Biofortification of vegetable crops

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2. Hunger Green revolution
Hidden Hunger
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5. Increase the production of nutritious food
to feed the hidden hungry planet
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6. University of horticultural sciences , bagalkot
k. R. C. college of horticulture , arabhavi
department of vegetable science
Biofortification of Vegetable crops
Basavaraj S Panjagal
Ph.D in Vegetable Science
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Seminar – I

7. INTRODUCTION
THE CONCEPT OF BIO FORTIFICATION
WHY BIOFORTIFICATION?
HOW IT DIFFER FROM ORDINARY FORTIFICATION
METHODS OF BIOFORTIFICATION
EXAMPLES FOR BIOFORTIFICATION IN VEGETABLE CROPS
CASE STUDIES
ACHIEVEMENTS OF BIOFORTIFICATION IN VEGETABLE CROPS
CONCLUSION
Topic Division
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8. HUMAN REQUIREMENT
Macro nutrient + micronutrient
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Figure 01 : World Risk Factors Causing Deaths
Malnutrition accounts of ≈ 30 million deaths
per year (≈ 1 death per second) (WHO estimate)
Risk
Factor

10. • Currently an estimated 92 million people are suffering from food and
nutrition insecurity (FAO, 2016).
• Malnutrition is responsible for 35% of all child deaths and 11% of the
global disease burden (FAO, 2016).
• “1 in every 5 people in the developing world is chronically undernourished
and more than half of 12 million child deaths each year are related to
malnutrition” (WHO,2016).
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11. Major Micronutrient deficiencies in human
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12. Welch and Graham, 2015; Field Crops Res
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14. Vitamin A deficiency
 A significant health problem in
world
Some VAD characteristics are
 Vision impairment
Night blindness{ treatable}
Permanent blindness{ if
untreated}
 Weakened immune system
Higher risk of sickness
Higher death rate
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16. Zinc Deficiency Symptoms
1) Illness from infectious
diseases, poor child growth;
2) pregnancy and childbirth
complications;
3) reduced birth weight
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17. Table-01 : Age and sex specific estimated average
requirements for vitamin A and iron
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18. Solution for Micronutrient deficiency
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19. Why Biofortification ?
 Fortification and supplementation are shorter term
public health interventions; mostly for acute cases
 Require infrastructure, sophisticated processing
technology, purchasing power, access to markets
 Not available in remote areas
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20. • Bios {Greek}=life
• Fortificare { Latin}= make strong
• Biofortification is the process of increasing the bio
available concentrations of an element in edible
portions of crop plants through traditional breeding
practices or modern biotechnology
White, 2006
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21. How it differs from fortification?
Biofortified
Bean rich in
Iron
Iodine
Fortified
Salt
Biofortification 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.
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22. Biofortification differ from Ordinary fortification
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23. Commercially available fortified foods
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24. Biofortification is Complementary to
Other Interventions
Improve Food
and Nutrition
Security
Fortified
Foods
Dietary
Diversity
Supplementation
Improved Crop
Productivity
Crop
Biofortification
Nutrition
Education
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25. Steps
• Identification of genetic variability
• Introgressing this variation into end-use quality attributes
• Testing the stability of micronutrient
• Large scale deployment of seed of improved cultivars to farmers
Biofortification requires a multidisciplinary research approach
- Direct linkages between agricultural researchers and various specialists like
nutritionists, public health officials, sociologists, political scientists, food
technologists and economists
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26. Methods of Biofortification
Agronomic
Biofortification
Conventional
breeding
Genetic engineering
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27. Table:2 SWOT analysis of bio fortification methods
Methods Agronomical Conventional Modern Method
STRENGTHS:
Comparatively simple
method than other methods
 suitable for immediate
results.
 Successful for minerals
and vitamins,
 one-off cost,
 Easier distribution,
 Successful for minerals and
vitamins,
 One-off cost,
 Easier distribution,
 Speed up process of
conventional plant breeding.
WEAKNESS
 Success limited to minerals
and dependent on several
factors,
 Needs regular application of
nutrients,
 Expensive difficult to
distribution.
 Long development
time.
Long development time,
Success limited to interactions
among transgenes.
OPPORTUNITIES
 Compliment to other
strategies
 Wide public acceptance,
 Simple legal frame
work
Fast ‘omics’developments
THREATS
 Negative environmental
impact, reverse exhaustion
(Eg: Se)
Requires genetic
variation
 Lack of awareness on GM crops.
Environmental impact(gene flow)
Prasad et al., 2015
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29. Objective
• Spirulina platensis has been used as biofortifying agent to enhance the iron
status in Amaranthus gangeticus plant.
Materials and methods:
• Experimentation was also carried out with different combinations of
Spirulina and fertilizers (Organic, Vermicompost, and Biofertilizer) in
different ratios.
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Kalpana et al.,2014

