1. Breeding Vegetables for Optimum Levels of Phytochemicals Kevin Crosby Texas A&M University

2. Plant Breeding Application of genetic theory, including molecular biology to develop improved plant cultivars Vegetables are high priority crops in Texas and USA- numerous cultivars from diverse breeding programs; $15 billion in USA

3. A.ANY HUMAN HANDLING OF PLANTS WHICH GENETICALLY ADAPTS THEM TO THE SERVICE OF MEN. B.THE ART OF SELECTION BASED UPON THE SCIENCE OF GENETICS. C. EVOLUTION DIRECTED BY THE WILL OF MAN. (Vavilov, 1935) Definitions

4. Wild Types vs. Improved

8. Foods for Health Elevated levels of flavonoids, carotenoids, ascorbic acid, minerals, pectins Traditional plant breeding- exploitation of naturally occurring genes in the germplasm Gene mapping and genetic transformation- increased Ca, vitamin C, β-carotene

9. Pepper Phytochemicals Peppers are good source of important human health related compounds Carotenoids, flavonoids, ascorbic acid, capsaicin Great genetic variation within pepper germplasm & at different maturity stages

10. Dietary Fiber in ½ cup Serving Green Peppers- raw2.4 Hot Red Peppers- raw 19.2 Hot Red Peppers- dried 57.6 Pinto Beans- cooked9.4 Prunes- dried3.8 Tomatoes- raw1.4 Spinach- cooked7.0 Squash- cooked3.0

12. Screening Germplasm Planted 60 diverse pepper lines at Weslaco and Uvalde Harvested green and red mature fruit Analyzed Vit C by HPLC for replicated samples

13. High Vitamin C Peproncinis- high at all locations, 557 ppm (green) to 2377 ppm (red) Pasilla, Anaheim, Marconi and Ancho also very high at red stage- 1900-2280 ppm TAM B36- Tropic Bell, highest at green stage: 600 ppm= 60 mg/100g

15. TAM Pasilla VR

16. Cultivar: B 36 (GR-RD) Vitamin C: 1758-2082 ppm Flavonoid: 6-13 ppm

17. High β-carotene Highest in red chile/paprika- C127 (2330 µg/100g) 38% RDA, Fidel orange chile (2366) Almost none in many wax and hot types TMJ2 (1242 µg/100g) > Grande (850)

18. Cultivar: C 127 Vitamin C: 1721 ppm Flavonoid: 45 ppm Beta-Carotene: 23.3 ppm

19. Habanero β-carotene TAM Mild- 760 µg/100gYucatan- 20 µg/100g

20. Vitamin C  -carotene Cultivar/line (mg·100g -1 ) x (µg·g -1 ) x Fruit type TAM Perlita 4521.7 a z 62.2 a z Western shipper- size 15 y TAM Uvalde15.2 b57.5 abWestern shipper- size 18 Explorer12.4 bc45.0 cdeWestern shipper- size 15 Cruiser12.1 bc36.3 eWestern shipper- size 12 Mission11.9 bc40.3 deWestern shipper- size 15 Green Ice11.8 bc4.70 fHoneydew- size 15 TAM Mayan11.2 bcd11.5 fCasaba- size 12 Gold Mark10.8 bcd41.4 cdeWestern shipper- size 12 TXC 201510.0 bcd49.8 bcWestern shipper- size 12 HMX 95839.4 cd46.0 cdWestern shipper- size 12 Valley Gold9.3 cd48.5 bcdWestern shipper- size 12 Mainpak9.2 cd43.5 cdeWestern shipper- size 12 Pronto8.0 cd43.0 cdeWestern shipper- size 12 TAM Dew Impr.7.2 cd4.70 fHoneydew- size 12 Primo7.0 cd56.7 abWestern shipper- size 12 z Mean separations by LSD, P ≤ 0.05. Values followed by the same letter are not significantly different. y Sizes based on number of fruit which fit into a standard melon packing box. x Based on fresh weight. Fresh fruit vitamin C and beta-carotene contents of melons from Weslaco

21. Selection Process Choose the best parents for target traits Controlled pollinations to create new families Selection for traits by combined quality analyses- field and lab Backcrossing or inbreeding to fix key genes

22. Controlled Pollination

23. Developing a New Melon Industry wants larger cantaloupes with good flavor and disease resistance We had excellent flavor, disease resistance, Vit C, carotene in TAM Uvalde So we crossed it with ‘Cruiser’ (very large, high yield but poor flavor, carotenoids)

24. ‘Chujuc’ Muskmelon Large, round fruit, Sz 9-12 High yield and resistant to PM High in beta-carotene Vitamin C, sugars

25. Genetic Linkage Mapping Develop population which segregates for gene(s) of interest Carefully measure phenotype of trait for each plant and parents Screen DNA for molecular markers which are linked to the trait (gene)

26. DNA Marker Linkage RAPD marker OAC09.900 expressing polymorphism between two DNA bulks from high and low beta-carotene F2 plants. 1 = Sunrise (low parent), 2 = TAM Uvalde (high parent), 3 = DNA bulk from low beta-carotene F2 plants, 4 = DNA bulk from high beta- carotene F2 plants, and 5 = a 100-bp DNA marker ladder. 1 2 3 4 5 OAC09.900 -1500 bp -600 bp

27. Figure 5. RAPD marker OAU02.600 expressing polymorphism between two DNA bulks from high and low ascorbic acid F 2 plants, and between ‘Dulce’ and TGR1551. 1=’Dulce’, 2=TGR1551, 3=DNA bulk from high ascorbic acid F 2 plants, 4=DNA bulk from low ascorbic acid F 2 plants, and 5=molecular size marker. -1500bp -600bp -300bp 1 2 3 4 5 OAU02.600

28. Molecular Markers in Pepper Family of high flavonoid CA377 x high vitamin C B22 F2 progeny, F1 progeny, Parents grown in two environments for mature fruit Screening with RAPD markers in bulks and genotyping each F2 progeny Constructing genetic linkage map

29. High Antioxidant Parents TAES B22CA 377 F1 Hybrid

30. -2072bp -600bp 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 M -1500bp -300bp OL07.2050 OL07.1100 OL07.1500 OL07.1700 OL07.800 OL07.480 OL07.520 Fig. 1. Segregation of seven RAPD markers in an F2 population derived from the pepper cross of CA 377 x B 22. 1 to 20=F2 plants of the CA 377 x B 22 cross and M=a 100-bp DNA marker ladder.

31. Current Progress

32. Flavor is Crucial Many people complain about tomatoes Long shelf life and green harvested fruit just do not taste very good Enhancing fresh market consumption and profitability for growers requires change

33. New Breeding Priorities Screen germplasm for best tasting tomato cultivars, lines, landraces Cross with TAMU heat-tolerant, disease resistant, high yielding breeding lines Developed better tasting, adapted cultivars

34. Improved Heirloom Types Introgressed virus, fusarium and heat resistance genes; developed more compact plants adapted to staked culture

35. Flavor Trumps Appearance

36. Why Vegetable Genetics? Vegetables are excellent source of key human health related phytochemicals $15 billion component of US agriculture Not all cultivars are equal- flavor and appearance are also crucial