1. Ministry of Agriculture, Livestock and Food Supply The First Scientific Meeting of The Global Cassava Partnership (GCP-1) July 21-25 2008 Ghent, Belgium Miguel Angel Dita Rodríguez Breeding cassava for enhancement of carotenoid, iron and zinc contents

2. Ministry of Agriculture, Livestock and Food Supply HARVEST PLUS TEAM MEMBERINSTITUTION Wânia Maria Gonçalves FukudaEmbrapa Cassava & Tropical Fruits Vanderlei da Silva SantosEmbrapa Cassava & Tropical Fruits Luciana Alves de OliveiraEmbrapa Cassava & Tropical Fruits Tatiane da Silva AmorimEmbrapa Cassava & Tropical Fruits Hernan CeballosInternational Center for Tropical Agriculture Marília NuttiEmbrapa Food Technology José Luis Carvalho VianaEmbrapa Food Technology Carlos Estevão Leite CardosoEmbrapa Cassava & Tropical Fruits Maurício Melo MascarenhasEmbrapa Cassava & Tropical Fruits Zara Maria FernandesEmbrapa Cassava & Tropical Fruits Marcio Canto PereiraEmbrapa Cassava & Tropical Fruits Miguel A. Dita RodriguezEmbrapa Cassava & Tropical Fruits Clóvis Manoel da SilvaEmbrapa Cassava & Tropical Fruits / EBDA

3. Ministry of Agriculture, Livestock and Food Supply INTRODUCTION Target site: Northeast of Brazil Vitamin A deficiency Cassava-dependent region Biofortified food Works must focus cassava BRAZILIAN GERMPLASM

4. Ministry of Agriculture, Livestock and Food Supply INTRODUCTION

5. Ministry of Agriculture, Livestock and Food Supply INTRODUCTION Target site: Northeast of Brazil Vitamin A deficiency Cassava-dependent region Biofortified food Works must focus cassava BRAZILIAN GERMPLASM

6. Ministry of Agriculture, Livestock and Food Supply Development cassava varieties improved for Carotene, Iron and Zinc contents Main Goal

7. Ministry of Agriculture, Livestock and Food Supply FIRST CRITICAL STEPS Characterization of 72 landraces for Total Carotenoid, Fe and Zn contents (for parent selection and varietal delivery) - Spectrophotometry, Atomic absorption and b-carotene quantification by HPLC Germplasm screening (CIAT Color Chart) - 1800 accessions from germplasm bank at Embrapa C&TF - 70 yellow and 2 pink landraces selected

8. Ministry of Agriculture, Livestock and Food Supply MAIN ACTIVITIES Parent Selection Crossing fields to obtain hybrids with higher Carotenoid, Fe and Zn contents Planting seedlings from crossings among yellow clones Harvest, selection and planting of seedlings Selection and planting clones for productivity trials Harvest, selection and planting of advanced productivity trials of clones with high carotenoid, Fe and Zn content and low HCN in roots

9. Ministry of Agriculture, Livestock and Food Supply RESULTS 6 Generations (Populations) developed - 2003 - 2004 - 2005 -2006 -2007 -2008 ( Parents from Landraces and elite hybrids )

10. Ministry of Agriculture, Livestock and Food Supply GENERATION 2003 18 crossing fields at Embrapa CNPMF Complete Diallele Selection of 12 progenitors with high carotenoid regardless of HCN contents in roots 33,000 seeds produced 10,000 seeds sent to CIAT/IITA 8,000 seeds at Embrapa CNPMF 5,500 seedlings transplantation Continue...

