Figure S1. Expression of S6-RNase and HT-proteins in S. lycopersicum.
Untransformed S. lycopersicum and plants expressing S6-RNase, HT-A, or HT-B
proteins are shown. (a) Protein blot analysis. Pistil protein extracts equivalent to
0.2 mg (S6-RNase) or 1.5 mg (HT-proteins) fresh weight were separated and
probed with antibodies, as shown. A silver stained blot is also shown to visualize
loading. The S6-RNase antibody detected the full-length polypeptide and lower
molecular weight fragments. Variable amounts of the S. lycopersicum 60-residue
HT-A truncated polypeptide (arrowhead) were detected in different accessions
and little or none of this peptide was detected in the Ailsa Craig background
expressing S6-RNase. (b) Pollen tube growth. Pistils were stained with aniline
blue fluorochrome 24 h after self-pollination. The brightly stained pollen tubes
penetrated to the ovary in each case (arrowheads), indicating a compatible
pollination. The plants shown, HTA-3 and HTB-3, displayed the highest level of
protein expression among all the transgenic plants tested. Scale bars, 1 mm.
Figure S2. HT-protein levels in HTA-3, HTB-3, and HT/S6 T2 pistil extracts
compared to dilutions of S. pennellii LA2560 extracts. Pistil extracts (fresh weigh,
FW, as shown) were separated and probed with specific antibodies, as shown.
Figure S3. Pollen tube growth in transgenic S. lycopersicum HTB/S6-4 T1 plant,
a plant showing a relatively weak UI response. Pistils were stained with aniline
blue fluorochrome 24 h after pollination with self-pollen or pollen from S.
lycopersicum cv. VF36 or S. pennellii LA0716. Arrow indicates pollen tubes that
reached the ovary or the site of pollen rejection. Insets show pollen tube at the
base of the style. Scale bars, 1 mm.
Figure S4. S. cheesmaniae LA0522 pollen tube growth in transgenic S.
lycopersicum plants. Pistils were stained with aniline blue fluorochrome 24 h after
pollination. Arrowhead indicates pollen tubes that reached the ovary or the site of
pollen rejection. Insets show pollen tube termini at the site of pollen rejection.
Scale bars, 1 mm.
Figure S5. S. galapagense LA0438 pollen tube growth in transgenic S.
lycopersicum plants. Pistils were stained with aniline blue fluorochrome 24 h after
pollination. Arrowhead indicates pollen tubes that reached the ovary or the site of
pollen rejection. Insets show pollen tube termini at the site of pollen rejection.
Scale bars, 1 mm.
Figure S6. S. pimpinellifolium LA3798 pollen tube growth in transgenic S.
lycopersicum plants. Pistils were stained with aniline blue fluorochrome 24 h after
pollination. Arrowhead indicates pollen tubes that reached the ovary or the site of
pollen rejection. Insets show pollen tube termini at the site of pollen rejection.
Scale bars, 1 mm.