesophyll tissue of the cla1-1 mutant. Transmission electron microscopic examination of plastids of wild-type (A and C) andcla1-1 (B and D) seedlings. Etioplasts were analyzed from cotyledons of seedlings that were dark-adapted for 4 d of wild type (A) and homozygous (B) cla1-1 mutants. Amyloplasts were analyzed from 10-d-old root seedlings of wild type (C) andcla1-1 (D) mutant. Transverse sections of the 10-d-old first leaf from plants of wild type (E), cla1-1 (F), andcla1-1 (G) mutant supplemented with 0.02% (w/v) DX.
Tables
Sample Chl a Chlb Chl Total Carotenoids mg pigment g−1 fresh wt Wild type 590 242 832 130 cla1-1DX 170 108 278 61 cla1-1 11 19 30 10 Pigments were extracted from 15-d-old plants grown in GM or GM supplemented during 10 d with 0.02% DX.
- Table II.
Identification of DX obtained by incubation of pyruvate/glyceraldehyde-3-P with CLA1 protein
Sample HPLC GC MSm/z (relative intensity) min % Product2-a 8.6 7.33 307 (21), 218 (100), 204 (27), 147 (91), 73 (57) DX2-b 8.5 7.30 307 (14), 218 (100), 204 (31), 147 (85), 73 (73) -
↵F2-a The data presented in this row correspond to those values obtained after the samples were treated with alkaline phosphatase in both crude enzyme extracts and an affinity-purified enzyme protein. The values are compared with those published for peppermint (Lange et al., 1998).
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↵F2-b DX corresponds to the authentic DX molecule. Molecular ion peaks of trimethylsilyl ether products (calculated as m/z 350) were not observed.
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