11. Ministry of Agriculture, Livestock and Food Supply Carotenoid evaluation (color chart - CIAT) of 3,265 seedlings Selection and planting of 330 clones with coloured roots Preliminar Trial 254 hybrids selected: 53% yellow, 31% orange, 22% pink 49% with low HCN (Photos: Maurício Mascarenhas 2005) GENERATION 2003

12. Ministry of Agriculture, Livestock and Food Supply 08 hybrids for yield and high Iron 12 hybrids for yield and high Zinc 51 hybrids for high carotenoids and low HCN IN FIELD (2008) On-farm advanced productivity trials GENERATION 2003

13. Ministry of Agriculture, Livestock and Food Supply GENERATION 2004 Policross Selection of 6 progenitors with high carotenoid and low HCN contents in roots 8.000 seeds produced 7.628 seeds planted in CNPMF 5.550 seedlings transplantation 128 hybrids with high carotenoid and low HCN in roots selectioned 100 hybrids with high carotenoid and high HCN in roots selected for processed food (Farinha) 636 clones selectioned Color chart

14. Ministry of Agriculture, Livestock and Food Supply GENERATION 2004 (IN FIELD, 2008) 15 hybrids for high carotenoids, Iron and Zinc 22 Hybrids for high carotenids and very low HCN 23 Hybrids for high carotenoids and Iron 20 Hybrids for high carotenoids and Zinc Carotenoids > 6,0  g/g Fresh W Iron > 13 mg/kg Zinc > 10 mg/kg Selection criteria

15. Ministry of Agriculture, Livestock and Food Supply GENERATION 2005 Policross Selection of 6 progenitors with high carotenoid and low HCN contents in roots 4.500 seeds produced 1561 seedlings transplanteds 35 hybrids selected for high carotenoids, Iron, Zinc and low HCN 211 hybrids selected for high carotenoids, Iron, Zinc and low HCN IN FIELD 2008

16. Ministry of Agriculture, Livestock and Food Supply Total carotenoids (μg.g -1 ) contents (TCC) in Landraces and hybrids (2003,2004, 2005) of cassava 8,41*0,443,17402005 12,412,766,21352004 10,470,874,42282003 15,510,636,672Landraces Maximum (μg.g -1 ) Minimum (μg.g -1 ) Average (μg.g -1 ) Number of genotypes Landraces/ hybrids Harvest time-points? Significant increment in the generations 2003 and 2004, but not in the 2005 TCC of the progenitors from these generations ranged from 1.50 to 4.0 μg.g-1

17. Ministry of Agriculture, Livestock and Food Supply 7 91 88 38 5 00 30 39 37 26 8 12 15 11 4 1 0 0 10 20 30 40 50 60 70 80 90 100 0 a 22 a 44 a 66 a 88 a 10> 10 Frequency 2003 2004 2005 cassava roots μg.g 1  Total carotenoid contents in hybrids (Families 2003, 2004 and 2005)

18. Ministry of Agriculture, Livestock and Food Supply HCN content in hybrids (families 2003, 2004 and e 2005) 0 5 10 15 20 25 0 a 2020 a 4040 a 5050 a 6060 a 7070 a 8080 a 9090 a 100 100 a 110 HCN-ppm (cassava roots) Frequency 2003 2004 2005 Considered as sweet cassava

19. Ministry of Agriculture, Livestock and Food Supply 30,6520,5423,80402005 77,51,013,11362004 51,10,08,21792003 56,50,09,272Landraces Maximum (mgKg -1 ) Minimum (mg Kg -1 ) Average (mg Kg -1 ) Number of genotypes Landraces/ Hybrids Iron (mg.Kg -1 ) contents in Landraces varieties and hybrids of cassava roots. (Families 2003, 2004 and e 2005) Zirconium mill Enviromental influence Soil pH Analysis are being repeated!

20. Ministry of Agriculture, Livestock and Food Supply 34,381,977,94402005 87.10.512.501362004 34.10.05.201792003 26.20.04.1472Landraces Maximum (mgKg -1 ) Minimum (mg Kg -1 ) Average (mg Kg -1 ) Number of genotypes Landraces/ Hybrids Zinc (mg.Kg -1 ) contents in Landraces varieties and hybrids of cassava roots. (Families 2003, 2004 and e 2005) Zirconium mill Enviromental influence Soil pH Analysis are being repeated!

