Merideth Bonierbale - First Global Conference on BiofortificationNovember 9-11, 2010, Washington, DC, USA

1. Challenges, Progress & State-of-
art in Breeding for Iron/Zinc
First Global Conference on Biofortification
November 9-11, 2010, Washington, DC
Merideth Bonierbale
International Potato Center
Wolfgang Pfeiffer
HarvestPlus
HarvestPlus Crop Leaders

2. 3.5 billion people in the developing world are Fe deficient
Women and children are most affected
Micronutrient Deficiencies ”Hidden
Hunger”
http://www.micronutrient.org
Prevalence of IDA among children
<5 years old• Maternal mortality
• Impaired mental
development &
capacity for labor
• Growth failure
• Susceptibility to
infections

3. Outline
• Genetic Variation in Crop Gene
pools
• Setting Breeding Targets
• Genetic Gains Achieved
• Molecular and Biotechnologies
• Product Delivery
• Research Needs

4. Genetic Variation in
Crop Gene pools:
Varieties, Landraces,
Breeding Populations,
Wild Relatives

5. Qualitative (HPLC)
Semi-
Quantitative
NIRS
XRF
Bench-top
Hand-held
Colorimetric
Image-Analyzer
Near-Infrared
Reflectance
Spectroscopy
X-Ray
Year 20102005
Samples
day-1
1000
500
$US Cost
sample-1
0.25
1.00
(0.5 - 3.0)
• High-throughput
• Fast & economical
• No contamination
> 30
5-10

6. e.g. Worldwide NIRS Quality Network
for Sweetpotato (and other crops)

7. Progress: Fe Variation Discovered in
Germplasm of 8 Staple Crops (ppm DW)
Wheat
Sweetpotato
Rice_unpolish
Rice_polish
Potato_Native
Potato_Adv
Maize
Cassava
6050403020100
Wheat
Sweetpotato
Rice_unpolish
Rice_polish
Potato_Native
Potato_Adv
Maize
Cassava
1501401301201101009080706050403020100
Beans
Pearl millet

8. Wheat
Sweetpotato
Rice_unpolish
Rice_polish
Potato_Native
Potato_Adv
Maize
Cassava
Beans
1101009080706050403020100
Pearl millet
Progress: Zn Variation Discovered in
Germplasm of 8 Staple Crops (ppm DW)

9. Beans
Pearl Millet
Polished_Rice
Potato_Adv
Potato_Native
Sweetpotato
Unpolished_Rice
Wheat
Fe vs Zn
Ord Crops N
Correlation
Coefficients
1 Pearl Millet 79 0.86
2 Sweetpotato 89 0.85
3 Unpolished_Rice 110 0.79
4 Wheat 176 0.64
5 Beans 215 0.63
6 Potato_Native 604 0.52
7 Polished_Rice 123 0.41
8 Potato_Adv 310 0.31
FEDW
ZnDW
160140120100806040200
120
100
80
60
40
20
0
Positive correlation between Fe & Zn allows
simultaneous improvement for both

10. The amount of Fe or Zn required in a
biofortified crop for significant impact
on nutritional status
Breeding Target
• ‘Baseline’ = amount obtained from
varieties consumed by
target population
+
• ‘Increment’ = amount to be added by
breeding
=

11. Consumption level
(g/day)
Increment of
nutrient required for
health impact
Concentration of
nutrient in the
crop as
consumed (after
storage, milling &
cooking )
Bioavailability
of nutrient in
the diet
Target
Nutrient
Content of
Staple
Foods
Influencial Factors

12. Primary Sources of Food for Populations at
Risk of Malnutrition
Intakeingrams
gen
Yams
Potato
Cassava
OFSP
Cowpea
Lentil
Bean
Maize
W
heat
Rice
Pearl M
illet
Barley
Sorghum
500
400
300
200
100
0
Grains
(100% DM)
Legumes
(100% DM)
Roots
(33% DM)
Tubers
(25-33% DM)
Intakeingrams
gen
Yams
Potato
Cassava
OFSP
Cowpea
Lentil
Bean
Maize
W
heat
Rice
Pearl M
illet
Barley
Sorghum
500
400
300
200
100
0
Grains
(100% DM)
Legumes
(100% DM)
Roots
(33% DM)
Tubers
(25-33% DM)
Consumption Levels of 13 Staple Food Crops
(DW) (DW) (FW) (FW)

13. Targeting Food & Production Systems
High Zn Wheat Targets in India
ME1: Temperate Irrigated
High Production
ME5: Irrigated High
Temperate Stress
42
38
46
36
22
19
12
11
12
14
9
8
6
4
9
8
4
2
3
2
4
7
5
18
0 10 20 30 40 50 60 70 80 90 100
Women
Children
Women
Children
AbundanceScarcity
Cereals Potato Vegetablesandfruits
Pulses Noodles Foodfronsocialprograms
Breadandcookies Sugars Meatandchicken
Other
Dietary Sources of Fe in Huancavelica, Peru

