DEV033647 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Response of AG::GUS reporter genes to LFY::LFY:VP16 demonstrating functionality of constructs. KB14 (A) and RH146 (B) in wild-type background. After crossing to LFY::LFY:VP16 plants, AG::GUS activity was expanded into sepals of KB14 (C) and became readily detectable in RH146 flowers (D).

• Supplemental Figure S2 -

  Fig. S2. Transactivation assays in yeast. Full-length cDNAs of the identified transcription factors without a GAL4 activation domain were inducibly expressed in yeast and tested for their capacity to activate transcription using the AAGAAT box either alone or in combination. Whereas the GARP transcription factors UNE16, At4g37180, At3g24120 and AtbZIP21 are potent transcriptional activators, TGA5 and TGA6 act as repressors. Interestingly, PAN does not exhibit activating potential when acting alone (red arrow), but is able to synergize with the activating GARP transcription factor At4g37180 (compare bars marked by gray and black arrows).

• Supplemental Figure S3 -

  Fig. S3. mRNA localization of a putative PAN co-factor. Expression of At4g37180 overlapped with PAN mRNA in the SAM and early floral meristems. (A) Sense control; (B) antisense probe.

• Supplemental Figure S4 -

  Fig. S4. Response of 5′AG enhancer sequences to loss of PAN function or mutations in the bZIP binding site. GUS-stained sections of LD-grown apices from (A) KB14, (B) pan KB14 and (C) sSST196 (AG::GUS reporter with mutated bZIP binding site).

• Supplemental Figure S5 -

  Fig. S5. Scanning electron micrographs of pan flowers from SD-grown plants. (A) Weak, (B) intermediate and (C) strong fruit phenotypes.

• Supplemental Figure S6 -

  Fig. S6. Response of 5′AG enhancer sequences to loss of PAN function in SD. Activity of KB14 AG::GUS in wild-type (A) and pan mutant (B) flowers from SD-grown plants.

• Supplemental Figure S7 -

  Fig. S7. Response of AG expression to loss of PAN function in LD and SD. (A-D) Expression of AG RNA in wild-type (A,C) and pan (B,D) inflorescences from plants grown in LD (A,B) and SD (C,D). Black arrowhead indicates high levels of AG RNA in wild type, whereas the white arrowhead points to reduced AG RNA accumulation in pan mutant flowers grown in SD.

• Supplemental Figure S8 -

  Fig. S8. Phenotype and AG::GUS activity of late stage pan KB14 flowers grown in SD. (A) Phenotype. (B) GUS staining.

• Supplemental Figure S9 -

  Fig. S9. Ectopic expression of WUS in various stages of indeterminate flowers of pan mutants grown in SD. Shown are sections from an early stage (A) and a higher magnification of a later stage flower (B) with phenotypes comparable to the ones shown in Fig. 4F and Fig. S5B.