1. HarvestPlus c/o CIAT A.A. 6713 Cali, Colombia Tel: +57(2)4450000 Fax: +57(2)4450073 HarvestPlus@cgiar.org www.HarvestPlus.org Biofortified MAIZE for Latinoamerica Natalia Palacios Rojas Thanda Dhliwayo, Felix San Vicente, Luis Narro, Babu Raman, Kevin Pixley, Kai Sonders

2. Outline  Biofortified maize work at CIMMYT  Agrosalud products  QPM and High Zn maize  ProA maize  Tortilla on the roast- Climate change impact in Mesoamerica  Biofortified maize in LA

3. Researchable Issues Genetic variation characterization/ allele mining Breeding Food technology: nutrient retention, food processing

4. Biofortification at CIMMYT WhiteYellowAdaptationCurrent effort Biofortification Project Quality protein maize (QPM) XXXTropical10% in LAH+ 60% in Ethiopia NuME Highland High ZnXXTropical5% in LAH+ ProAXXSubtropical70% SAH+

5. CountryInstitutionCultivarYear Colombia CORPOICA S06TLWQ AB-22013 FENALCE FNC31AC, FNC32AC 2010 Guatemala ICTA ICTA MAYA2011 PROSEMILLAS HRQ-596, HRQ- 2988 2010- 2011 NicaraguaINTA MAZORCA DE ORO, S06TLWQ AB-2 2013 PanamaIDIAP IDIAPMQ-02, IDIAPMQ-07, IDIAPMQ-12, IDIAPMQ14, S06TLWQ AB-2 2009- 2010

6. QPM is not always conducive to high Zn. However, high Zn has been mainly observed in QPM background. InbredMean Zn (ppm) CML26429.86 b,a,c CML264Q34.25 a CML45131.46 b,a CML451Q31.53 b,a CML24730.63 b,a,c CML247Q26.61 d,c CLO245027.55 b,d,c CLO2450Q30.46 b,a,c CML27323.98 d CML273Q29.41 b,d CML25427.24 b,d,c CML254Q28.56 b,d,c Currently doing some proteomic studies to understand the mechanism (Instituto de Biotecnologia-UNAM)

7. High Zn tropical maize lines PedigreeZn (±2 ppm) (CLQRCWQ97/CLQRCWQ38)-B-6-1-1-1-1-1-1-1 50.49 (CLQRCWQ97/CLQRCWQ38)-B-2-1-1-3-1-1-1-1 49.03 (((CML491/DTPW-C9-F104-5-4-1-1-B-B)/CML491)-B-103/CML491)-15-1-1-1-1 48.29 ((CML491/LAPOSTASEQ-C7-F64-2-6-2-1-B-B)/CML491)-B-92-1-1-1-1 47.49 ((CML491/LAPOSTASEQ-C7-F103-2-2-2-1-B*3)/CML491)-B-21-1-1-1-1-1 47.45 ((CML491/LAPOSTASEQ-C7-F64-2-6-2-2-B*3)/CML491)-B-37-1-1-1-1 47.35 ((CML491/((((CML176/R1)/R1)-73-1-3-1/R1)-79/R1)-36-1-B)/CML491)-B-18- 2-2-1-1 47.18 ((CML491/LAPOSTASEQ-C7-F64-2-6-2-2-B*3)/CML491)-B-25-1-2-1-1 46.70 ((CML444/CLQRCWQ26)/CLQRCWQ26)-B-6-1-1-1-1-1 46.58 ((CML247/((((CML176/R1)/R1)-73-1-3-1/R1)-79/R1)-36-1-B)/CML247)-B-14- 2-1-1-1 50.44 (CLRCW79/CLRCW98)-B-22-3-1-1 44.18 ((CML247/((((CML176/R1)/R1)-73-1-3-1/R1)-79/R1)-36-1-B)/CML247)-B-14- 2-2-1-1 40.17 (CLRCW79/CLRCW98)-B-22-2-1-1 39.79 ((CML247/((((CML176/R1)/R1)-73-1-3-1/R1)-79/R1)-36-1-B)/CML247)-B-13- 1-5-1-1 39.74 (CLRCW79/CLRCW98)-B-27-2-1-1 39.61 (CLRCW79/CLRCW98)-B-26-1-1-1 39.15 ((CML247/((((CML176/R1)/R1)-73-1-3-1/R1)-79/R1)-36-1-B)/CML247)-B-18- 1-1-1-1 38.82