21. Ministry of Agriculture, Livestock and Food Supply Contents of total carotenoids (  g.g -1 ) on fresh root weight (Generation 2003 ) 146%- 42%58.1%Gain 10,470,874,49Hybrids 4,001,502,84Progenitors MaxMinMean 2003 family (228 genotypes)

22. Ministry of Agriculture, Livestock and Food Supply Contents of total carotenoids (  g.g -1 ) on fresh root weight (Generation 2004) 209,4%28,9%84.2 %Gain 12,41*2,766,21Hybrids 4,012,143,37Progenitors MaxMinMean 2004 family (135 genotypes)

23. Ministry of Agriculture, Livestock and Food Supply HYBRIDS (GENERATION 2003) SELECTED (Carotenoid, Zn, Fe, and low HCN, Agronomic traits) Participatory trials in communities of 3 Northeast States (CE, PE BA) Are being harvesting this week!

24. Ministry of Agriculture, Livestock and Food Supply RELEASING VARIETIES On-farm Participatory Breeding

25. Ministry of Agriculture, Livestock and Food Supply CASSAVA: Participatory Breeding (Romero-Hernandez, 1993,2002, 2003 ) Selection and multiplication of 9 landraces (7 yellow + 2 pink) for fresh consumption Why? 1.Acceptance tests of yellow root genotypes by smallholder in the Northeast 2.Preparation of ‘consumers’ for the next generations of elite hybrids to be released from the 2003, 2004… generations 3.Supply well-characterized cassava varieties with more carotenoid contents than local varieties (commonly white-pulp)

26. Ministry of Agriculture, Livestock and Food Supply Beta-carotene content in landraces selected with low HCN selected (Parents) Collaboration with Dr. Délia Rodriguez-Amaya - UNICAMP

27. Ministry of Agriculture, Livestock and Food Supply On-Farm Participatory trials : -35 trials in: -28 farmers communities at: - 4 States of the Bazilian Northeast: (Pernambuco, Piauí, Bahia and Maranhão) Growth curve (from 6 to 13 months): HCN in roots, fresh and dry weight, root pigmentation, yield. Cassava: participatory breeding

28. Ministry of Agriculture, Livestock and Food Supply Acceptance frequency distribution of the cassava varieties in participaty tests in Northesast Brazil. Classes. 1: “Outstanding” to… 10: very bad

29. Ministry of Agriculture, Livestock and Food Supply Acceptance probability of cassava varieties evaluated by farmers in 35 participatory trials in Northeast Brazil. Logistic Regression curves DendéKlainasikAipim cacauCacau amarelo

30. Ministry of Agriculture, Livestock and Food Supply Dendê: BRS Dourada “Golden” Klainasik: BRS Gema de Ovo “Egg Yolk” Varieties Released by Embrapa

31. Ministry of Agriculture, Livestock and Food Supply Cenoura Rosada = BRS Rosada “Pink” Varieties Released by Embrapa

32. Ministry of Agriculture, Livestock and Food Supply CAROTENOIDS CONTENT IN LANDRACES 15,51 mg.g -1 BRS Gema de ovo BRS Dourada 72 landraces 0 5 10 15 Total carotenoid content (  g/g root pulp - fresh weight) White

33. Ministry of Agriculture, Livestock and Food Supply CONSIDERATION - I Results indicates that exploring biodiversity and using conventional breeding approaches: Carotenoids, Iron and Zinc contents in cassava roots can be improved.

34. Ministry of Agriculture, Livestock and Food Supply CONSIDERATION- II One key issue of this selection is that about half of the of the high-TCC hybrids evaluated presented low cyanogenic content in the roots and, therefore, are suitable to be consumed boiled, which makes carotenoids more bioavailable than other ways of consumption.

35. Ministry of Agriculture, Livestock and Food Supply CONSIDERATION - III The high acceptance probability of de varieties BRS Dourada and BRS Gema de Ovo is very important to introduce the new cassava varieties improved to high, betacarotene, Iron and Zinc content in roots: The coloured roots varieties were not rejected by consumers!

36. Ministry of Agriculture, Livestock and Food Supply Thank You!