1. Enhancing Food Nutritional Quality

2. Fe I ↑ WtPEMVit. A Estimates of World Wide Malnutrition

3. Micro-Nutrient Deficiency Diseases in the Developing World ↓ Vit. A impairs immune system, increasing the risk of illness and death. ↑ blindness. ↓ Vit. A impairs immune system, increasing the risk of illness and death. ↑ blindness. ↓ Iodine causes reduced mental capacity and irreversible brain damage, ↓ Iodine causes reduced mental capacity and irreversible brain damage, ↓ Iron results in extreme fatigue with adult diminished work capacity (15%). Children are less able to concentrate and learn. Pregnant women have an ↑ risk of death during childbirth (20 % in Asia and Africa), ↓ Iron results in extreme fatigue with adult diminished work capacity (15%). Children are less able to concentrate and learn. Pregnant women have an ↑ risk of death during childbirth (20 % in Asia and Africa),

4. Chronic Disease in the Developed World Heart disease and Cancer cause most deaths Heart disease and Cancer cause most deaths Chronic diseases are associated with…….. Chronic diseases are associated with…….. – Low activity levels – Excess kilocalories – Excess and types of dietary fat – Low micro-nutrients – Low phytochemicals?

5. "Saber" A New High Yielding Rice Cultivar for Producers in the Southern U.S. Anna McClung Toni Marchetti Christine Bergman Robert Fjellstrom USDA-ARS

6. Why? World Food Supply World Food Supply – FAO (2001) – 2,807 Kcal/person/d – 76 g protein/person/d (PEM)

8. How Can We Decrease Malnutrition? Vitamin and Mineral Supplements (acute problems) Vitamin and Mineral Supplements (acute problems) Food Fortification (reaches only some) Food Fortification (reaches only some) Biotechnology (sustainable solutions?) Biotechnology (sustainable solutions?) Traditional, Molecular assisted and Mutation Plant Breeding (sustainable solutions) Traditional, Molecular assisted and Mutation Plant Breeding (sustainable solutions)

9. Food Fortification Add nutrients to staple foods Add nutrients to staple foods Add nutrients to processed foods Add nutrients to processed foods Blend more than one staple food together Blend more than one staple food together

10. Pasta Fortification Excess wheat not suitable for use in bread or pasta Wheat is low in lysine Wheat + legumes = a complete protein (Soft wheat + 30% cowpea) x special processing method = high protein pasta

11. Thai Poverty and Hunger Reduction Program (1984) 1.Production of diversified foods for home consumption 2.Skill development and credit for commercial food processing 3.Fortification of Thai instant noodles with vitamin A, iron and iodine 4.Mandatory nutrition labeling of food products 5.Promotion of 9 healthy diets for age-specific groups such as infants and young children, adolescent girls and pregnant women 6.Free or highly subsidized health care 7.Surveillance of children under five years and children in primary school Progress in Reducing Underweight Children (< five) Source: Bureau of Health Promotion, Ministry of Public Health, Government of Thailand, 1998

12. Foods being fortified with………. Vitamin AIron BoliviaSugar-- BotswanaTsabana-- Brazil*Sugar-- Chile--Wheat flour Egypt--Wheat flour El SalvadorSugarWheat flour Grenada--Wheat flour GuatemalaSugarWheat flour HondurasSugar-- IndiaCow's milk--

13. Crop Genetic Modification Over, under or blocked expression of native genes Over, under or blocked expression of native genes Insertion of non-native genes Insertion of non-native genes Traits in commercial cultivars today = herbicide resistance Traits in commercial cultivars today = herbicide resistance

14. Genetically Modified Crops of Today and Tomorrow

15. Rice Accounts for 80% of the total kilocalories and up to 20% of the protein consumed by ~2.7 billion Asians, or half the world's population. Accounts for 80% of the total kilocalories and up to 20% of the protein consumed by ~2.7 billion Asians, or half the world's population. The most rapidly growing food source in Africa The most rapidly growing food source in Africa Fortification of wheat flour with iron etc. has been a successful strategy to decrease deficiencies in developed countries. Similar fortification of rice is not as practical. Fortification of wheat flour with iron etc. has been a successful strategy to decrease deficiencies in developed countries. Similar fortification of rice is not as practical.

16. “Golden Rice” Rice does not contain beta-carotene Genes from daffodils and bacteria isoprenoid pathways were cloned Rice was transformed with genes that code for: 1.Phytoene synthase 2.Phytoene desaturase 3.Lycopene cyclase 4.Carotene desaturase

17. Color due to beta carotene Could provide 15 - 100% of daily requirements Biotechnology can increase crop nutrients and controversy Significantly reduce the incidence and severity of vitamin A deficiency? Bioavailability? Yield drag? Seed cost? Consumer acceptance? "Golden Rice"

18. High-Iron Rice? Ferritin gene from Phaseolus = 2.5-fold ↑ in Fe content Cysteine-rich polypeptides enhance iron absorption → a metallothionin gene from Oryza has been overexpressed = a 7.0- fold increase in endosperm cysteine A thermotolerant phytase from Aspergillus fumigatus To prevent activity which could interfere with germination, the enzyme was excreted into the extracellular space In small intestine simulation experiments the phytase degraded phytate to zero within 1.0 hour However, in transgenic rice the enzyme did not refold properly after cooking, i.e., it lost it’s thermotolerance The enzyme has been targeted to the phytase storage vesicles to reduce the phytate content directly. Effect on germination?