8. Multilocation evaluation of stage II hybrids in Mesoamerica- 2012 PedigreeGYpERADEHEarAspPHPltAsp CLWQ446/CML4918.946.361.1155.42.2269.72.5 CLWQ415/CLQRCWQ1318.8213.561.0170.32.9293.32.6 Commercial Check 1 (Monsanto DK357)8.706.458.0144.62.7270.42.9 CLWQ373/CLQRCWQ488.613.661.5166.92.5285.62.6 CLWQ425/CML4918.548.859.5148.32.6259.22.6 CLWQ326/CLQRCWQ487.768.761.0137.12.3257.02.5 CLWQ356/CLQRCWQ487.637.260.3138.32.3252.62.5 CLWQ365/CLQRCWQ487.608.359.7135.32.4261.92.6 CLWQ337/CLQRCWQ487.598.662.2148.22.2269.32.8 CLWQ363/CLQRCWQ487.548.361.2144.22.3261.12.9 Commercial Check 17.4510.659.6132.82.9249.72.6 2013: Twenty 3-way high Zn hybrids in evaluation

9. QPM screening Colorimetric assays: Tryptophan Lysine NIR: Protein Tryptophan Lysine Nurit et al., JAFC, 2009; Rosales et al., JAFC, 2011; Galicia et al., Fitotecnia Mexicana, 2011 SNP1(315) – Uni-plex Ensayo para O2

10. Screening tools for micronutrients Fe Zn RSQ modelRSQ internal validation RSQ external validation (n=274) Fe0.89 Zn0.96 0.90

11. Preliminary data for Fe and Znd association mapping Max21.337.8 Min10.122.1 Avg15.3525.2 Fe Zn Association mapping panel: 532 lines

12.  UPLC  Improvement of HPLC  ATR FT-IR (in validation) Molecular Markers  LYCE  HYDB1 Biochemical Analysis Screening Tools 0.0 0.2 0246810 AUAU 0.1 1 2 3 4 5 6 7 B Babu et al, TAG 2013

13. ProA association mapping Association mapping panel: 436 lines- 3 cycles of phenotyping Swarno et al, In preparation

14.  Competitive agronomically  Provitamin A concentration varying from 6-8 ppm  Second wave is building momentum… Mg ha -1 3 high proA hybrids released in Zambia in September 2012 Mean of 8 commercial hybrids Approx. LSD (P<0.05)

15. Pedigree Type germplasm/maturity pVAC ug/gDW CrtB1 favorable allele KUI carotenoid syn-FS11-1-1-B-B-B-B-B-B-B-B-B Sub-trop/Early 6.81 (KUI carotenoid syn-FS11-1-1-B-B-B/(KU1409/ DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-1-B-B Sub-trop/Early16.5+ (KUI carotenoid syn-FS11-1-1-B-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B- 3(MAS:L4H1)-1-B-B-B Sub-trop/Early 35.72+ KUI carotenoid syn-FS17-3-2-B-B-B-B-B-B 6.72 (KUI carotenoid syn-FS17-3-2-B-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B- 1(MAS:L4H1)-5-B-B-B Sub-trop/Early 29.02+ (KUI carotenoid syn-FS17-3-2-B-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B- 4(MAS:L4H1)-2-B-B-B Sub-trop/Early 32.56+ CML297Sub-trop/V Late6.3 (CML297/(KU1409/DE3/KU1409)S2-18-2-B)-B- 2(MAS:L4H1)-4-B-B-B Subtropical/late15.55+ Florida A plus Syn-FS2-2-1-B-B-B-B-B-B Sub-trop/Early 8.47 (Florida A plus Syn-FS2-2-1-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B- 3(MAS:L4H1)-2-B-B-B Sub-trop/Early10.78+

