DEV007310 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. DAPI-stained glc mature pollen grains. Scale bar: 20 μm.

• Supplemental Figure S2 -

  Fig. S2. Genome organization of the glc locus confirmed by Southern blot analysis. Genomic DNA was extracted from individual wild-type Columbia, wild-type Landsberg, and glc/+ plants, digested with BglII (B) or XbaI (X), and transferred to nylon filters. The filters were hybridized with probes NPTII, 3flank, or 5flank. The BglII and XbaI sites in black on the wild-type allele are of Columbia sequence. The BglII site in blue for the wild-type Landsberg allele is inferred from the band patterns on the blot with the 3flank probe. Only the green chromosome segment in the wild-type allele is not drawn to scale.

• Supplemental Figure S3 -

  Fig. S3. The glc locus in type I (rescued to wild-type phenotype) and type II recombinants (mutant phenotype). (A) Southern blot of XbaI-digested genomic DNA with a probe hybridized to the NPTII gene inside the Ds element. Arrow points to the band in glc mutant. (B) CAPS markers analysis for the recombinant lines in Ler background and in hybrid Ler/Col background. (C) Southern blots of BglII-digested genomic DNA with a probe hybridized in the deletion region (At1g65380) and another probe hybridized outside the Ds as loading control (At1g66045). After the signal intensities of bands with At1g65380 probe were quantified, the filter was rehybridized with At1g66045 probe and signal quantification was repeated for At1g66045. The raw signal intensities of the bands on each blot are in the second and third rows of the table. The final relative DNA amount for At1g65380 of each plant line (At1g65380/At1g66045) was normalized by dividing the signal intensity with At1g65380 probe by the signal intensity with At1g66045 probe. R1, R2 and H1, H2 are type I recombinants in Ler/Ler and Ler/Col background, respectively. r3, r4 and h3, h4 are type II recombinants in Ler/Ler and Ler/Col background, respectively. M, original glc mutant line. L, wild-type Ler. Marker sizes are in kb.

• Supplemental Figure S4 -

  Fig. S4. A model for the creation of glc mutation and the two types of recovered recombinants. The chromosomes are not drawn to scale. (1) The Ds element was inserted into the blue chromosomal region. This insertion deleted this part of the chromosome and transposed the adjacent chromosomal region (yellow) to the centromeric side of the deletion. (2) The mutant chromosome harboring glc mutation was created from the Ds insertion event. (3) Mutant and the wild-type chromosomes paired during meiosis. Cross-over occurred between the homologous regions (yellow). (4) Resolution of the cross-over produced two types of recombinants: (a) Type I retained the original Ds insertion but recovered the deleted DNA (blue), reverting the mutant phenotype to wild-type while maintaining kanamycin resistance of the plants; (b) Type II lost the Ds insertion together with the associated original deletion from step (2), maintaining the mutant phenotype but rendering the plants kanamycin sensitive.