30. Kalpana et al., 2014
Sl. No. Sample Control Biofertilizer Iron(mg/100g)
1 A1 A4
(17.92)
S:B (25:75) 31.80
2 A2 S:B (50:50) 21.65
3 A3 S:B (75:25) 44.85
Sl. No. Sample Control Vermicompost Iron(mg/100g)
1 A1 A4
(2.35)
S:V (25:75) 43.99
2 A2 S:V (50:50) 2.76
3 A3 S:V (75:25) 2.35
Table: 3 Iron Content of Different Ratios of Spirulina vs Bio-Fertilizer Treated
Plants
Table: 4 Iron Content of Different Ratios of Spirulina vs Vermicompost Treated Plants
*(S:B) Spirulina: Biofertilizer (Azolla)
*(S:V) Spirulina: Vermicompost
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31. Sl. No. Sample Control Organic manure Iron(mg/100g)
1 A1 A4
(2.43)
S:O (25:75) 4.1
2 A2 S:O (50:50) 4.0
3 A3 S:O (75:25) 3.39
Kalpana et al., 2014
Table: 5 Iron Content of Different Ratios of Spirulina vs Organic manure treated
Plants
*(S:O) Spirulina: Organic manure
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32. Objective: To increase the carotenoids and amino acid content (Protein) in
cassava roots by inter-specific hybridization
Material and methods:
• ICB 300 (inter-specific hybrid): Cassava UNB 01 x M. oligantha
• ICB 300- derived offspring (Progeny 4, Progeny 9 and Progeny 10)
• UnB-400
• UnB-500
• Lycopene and Aminoacid analysis: Spectrometer And HPLC.
Nagib et al., 2009
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Table :6 Quantification (μg /g of tissue) trans-β-carotent and cis-β Carotene of some
Manihot cultivar organs
Roots trans - β carotene cis - β carotene
UNB 01 0.16 0.09
ICB 300 1.24 0.96
ICB- 400 0.19 0.12
ICB- 300
UnB- 400 UnB-01
Nagib et al., 2009

34. Table 7. Amino acid (AA) profile (g/100g) in peeled roots of Cassava cultivar UnB, its
Interspecific hybrid with M. oligantha ICB 300 (sample3 & Diploid) and ICB 300-Derived
offspring (Progeny 10)
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Nagib et al., 2009

35. Aim:
• To improve the nutritional value of potato, the AmA1 coding sequence was
successfully introduced and expressed in tuber.
• To increase in the growth and production of tubers in transgenic
populations and also of the total protein content with an increase in most
essential amino acids.
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Veg.Dept Chakraborty et al., 2000

36. Protein rich potato
Amaranthus
hypochondriacus
VECTOR
Protato ( >60% in
total protein )
Potato
AmA1gene
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Veg.Dept Chakraborty et al., 2000

37. Schematic representation of AmA1 expression
plasmids containing AmA1 coding sequence
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Veg.Dept Chakraborty et al., 2000

38. Table: 8 amino acid profile of tubers of wild-type and transgenic lines
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Chakraborty et al., 2000

39. Objective:-
The feasibility of developing crops with enough folate to supply the adult
recommended dietary allowance in a single standard serving (100 g/serving).
13-Jun-21 39
Veg.Dept Rocio et al., 2007

40. Introduction:-
• Folate deficiency is associated with spina bifida and other birth
defects and some cancers.
• Folates are tripartite molecules consisting of pteridine, p-
aminobenzoate (PABA), and glutamate moieties.
Materials & Methods:-
• Expression Vector Construction- AtADCS coding sequence,
obtained by PCR by using as template the cDNA clone RAFL09-
32-D4.
• Transgenic Plants- Tomato was transformed by A. tumefaciens harboring
the AtADCS construct.
• Folate Analysis , Pteridine and PABA Analysis.
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Rocio et al., 2007

41. 13-Jun-21 41
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42. Accumulation of folate, PABA, and pteridines in G x A (GCHI/
AtADCS) late red ripe tomatoes
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Rocio et al., 2007

