Breeding cereals for biofortification can help address widespread micronutrient deficiencies. Variability exists among crop varieties for iron and zinc content. Pearl millet varieties with 10-30% higher iron and zinc have been developed through breeding. For rice, high zinc varieties with 35-40 μg/g zinc in polished grains have been identified. Golden rice has been developed through genetic engineering to produce beta-carotene and address vitamin A deficiency. Wheat breeding draws on wild relatives and landraces to introgress genes for higher iron and zinc into elite varieties. Ongoing biofortification research and new varieties developed through conventional and molecular breeding aim to make staple crops more nutritious.

1. Breeding For Biofortification in
Cereals
Presented
By
Ashwani Kumar
Regd. No. – J-13-D-180-A
Division of Plant Breeding And Genetics
Sher-e-Kashmir University of Agricultural Sciences & Technology,
Jammu

2. 3 billion people worldwide suffer micronutrients
deficiency
2.5 billion world population suffer from Zinc
deficiency
1.6 billion population suffer from Iron deficiency
1 billion people reside in iodine deficient regions
400 million people have vitamin A deficiency
 Malnutrition accounts ~30 million death/year
Malnutrition Problem
Source : WHO, 2012

3. Nearly Half of The World Population is Affected From
Iron & Vitamin A Deficiency
Source:- Welch and Graham, 2010; Field Crops Res.

4. Wide Spread Zinc Deficiency
(Alloway, 2014, In: Zinc in soils and Crop Nutrition. IZA Publications, Brussels)

5. India is one of the countries having
problem of malnutrition
More than 50% of women, 46% of
children below 3 years are
underweight and 38% are stunted
As per India state hunger index, all
the states are with serious to alarming
indices with M.P. most alarming.
In India
Source : World Bank

6. Food availability is not a problem, nor it like to
be….
More important is what kind of food will be
available
- Nutritious crops
- Biofortified crops – staple crops breed for
additional micronutrients
How can we Nourish 1.2 Billion People

7. Bio-fortification:
Greek word “bios” means “life” and Latin word “fortificare” means
“make strong”.
Bio-fortification:
Biofortification is a method of breeding crops to increase their
nutritional value
Bio-fortification refers to increasing genetically the bio-available
mineral content of food crops (Brinch-Pederson et al., 2007).
Bio-fortification differs from ordinary fortification because it
focuses on making plant foods more nutritious as the plants
are growing, rather than having nutrients added to the foods
when they are being processed.
What is Bio-Fortification

8. Bio-fortification Differs Ordinary Fortification
More
nutrients
consumed
Dietry
supplements
Varied, plant-
based diet

9. Some points present here to clearly identified role of crop bio
fortification …….
To overcome the mal-nutritions in human beings
To increment of nutritional quality in daily diets
To improvement of plant or crop quality and increment of
variability in germplasm
Biofortification for important crop plants through biotechnological
applications is a cost-effective and sustainable solution for
alleviating VAD, etc.,.
Importance of crop Biofortification

10. India Biofortification
 Indian Parliament recenttly has passed a budget which
includes $15 million for biofortification (DBT) for
rice, wheat and maize over five years.
 Crop leaders appointed for each crops; traget nutrients
are iron, zinc and vitamin A.
 Joint meetings held every years
 MOU has been signed
Source : MoA, Govt. of India

11. Genetic Bio-fortified Crops
Source : Harvest Plus Programme

12. Discovery
Identify target population
Set nutrient target level
Screen germplasm & gene discovery
Development
Breed bio-fortified crops
Test the performance of New crop varieties
Measure Nutrient retention in crop
Evaluate Nutrient Absorption & Impact
Dissemination
Develop strategies to disseminate the seed
Promote marketing & Consumption of Bio-fortified crops
Improve Nutritional Status of Target Population
Pathway for Biofortification
Source : HarvestPlus, 2009

13. The amount of Fe, Zn and Vit A 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
Breeding Target

14. Iron Biofortification in Cereals
Germplasm still below the 100% traget levels by 2013 for the
three main cereals even if breeding would concentrate on
increasing iron levels
No direct breeding efforts for iron for rice, wheat and maize
under HarvestPlus II
Transgenic approach is only option

15. Variation for Fe content in major cereals
crops documented in various studies
Source : Goudia & Hash, 2015
30

16. Variation for Zn content in major cereals
crops documented in various studies
Source : Goudia & Hash, 2015
30

17.  In this study 122 hybrids (21 hybrids from 9 public sector research
organizations, including ICRISAT; and 101 hybrids from 33 seed companies)
was used.
 This study showed the existence of about two fold variability for Fe density
(31–61 ppm) and zinc density (32–54 ppm) among 122 commercial and
pipeline hybrids developed in India.
Pearl Millet
 India, which has the largest pearl millet area (>9 mh) in the world.
 Pearl Millet, as a species, has higher levels of Fe and Zn densities than other
major cereal crops.

