Molecular Breeding for Development of Biofortified Maize Hybrids in India

IndianAgriculturalResearchInstitute
Molecular Breeding for Development of
Biofortified Maize Hybrids in India
Firoz Hossain
Maize Programme Leader
ICAR-Indian Agricultural Research Institute, New Delhi

Malnutrition: Global status
• 2 billion people are malnourished worldwide.
• 815 million people are undernourished worldwide.
• Children (<5 yrs): 151 million stunted. while 51 million
wasted, and 17 million severely wasted.
• Malnutrition contributes to loss in 11% GDP in Asia and
Africa.
Source: Global Nutrition Report-2017
UNICEF/WHO/World Bank Group - Joint Child Malnutrition Estimates-2018
DivisionofGenetics,ICAR-IARI
Stunting WastingOver-weight

Fe, Zn & Vitamin-A deficiency
Source: www.harvestplus.org
Anemia in infants and children
Vitamin A deficiency in pre-school children
Severity of Zn deficiency

Loss in GDP due to malnutrition
Source: www.harvestplus.org
$16
$1
$1 invested in proven nutrition programme offers
benefits worth $16 (IFPRI 2016)

Source: IFPRI, 2016
• Global community committed to a new set of objectives in 2015
• 17 goals anchor the global development agenda till 2030.
• Core is to eliminate extreme poverty, hunger, and malnutrition.
• 12 of the 17 goal-indicators related to nutrition
Sustainable Development Goals (SDGs)DivisionofGenetics,ICAR-IARI

Maize – As a source of livelihood
Regions Area Production Productivity
World 187 mha 1060 mt 5.67 t/ha
Asia 63 mha 324mt 5.14 t/ha
India 9.5 mha 24.3 mt 2.45 t/ha
 Important source of food and
nutritional security in the developing
world, especially in Africa and Latin
America.
 Maize together with rice and wheat
provides at least 30% of the food
calories to more than 4.5 billion people
in 94 developing countries.
 By 2050, the demand for maize in the
developing world will be doubled.
FAOSTAT 2016
Utilization of maize in India

Journey of maize biofortification in IndiaJourney of maize biofortification in IndiaDivisionofGenetics,ICAR-IARI
1957
1997
2001
2005
2007
2010
2017
1971
1998
2004
2006
2008
2013
AICRP-Maize
Hard o2
Composite (1):
Shakti-1
First QPM
Hybrid (1):
Shaktiman-1
QPM
Hybrid (1):
HQPM-1
QPM
Hybrid (1):
HQPM-5
QPM
Hybrid (1):
HQPM-4
Soft o2
Composites (3):
Shakti
Rattan
Protina
1998
AMBIONET
QPM hybrid (1):
Shaktiman-2
QPM
Hybrids (2):
Shaktiman-3,
Shaktiman-4
QPM
Hybrid (1):
HQPM-7
MAS-QPM
Hybrid (1):
Vivek QPM-9
QPM hybrids (2):
Pratap QPM-1,
Shaktiman-5
MAS-ProA
Hybrid (1):
Pusa VQ9 Imp
MAS-QPM
hybrids (3):
Pusa HM4 Imp
Pusa HM8 Imp
Pusa HM9 Imp
99hybrids : 84 normal, 14 QPM, 1 ProA
94 conventionally bred, 5 MAS-bred

• Identified for release
for commercial
cultivation in zone I &
IV; notified in 2008
from CVRC
• Released for
commercial
cultivation by the
State Varietal Release
Committee of
Uttarakhand in 2007
30% lysine
40% tryptophan
First MAS-based hybrid released in 2008

QPM hybrid released in 2017
Pusa HM4 Improved: MAS-based hybrid
North Western Plain Zone
(NHZ):
Punjab, Haryana, Delhi,
Uttarakhand (Plain), UP
(Western region)
Characters AQH4
% tryptophan 0.91
% lysine 3.62
Avg. grain yield
(kg/ha)
6419
Potential yield (kg/ha) 8568
Duration (75% dry
husk)
87

