Breeding For Biofortification in Cereals

Presented By

Ashwani KumarRegd. No. – J-13-D-180-A

Division of Plant Breeding And GeneticsSher-e-Kashmir University of Agricultural Sciences & Technology,

Jammu

3 billion people worldwide suffer micronutrients deficiency

2.5 billion world population suffer from Zinc deficiency

1.6 billion population suffer from Iron deficiency1 billion people reside in iodine deficient regions400 million people have vitamin A deficiency Malnutrition accounts ~30 million death/year

Malnutrition Problem

Source : WHO, 2012

Nearly Half of The World Population is Affected From Iron & Vitamin A Deficiency

Source:- Welch and Graham, 2010; Field Crops Res.

Wide Spread Zinc Deficiency

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

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

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

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

Bio-fortification Differs Ordinary Fortification

More nutrients

consumed

Dietry supplements

Varied, plant-based diet

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

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

Genetic Bio-fortified Crops

Source : Harvest Plus Programme

Discovery Identify target population Set nutrient target level

Screen germplasm & gene discovery

Development

Breed bio-fortified cropsTest the performance of New crop varietiesMeasure Nutrient retention in cropEvaluate Nutrient Absorption & Impact

Dissemination

Develop strategies to disseminate the seedPromote marketing & Consumption of Bio-fortified crops

Improve Nutritional Status of Target Population

Pathway for Biofortification

Source : HarvestPlus, 2009

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

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

Variation for Fe content in major cereals crops documented in various studies

Source : Goudia & Hash, 2015

30

Variation for Zn content in major cereals crops documented in various studies

Source : Goudia & Hash, 2015

30

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.

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).

Biofortification Through Breeding High Iron Pearl Millet

ICTP8203ICRISAT Bred OPV

(70-74 ppm Fe)With 10% Higher Yield

Marketed by NIRMAL Seeds

86M86Pioneer Hybrid (54-63ppm Fe)

Pearl Millet Cultivar Commercialized In India

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

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

World’s first high-zinc rice released in Bangladesh

Released Varieties

Source : HarvestPlus, 2014

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

Fe

Zn

Wild Type Transgenic

Genetic Engineering For Bio-FortificationGenetic 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

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 APathway

is completeand functional

GoldenRice

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?

Genetic Modification: Golden RiceBC derived lines in Swarna background using

Kaybonet-GR2-R event as donor

Swarna Golden Swarna

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

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

S.No. Species Numberof accessions

Genome Iron mg/kg Zinc mg/kgRange Mean Range Mean

1 T. aestivum 13 ABD 21.26- 30.59 27.69 14.88 - 19.33 22.152 T. durum 2 AB 21.91 – 25.60 23.58 13.68- 19.60 18.793 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 Sl59.12 – 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

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

Released Varieties

Sources : HarvestPlus Wheat for Zinc

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.

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.

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

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

Released Varieties

Sources : HarvestPlus Mazie for Vit A

Source : Goudia & Hash, 2015

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