Figure S1. Chloroplast haplotype network location and karyotypes of Hieracium
subgenus Pilosella plants used in this study.
(a) Plants used in this study were mapped to a previously established haplotype
network of chloroplast trnT-trnL sequences in indicated Hieraciinae genera (Fehrer
et al., 2007a; b). Haplotypes are represented as coloured circles, sizes corresponding
to the number of species sharing that haplotype. Extinct haplotypes are shown as
open circles and the extinct haplotype introgressing Pilosella 2 is indicated by a solid
black circle. (b) Karyotypes of plants obtained from root tip squashes.
Figure S2. Defects in the progression of apomixis in mutants with structurally
defective ovules. Plant identity and stage are indicated in the bottom right corner.
Abbreviations: ai, aposporous initial cell; en, endosperm; end, endothelium; es,
embryo sac; f, funiculus; int, integument; ov, ovule. Single headed arrows indicate
embryos; double headed arrows show the space between the ovary and the fruit
wall. The white dotted lines show the outline of the funiculus. Bars in (a), (b) and (d)
= 400 μm and in (c) and (e) = 200 μm. Bars in insets in (b)-(d) = 50 μm.
Figure S3. Morphology of Hieracium mutant plants and their progeny.
(a) Hybrid progeny from a cross between mutant 179 (LOAlop; in the R35
background (R35) with yellow petals) and H. aurantiacum (A35 with red petals)
have dual colour petals respectively (inset). (b) Unpollinated mutant 134 (loaLOP)
plants give rise to small plants with half the relative ploidy (0.5x) of the R35 parent.
Capitula and floret size is also reduced (inset). (c) Hybrid arising from a cross
between mutant 115 (loalop) and sexual P36. Scale bars in (a)-(c) are 10 cm, insets
5 mm.
Figure S4. Expression of pFM-GUS in the ovule of apomictic H. praealtum R35.
Bright field (a) and DIC (b) images for the same ovule are presented. Expression of
the marker was observed in degenerating megaspores (dm) and not in the AI cell
(ai) in apomict R35 or selected megaspore of sexual P36 (not shown). Bars in (a)
and (b) = 50 µm.
Figure S5. DNA gel blot analysis of apomicts and irradiation mutants with defective
LOA and LOP loci to examine linkage of HFIE to the LOP locus. Genomic DNA from
sexual H. pilosella P36, apomict H. praealtum R35, six loaLOP mutants, seven LOAlop
mutants, and a loalop mutant were digested with the restriction enzyme Spe I and
probed with HFIE cDNA (Rodrigues et al. 2008). HFIE, a single copy gene (Rodrigues
et al., 2008) is present in all plants and not lost in mutants defective in LOP function
that also carry large deletions in the locus.