Peninsular Zone (PZ):
Maharashtra, Karnataka,
AP, Telangana &TN
Characters AQH8
% tryptophan 1.06
% lysine 4.18
Avg. grain yield
(kg/ha)
6258
Duration (75% dry
husk)
95

North Eastern Plain Zone
(NEPZ):
Bihar, Jharkhand, Odisha,
UP (Eastern region), West
Bengal
Characters AQH9
% tryptophan 0.68
% lysine 2.97
Avg. grain yield
(kg/ha)
5201
Duration (75% dry
husk)
89

Grain yield in QPM and NormalGrain yield in QPM and Normal
hybridshybrids
AICRP-2016

New mutant:New mutant: opaque16opaque16
• A recessive mutant
• Located on chromosome 8
• Gene is not cloned
• Not available in Indian
germplasm
Dr. Wenpeng Yang, GAAS, China
4F1:
CIMMYT inbred x Chinese inbred

Development of novelDevelopment of novel o16o16o16o16 inbredsinbreds
Normal o16o16 F2 populationsx
CML533 x o16o16
umc1149
CML533 x o16o16
umc1149
S. No. Pedigree Genotype
% Lysine in sample % Tryptophan in sample
Range Mean Range Mean
1. CML533 ×
QCL3024
o16o16 0.111-0.376 0.269 0.027-0.117 0.074
Wild type 0.097-0.197 0.134 0.034-0.055 0.044
2. CML537 ×
QCL3024
o16o16 0.128-0.317 0.223 0.032-0.108 0.069
Wild type 0.107-0.124 0.117 0.010-0.045 0.026
o16o16 o16o16 o16o16 o16o16

Effects ofEffects of opaque16opaque16
o2o2-soft Normal o2o2-modified (QPM)
o2o2/o16o16 o16o16 o16o16
Normal o2o2-soft o2o2-modified (QPM) o16o16 o2o2/o16o16

S. No Hybrid Parentage Area of adaptation
1. HQPM-1 HKI193-1 × HKI163 Across country
2. HQPM-4 HKI193-2 × HKI161 Across country
3. HQPM-5 HKI163 × HKI161 Across country
4. HQPM-7 HKI193-1 × HKI161 Karnataka, AP, Telengana, TN & Maharashtra
opaque16
opaque16
Lysine: Avg. 50% increaseTryptophan: Avg. 87% increase
0.06
0.25
PyramidingPyramiding opaque2opaque2 andand opaque16opaque16
o2
o2
o2
o2
o2
o2
o2
o2
o2+o16
o2+o16
o2+o16
o2+o16
o2+o16
o2+o16
o2+o16
o2+o16

White QPM hybrids with o2 +o16
S. No Hybrid Parentage Area of adaptation
1. HM-5 HKI1344 × HKI1348-6-2 Haryana
2. HM-12 HKI1344 × HKI1378 North Eastern Plain Zone
Donor (o2 + o16) HKI1344 HKI1378 HKI1348-6-2
BC2F2-derived progenies with o2 + o16 are now developed

Variation of carotenoidsVariation of carotenoids
Indian inbreds
CIMMYT-HarvestPlus inbreds
• Indian inbreds are low in β-carotene
• Kernel colour: positively correlated with
lutein, zeaxanthin and β-cryptoxanthin
• Not correlated with β-carotene
• Thus kernel colour can not be used as a
criterion for selection of high β-carotene

Provitamin-AProvitamin-A
enrichment:enrichment:crtRB1crtRB1
Carotenoid biosynthesis pathway
S.
No.
Hybrid Maturity
group
Area of
adaptatio
n
Yield
potential
1. Vivek QPM 9 Extra early NHZ, PZ 55 q/ha
2. Vivek Hybrid 27 Extra early
NEPZ, PZ
55 q/ha
3. HM 4 Medium NWPZ 68 q/ha
4. HM 8 Medium PZ 68 q/ha
ProA at harvest