43. Objective:-
To evaluate the inheritance of seed iron and zinc concentrations and contents in an intra-
gene pool Mesoamerican × Mesoamerican RIL population grown over three sites and to
identify QTL for each mineral.
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Veg.Dept Matthew et al., 2010

44. Introduction
• Common bean var. can be classified into 2 major gene pools within the species
based on seed size and origin (i.e. domestication and center of diversity) (Singh
et al. 1991).
• The inheritance of nutrition traits is mostly quantitative varies depending on
the source genotype.
Material & Methods:-
• Plant material-
– 110 RILs from the cross G14519 × G4825.
– Both parents are from the Mesoamerican intra-genepool and the cross was classified by Blair
et al. (2006a) based on a molecular marker survey.
• Experimental Sites- 3
• Mineral Analysis- Based on AAS(Atomic absorption Spectroscopy)
• QTL analysis- with composite interval mapping (CIM) analysis.
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Veg.Dept Matthew et al., 2010

45. Figure.2. Population distributions for Fe and Zn conc. measured in ppm with AAS and seed weight
measured in grams for 100 seed in the G14519 × G4825 recombinant inbred lines grown over three
locations. mineral values indicated by arrows
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Matthew et al., 2010

46. Genetic map for G14519 × G4825 RIL showing linkage groups b01 through b11 with QTL for
the conc. and content of Fe &Zn
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Veg.Dept Matthew et al., 2010

47. (b). Highlighting the cluster of QTL
on linkage group b06
 Vertical lines represents
the region in which the
marker-phenotype
associations are above the
LOD threshold for CIM
analysis with iron QTL (red)
and zinc QTL (blue).
 Horizontal marks indicate
the LOD peak for the QTL.
Small boxes represent
markers that were significant
in single point analysis (SPA)
for the analysis of Fe & Zn
content
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Veg.Dept Matthew et al., 2010

48. • The parents were highly contrasting with G14519 avg. 80 ppm and 34 ppm ,
respectively for seed Fe & Zn and G4825 averaging 46 ppm Fe and 25 ppm Zn over all
sites.
• 3 QTL for Zn were found near the same position on linkage group b06 In all the
QTL,for this part of linkage group b06, the +ve allele for higher mineral conc. was
from the high mineral parent, G14519.
• The QTL near BM158 could be a single major gene; or a tight cluster of genes
controlling the conc. of both minerals showing a stable, cross location QTL for seed
iron and zinc levels.
• The co-localization of QTL for seed iron and zinc and the major locus on linkage
group b06 could be useful for MAS selection and allow the improvement of various
classes through backcrossing for example.
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Veg.Dept Matthew et al., 2010

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Objective
• Improve TCC in cassava roots by reducing standard length of each cycle.
Materials and methods:
Site- CIAT Colombia 1000-2000genotypes produce yellow roots.
Extraction and quantification of carotenoids- HPLC
Ceballos et al., 2013

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Figure 3: Illustration of the chronology of rapid cycling recurrent selection in cassava for
enhanced carotenoids content
Ceballos et al., 2013

51. Figure 7. Illustration of the variation observed for root color intensity in genotypes from a full-sib
family, whose total carotenoids content ranged from < 1.0 to 25.8 μg g-1 (fresh weight basis).
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Ceballos et al., 2013

52. Table: 9 Summary of the data generated over the years
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Ceballos et al., 2013

