Supporting information
Figure S1. Molecular characterisation of the rad51d mutant. The SAIL_564_A06 line
carrying a T-DNA insertion in the Arabidopsis RAD51D coding sequence was found by
interrogating the SIGnAL T-DNA express database (Alonso et al., 2003). To clarify the
nomenclature we have named the corresponding allele rad51d-3 with rad51d-1 and rad51d-2
being the alleles previously characterised by Durrant et al. (2007) and Inagaki et al. (2009)
respectively. Plants homozygous for rad51d-3 T-DNA insertions were identified by PCR in
the T3 seeds provided by the Nottingham Arabidopsis Stock Centre. As for other xrcc2 and
rad51b mutants, no obvious growth or fertility defects were observed in plants homozygous
for rad51d-3. Molecular characterisation of the insertion was done by amplifying the T-DNA
junctions and sequencing of the corresponding PCR products. The results are summarised in
panel (A). The rad51d-3 allele T-DNA is inserted in exon 8 of the RAD51D gene and the
insertion is associated with a deletion of 11bp of the exon. As frequently observed, the
inserted T-DNA is flanked by two left borders in opposite orientations. Exons are shown as
boxes and PCR primers as arrowheads. (B) Plants homozygous for the T-DNA insertion
were selected by PCR, and semi-quantitative RT-PCR analysis performed to assess the
presence of AtRAD51D transcripts in total RNA isolated from wild-type and mutant leaves.
AtRAD51D transcripts were detected in the wild-type but no AtRAD51D mRNA could be
detected using primers flanking the T-DNA insertion in RNA from rad51d-3 plants. In
contrast, truncated AtRAD51D mRNAs could be detected upstream of the T-DNA insertion.
The T-DNA insertion in the atrad51d-3 mutants thus prevents the production of the full-length
mRNAs (and hence proteins) of AtRAD51D. (C). Mutation in Arabidopsis RAD51D confers
hypersensitivity to the DNA cross-linking agent, Mitomycin C (MMC). Percentages of wildtype and rad51d sensitive plants at different MMC doses (0, 20, and 40 µM MMC). 50
plantlets were analysed in each case. In the absence of MMC, wild-type plants develop at
least four true leaves (excluding the cotyledons), thus plants with three leaves or less were
considered as sensitive.
Figure S2. Crossing scheme for the introgression of Landsberg markers into the xrcc2
mutant background. Landsberg erecta plants were crossed with xrcc2 mutants (Col0
background). F2 plants were tested for heterozygosity of the markers and XRCC2 genotype.
Marker segregation analyses were done on progeny of these F2 plants.
Table S1. List of PCR markers between Col and Ler. For each marker, the corresponding
Bacterial Artificial Chromosome, primers, size of the Columbia and Landsberg products, and
the physical distance of the intervals are listed.