18. Objective: To compare the capacity of iron (Fe) biofortified and
standard pearl millet (Pennisetum glaucum L.) to deliver Fe for
hemoglobin (Hb)-synthesis.
Methods: Two isolines of PM, a low-Fe-control (“DG-9444”, Low-
Fe) and biofortified (“ICTP-8203 Fe”,High-Fe) in Fe (26 μg and 85
μg-Fe/g, respectively) were used.
Results: Improved Fe-status was observed in the High-Fe
group, as suggested by total-Hb-Fe values (15.5±0.8 and
26.7±1.4 mg, Low-Fe and High-Fe respectively, P<0.05).

19. Biofortification Through Breeding High Iron
Pearl Millet
ICTP8203
ICRISAT Bred OPV
(70-74 ppm Fe)
With 10% Higher Yield
Marketed by NIRMAL Seeds
86M86
Pioneer Hybrid (54-63ppm Fe)

20. Pearl Millet Cultivar
Commercialized In India

21.  Rice is a staple food crop for more than 1 billion poor people.
 The Rice endosperm is deficient in many nutrients
including vitamins, proteins, micronutrients, EAAs, etc.
 The Aleurone layer of dehusked rice grains is nutrient rich
but is lost during milling and polishing.
 Rice plants produce β-carotene (provitamin A) in green
tissues but not in the endosperm (the edible part of the seed).
 To overcome the deficiency of vit A in human beings.
Rice Biofortification

22. 3500 rice assessions, 100 popular lines have been
screened
14 genotypes with high Zn content in polished
grains with 35-40ug/g have been identified.
Selection and phenotyping of 40 rice genotypes
are under multi-location trails.
Breeding for High Zinc Rice
Source: MSSRF & IGAU, Raipur

23. World’s first high-zinc rice released in Bangladesh

24. Released Varieties
Source : HarvestPlus, 2014

25. Methods used for Rice Biofortification
Marker Assisted Selection
 Five Mapping population have
been developed and purified
 Molecular marker for genes
associated with iron uptake,
transport and accumulation
have been designated
 Marker Assisted Selection is
eligible for organic certification

26. Fe
Zn
Wild Type Transgenic

27. Genetic Engineering For Bio-Fortification
Genetic engineering is the obvious alternative to enhance the β-
carotene levels in crop plants.
The development of the ‘golden rice’ proved that, it is possible to
redirect a complete biosynthetic pathway of carotenoids by genetic
engineering of multiple genes encoding key enzymes of the
pathway.
 So, Golden Rice is such a bio-fortified crop.
A example of Golden Rice was developed in the year 2000

28. The Golden Rice Solution
IPP (Isopentenyl pyrophosphate)
Geranylgeranyl diphosphate
Phytoene
Lycopene
 -carotene
(vitamin A precursor)
Phytoene synthase
Phytoene desaturase
Lycopene-beta-cyclase
ξ-carotene desaturase
Daffodil gene
Single bacterial gene;
performs both functions
Daffodil gene
-Carotene Pathway Genes Added
Vitamin A
Pathway
is complete
and functional
Golden
Rice

29. Addition of 2 genes in rice genome will complete the biosynthetic
pathway:
1. Phytoene synthase (psy): derived from daffodils (Narcissus
pseudonarcissus). Psy is a transferase enzyme involved in
the biosynthesis of carotenioids. It catalyzes the
conversion of GGPP to phytoene.
2. Lycopene cyclase (crt1)- isolated from soil bacteria Erwina
uredovora.
3. Produce enzymes and catalysts for the synthesis of carotenoids in
the endosperm of rice.
How Does It Work?

30. Swarna Golden Swarna

31. Breeding strategy for Wheat
Low genetic variation in cultivated wheat for Zn/Fe
Wild relatives (T. spelts, Ae. tauschii, emmer wheat and
landraces) known to have upto 190 ppm
Recreated synthetics, wild and landraces are being used as
Progenitor for high Zn/Fe
Limited backcross approach to introgress high Zn genes into
elite wheats
Selected bulk scheme- Most effective method
2nd round of breeding using wide-cross derived lines with
better yielding parents
A rapid, High-throughput, non-destructive XRF machine being
used for fast-track Zn/Fe analysis

