S5.1 Maize HarvestPlus: Biofortifying maize with Provitamin A carotenoids

Maize HarvestPlus:
Biofortifying maize
with Provitamin A carotenoids
Kevin Pixley, Natalia Palacios, Raman Babu
& Abebe Menkir (CIMMYT & IITA)

11th Asian Maize Conference
November 7-11, 2011; Nanning, China

Dietary sources
 Vitamin A
– Meat (esp. liver)
– Vegetables (carrot, sweet potato, spinach)
 Iron
– Red meat, fish, poultry
– Lentils, beans, leafy vegetables
 Zinc
– Oysters, animal proteins,
– Beans, nuts, whole grains
$$ $

$$$$ $$$

Business
5 January 2011 Last updated at 14:42 ET

World food prices at fresh high, says UN
The FAO's Abdolreza Abbassian warned
over low level food reserves

Global food prices rose to a fresh high in
The FAO's Abdolreza
December, according to the UN's Food Abbassian warned over
and Agricultural Organisation (FAO). low level food reserves

Its Food Price Index went above the
previous record of 2008 that saw prices
spark riots in several countries.

As India‟s economy grows by
some 9% a year, food prices
are soaring.

In late December …food
inflation had reached 18.3%,
with pricey vegetables most to
blame.
“…he offers pyaz, a staple
for much Indian cooking, for
60 rupees ($1.33) a kilo, the
most he can remember.”

Jan 6th 2011

The “Tortillazo” is coming…
– 50% increase to price of tortillas

Middle East

Egypt protests: Hosni Mubarak faces fifth
'day of rage'
The unrest in Egypt follows
an uprising in Tunisia two
weeks ago, in which
President Zine al-Abidine
Ben Ali was toppled after 23
years in power.

The Tunisian upheaval began
with anger over rising food
prices, high unemployment
and anger at official corruption - problems which have also left many
people Egypt feeling frustrated and resentful of their leadership.

Biofortification of staple food crops
• Micronutrients available in staple foods
– Sustainable, affordable
– Accompanied by dietary/nutrition information
– Complemented by supplementation and fortification
• Acute malnutrition
• Equal or better agronomic performance of biofortified
crops
– Yield, disease resistance, drought tolerance…

Yan et al., Nature Genetics 2010

ProA (ug/g) in 9 genotypic classes of 6
crosses
LycE HydB Pop 1 Pop 2 Pop 3 Pop 4 Pop 5 Pop 6
4 1 12.03 9.97 6.15 5.74
11.94 9.70 6.12 5.75
4 2 4.27 3.46 3.99 3.90
4.09 3.57 4.08 3.74
4 H 5.60 6.17 4.78 7.42
5.74 4.73 7.51
2 1 7.41 11.11 7.19 6.25 5.10
7.40 10.74 6.92 6.19 5.09
2 2 2.08 3.36 3.73 2.60 4.28 3.31
2.06 3.34 3.71 2.50 4.33 3.29
2 H 4.16 4.07 5.30 3.88 4.42 3.88
4.11 4.07 5.40 3.89 4.43 3.93
H 1 9.45 12.96
9.47 13.20
H 2 3.58 4.14 3.95
3.59 4.21 4.13
H H 4.83 6.82 5.08 5.41 3.64
4.87 6.80 4.98 5.41 3.60

Seed genotyping pre-planting

 Dry chipping using dog nail clippers
 ≈10,000 seeds genotyped pre-planting each season

Seedling genotyping
 Leaf genotyping and
transplanting allows
more time to
complete the work,
without delaying the
season
 Plants can be also
genotyped in the
field
 Incurs costs for 50%
or >>50% of plants
to be discarded

Favorable CrtB1 allele  2x - 3x ProA
CrtB1
ProA
Pedigree Adaptation favorable
(ug/g DW)
allele

KUI carotenoid syn-FS11-1-1-B-B-B-B-B-B-B-B Sub-trop/Early 5.3

Carotenoid Syn3-FS8-4-3-B-B-B-B-B-B-B-B Sub-trop/Early 5.5

(KUI carotenoid syn-FS17-3-2-B-B-B/(KU1409/
Sub-trop/Early 17.3 +
DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-5-B-B

(KUI carotenoid syn-FS11-1-1-B-B-B/(KU1409/
DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-1-B-B

