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Open Access Research article

Requirement of proline synthesis during Arabidopsis reproductive development

Dietmar Funck1*, Gudrun Winter1, Lukas Baumgarten1 and Giuseppe Forlani2

Author Affiliations

1 Department of Plant Physiology and Biochemistry Biology Section, University of Konstanz, Universitätsstraße 10, 78464, Konstanz, Germany

2 Department of Life Science and Biotechnology, University of Ferrara, via L. Borsari 46, 44121, Ferrara, , Italy

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BMC Plant Biology 2012, 12:191  doi:10.1186/1471-2229-12-191

Published: 13 October 2012

Additional files

Additional file 1:

Figure S1. Developmental defects of homozygous p5cs2 mutants. A: Five-week-old plants cultivated in short-day conditions. B: Two-week-old seedlings cultivated axenically on half strength MS medium with 30 mM sucrose.

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Additional file 2:

Figure S2. Heredity diagram for p5cs1/p5cs2 double heterozygous mutants. A indicates wildtype P5CS2 allele; a indicates p5cs2 mutant allele; B indicates wildtype P5CS1 allele; b indicates p5cs1 mutant allele; green shading indicates herbicide resistance mediated by the T-DNA insertion in p5cs2 mutant alleles; pink shading indicates non-viable gamete or embryo; hatched pink shading indicates that the respective progeny will not be observed because of non-viable pollen. Note that the thick black lines delimit the heredity diagram for a heterozygous p5cs2 single mutant, while the red lines delimit the progeny of a heterozygous p5cs2 mutant that is additionally homozygous for a p5cs1 mutation.

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Additional file 3:

Figure S3. Expression of P5CR-GFP in embryos. Epifluorescense and brightfield images of isolated embryos displayed in false colour. A-D: Embryo of a homozygous p5cr-2 mutant plant complemented with pGWB4-P5CR, containing the native P5Cpromoter and gene fused with GFP. E-H: Embryo of a Col-8 wildtype plant transformed with pGWB5-P5CR, expressing the P5CR-cDNA fused to GFP under control of the CaMV-35S promoter. I-L: Embryo of a Col-8 wildtype plant. Scale bar = 20 μm.

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Additional file 4:

Figure S4. Subcellular localisation of P5CR-GFP and complementation of the p5cr-1 mutant. A-D: Epifluorescence images of homozygous p5cr-1 mutants expressing a P5CR-GFP fusion protein under control of the native P5CR promoter. A-C: Epidermal and spongy parenchyma cells; scale bar = 20 μm. A: GFP fluorescence. B: Overlay of GFP and chlorophyll fluorescence. C: Brightfield image of the same area. D: Comparison of a P5CR-GFP expressing root tip (left) and a non-transgenic root tip, overlay of a brightfield image with GFP fluorescence, scale bar = 50 μm. E: Schematic drawing of the DNA construct in pGWB4 that was used to complement the p5cr mutants in comparison to the wildtype P5CR gene. Arrows indicate the binding sites of the primers that were used to determine the genotype of the complemented mutants (see panel I). F-H: Epifluorescence images of epidermal and spongy parenchyma cells of a wildtype plant transformed with pEG103-P5CR. Note that overexpression with this construct resulted in the formation of cytosolic protein aggregates; scale bar = 20 μm. F: GFP fluorescence. G: Overlay of GFP and chlorophyll fluorescence. H: Brightfield image of the same area. I: PCR-genotyping of p5cr-1 mutants complemented by transformation with pGWB4-P5CR. See panel E for the primer binding sites in the genomic and the transgenic copy of P5CR. J: Schematic drawing of the constructs in pEG103 and pGWB5 for overexpression of P5CR-GFP.

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Additional file 5:

Table S1. PCR-primers used in this study.

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