1. Transcription factors in tomato fruit development

2. EU-SOL WP1.2 Relationship between early fruit development and primary fruit composition Identification of transcription factors involved in (early) fruit development

3. What target genes to choose? Differential expression during fruit development is a lead……..

4. Cornell fruit development experiments Gene expression analysis using a 12,000 element (representing app. 8,000 genes) cDNA microarray

5. Expression of 192 transcription factors DAP 7-57 2 log ratios of -4 to +4

6. Transcription factor AP2 function in A. thaliana ap2 mutant

7. Transcription factor AP2 has additional functions in tomato (l: wt; r: AP2 knockdown)

8. AP2 functions in fruit development as well

9. TCP transcription factors

10. TCP functions Known functions indicate that TCPs are regulators of growth: Outgrowth of branches in maize (tb1) Limiting growth of flower parts, causing flower asymmetry (cycloidea and dichotoma in Antirrhinum) Control of leaf curvature (cincinnata in Antirrhinum) Suggested to regulate transition from mitosis to expansion by interaction with cell cycle gene promoters

11. Phylogenetic tree of non-tomato TCP proteins Class I Class II cin cyc;dich tb1 jaw-D (down TCP2,3,4,10,24)

12. Tomato TCP gene expression SlTCP12 SlTCP18 SlTCP15 SlTCP21 SlTCP07 SlTCP13 SlTCP06 SlTCP05 SlTCP09 SlTCP11 SlTCP16 SlTCP17 SlTCP14 SlTCP19 SlTCP01 SlTCP08 SlTCP03 SlTCP10 SlTCP04 SlTCP02 SlTCP20 Seedling Leaf Root Flower Fruit I Fruit II FruitIV M. Green Breaker Turning Red Up late in fruit Off late in fruit

13. Study of TCP gene function in fruit development Modification of expression of selected genes by (early fruit specific) ectopic expression or knock- down by RNAi/amiRNA Mapping of interactions between TCP proteins as well as between TCP proteins and other TFs (targetted/non-targetted)

14. Activity of the TPRP promoter in fruit TPRP promoter works well untill at least mature green stage; 2A11 promoter is considered as alternative for later stages Early stages Mature greenBreakerRipe fruit

15. Constructs for fruit-specific over-expression or for suppression by RNAi and amiRNA XX TPRP-F1 promoterT NOSatt R1 att R2Gateway cassette att L1 att L2ORF of interest NPT II XX TPRP-F1 promoterT OCSatt R1 att R2Gw cassette NPT II att R2 att R1Gw cassette XX intron att L1 att L2 partial mRNA att L2 att L1 partial mRNA att L1 att L2 miR miR*

16. Characterization of tomato TCP-dimerization in Y2H assays All 21 orf’s recombined into the BD- and AD- vectors Autoactivation of BD-constructs checked on –LTH with increasing AT: 7 with some activity, of which 4 strong Tested interactions in a matrix setup on –LTH (+5, 10, 15 mM AT) and –LTA