2. OUTLINE OF PRESENTATION  Phenylpropanoid/avenanthramide biosynthesis  Health benefits of avenanthramides  Application of suspension cultures to investigate avenanthramide biosynthesis

3. General Phenylpropaniod Pathway + ProteinTryptophan/protein PAL

4. Phenylpropanoid biosynthesis

5. Reaction of Phenylpropanoid and 5- Hydroxyanthranilic Acid Catalyzed by HHT Form R phenylpropanoid A (Bp) H p-Coumaric B (Bf) OH caffeic C (Bc) OCH 3 ferulate

6. Grain Avenanthramide Content Shows Strong Genotype X Environment Interaction Emmons & Peterson, Crop Sci 41:1676 (2001)

7. Grain Avenanthramide Content Shows Strong Genotype X Environment Interaction Emmons & Peterson, Crop Sci 41:1676 (2001)

8. Effect of Avenanthramide C on VSMC Proliferation [ 3 H] thymidne incorporation (% of control) Nie, Wise, Peterson, Meydani, Atherosclerosis, 186, 260-266 (2006).

9. Avn C inhibition of Restinoblastosis protein (pp-Rb) polyphosphorylation in VSMC Nie, Wise, Peterson, Meydani, Free Rad. Biol. Med, in press

10. Effect of Dietary Avenanthramide C on Exercise Stressed Rats Reduction of oxidized glutathione levels and lower ratio of reduced to oxidized form of glutathione. O’Moore and Ji, (2006) unpublished results.

11. Summary of Health Effects  Reduces vascular smooth muscle cell proliferation  Increased NO production  Inhibition of ppRb, arrest G1-S1 cell cycle progression  Elevation in anti-oxidant enzyme system in rats

12. Oat (cv Belle) Suspension Cultures

13. HPLC Trace of Media Extract Subsequent to Chitin Elicitation

14. Avenanthramide A in Culture Media After Elicitation with Chitin

15. Avenanthramide A in Tissue Extracts Subsequent to Chitin Elicitation

16. Northern Analysis of Suspension Cells for Barley PR-1 and HHT Subsequent to Elicitation 18S RNA bPR-1 HHT

17. HHT Enzyme Activity Post Induction

18. Conclusions/Future Directions  Avenanthramides are a “value added” phytonutrient unique to oat  Avn content in the grain is highly variable  Basis for variability is unknown  Suspension cultures offer a model to study avenanthramide biosynthesis at the molecular level  SSH cDNA library prepared  What is the involvement of the various isozymes (HHT, 4-CL, PAL)?

19. ACKNOWLEDGEMENTS Jean Mayer Human Nutrition Research Center MOHSEN MEYDANI LIN NIE Univ Wisconsin, Dept Kinesiology LI LI JI KATHLEEN O’MOORE Cereal Crops Research RACHEL ANGEL LAURI HERRIN NICKOLAS PARTRIDGE RON SKADSEN JOHN HERBST