1. Homeobox leucine zipper protein 9 (HAT9) -AT2G22800 -Homeodomain(s) -Leucine Zipper Motif -DNA Binding -Dimerization ?? ~ 1400 -Helix-turn-helix 5’3’ FWRV Exon 1 UTR Exon 3Exon 2 Intron 1Intron 2 102513618697869120314351 5’3’ Research Talk HC70AL Spring 2004 by Bobby Fam

2. Evolutionary Relationships of HAT Proteins -Alignment of HAT9 and HAT22 reveals 90% identity in the homeodomain-leucine zipper region and 71% identity overall. -Little homology is seen outside of the homeodomain- leucine zipper region for other proteins.

3. Gene Activity of AT2G22800 1kb 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1kb Leaf: 1,2 Stem: 3,4 Flower: 5,6 Ovule: 7,8 Embryo: 9,10 Axis: 11,12 Leaf: 13,14 Flower: 15,16 -Gene specific bands for leaf, stem, flower, unfertilized ovules, and 30 DAP axis. -Arabidopsis: Activity in WT mature green seeds, WT Post- Maturation Green Seeds, and WT Roots -Possible Alternative Splicing 14 DAP embryo (lane 9).

4. Madison Screen Round 1 1022 27 Control 1600 bp 1800 bp 4500 bp Round 2 1 1 2 2 3 3 4 4 5 5 6 6 7 7 8 8 9 10 C 1kb -T-DNA insert in either superpool 10 or 22 -Sequencing data matches HAT9 but has no T-DNA -Reamplification yielded no matching bands

5. SALK T-DNA Insertion Gene: AT5G67300SALK Line: 039074 1 2 3 4 5 6 7 8 9 WT H 2 0 1kb1kb 1 2 3 4 5 6 7 8 9 WT H 2 0 FW/RV Gene-specific RV Gene-specifc & LBb1 primer -Plant 5 represents a homozygous T- DNA genotype -Plants 1,2,3,4,6,7,8, & 9 represent

6. Phenotyping SALK Insert Plant Mutant FlowerWT leaf on left; Mutant leaf on right WT Silique top; Mutant Silique bottom Mutant Trichome -Mutant plant does not seem to differ in phenotype from the Wild-type in any significant manner

7. To the Future… -Sequence Madison Inserts -Identify DNA Pool (Reorder) -Identify Seed Pool -Identify Knock-out Line -Grow Plants & Observe AT2G22800 AT5G67300 -Investigate other knockout lines -Study other genes and their effects on seed development -Determine specific role of genes on seed development -Engineer the 21st Century Crop