Release of first proA hybrid in
2017
Pusa Vivek QPM9 Improved: MAS-derived proA maize
Characters APQH9
Provitamin-A (µg/g) 8.15
% tryptophan in protein 0.74
% lysine in protein 2.67
Avg. grain yield-NHZ
(kg/ha)
5588
Potential yield-NHZ (kg/ha) 7968
Avg. grain yield-PZ (kg/ha) 5916
Potential yield-PZ (kg/ha) 9368
Duration (75% dry husk) NHZ: 93
PZ: 83
• Northern Hill Zone (NHZ): J&K, HP, Uttarakhand (Hills) & NEH states
• Peninsular Zone (PZ): Maharashtra, Karnataka, AP, Telengana &TN

Pyramiding ofPyramiding of lcyElcyE andand crtRB1crtRB1 inin
QPM hybridsQPM hybrids
Phytoene synthase
Lycopene cyclase
Carotenoid biosynthesis pathway
S.
No.
Hybrid Parental inbreds Adaptation
1. HQPM-1 HKI193-1 x HKI163 Across the country
2. HQPM-4 HKI193-2 x HKI161 Across the country
3. HQPM-5 HKI163 x HKI161 Across the country
4. HQPM-7 HKI193-1 x HKI161
AP, TN, Telengana,
Karnataka, Maharashtra
QPM hybrids targeted for provitamin-A enrichment
x
HQPM-1 HQPM-1-ProA
Provitamin-A: 2.3 ppm Provitamin-A: 9.95 ppm
Three proA version are in National Testing

ICAR-MaizeBiofortificationNetwork
Newly derived proA rich Indian maize germplasm
• ProA among the elite inbreds: 1.2-2.2 ppm,
• New crtRB1-based progenies: mean proA: 15.9 ppm, range: 9.4-21.5 ppm
M F2 individuals
F2 populations (>100 individuals): 10 elite inbreds x HP704-22 (crtRB1 favourable)

Enrichment of vitamin EEnrichment of vitamin E
• Maize kernel is rich in total tocopherol, γ-tocopherol: ~80%; α-tocopherol: ~20%.
• α-tocopherol possesses higher vitamin E activity.
• In humans, it quenches free radicals and protects PUFA from damage.
• It protects from cardiovascular-, Alzheimer's-, neurological- and age related- disorders.
• 20% people possess suboptimal α-tocopherol – requires urgent attention.
• Favourable VTE4 allele can increase α-tocopherol by 3.2 fold

S.
N.
Hybrid Parental inbreds Adaptation Genes targeted
1. HQPM-1-PV HKI193-1 x HKI163 Across the country o2+crtRB1+lcyE+VTE4
2. HQPM-4-PV HKI193-2 x HKI161 Across the country o2+crtRB1+lcyE+VTE4
3. HQPM-5-PV HKI163 x HKI161 Across the country o2+crtRB1+lcyE+VTE4
4. HQPM-7-PV HKI193-1 x HKI161 AP, TN, Kar, Tel, MH o2+crtRB1+lcyE+VTE4
HKI161-VTE HKI163-VTE
Triple-biofortified maize hybridsTriple-biofortified maize hybrids
[QPM+Provitamin-A+vitamin-E][QPM+Provitamin-A+vitamin-E]

Kernel -Fe and -ZnKernel -Fe and -Zn
Kernel –Fe and –Zn:
Governed by polygenes
Effect of environment is too high
30 ppm Fe
5% bioavailability
3.0 ppm
30 ppm Fe +
lpa1/ lpa2 10% bioavailability
3.0 ppm
Direct approach
Indirect approach
lpa1 & lpa2 contains 50-60%
reduced phytic acid
 Reduced phytic acid (lpa) - enhances the bioavailability of -Fe and -Zn
 Increase in Lysine and ProA - enhances the bioavailability of -Fe and -Zn
60 ppm Fe
Baseline
Target