14. 0
20
40
60
80
100
120
140
Pearl Millet Bean Bean Rice Rice Maiz Maiz Wheat Wheat Potato Sweetpotato
Fe Fe Zn Fe Zn Fe Zn Fe Zn Fe Fe
ppm
Baseline & Target Levels for 7 Crops
8
44
8
8
32
64
8
11
22 22
Non-Biofortified
Avg. Baseline
BreedingTarget
Genetic Variation
Discovered
30 Target
Increment

15. Breeding Focus by Crop
and Micronutrient
HarvestPlus investment level
I. Zn Rice, Zn Wheat
II. Fe Pearl Millet, Fe Bean
III. Fe Potato, Fe Sorghum,
Fe Lentil.
Zn Maize – initially
HarvestPlus, then
AgroSalud
Fe & Zn Sweetpotato in
SASHA

16. Fe (ppm)
P
(ppm)
Rice polished
Maize
Wheat
Bean
Cassava
Potato
Lentil
Pearl Millet
Yams
Phytate: Inhibitor of Fe Absorption
5%
SweetPotato10%

17. Iron Bioavailability in Landrace Potatoes
0
10
20
30
40
50
60
704393 703168 705543 702464 700234 700787 701997 703274 703488
Caco2cellsferritinformation(ng
ferritin/mgprotein)
Caco-2 Cell Ferritin Formation
USDA/SRS & CIP

18. 0,0
5,0
10,0
15,0
20,0
25,0
30,0
15,5 23,3 31,1 38,8 46,6 54,4 62,1 69,9 77,7 85,4 93,2
mg Ac Ascorbico/ 100g DW
Porcetajedepoblación
Base Population
k = 5%
Progeny of selected
individuals
mg Vitamin C/ 100g
Percent
Genetic Gains from Selection
Gs = k * δp * H2
Selection
Intensity
HeritabilityGenetic
Variation

19. Non-negotiable
Core Traits
Product Attributes - Breeding Objectives

20. Fe Zn
FV
N=89
PBF
N=64
FV
N=89
PBF
N=64
Genetic Gains for Fe and Zn in OFSP
African farmers’ varieties (FV) vs. Pre-breeding families (PBF
(ppmindrymatter)

21. Cycle 1 Families: Source of superior
clones
Selected genotypes with high micronutrient concentration
Fe(mg/kg)
32.0
31.0
30.0
29.0
28.0
27.0
26.0
25.0
24.0
23.0
22.0
21.0
20.0
19.0
18.0
17.0
16.5
16.0
15.0
14.0
12.5
12.0
11.0
9.0
8.0
7.0
6.0
5.0
4.0
3.0
2.0
1.0
45
40
35
30
25
20
15
10
5
Boxplot Ranges of Fe (mg/kg) in diploid potato families (NCD-II)
Base line
(17.5 mg/kg

22. Biotechnologies
–Reduce time and cost to meet breeding
goals
–Exceed levels feasible by conventional
breeding

23. Identification of genes affecting variation
in Fe & Zn in Bean by QTL analysis
• Overlapping Fe and Zn
QTL suggest common
transcriptional control of
uptake
• Identification of allelic
variation for FRO to
facilitate transfer of high
mineral traits among
genepools and varieties
Blair et al., 2010

24. Promoter Genomic Wheat Ferritin clone
1DX5 Glutenin Nos
A
B
C
• Introduce extra copies of
the most active wheat
allele of the TaFer1 gene
into wheat.
• Biolistic transformation
using glutenine1Dx5
promoter to drive
endosperm specific
expression.
Transgenic WT- Bob white
Prussian blue stain
Targeting Ferritin to the Wheat Endosperm
Exon Intron
(Aarhus University/Denmark)

25. Research Agreements & Contracts
with 7 Private Companies - # will
increase
• GxE Performance Testing
• Joint Development of Fe/Zn-dense
Hybrids (private NARS) & OPVs
(public NARS) by Consortium
PartnersPartners in Deployment
Variation for Fe in
Inbred lines & Hybrids
Product Delivery: Case of Pearl Millet

26. Breeding cycles
Iron(ppm)
2005 2007 2009 2011
50
65
80
100
Lines in release
process
Families in
pipeline
2010
Baseline
Target
Incremental gains toward breeding goal for
Fe in Bean

27. Wolfgang H PFEIFFER
Product Development & Release Schedule

28. What Do We Have in Hand to
Succeed in Biofortification ?
 Micronutrient-enrichment traits are available
within the genomes of staple food crops.
 Target levels have been defined.
 Breeding schemes established.
 First-ever genetic gains for mineral traits
realized
 Mainstreamed into adapted populations.
 Product pipelines defined.
 Partners engaged
 Early-stage efficacy trials: looking good!