19. "Golden Rice" was possible because….. (I. Potrykus) I had stable, public funding over a long period of time which I could use independent of the opinion of others. I had with Peter Beyer the perfect partner, who understood the underlying science and provided the necessary genes and analytical expertise. The Rockefelller Foundation was willing to add substantial financial support over a long time period. The Swiss Federal Institute of Technology supported the concept of strategic research for developing countries. The project was embedded in an enthusiastic group of coworkers (over 60), all motivated to contribute to food security with their work. I was naive enough to believe in it’s success……….

20. What Do You Think? "If anyone tells you that GMOs are going to feed the world, tell them that it is not… To feed the world takes political and financial will – it’s not about production and distribution.“ Steve Smith, Novartis

21. Plant Breeding Since the origins of agriculture selection pressure has created new crops Since the origins of agriculture selection pressure has created new crops Selection is directed evolution. Selection is directed evolution. A plant product (a “phenotype”) is the result of the plant’s genotype interacting with the environment. A plant product (a “phenotype”) is the result of the plant’s genotype interacting with the environment. Plant breeders develop varieties (“packages of genes) that are specifically adapted to certain environments, and which have specific traits that consumers/users/processors demand. Plant breeders develop varieties (“packages of genes) that are specifically adapted to certain environments, and which have specific traits that consumers/users/processors demand.

22. Rice Cultivars CH 1007EAS 3Romeno N2404Taipei 177 Yachikogane

23. Barbra McClintock (1902 – 1992) Nobel Prize Winner (1983) Nobel Prize Winner (1983) "Over the many years, I truly enjoyed not being required to defend my interpretations. I could just work with the greatest of pleasure. I never felt the need nor the desire to defend my views. If I turned out to be wrong, I just forgot that I ever held such a view. It didn't matter." -- Barbara McClintock, 1983 "Over the many years, I truly enjoyed not being required to defend my interpretations. I could just work with the greatest of pleasure. I never felt the need nor the desire to defend my views. If I turned out to be wrong, I just forgot that I ever held such a view. It didn't matter." -- Barbara McClintock, 1983

24. Oilseed In Canada rapeseed was made into a new crop: "canola." (Canadian + oil) In Canada rapeseed was made into a new crop: "canola." (Canadian + oil) Canola is essentially free of erucic acid (C 22:1) (linked w/ HRT DZ) and glucosinolates Canola is essentially free of erucic acid (C 22:1) (linked w/ HRT DZ) and glucosinolates – ~ 5% saturated, 60% mono-unsaturated, 30% polyunsaturated lipids – Meal is palatable to livestock World's 2 nd – 3 rd most important food oil source World's 2 nd – 3 rd most important food oil source Baldur Rosmund Stefansson, PhD (1917 – 2002): “Father of Canola"

25. Rice White versus brown? Why? White versus brown? Why? Rancidity = TG → Free FA → aldehydes + ketones Rancidity = TG → Free FA → aldehydes + ketones 40 M metric T/year 40 M metric T/year Source of oil, vitamins, minerals, dietary fiber, protein, phytochemicals Source of oil, vitamins, minerals, dietary fiber, protein, phytochemicals $

26. Distribution of rice bran fatty acid and total oil lipid content across a set of U.S. and exotic Oryza sativa germplasm grown in the same environment.

27. r= -0.01

28. r = 0.89

29. A1A1 A2A2 C B

32. Tocopherols Tocotrienols Vitamin E Complex 8 homologs: , , ,  Found in plant oils Rice bran oil and palm are rich sources of tocotrienols Tocotrienols inhabit cholesterol synthesis in laboratory animals Lower serum total and LDL cholesterol in humans

34. Gamma Oryzanol Fraction (ferulate esters of triterpene alcohol) Plant sterols (phytosterols) Structure similar to cholesterol 10 Compounds Health Effects – UV protectant – Anti-inflammatory – Cholesterol lowering

35. Distribution of rice bran tocol and gamma- oryzanol contents across a set of U.S. and exotic Oryza sativa germplasm grown in the same environment.