32. Wheat Biofortification Initiatives
CGIAR’s HarvestPlus Challenge program to breed
nutrient dense staple foods
 Synthetic hexaploid wheat from T. dicocicon and Aegilops taushii with high
micronutreint were used in CIMMYT wheat breeding program.
 Developed agronomically superior wheat with 100% more Zinc and 30% more
Iron than the morden cultivars.
 Zn intake was 72% higher from the biofortified wheat with 95% extraction and
0.5mg/d higher absorption than the control wheat.
Department of Biotechnology, Govt. India “Biofortification of Wheat
for enhanced micronutrients using conventional and molecular
breeding" Phase I (2005) and Phase II (2011)
 PAU, Ludhiana using progenitor A and B genomes and related species
 IARI, New Delhi using progenitor D genome
 IIT Roorkee; Eternal University, Baru Sahib; G.B.P.U.A.&T. Pantnagar using non-
progenitor species with S, U and M genomes

33. S.No. Species Number of
accessions
Genome Iron mg/kg Zinc mg/kg
Range Mean Range Mean
1 T.aestivum 13 ABD 21.26- 30.59 27.69 14.88 - 19.33 22.15
2 T.durum 2 AB 21.91 – 25.60 23.58 13.68- 19.60 18.79
3 T.boeoticum 19 Am
23.88 – 40.50 30.91 22.12 - 39.06 29.27
4 T. dicoccoides 17 AB 27.67 – 42.67 32.98 22.50 – 66.51 35.33
5 T.arraraticum 6 AG 23.10 – 59.06 29.85 19.27 – 30.54 23.52
6 Ae longissima 5 Sl
59.12 – 81.59 73.24** 24.99 – 50.52 41.66
7 Ae. kotschyi 14 US 22.89 – 90.96 67.46** 22.29 – 58.61 49.27
8 Ae. peregrina 10 US 34.37 – 82.32 52.85** 33.13 – 49.49 39.54
9 Ae. cylindrica 3 CD 52.21-93.27 66.76** 32.38 – 52.18 38.51
10 Ae. ventricosa 3 DN 55.41 – 93.52 65.75** 24.01 – 39.08 33.81
11 Ae. ovata 3 UM 52.25 – 81.97 69.95** 31.93- 40.81 37.7
Range and mean of grain iron and zinc content of wheat and durum cultivars
and wild Triticum and Aegilops species

34.  Screening of several 100 wheat accessions
 Showed 4-5 fold variability for grain Fe & Zn
 Range of concentration in hexaploid wheat,
T. dicoccon & landraces
Range Mean
Fe 25-56 mg/kg 37 mg/kg
Zn 25-65 mg/kg 35 mg/kg

35. Released Varieties
Sources : HarvestPlus Wheat for Zinc

36.  Fe and Zn concentrations were evaluated in a set of 30 diverse maize genotypes .
 Ranges of Fe and Zn concentrations were 11.28–60.11 mg/kg and 15.14–52.95
mg/kg, respectively.
 Based on the performance 4 highly promising inbreds and 3 landrace accessions
were identified as highly promising for Fe concentration, including a HarvestPlus
line, HP2 (42.21 mg/kg).
 Similarly, for Zn concentration, three inbreds and one landrace were identified as
highly promising, including V340 (43.33 mg/kg).
 Study identified HP2 and BAJIM 06-17 for Fe concentration and IML467 for Zn
concentration as the most stable genotypes across the environments.

37. Development of vitamin A-rich cereals can help in alleviating the
widespread problem of vitamin A deficiency.
 A favourable allele of the b-carotene hydroxylase (crtRB1) gene
was introgressed in the seven elite inbred parents, which were low
(1.4 mg/g) in kernel b-carotene.
Concentration of b-carotene among the crtRB1-introgressed inbreds
varied from 8.6 to 17.5 mg/g – a maximum increase up to 12.6-fold
over recurrent parent.
The reconstituted hybrids developed from improved parental
inbreds also showed enhanced kernel b-carotene as high as 21.7
mg/g, compared to 2.6 mg/g in the original hybrid.

39. Maize lacks lysine and tryptophan necessary for
protein synthesis
QPM contains a naturally-occurring mutant (opaque2)
maize gene that increses the amiunt of those two
essential amino acids
Two studies shows that children consuming QPM had a
growth rate in height 15% greater than that of children
who ate conventional maize
Quality Protein Maize (QPM) Improve
growth rate of children
Sourse :CIMMYT, 2014

40. Few seed companies are willing to invest as there is little
profit incentive
- Research and maintenance costs and lack of market
premium
QPM must be grown separately from conventional maize
- To prevent dillution from natural gene flow
- Labelling and consumer education are necessary
- Framer are unable to distinguish QPM with other
varieties
Obstacles for QPM

42. Released Varieties
Sources : HarvestPlus Mazie for Vit A

43. Source : Goudia & Hash, 2015

44. Thank you
Food is the moral right of all who are
born into this world -- Borlaug
Nutritious food is the moral right of all
who are born into this world