Open Flower. Juvenile leaf Flowerbud. Carpel 35 NA NA NA NA 61 NA 95 NA NA 15 NA 41 3 NA

Transcription

1 PaxDB Root Juvenile leaf Flowerbud Open Flower Carpel Mature Pollen Silique Seed Sec3a Sec3b Sec5a Sec5b Sec6 Sec8 Sec10a/b Sec15a Sec15b Exo84a Exo84b Exo84c Exo70A1 Exo70A2 Exo70A Scale > Supplemental Figure S1. Abundance of each Arabidopsis exocyst subunit in different tissues from the Protein Abundance Across Organisms database (Wang et al., 2012). In PaxDB, protein abundance is listed as a value in parts per million (ppm) for each protein identified in a tissue-specific proteomics dataset. = Not Available. Previously we have shown by microarray expression analyses that SEC3a, SEC3b, SEC5a, SEC5b, SEC6, SEC8, SEC10a/b, SEC15a, SEC15b, EXO70A1, EXO84b and EXO84c are all are expressed in the stigma (Safavian et al., 2014). EXO84a was not represented on the microarray, but EXO84a is shown to be pollen-specific in this dataset.

2 sec3ab sec5ab sec6 sec8 sec10ab sec15b sec15ab exo84b DIC Aniline blue DIC Aniline blue DIC Aniline blue Wildtype Col-0 DIC Aniline blue DIC Aniline blue Supplemental Figure S2. Pollen grain attachment and pollen tube growth following pollination of transgenic pistils with wild-type Col-0 pollen. Following 2 hour pollinations, the transgenic pistils were stained with aniline blue to visualize the pollen tubes. All transgenic pistils from the exocyst subunit R silencing/knockout lines showed reduced compatible pollen responses with reduced pollen grain adhesion (DIC images) and pollen tube growth (aniline blue images). Scale bars = 50 µm.

3 sec3ab sec5ab sec6 sec8 sec10ab sec15ab exo84b DIC Aniline blue DIC Aniline blue DIC Aniline blue Wildtype Col-0 DIC Aniline blue DIC Aniline blue Supplemental Figure S3. Pollen grain attachment and pollen tube growth following pollination of wild-type Col-0 pistils with transgenic pollen. Following 2 hour pollinations, pistils were stained with aniline blue to visualize the pollen tubes. All the transgenic pollen from the exocyst subunit R silencing/knockout lines showed wild-type compatible responses with abundant transgenic pollen grain adhesion (DIC images) and pollen tube growth (aniline blue images) on wild-type Col-0 pistils. Scale bars = 50 µm.

4 Col-0 sec3ab line 1 sec5ab line 2 sec6 line 1 sec8 line 3 sec10ab line 3 sec15ab line 2 exo84b line 3 Supplemental Figure S4. Phenotypes of flowering plants from the exocyst subunit R silencing/knockout lines. One representative R silencing/knockout line is shown for each exocyst subunit. In comparison to wild-type Col-0, normal growth and flowering was generally observed for all the transgenic lines, with the exception of smaller siliques due to reduced seed set. Scale bars = 3 cm.

5 A B C Col-0 sec3ab D E F sec5ab sec6 sec8 sec10ab G H I sec15ab exo84b Supplemental Figure S5. Phenotypes of stigmatic papillae from the exocyst subunit R silencing/knockout lines. Elongated stigmatic papillae are present in late stage 12 flower buds (this is the final stage prior to anthesis and anther dehiscence; Smyth et al., 1990; Sanders et al., 1999) for wild type Col-0 and all the R silencing/knockout lines. For each line, the late stage 12 flower bud is shown in the top left panel while a close-up of the stigma is shown in the top right panel (n>30 for each line). For comparison, a freshly-opened flower (Stage 13) in shown in the bottom left panel while a close-up of the stigma is shown in the bottom right panel. Representative images are shown from R silencing/knockout lines for each exocyst subunit. Fully elongated stigmatic papillae were observed in all transgenic lines. Scale bars = 150 µm.

6 sec3ab line 2 sec6 line 1 sec8 line 2 sec15ab line 1 Col-0 Supplemental Figure S6. Phenotypes of tracheary elements in pistils from the exocyst subunit R silencing/knockout lines. The Arabidopsis exo70a1 mutant was previously found to have disrupted tracheary elements (TEs) in the pistil, potentially implicating other exocyst subunits in TE development (Li et al, 2013). This study used the SLR1 promoter to direct strong expression at late stages of stigma development, and this strategy was predicted to allow normal TE differentiation in the pistil. Intact TEs are clearly visible in dark field microscopy images (marked by yellow arrows) for representative pistils from select R silencing/knockout lines. Scale bar = 400 μm.