38. Phenolics Phenolics are one of the most diverse groups of substances in the plant kingdom (> 8,000 compounds) They are products of plant secondary metabolism, arising from the shikimate pathway Their chemical structure ranges from simple compounds to highly polymerized substances like tannins

39. Phenolic Compounds: Properties Chemical Properties Natural antioxidants Radical scavengers Metal chelators Nutrition and Health Properties Prevention of cancer and cardiovascular diseases Antimicrobial, anti-diarrheal, anti-viral, and anti-inflammatory effects Food sensory quality: astringency and bitterness Some are anti-nutritive, i.e. tannins Ecological Properties Resistance to pathogenes and predators UV-B tolerance Pigmentation Allelopathic effects Antifungical and antibacterial effects Industrial Applications Food additives: natural colorants and preservatives Production of paints, paper, cosmetics Tanning agents

40. Plant Phenolics: Classification Class Basic skeleton Examples Monophenols Simple phenols Simple phenols C 6 C 6 Resorcinol, thymol Phenolic acids Phenolic acids C 6- C 1 C 6- C 1 Salicylic acid, p-hydroxybenzoic acid Cinnamic acids Cinnamic acids C 6- C 3 C 6- C 3 Caffeic acid, ferulic acid Polyphenols Flavonoids Flavonoids C 6- C 3- C 6 C 6- C 3- C 6 Flavones: isovitexin, luteolin Flavonols: quercetin, kaempferol Anthocyanins: cyanidin glycosides Flavanols: (+)-catechin, (-)-epicatechin Tannins Tannins (C 6- C 3- C 6 ) n (C 6- C 3- C 6 ) n Condensed tannins or proanthocyanidins: Oligomers and polymers of flavanols

41. Phenolic Content in Rice Fractions Total phenolic content (mg PhlE/100 g) † (mg PhlE/100 g) † Brown Rice 27.6 Milled Rice 14.8 Husk128.4 Bran272.2 Source: Singaravadivel and Raj, 1979 Phloroglucinol equivalents per 100 g (dry weight basis) † Phloroglucinol equivalents per 100 g (dry weight basis)

42. Light-Brown Brown Purple Red Percent (%) GAE/g whole grain*

45. † Purple rice bran Red rice bran Brown rice bran Light-brown rice bran ‡ Dried fruits Common berries Whole grain wheat cereal/raisins Whole grain wheat cereal Whole grain oat cereal 100% whole grain bread Corn cereal Rice cereal Common fruits White bread Common vegetables Melons Antioxidant Activity of Foods versus Rice Bran † Our data ( , n = 17), with error bars; ‡ Source: Miller et al 2000 (  ) * TE/100 g, µM trolox equivalents per 100 gram; Antioxidative activity measured using DPPH 050001000015000200002500030000350004000045000 TE/100g *

46. CultivarColorFractionTE/g ‡ Dixiebellelight-brownlow-P † 29 tannins 148 Jacintolight-brownlow-P 46 tannins 201 EAS 3redlow-P 47 tannins2731 Paddy Vanhaliredlow-P 48 tannins2777 Pai Chiao pairedlow-P 51 tannins2674 IAC blackpurplelow-P 825 tannins1467 HB-1purplelow-P 79 tannins 669 Antiradical Effect of Isolated Fractions † Low-P, low-molecular phenolics ‡ ‡ TE/g, trolox equivalents in µM trolox per g (dry weight basis)

47. HPLC Analysis of Phenolic Standards HPLC Analysis of Low-Molecular Phenolics from EAS 3 (Red Bran) Non-identified compounds 280 nm 350 nm 280 nm 350 nm

48. Polymeric Phenolics Simple Phenolics A B HPLC-RP Separations of Rice Bran Ethyl Acetate Extracts. A: ‘EAS-3’ (Peru). B: ‘Cypress’ (U.S.). A B

49. Future Collaborative Research Fractionate individual phenolic compounds Fractionate individual phenolic compounds Identify their structures using MS Identify their structures using MS Use preparative HPLC to collect large quantities of the fractions Use preparative HPLC to collect large quantities of the fractions Evaluate their individual radical scavenging abilities and anti-cancer properties Evaluate their individual radical scavenging abilities and anti-cancer properties Evaluate ability to prevent rice bran oil rancidity Evaluate ability to prevent rice bran oil rancidity

50. Future Collaborative Research Reducing rice bran rancidity Reducing rice bran rancidity Cross low esterase activity rice x High yielding U.S. cultivar Cross low esterase activity rice x High yielding U.S. cultivar Use anti-sense technology to knock out lipase and lipoxygenase activity Use anti-sense technology to knock out lipase and lipoxygenase activity Sensory evaluation of products Sensory evaluation of products

51. Gene Expression Rice bran phytochemicals Are they modulating gene expression rates Microarray technology allows the evaluation of the expression of many genes from a genome at the same time Tocotrienol effects on enzyme activity – gene x nutrient interaction?

52. Goal = Improve the Nutritional Content of Rice High phytochemical x Low rancidity cultivar x High Yielding cultivar High Yielding, Low rancidity, High Phytochemical