s5.1 maize harvestplus: biofortifying maize with provitamin a carotenoids

Maize HarvestPlus:

Biofortifying maize

with Provitamin A carotenoids

11th Asian Maize Conference

November 7-11, 2011; Nanning, China

Kevin Pixley, Natalia Palacios, Raman Babu

& Abebe Menkir (CIMMYT & IITA)

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

December, according to the UN's Food

and Agricultural Organisation (FAO).

Its Food Price Index went above the

previous record of 2008 that saw prices

spark riots in several countries.

The FAO's Abdolreza

Abbassian warned over

low level food reserves

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


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.

Middle East


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

Pedigree Adaptation ProA

(ug/g DW)

CrtB1 favorable

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/ DE3/KU1409)S2-18-2-B)-B-1(MAS:L4H1)-5-B-B

Sub-trop/Early 17.3 +

(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-1(MAS:L4H1)-1-B-B

Sub-trop/Late 15.7 +

(CML297/(KU1409/DE3/KU1409)S2-18-2-B)-B-2(MAS:L4H1)-4-B-B

Sub-trop/Late 11.0 +

(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

Favorable CrtB1 allele 2x - 3x ProA

Methods for carotenoid screening

NIR (?)

HPLC 30/d 0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

ug

/g (

pp

m)

BC1 S2 Lines

β-cryptoxanthin

β-carotene

UPLC >100/d

Cross high proA x good drought tolerance…

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

X X X

Photo: N. Palacios

Photo: H. De Groote

What happens to provitamins A during

cooking? 9th ARMW,

Beijing „05

≈25-35% loss

What happens to provitamins A after

we eat them?

Parker, FASEB J, 1996

Mark Failla

Department of

Human

Nutrition

In vitro assessment of

bioaccessibilty of

carotenoids from foods

Bioavailability promoters in plant foods

Substance Nutrient Major Sources

Certain amino acids (e.g., cysteine,

histidine)

Fe/Zn Animal meats

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.,

inulin, resistant starch, raffinose)

Ca, Mg, Fe, Zn Onion, garlic, whole wheat grain,

common beans

Vitamin E Vitamin A Vegetable oils, green leafy

vegetables

Ross Welch

Fe bioavailability as affected by ß-

carotene supplement

Control with no ß-carotene added

(Garcia-Casal et al., 1998)

9th ARMW,

Beijing „05

Only 5% of Fe

is bioavailable

Fe bioavailability: 3 trials @ 2-3 sites

• Significant genotypic effects

– Significant GCA effects

– Non-significant SCA effects

• Large environmental effects

• Small GxE effects

Fe GrYld

Trial 1 FerFe -0.53* 0.16

Trial 2 -0.71** -0.05

Trial 3 -0.39 -0.29

Trial 1 FeraaFe -0.71** 0.26

Trial 2 -0.80** -0.07

Trial 3 -0.59* -0.22

GrYld -0.63*

-0.43

0.02Pixley et al., 2011

(Field Crop Res 123:153-160)

dehydrogenase

Vision retinol retinal

Zn

monooxygenase

Digestion -carotene 2 retinal

Zn

Protein synthesis

Zn retinol binding protein retinol:RBP in blood

(RBP)

Zn in the action of retinol dehydrogenase

(retinol to retinal); essential pigment for

vision

Zn is a probable co-factor for b-carotene

mono-oxygenase (cleaves proA to vitA)

Zn deficiency depresses synthesis of the

carrier protein of vitA => lower plasma

retinol concentrations

Pedigree Fe Zn Al

ppm ppm ppm

KUI carotenoid syn-FS17-3-1-B-B/CML353-B 18.1 33.3 0.6

KUI carotenoid syn-FS17-3-1-B-B/(CML-239 x GWIC) -1-7TL-1-1-1-B 17.9 34.8 0.3


KUI carotenoid syn-FS17-3-1-B-B/(CML-356 x GWIB) -1-23TL-1-2-1-B 21.1 37.7 0.5








KUI carotenoid syn-FS25-3-2-B-B/P903 C0 H364-1-8TL-3-2-1-1-B-B-B-B-B -B 19.8 29.3 0.3


Carotenoid Syn3-FS5-1-5-B-B/CML353-B 20.5 28.6 0.6


CML-305-B-B/CML356-B 17.2 23.1 0.4

CML-304-B-B/CML353-B 14.5 26.0 0.3

Average 20.4 31.6 0.8

Combining proA and Zn

Tanumihardjo et

al., JAFC 2010

Pedigree Fe Zn Al

ppm ppm ppm












KUI carotenoid syn-FS25-3-2-B-B/P903 C0 H364-1-8TL-3-2-1-1-B-B-B-B-B -B 19.8 29.3 0.3




CML-305-B-B/CML356-B 17.2 23.1 0.4

CML-304-B-B/CML353-B 14.5 26.0 0.3

Average 20.4 31.6 0.8

Obatanpa-ProA

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

Best hybrid

check

To

ns

per

he

cta

re

Hybrid 1: 8.9

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

ProA

(ug/g)

Mg h

a-1

Ch

iaw

a

Masu

mb

a

GV

alley

Mu

tan

da

Mis

am

fu

Ms

ekera

SC

CI

Zambia NPT results, seven sites 2010-2011

Site

Mean of 8

commercial

hybrids

Approx. LSD (P<0.05)

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

Marx Mbunji, SeedCo

– Crop needs to be high yielding; don‟t

focus on “micro” things…

Photo

credit: H.

DeGroote

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 &

NUTRITIOUS!!! A. M. Ball

Mr. M. Ililonga, Zambia Consumer Association

– Consumers are very sensitive about cost (should be affordable);

cost of staple food is of concern to people

Will consumers buy & consume

biofortified maize?

$$

$$$$ $$$

$

Molecular

biology Plant breeding

Multi-disciplinary approaches

+ +

Agronomy

+ + +

Food technology Nutrition Socio-economics Education &

marketing

Healthy

children

AU

-0.02

0.00

0.02

0.04

0.06

0.08

0.10

0.12

Minutes

2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00

3.3

32

3.9

61

4.4

85

4.7

50

5.4

73

6.7

17

7.0

24

7.2

17

7.9

57

9.1

83 10.5

78

11.5

17

13.6

98

16.2

46

18.8

59

+ +

Plant biochemistry

+

+ +

Farmer participation

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

Xie xie!