16. HybridStage ProA (ug/gDW)Agron Comments SCBTester11B01//((MAS[206/312]-23-2-1-1-B-B- B/[BETASYN]BC1-10-2-1-#/CML-305-B)-B-11-1-B-B)-B III 11.21Average SCBTester11B01 // (CML297/(P72c1xCML297 x CL- 02410-3-1-1-B/CML297)-B-1)-3-1-1-B III 11.78Average SCBTester11B01 // (CML297/(CML489/[BETASYN]BC1- 2-#/CML297)-B-24)-13-2-B III 13.91Top 1/3 of trial SCBTester11B01 // (CML297/(P72c1xCML297 x CL- 02410-3-1-1-B/CML297)-B-16)-13-4-1-B III 13.97Top 1/3 of trial SCBTester11B01 // (CML297/(P72c1xCML297 x CL- 02410-3-1-1-B/CML297)-B-1)-3-2-1-B III 14.68Top 1/3 of trial SCATester11B04//(((KUI carotenoid syn-FS17-3-1-B-B- B/(KU1409/DE3/KU1409)S2-18-2- B)//[[[NAW5867/P30SR]-40-1/[NAW5867/P30SR]-114- 2]-16-2-2-B-2-B/CML395-6]-B-20-1-B-3- #/[BETASYN]BC1-3-1-1-#-B-B-B)-B(MAS:HydB1)-2)-B II 11.96Top 1/3 of trial SCATester11B04//((CML297/(KU1409/DE3/KU1409)S2- 18-2-B)-B-2(MAS:L4H1)-1-1-1)-B II 15.53Average SCATester11B04//((KUI carotenoid syn-FS11-1-1-B-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-2- 1)-B II 18.47Top 1/3 of trial SCATester11B04//((KUI carotenoid syn-FS11-1-1-B-B- B/(KU1409/DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-2- 1)-B II 18.65Top 1/3 of trial New wave High Provitamin A hybrids (Subtropical maize) 66 hybrids in trial 99 hybrids in trial

17. Step pVAC decay Maize drying None Maize storage Yes.  Higher for flour, Less in kernels  At higher temperature, higher degradation  pVAC decay varies among genotypes  pVAC more stable after the first 6 weeks of storage Palacios-Rojas et al, in preparation

18. Sample Lut (µg/g) Zeax (µg/g) βCX (µg/g) βC (µg/g) Prov A (µg/g) Grain3.9 a 5.2 ab 3.4 bc 1.1 bcd 2.8 bc Lime cooked grain 2.7 ab 2.7 ed 3.1 cd 1.4 bcd 2.9 bc Dough3.4 ab 5.5 a 4.2 a 1.6 bc 3.7 b Flour2.9 ab 4.3 ab 3.9 ab 1.9 b 3.8 b P<0.05 No significan carotenoid losses during nixtamalization Palacios-Rojas et al, in preparation

19. 40% loss of ProA during sweet tamal preparation Tamal dough1.1 cd 0.7 gf 2.6 de 0.8 cd 2.2 cd Tamal dulce1.1 cd 0.7 gf 2.2 e 0.6 ed 1.7 d Sample Lut (µg/g) Zeax (µg/g) βCX (µg/g) βC (µg/g) Prov A (µg/g) Grain3.9 a 5.2 ab 3.4 bc 1.1 bcd 2.8 bc Hernandez-Montes et al, in preparation

20. LutZeaxBCXBCProA Raw5.10 a13.42 b5.43 b2.61 b5.32 b Boiled (African style) 4.46 a12.06 b4.50 c2.13 c4.38 c Bolied (Mexica n style) 4.95 a17.14 a6.22 a3.27 a6.38 a Roasted4.57 a15.34 a5.36 b2.64 b5.32 b No significant losses of ProA in boiled and roasted green maize Rosales et al, in preparation

21. Climate change preditions Source: Report ToR, 2012

22. Maize yield differences between the current climate and 2020s predicted Poor soil conditions Good soil conditions Source: Report ToR, 2012

23. Biofortified maize for LA NicaraguaGuatemalaPanamaColombiaMexico High ZnXXXXX ProAXX DroughtXXXXX HeatXXXX TarspotXXXX GLSX Ear rotXXXXX Early material XXX Intercropping: Maize + Beans in the hills CA Retention studies on nixtamalized products Understanding on ProA degradation (role of antioxidants- vitamin E; biodisponibilidad-oil) Potentially: storage pest and mycotoxin evaluation Capacity building- Quality monitoring of current released cultivars

24. Partners Colombia: Corpoica, Fenalce Guatemala: INTA Nicaragua: ICTA Panama: IDIAP Mexico: INIFAP, TEC-Monterrey

25. From the Field to the Plate Pre-breedingBreeding Seed supplier FarmerGrain Grain Market Miller Food Processing RetainerConsumer