Phytate biosynthesisPhytate biosynthesis
 lpa mutants in maize: lpa1, lpa2-1, lpa2-2, lpa3, lpa241
 Mutants viz. lpa1 and lpa2-1 used in our programme

Introgression of lpa allele
S. No. Hybrid Parentage
Maturity
group
Area of adaptation
GY
(t/ha)
1 HQPM-1-PV HKI193-1-PV × HKI163-PV Late NWPZ, NEPZ, CWZ, PZ 6.2
2 HQPM-4-PV HKI193-2-PV × HKI161-PV Late NWPZ, NEPZ, CWZ, PZ 6.0
3 HQPM-5-PV HKI163-PV × HKI161-PV Late NWPZ, NEPZ, CWZ, PZ 5.8
4 HQPM-7-PV HKI193-1-PV × HKI161-PV Late PZ 7.2
5 AQH4 HKI1105Q × HKI323Q Medium NWPZ 6.8
6 AQH8 HKI1105Q × HKI161 Medium PZ 6.8
7 AQH9 HKI1105Q × HKI1128Q Medium NEPZ 6.0
8 AQH10 HKI-193-2 x HKI-1128Q Medium NWPZ, PZ, CWZ 7.2
9 AQH11 HKI1128Q x HKI163 Late NWPZ, NEPZ, CWZ, PZ 5.5
• High phytate causes deficiency in P, Ca, Fe and Zn
• Phosphate is supplemented in the diet - adds to cost
• Phytase is used in diet which is costly - adds to cost
Feed specific maize hybrids!

Pusa Super Sweet Corn-1 (sh2sh2) [2018]
• Northern Hill Zone (NHZ)
• North Western Plain
Zone (NWPZ)
• North Eastern Plain Zone
(NEPZ)
• Peninsular Zone (PZ)
Characters ASKH-4
% avg. brix 15.9
Avg. green cob yield
(t/ha)
13.0
Avg. dehusked cob
yield (t/ha)
9.3
Potential dehusked
cob yield (t/ha)
10.2
Fodder yield (t/ha) 16.2
Avg. maturity (DAS) 77.6
Biofortification of sweet cornBiofortification of sweet corn

Sweet corn hybrids in AternaSweet corn hybrids in AternaDivisionofGenetics,ICAR-IARI
Dehusked cob
yield:
ASKH-1: 10.8 t/ha
ASKH-2: 9.8 t/ha
ASKH-4: 10.2 t/ha
Sweetness in
Kernel:
20-22% brix
ASKH-1 & ASKH-
4 evaluated in
NIVT & AVT-I
ASKH-1
ASKH-4
ASKH-2 ASKH-2
ASKH-1
&
ASKH-4:
AVT-II
(K-2017)

Sweet Corn + ProA + QPMSweet Corn + ProA + QPMDivisionofGenetics,ICAR-IARI
S.
No.
Hybrid Parental inbreds Adaptation Genes targeted
3. HQPM-1 HKI193-1 x HKI163 Across the country sh2+o2+crtRB1+lcyE
4. HQPM-4 HKI193-2 x HKI161 Across the country sh2+o2+crtRB1+lcyE
5. HQPM-5 HKI163 x HKI161 Across the country sh2+o2+crtRB1+lcyE
6. HQPM-7 HKI193-1 x HKI161
AP, TN, Telangana,
Karnataka, Maharashtra
sh2+o2+crtRB1+lcyE
HKI161* HKI163* HKI193-1* HKI193-2*
*BC2F1 cobs:
Homozygous for =
o2+crtRB1+lcyE
sh2sh2 sh2sh2 sh2sh2 sh2sh2

543bp
296bp
491bp
373bp
Sweet Corn + ProA + VitESweet Corn + ProA + VitE
crtRB1
VTE4

