Article Figures & Data
Figures
- Figure 1.
Genetic and Physical Mapping of CER6 on Chromosome 1 and T26J14.
cer6-2 was positionally mapped between mi185 and AP1 on chromosome 1 by scoring PCR-based markers; the indicated positions in centimorgans correspond to the map generated from recombinant inbred lines (http://nasc.nott.ac.uk/new_ri_map.html). New PCR-based markers were generated at ETR and AP1; the centimorgan positions correspond to the phenotypic marker map (Koornneef, 1994) and within the BAC clones T2E12, T26J14, and F24J5 (AF-1 through AF-6). Circled numbers indicate the number of recombination events detected between each marker and CER6. The 45-kb region between AF-4 and AF-5 (open box) defines the boundaries of the region containing CER6; annotated genes (gray boxes), some of which correspond to ESTs (filled circles), are indicated. The white line within the T26J14 BAC indicates the fragment used for complementation experiments.
- Figure 2.
Sequence Analysis of CER6 and CER60.
(A) Scale drawing of the CER6 and CER60 genes, indicating the percentage of DNA sequence identity for each exon.
(B) Sequence alignment of CER6 and CER60, generated by using ClustalW 1.8 (http://dot.imgen.bcm.tmc.edu:9331/multi-align/multi-align.html) and Boxplot (http://www.ch.embnet.org/software/BOX_form.html). (*), residues altered by the mutations; black shading, identical residues; gray shading, similar residues; (†), position of the exon1/exon2 splice junction. Dashes were introduced to optimize alignment.
- Figure 3.
Morphology of the cer6-2R Pollen Coat.
Transmission electron microscopy of wild-type (WT), cer6-2, and cer6-2R pollen grains, showing the resemblance between cer6-2R pollen grains and the wild type. C, cytoplasm; E, exine; I, intine; L, lipid droplet; PC, pollen coat. Bars = 1 μm.
- Figure 4.
Lipid Contents in the Pollen Coats of Wild-Type, cer6-2, and cer6-2R Pollen.
Chloroform-extracted lipids were analyzed by gas chromatography/mass spectrometry. Lipids constituting >1% of wild-type pollen coat lipids are reported. Quantities were normalized to an internal control. All classes of lipids for a given carbon chain length—18, 24, or 28—were pooled for that length. Error bars indicate standard deviation; for the wild type (derived from Landsberg and Columbia), n = 2; for cer6-2, n = 2; and for cer6-2R, n = 8. (*), C29 lipids; (#), C30 lipids.
- Figure 5.
Presence of Lipid Crystals on cer6-2R Stems.
Scanning electron microscopy of wild-type (WT), cer6-2, and cer6-2R stems shows fewer crystals on cer6-2R stems than on those of the wild type. Bars = 5 μm.
Tables
- Table 1.
Complementation of Fertility and Stem Cuticle Phenotype of cer6-2 Mutants with CER6
Linea Stem Epicuticular Waxb Seeds (n)c Kan Resistanceb,d WT CER6b,e Ler +++ 27.2 ± 2.9 (23) NA + cer6-2 – 0.3 ± 0.6 (14) NA – 6-2 FJ +++ 29.5 ± 2.0 (12) + + 6-2 FG +++ 27.4 ± 4.7 (15) + + 6-2 FI +++ 24.4 ± 6.0 (18) + + 6-2 R1 +++ 22.2 ± 2.9 (15) + + 6-2 FF +++ 20.8 ± 2.7 (18) + + 6-2 RA ++ 28.3 ± 3.3 (19) + + 6-2 F4 ++ 21.6 ± 4.1 (19) + + 6-2 FK + 21.8 ± 5.1 (8) + + 6-2 F5 + 11.5 ± 3.0 (13) + + 6-2 FH – /+ 26.1 ± 3.3 (19) + + 6-2 F3 – /+ 23.1 ± 3.1 (16) + + 6-2 RB – 26.3 ± 2.2 (16) + + 6-2 FD – 15.9 ± 3.3 (14) + + 6-2 FA – 0.4 ± 1.0 (10) + – 6-2 FB – 0.4 ± 0.7 (10) + – -
↵a Lines 6-2 FJ through 6-2 FD denote cer6-2 lines transgenic for a wild-type copy of CER6, ranked by their stem cuticle phenotypes. Lines 6-2 FA and 6-2 FB contain vector sequences but lack CER6. Ler, Landsberg erecta.
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↵b (+ + +), wild-type wax levels; (++), (+), (–/+), intermediate wax; (–), no wax.
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↵c Mature seeds per half seed pod and standard deviation.
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↵d The presence of the kanamycin resistance gene was determined by PCR. NA, not applicable.
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↵e The presence of wild-type (WT) CER6 was assessed using two dCAPS markers.
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