53. ACHIEVEMENTS
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54. 54
Introduction
Sweet potato
CIP-440127 - from CIP - Carotene 6.2-7.6 mg/100g
ST -14 – from japan – Carotene 13.2-14.4 mg/100g
ST - 14
CIP-440127
Beet root
Detroit dark red – deep red
Introduced from USA
H.P. Singh, Vegetable Varieties of India, 2011
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55. 55
Crop Variety Colour pigments
Watermelon Durgapura kesar Yellow Carotene
Palak Punjab Green Purple (stem) Anthocyanin
Beta carotene
Pusa Bharathi Green Beta carotene
Amaranthus Pusa Lal Chaulai Red(Magenta) Anthocyanins
Arka Arunima Purple Anthocyanins
Basella Local Red Red Carotenoids
Local Green green Leutin
Pumpkin Arka chandan Bright orange Carotene
carrot Ooty 1- half sib
progeny selection
of DC-3
Deep
orange
Carotene
Shalimar -1 Orange Carotene -56.1
mg/100g
Local selection
Durgapura Kesar
Arka Chandan
H.P. Singh, Vegetable Varieties of India, 2011
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56. 56
Clonal selection
Pure line selection
Crop Variety Colour Pigment
Chilli Arka Abhir (Paprika)
- PLS - Devanur
dubha
Red Capsanthin
KTPL- 19 (IARI)
- PLS - P12
Red Capsanthin
• Sweet potato variety Co-5 is from CIP 440038
• Orange fleshed variety
H.P. Singh, Vegetable Varieties of India, 2011
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57.  Solanum chilense,
 Solanum hirsutum,
Solanum cheesmanii,
Solanum lycopersicoides
 Anthocyanin fruit (Aft) - S. chilense
 Aubergine (Abg) - S. lycopersicoides
 Atroviolacium (atv) - S. cheesmanii
Tomato
Inter specific hybridization
× Solanum lycopersicon
Carotenoid (lycopene)
rich varieties
Anthocyanin rich varities
× Solanum lycopersicon
Rick and Stevens (1986)
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58. 58
Crop Variety Colour Amount
Brinjal Punjab Sadabahar
- Jap Long x R-34
Blackish
purple
Anthocyanin
Watermelon
Arka Jyoti - IIHR -20
×Crimson Sweet
Crimson
red
Carotene
Durgapura lal –
Sugar baby × K3566
Dark red Carotene
Tapioca Sree Visakam Yellow Beta carotene
Sweet
Potato
Sree Rethna Purple
skin &
Yellow
flesh
Carotene
Intervarietal hybridization
Durgapura lal
Sree Visakam
H.P. Singh, Vegetable Varieties of India, 2011
13-Jun-21 Veg.Dept
Punjab Sadabahar

59. IARI -biofortified Varieties
Cauliflower : Pusa Betakesari
First ever indigenously bred bio-fortified beta
carotene (800 – 1000 µg/100 g)
Released-2015-16
by Marker assisted backcross method
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60. Pusa Kiran- Fe rich
Pusa Meghali- carotene rich
Pusa Hybrid 1- Fe rich
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61. Orange-fleshed sweet potatoes(OFSP) in
Orissa
Some var:-
•Sree kanaka
•Sree Rethna
•Sree Vardhini
•Varsha
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63. CROP VARIETY ATTRIBUTES
Tomato Pusa Uphar, Pusa Rohini
Pusa Hybrid 2, Pusa Red Plum
Vitamin C & Lycopene
Paprika KTPL-19 Capsanthin
Carrot
Pusa Asita
Pusa Rudhira
Anthocyanin
Lycopene
Pusa Vrishti ,Pusa Yamdagni
Pusa Meghali ,Pusa Nayanjyoti
Beta carotene
Radish Pusa Jamuni Anthocyanin
Pusa Gulabi Lycopene
Released vegetable varieties
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64. CROP VARIETY ATTRIBUTES
Red cabbage Red Acre Anthocyanin
Beetroot Pusa Bharati
Pusa Jyoti
Ascorbic acid Vitamin A ,C, Fe, Ca
Bitter gourd Pusa Aushadi
Pusa Vishesh
Pusa Hybrid -2
Beta carotene
Ca & Fe
Ca &Fe
Pumpkin Arka Chandan Beta-carotene
Tapioca Sree Visakam Beta-carotene
Sweet potato Sree Kanaka ,
Pusa Bharati
Beta -carotene
Amaranthus Amt. 105 Carotene and protein
Palak Pusa Bharati Beta -carotene
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65. Advantages and Disadvantages for
Biofortification of vegetable crops
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Vishal and Borkar et.al 2014.

66. Brazil
India
Biofortification
Program
Who Promotes Biofortification?
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67. Government programmes
 Balwadi nutrition programme
 Special nutrition programme
 Integrated child development service(ICDS) scheme.
 Wheat based nutrition programme
 Nutrition programme for Adolescent Girls
 National nutritional anemia prophylaxis programme
 Weekly iron and folic acid supplementation programme for adolescents.
 National prophylaxis programme against nutritional blindness due to Vitamin
A deficiency.
 Mid-day meal programme
 Chiranjivi Yojana
 Akshaya Patra
 Annapurna Scheme
 Antyodaya Anna yojna
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68. • Biofortification help in overcoming nutrient
deficiency economically especially in rural areas.
• Application of biofortified crops would benefit
farmers by increasing their income in the long run .
• Functional crops can play an important role in
fighting against different types of nutrition deficiency
and malnutrition.
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