(CML297/(KU1409/DE3/KU1409)S2-18-2-B)-B-
Sub-trop/Late 15.7 +
1(MAS:L4H1)-1-B-B

(CML297/(KU1409/DE3/KU1409)S2-18-2-B)-B-
Sub-trop/Late 11.0 +
2(MAS:L4H1)-4-B-B

(Florida A plus Syn-FS2-2-1-B-B/(KU1409/
DE3/KU1409)S2-18-2-B)-B-3(MAS:L4H1)-2-B-B

CML300 Sub-trop/Late 7.3
CML297 Sub-trop/V Late 6.3

Methods for carotenoid screening
14.00

12.00

10.00
β-cryptoxanthin

ug/g (ppm)
8.00

6.00

4.00

HPLC 2.00
β-carotene
30/d 0.00
BC1 S2 Lines

UPLC
>100/d

NIR (?)

Cross high proA x good drought tolerance…

2nd Dose (of high proA) F1’s
2nd Dose S1‟s

X XX

What happens to provitamins A during
cooking? 9th ARMW,
Beijing „05

Photo: N. Palacios
≈25-35% loss

Photo: H. De Groote

What happens to provitamins A after
we eat them?

In vitro assessment of
bioaccessibilty of
carotenoids from foods

Mark Failla
Department of
Human
Nutrition
Parker, FASEB J, 1996

Bioavailability promoters in plant foods
Substance Nutrient Major Sources

Certain amino acids (e.g., cysteine, Fe/Zn Animal meats
histidine)

Fats & lipids Vitamin A Animal fats, vegetable oils

Zn, Fe Vitamin A Animal meats

-carotene, provitamin A carotenoids Fe/Zn Orange & green vegetables

Non-digestible carbohydrates (e.g., Ca, Mg, Fe, Zn Onion, garlic, whole wheat grain,
inulin, resistant starch, raffinose) common beans

Vitamin E Vitamin A Vegetable oils, green leafy
vegetables

Ross Welch

Fe bioavailability as affected by ß-
carotene supplement
9th ARMW,
Control with no ß-carotene added Beijing „05

Only 5% of Fe (Garcia-Casal et al., 1998)
is bioavailable

Fe bioavailability: 3 trials @ 2-3 sites
Fe GrYld
• Significant genotypic effects
Trial 1 FerFe -0.53* 0.16
– Significant GCA effects
Trial 2 -0.71** -0.05
– Non-significant SCA effects
Trial 3 -0.39 -0.29
• Large environmental effects
• Small GxE effects Trial 1 FeraaFe -0.71** 0.26
Trial 2 -0.80** -0.07
Trial 3 -0.59* -0.22

GrYld -0.63*
-0.43
0.02
Pixley et al., 2011
(Field Crop Res 123:153-160)

KUI carotenoid syn-FS17-3-1-B-B/(CML-239 x GWIC) -1-7TL-1-1-1-B
17.9 34.8 0.3
Combining proA and Zn
KUI carotenoid syn-FS17-3-1-B-B/CML356-B
20.5 41.1 0.5
KUI carotenoid syn-FS17-3-1-B-B/(CML-356 x GWIB) -1-23TL-1-2-1-B
dehydrogenase 21.1 37.7 0.5
Vision retinol retinal  Zn in the action of retinol dehydrogenase
Zn carotenoid syn-FS17-3-2-B-B/CML353-B
KUI
19.7 32.0 0.3
monooxygenase
(retinol to retinal); essential pigment for
Digestion -carotene 2 retinal vision 18.7 32.5 0.5
Zn
 Zn is a probable co-factor for b-carotene0.2
15.8 29.8
mono-oxygenase (cleaves proA to vitA) 2.0
22.4 35.2
Protein synthesis  Zn deficiency depresses synthesis of the
KUI carotenoid syn-FS17-3-2-B-B/(CML-356 x GWIB) -1-23TL-1-2-1-B
23.1 33.2 2.0
Zn retinol binding protein
(RBP)
retinol:RBP in blood
carrier protein of vitA => lower plasma 2.0
22.9 33.0
KUI carotenoid syn-FS25-3-2-B-B/(CML-239concentrations
retinol x GWIC) -1-7TL-1-1-1-B
22.0 31.9 2.1
KUI carotenoid syn-FS25-3-2-B-B/P903 C0 H364-1-8TL-3-2-1-1-B-B-B-B-B -B
Pedigree
19.8
Fe
29.3
Zn
0.3
Al
KUI carotenoid syn-FS25-3-2-B-B/(CML-356 x GWIB) -1-23TL-1-2-1-B ppm ppm ppm
Tanumihardjo et 22.6 29.8 0.5
al., JAFC 2010 Carotenoid Syn3-FS5-1-5-B-B/CML353-B 18.1 33.3 0.6
20.5 28.6 0.6
17.9 34.8 0.3
Carotenoid Syn3-FS5-1-5-B-B/CML355-B
KUI carotenoid syn-FS17-3-1-B-B/CML356-B 32.0 32.9 0.6
20.5 41.1 0.5
CML-305-B-B/CML356-B
KUI carotenoid syn-FS17-3-1-B-B/(CML-356 x GWIB) -1-23TL-1-2-1-B 17.2
21.1 23.1
37.7 0.4
0.5
CML-304-B-B/CML353-B
KUI carotenoid syn-FS17-3-2-B-B/CML353-B 14.5
19.7 26.0
32.0 0.3
0.3
Average
18.7 31.6 0.8
20.4 32.5 0.5
15.8 29.8 0.2

