1. Cells and cell growth Membranes and cell walls Fertilization and embryogenesis Seed development and dormancy Germination Water transport Water relations Plant nutrition Solute transport Gene function

2. Three learning goals : - what is a gene structural genetic elements of hertitable units requirements: promoter intron, exons, ORF 5’ and 3’ UTR - transcription and translation biosynthesis of RNA and proteins requirements: RNA polymerase, transcription factors mRNA, splicing ribosomes, tRNA, polypeptides - gene identification and cloning correlation of genetype and phenotype requirements: mutagenesis, insertion, gene mapping cloning and sequencing, EST, expression studies

3. Hunting Genes

4. 7 “characters” equal seven genes

5. What is a Gene? A locatable region of genomic sequence, corresponding to a unit of inheritance, which is associated with regulatory regions, transcribed Regions and/or other functional sequence regions. Sequence ontology consortium, Berkeley 2006 Dystrophin (human) 2.5 mio bp 11.000 coding bp ycf1 (chloroplast) 6800 bp6800 coding bp MRP1 (plant) 13400 bp4870 coding bp Plants 25,000 Humans 25,000 Fly 12,000 Yeast 6,000 E.coli 5,000 DNA virus 10-300

6. Nomenclature: Geneunit of inheritance Mutationchanges in gene DNA sequence that may lead to changes in phenotype Phenotypeobservable (measurable) characteristics of a plant Genotypeall genes or alleles that determine a plant’s phenotype recessivephenotype not visible in heterozygous stage dominantphenotype visible in heterozygous stage Wildtypephenotype of the majority in a population

7. RNA protein AAAAAAAm 7 G cap

8. Gene regulation and signal transduction What is a gene?

9. Transposons in petunia control flower color transposon activity revertant sector revertant flower mutant flower

10. Heliotron

11. Jumping genes in corn

12. Transposable elements all over the place

13. Transposition event: change from white to colored No transposon activity: white background color Transposons in maize kernels Small spots: frequent, late transposition events Revertant: transposon excised in germline

14. Plant transposable elements

15. Transposon tagging and gene cloning

17. Gene of interest T-DNA LBRB ProteinmRNA

18. T-DNA LBRB Gene o f interest T-DNA LBRB Gene o f interest

19. T-DNA LBRB Gene o f interest mutant wildtype

20. Hunting a maize dwarf gene N. R. Fuller, NSF  Plant architecture:  one of the greatest improvements of crop yield in the history of agriculture Important regulators influencing yield in maize  Plant growth – gibberellic acid (GA): – auxin: – brassinosteroids (BR):  Sex determination: – jasmonic acid (JA): 20 teosinte corn

21. Phenotype of na1 mutants na1WTna1WT 21

22. GA inhibitor reduces na1 mesocytl elongation in the dark 22 WTna1d5 scale bar 10 cm; uniconazole (UCZ) UCZ [µM]0350 0

23. Alleles of na1 5’UTRATGTAG Exon1Exon2 3’UTR B73 reference 5’UTRATGTAG Exon1Exon2 3’UTR unknown insertion in na1-1 Mu8 insertion in na1 Mu4 insertion in na1-3 Mu8 insertion in na1-4 EMS induced G to A exchange in na1-2 +489 STOP 3-oxo-5-  -steroid 4-dehydrogenase ? ? ubiquitin DET2 WTbrts1-1na1-1na1-2H2OH2O RT-PCR na1 na1-1

24. H 2 O 5 µM PCZ WT na1d5 H 2 O 5 µM PCZ propiconazole (PCZ) [µM] height (%) propiconazole treatment mimic na1

25. propiconazole induces „tasselseed“ 25 WT PCZ-- na1WTna1 ++ TS-++++ propiconazole (PCZ), tasselseed phenotype (TS)

26. Expression analysis through microarrays

27. Gene regulation and signal transduction What is a reporter gene reporting? Here we are!!!!

28. Gene regulation and signal transduction What is a reporter gene reporting?