Biofortified maize hybrids at IARIBiofortified maize hybrids at IARI
S. N. Traits Gene combination
1. QPM opaque2
2. QPM-High AA opaque2 + opaque16
3. ProA crtRB1
4. ProA + QPM opaque2 + crtRB1/lcyE
6. ProA + QPM + VitaminE opaque2 + crtRB1 + lcyE + VTE4
7. ProA /QPM + low phytate opaque2 /crtRB1 + lpa1/lpa2
8. Fe and Zn lpa1/lpa2
9. Sweet corn + ProA + VitaminE shrunken2 + crtRB1 + VTE4
10. Sweet corn + ProA + QPM shrunken2 + crtRB1 + opaque2
35 hybrids with one or multiple trait combination available

ICAR-MaizeBiofortificationNetwork
Effects of QPM on ‘Vanaraja’
Diet 3 week 6 week 9 week
BWG FCR BWG FCR BWG FCR
Diet 1 Yellow Maize 205.3 2.023 529.7 2.181ab 909.2 2.361
Diet 2 Vivek Hyb 9 213.0 1.979 586.9 2.202a 956.1 2.404
Diet 3 Pusa VQ9 Improved 210.6 1.980 583.4 2.034c 972.4 2.339
Diet 4 Vivek QPM 9 186.6 2.182 584.8 2.131bc 960.7 2.357
Diet 5 White Maize 191.4 2.096 563.6 2.177ab 962.0 2.252
N 7 7 7 7 7 7
1 2 3 4 5
2.0
2.1
2.2
2.3
BWG
FCR
500
550
600
650
Diets
BWGFCR (FI/BWG)
1: DPR Maize, 2: Vivek Hyb9, 3: APQH9, 4: Vivek QPM 9, 5: White maize
Effect of feeding different source of maize on
performance in Vanaraja birds during 6 weeks of age
QPM Normal
QPM
+
ProA
Normal Normal
VH9 PVQ9I VQ9
Vanaraja

ICAR-MaizeBiofortificationNetwork Effects of biofortified maize on poultryDivisionofGenetics,ICAR-IARI
Díaz-Gómez et
al. 2015
Moreno et al. 2016

Bioavailability of proA using in vitro model
Consumption of 200 g of proA hybrids per day:
•Pusa-PV-16-3: 52% RDA
•Pusa-APQH8: 64% RDA
in adult Indian men after adjusting for cooking losses and conversion
factors.
•Human trial using QPM + ProA maize to be undertaken in 2018-19 by
National Institute of Nutrition, Hyderabad: Grains of ‘Pusa Vivek
QPM9 Improved’ have been generated in isolation

Biofortification Network-India
1. CRP-Biofortification in Maize
for Nutritional Security (5):
IIMR, IARI, VPKAS, HAU &
TNAU
2. CRP on Molecular Breeding
(3): IARI, VPKAS & IIMR
3. DBT-Biofortification Network
(3): IARI, PAU & TNAU
4. Other institution: JNKVV,
ICAR-RC-NEH, HPKV &
PJATSAU/ANGRAU
5. Poultry research: DPR
6. Human nutrition: NIN

India’s National Nutrition Strategy-2017
• Launched on September 5, 2017 by NITI
Aayog
• Four proximate determinants of nutrition:
Nutritional
Security
Health
service
Food
Drinking
water &
sanitation
Income &
livelihoods
The Nutrition Strategy framework
envisages a Malnutrition Free India
Development + Awareness + Policy = Successful Implementation

Acknowledgements
• ICAR
• Director, IARI, New Delhi
• Director, IIMR, Ludhiana
• Dr. BM Prasanna
• Dr. Sain Dass
• IIMR-Winter Nursery
• CCS-HAU, Uchani Centre
• VPKAS, Almora
• AICRP-Maize Centres
• ICAR-CRP
• DBT & DST, Govt. of India
• CIMMYT-HarvestPlus
• USDA & GAAS
• Drs. Vignesh & Rajkumar
• Collaborators
• Students & Research Fellows

Molecular Breeding for Development of Biofortified Maize Hybrids in India

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