Trials
2009-10
Mpungwe, SeedCo
Msekera, ZARI
G.Valley, ZARI
Lusaka N, ZamSeed
Lusaka W, SeedCo

Harare, CIMMYT
ART, CIMMYT

8 promising proA hybrids: 5 sites in
Zambia + 2 sites in Zimbabwe
ProA
(ug/g)
Best hybrid
Tons per hectare

Hybrid 1: 8.9
check
Hybrid 2: 7.1
Hybrid 3: 6.3
Hybrid 4: 6.5
Hybrid 5: 7.4
Hybrid 6: 5.7
Hybrid 7: 6.9
Hybrid 8: 5.9

Zambia NPT results, seven sites 2010-2011

Approx. LSD (P<0.05)
Mg ha-1

Mean of 8
commercial
hybrids
Masumba

Mutanda

Msekera

SCCI
Misamfu
GValley
Chiawa

Site

Will consumers and farmers
consume/grow biofortified crops?
• ProA maize is orange; consumers
prefer white
• Will farmers choose to grow orange
maize varieties?
• Will seed companies market the
new orange varieties?

H+ stakeholder consultation, Zambia

Madson Chisi, Zambia Agricultural
Research Institute
– Nshima from finger millet is red; from
sorghum is grey… and people eat
them; color is not a big problem.
– Technology should be neutral or labor
reducing (cooking, processing, etc.)
– Find ways to involve farmers as soon
as possible
Photo
Marx Mbunji, SeedCo credit: H.
– Crop needs to be high yielding; don‟t DeGroote
focus on “micro” things…

H+ stakeholder consultation, Zambia
Matthias Kanyemba, Ministry of Agriculture
– We need to advise the health benefits of orange maize to make it
accepted
– MoA has nutrition workers at all levels – Provincial to Village
Fordson Nyerenda, Ministry of Health:
– Use nutrition, education and communication unit from the
Ministries to promote the orange maize
– Training of partners on Biofortification: Need information and
education for policy makers, extensionists, nutritionists,
farmers...
Beatrice Kawana National Food and Nutrition Commission:
– Nutrition education and communications unit which helps to
produce documentaries, posters, and other information

Raising awareness; creating demand

DELICIOUS &
A. M. Ball NUTRITIOUS!!!

Will consumers buy & consume
biofortified maize?
Mr. M. Ililonga, Zambia Consumer Association
– Consumers are very sensitive about cost (should be affordable);
cost of staple food is of concern to people

$$ $

$$$$ $$$

Multi-disciplinary approaches
0.12

5.473
0.10

16.246
0.08

10.578
7.024
0.06

7.957
AU
+

3.961
+

7.217
++

18.859
4.485

11.517

13.698
6.717
4.750

9.183
0.04

3.332
0.02
0.00
-0.02
2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00
Minutes

Molecular Plant biochemistry
Plant breeding Agronomy
biology
+

+ + + + +

Education & Socio-economics Nutrition Food technology
Farmer participation
marketing

Healthy
children

Bonus Slide: Research question
 Carotenoids and mycotoxins
– Carotenoids in grain  less mycotoxin
production
• The results indicate that the presence of
carotenoids in endosperm may decrease the
amount of aflatoxin produced by A.flavus.
– Carotenoids in food  lesser carcinogenic effect
of toxins
• Carotenoids and vitamin A work via multiple
mechanisms that provide protective support
against cancer initiation and pathogenesis of
other mutation-related diseases.

Norton, Phytopath, 1997
Flora et al., Mutagenesis, 1999

S5.1 Maize HarvestPlus: Biofortifying maize with Provitamin A carotenoids

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